CHAPTER 11 – TRANSPORT IN PLANTS

Chapter 11

Transport in Plants

  • Plants need to move molecules over very long distances, much more than animals do; they also do not have a circulatory system in place.
  • Water taken up by the roots has to reach all parts of the plant, up to the very tip of the growing stem. The photosynthates or food synthesised by the leaves have also to be moved to all parts including the root tips embedded deep inside the soil.
  • Movement across short distances, say within the cell, across the membranes and from cell to cell within the tissue has also to take place.
  • In a flowering plant the substances that would need to be transported are water, mineral nutrients, organic nutrients and plant growth regulators.
  • Small distance transport – by diffusion and by cytoplasmic streaming supplemented by active transport.
  • Long distance Transport – through the vascular system (the xylem and the phloem) and is called translocation.
  • In rooted plants, transport in xylem (of water and minerals) is essentially unidirectional, from roots to the stems.
  • Organic and mineral nutrients however, undergo multidirectional transport.
  • Organic compounds synthesised in the photosynthetic leaves are exported to all other parts of the plant Including storage organs. From the storage organs they are later re-exported.
  • The mineral nutrients are taken up by the roots and transported upwards into the stem, leaves and the growing regions. When any plant part undergoes senescence, nutrients may be withdrawn from such regions and moved to the growing parts.
  • Hormones or plant growth regulators and other chemical stimuli are also transported, though in very small amounts, sometimes in a strictly polarised or unidirectional manner from where they are synthesised to other parts.

Means of Transport

Diffusion

  • Passive movement and may be from one part of the cell to the other, or from cell to cell, or over short distances, say, from the intercellular spaces of the leaf to the outside.
  • No energy expenditure takes place.
  • Molecules move in a random fashion, the net result being substances moving from regions of higher concentration to regions of lower concentration.
  • Diffusion is a slow process and is not dependent on a ‘living system’.
  • Diffusion is obvious in gases and liquids, but diffusion in solids rather than of solids is more likely.
  • Diffusion is very important to plants since it the only means for gaseous movement within the plant body.
  • Diffusion rates are affected by the gradient of concentration, the permeability of the membrane separating them, size of the substances, temperature and pressure.

Facilitated Diffusion

  • The diffusion of any substance across a membrane also depends on its solubility in lipids, the major constituent of the membrane.
  • Substances soluble in lipids diffuse through the membrane faster.
  • Substances that have a hydrophilic moiety, find it difficult to pass through the membrane; their movement has to be facilitated. Membrane proteins provide sites at which such molecules cross the membrane.
  • They do not set up a concentration gradient: a concentration gradient must already be present for molecules to diffuse even if facilitated by the proteins. This process is called facilitated diffusion.
  • In facilitated diffusion special proteins help move substances across membranes without expenditure of ATP energy.
  • Facilitated diffusion cannot cause net transport of molecules from a low to a high concentration – this would require input of energy.
  • Transport rate reaches a maximum when all of the protein transporters are being used (saturation).
  • Facilitated diffusion is very specific: it allows cell to select substances for uptake.
  • It is sensitive to inhibitors which react with protein side chains.
  • The proteins form channels in the membrane for molecules to pass through. Some channels are always open; others can be controlled. Some are large, allowing a variety of molecules to cross.
  • The porins are proteins that form huge pores in the outer membranes of the plastids, mitochondria and some bacteria allowing molecules up to the size of small proteins to pass through.
  • When an extracellular molecule bound to the transport protein; the transport protein then rotates and releases the molecule inside the cell, e.g., water channels (made up of eight different types of aquaporins.)

Passive symports and antiports

  • Some carrier or transport proteins allow diffusion only if two types of molecules move together.
  • In a symport, both molecules cross the membrane in the same direction; in an antiport, they move in opposite directions.
  • When a molecule moves across a membrane independent of other molecules, the process is called uniport.

Active Transport

  • Active transport uses energy to pump molecules against a concentration gradient.
  • Active transport is carried out by membrane-proteins. Hence different proteins in the membrane play a major role in both active as well as passive transport.
  • Pumps are proteins that use energy to carry substances across the cell membrane. These pumps can transport substances from a low concentration to a high concentration (‘uphill’ transport).
  • Transport rate reaches a maximum when all the protein transporters are being used or are saturated.
  • Like enzymes the carrier protein is very specific in what it carries across the membrane.
  • These proteins are sensitive to inhibitors that react with protein side chains.

Comparison of Different Transport Processes

  • Proteins in the membrane are responsible for facilitated diffusion and active transport and hence show common characterstics of being highly selective; they are liable to saturate, respond to inhibitors and are under hormonal regulation.
  • But diffusion whether facilitated or not – take place only along a gradient and do not use energy.

Plant-Water Relations

  • Water is essential for all physiological activities of the plant and plays a very important role in all living organisms.
  • It provides the medium in which most substances are dissolved. The protoplasm of the cells is nothing but water in which different molecules are dissolved and (several particles) suspended. A watermelon has over 92 per cent water; most herbaceous plants have only about 10 to 15 per cent of its fresh weight as dry matter.
  • Of course, distribution of water within a plant varies – woody parts have relatively very little water, while soft parts mostly contain water. A seed may appear dry but it still has water – otherwise it would not be alive and respiring!
  • Terrestrial plants take up huge amount water daily but most of it is lost to the air through evaporation from the leaves, i.e., transpiration. A mature corn plant absorbs almost three litres of water in a day, while a mustard plant absorbs water equal to its own weight in about 5 hours. Because of this high demand for water, it is not surprising that water is often the limiting factor for plant growth and productivity in both agricultural and natural environments.

Water Potential

  • Water potential (Psi w) is a concept fundamental to understanding water movement. Solute potential (Psi s) and pressure potential (Psi p) are the two main components that determine water potential.
  • Water molecules possess kinetic energy. In liquid and gaseous form they are in random motion that is both rapid and constant. The greater the concentration of water in a system, the greater is its kinetic energy or ‘water potential’. Hence, it is obvious that pure water will have the greatest water potential.
  • If two systems containing water are in contact, random movement of water molecules will result in net movement of water molecules from the system with higher energy to the one with lower energy. Thus water will move from the system containing water at higher water potential to the one having low water potential.
  • This process of movement of substances down a gradient of free energy is called diffusion.
  • Water potential is denoted by the Greek symbol Psi and is expressed in pressure units such as pascals (Pa).
  • By convention, the water potential of pure water at standard temperatures, which is not under any pressure, is taken to be zero.
  • If some solute is dissolved in pure water, the solution has fewer free water and the concentration of water decreases, reducing its water potential. Hence, all solutions have a lower water potential than pure water; the magnitude of this lowering due to dissolution of a solute is called solute potential or Psi s. Psi s is always negative. The more the solute molecules, the lower (more negative) is the Psi s.
  • For a solution at atmospheric pressure (water potential) Psi w = (solute potential) Psi s.
  • If a pressure greater than atmospheric pressure is applied to pure water or a solution, its water potential increases. It is equivalent to pumping water from one place to another.
  • Pressure can build up in a plant system when water enters a plant cell due to diffusion causing a pressure built up against the cell wall, it makes the cell turgid, this increases the pressure potential. Pressure potential is usually positive.
  • Though in plants negative potential or tension in the water column in the xylem plays a major role in water transport up a stem. Pressure potential is denoted as (p.
  • Water potential of a cell is affected by both solute and pressure potential. The relationship between them is as follows:

Osmosis

  • The plant cell is surrounded by a cell membrane and a cell wall. The cell wall is freely permeable to water and substances in solution hence is not a barrier to movement.
  • In plants the cells usually contain a large central vacuole, whose contents, the vacuolar sap, contribute to the solute potential of the cell.
  • In plant cells, the cell membrane and the membrane of the vacuole, the tonoplast together are important determinants of movement of molecules in or out of the cell.
  • Osmosis is the term used to refer specifically to the diffusion of water across a differentially- or semi-permeable membrane. Osmosis occurs spontaneously in response to a driving force.
  • The net direction and rate of osmosis depends on both the pressure gradient and concentration gradient.
  • Water will move from its region of higher chemical potential (or concentration) to its region of lower chemical potential until equilibrium is reached. At equilibrium the two chambers should have the same water potential.


In above Fig two chambers, A and B, containing solutions are separated by a semi-permeable membrane.

  • Solution of which chamber has a lower water potential? – B
  • Solution of which chamber has a lower solute potential? – B
  • In which direction will osmosis occur? – A(B
  • Which solution has a higher solute potential?- A
  • At equilibrium which chamber will have lower water potential?- Equal
  • If one chamber has a psi of -2000 kPa, and the other -1000 kPa, which is the chamber that has the higher psi ?- second with -1000kP.

  • Experiment – a solution of sucrose in water taken in a funnel is separated from pure water in a beaker through a semi-permeable membrane (Egg membrane – Can be obtained by removing the yolk and albumin through a small hole at one end of the egg, and placing the shell in dilute solution of hydrochloric acid for a few hours. The egg shell dissolves leaving the membrane intact). Water will move into the funnel, resulting in rise in the level of the solution in the funnel. This will continue till the equilibrium is reached. External pressure can be applied from the upper part of the funnel such that no water diffuses into the funnel through the membrane. This pressure required to prevent water from diffusing is the osmotic pressure and this is the function of the solute concentration; more the solute concentration, greater will be the pressure required to prevent water from diffusing in. Numerically osmotic pressure is equivalent to the osmotic potential, but the sign is opposite. Osmotic pressure is the positive pressure applied, while osmotic potential is negative.

Plasmolysis

    • The behaviour of the plant cells (or tissues) with regard to water movement depends on the surrounding solution.
    • If the external solution balances the osmotic pressure of the cytoplasm, it is said to be isotonic.
    • If the external solution is more dilute than the cytoplasm, it is hypotonic and if the external solution is more concentrated, it is hypertonic.
    • Cells swell in hypotonic solutions and shrink in hypertonic ones.
    • Plasmolysis occurs when water moves out of the cell and the cell membrane of a plant cell shrinks away from its cell wall. This occurs when the cell (or tissue) is placed in a solution that is hypertonic (has more solutes) to the protoplasm. Water moves out; it is first lost from the cytoplasm and then from the vacuole.
    • The water when drawn out of the cell through diffusion into the extracellular (outside cell) fluid causes the protoplast to shrink away from the walls. The cell is said to be plasmolysed.
    • The movement of water occurred across the membrane moving from an area of high water potential (i.e., the cell) to an area of lower water potential outside the cell.
    • The process of plamolysis is usually reversible.
    • When the cells are placed in a hypotonic solution (higher water potential or dilute solution as compared to the cytoplasm), water diffuses into the cell causing the cytoplasm to build up a pressure against the wall, that is called turgor pressure. The pressure exerted by the protoplasts due to entry of water against the rigid walls is called pressure potential. Because of the rigidity of the cell wall, the cell does not rupture.
    • This turgor pressure is ultimately responsible for enlargement and extension growth of cells.

Imbibition

  • Imbibition is a special type of diffusion when water is absorbed by solids – colloids – causing them to enormously increase in volume. e.g., absorption of water by seeds and dry wood.
  • The pressure that is produced by the swelling of wood had been used by prehistoric man to split rocks and boulders.
  • If it were not for the pressure due to imbibition, seedlings would not have been able to emerge out of the soil into the open.
  • Imbibition is also diffusion since water movement is along a concentration gradient; the seeds and other such materials have almost no water hence they absorb water easily.
  • Water potential gradient between the absorbent and the liquid imbibed is essential for imbibition.
  • In addition, for any substance to imbibe any liquid, affinity between the adsorbant and the liquid is also a pre-requisite.

Long Distance Transport of Water

  • Long distance transport of substances within a plant cannot be by diffusion alone. Diffusion is a slow process. It can account for only short distance movement of molecules. For example, the movement of a molecule across a typical plant cell (about 50µm) takes approximately 2.5 s.
  • In large and complex organisms, often substances have to be moved across very large distances. sometimes the sites of production or absorption and sites of storage are too far from each other; diffusion or active transport would not suffice. Special long distance transport systems become necessary so as to move substances across long distances and at a much faster rate.
  • Water and minerals, and food are generally moved by a mass or bulk flow system.
  • Mass flow is the movement of substances in bulk or en masse from one point to another as a result of pressure differences between the two points. It is a characteristic of mass flow that substances, whether in solution or in suspension, are swept along at the same pace, as in a flowing river.
  • This is unlike diffusion where different substances move independently depending on their concentration gradients.
  • Bulk flow can be achieved either through a positive hydrostatic pressure gradient (e.g., a garden hose) or a negative hydrostatic pressure gradient (e.g., suction through a straw).
  • The bulk movement of substances through the conducting or vascular tissues of plants is called translocation.
  • The higher plants have highly specialised vascular tissues – xylem and phloem.
  • Xylem is associated with translocation of mainly water, mineral salts, some organic nitrogen and hormones, from roots to the aerial parts of the plants.
  • The phloem translocates a variety of organic and inorganic solutes, mainly from the leaves to other parts of the plants.

How do Plants Absorb Water?

  • The responsibility of absorption of water and minerals is of the root hairs.
  • Root hairs are thin-walled slender extensions of root epidermal cells that greatly increase the surface area for absorption.
  • Water is absorbed along with mineral solutes, by the root hairs, purely by diffusion.
  • once water is absorbed by the root hairs, it can move deeper into root layers by two distinct pathways: Apoplast pathway, Symplast pathway.
  • Apoplast is the system of adjacent cell walls that is continuous throughout the plant, except at the casparian strips of the endodermis in the roots.
  • The apoplastic movement of water occurs exclusively through the intercellular spaces and the walls of the cells.
  • Movement through the apoplast does not involve crossing the cell membrane.
  • This movement is dependent on the gradient.
  • The apoplast does not provide any barrier to water movement and water movement is through mass flow.
  • As water evaporates into the intercellular spaces or the atmosphere, tension develop in the continuous stream of water in the apoplast, hence mass flow of water occurs due to the adhesive and cohesive properties of water.
  • The symplastic system is the system of interconnected protoplasts. Neighbouring cells are connected through cytoplasmic strands that extend through plasmodesmata.
  • During symplastic movement, the water travels through the cells – their cytoplasm; intercellular movement is through the plasmodesmata.
  • Water has to enter the cells through the cell membrane, hence the movement is relatively slower.
  • Movement is again down a potential gradient.
  • symplastic movement may be aided by cytoplasmic streaming.

  • Most of the water flow in the roots occurs via the apoplast since the cortical cells are loosely packed, and hence offer no resistance to water movement. However, the inner boundary of the cortex, the endodermis, is impervious to water because of a band of suberised matrix called the casparian strip.
  • Water molecules are unable to penetrate the layer, so they are directed to wall regions that are not suberised, into the cells proper through the membranes.
  • The water then moves through the symplast and again crosses a membrane to reach the cells of the xylem.
  • The movement of water through the root layers is ultimately symplastic in the endodermis. This is the only way water and other solutes can enter the vascular cylinder.
  • Once inside the xylem, water is again free to move between cells as well as through them. In young roots, water enters directly into the xylem vessels and/or tracheids. These are non-living conduits and so are parts of the apoplast.

  • Some plants have additional structures associated with them that help in water (and mineral) absorption. A mycorrhiza is a symbiotic association of a fungus with a root system.
  • The fungal filaments form a network around the young root or they penetrate the root cells. The hyphae have a very large surface area that absorb mineral ions and water from the soil from a much larger volume of soil that perhaps a root cannot do.
  • The fungus provides minerals and water to the roots, in turn the roots provide sugars and N-containing compounds to the mycorrhizae.
  • Some plants have an obligate association with the mycorrhizae. e.g., Pinus seeds cannot germinate and establish without the presence of mycorrhizae.

Water Movement up a Plant

Root Pressure

  • As various ions from the soil are actively transported into the vascular tissues of the roots, water follows (its potential gradient) and increases the pressure inside the xylem. This positive pressure is called root pressure, and can be responsible for pushing up water to small heights in the stem.
  • Effects of root pressure is also observable at night and early morning when evaporation is low, and excess water collects in the form of droplets around special openings of veins near the tip of grass blades, and leaves of many herbaceous parts. Such water loss in its liquid phase is known as guttation.
  • Root pressure can, at best, only provide a modest push in the overall process of water transport. They obviously do not play a major role in water movement up tall trees.
  • The greatest contribution of root pressure may be to re-establish the continuous chains of water molecules in the xylem which often break under the enormous tensions created by transpiration.
  • Root pressure does not account for the majority of water transport; most plants meet their need by transpiratory pull.

Transpiration pull

  • The flow of water upward through the xylem in plants can achieve fairly high rates, up to 15 metres per hour.
  • Most researchers agree that water is mainly ‘pulled’ through the plant, and that the driving force for this process is transpiration from the leaves. This is referred to as the cohesion-tension-transpiration pull model of water transport. But, what generates this transpirational pull?
  • Water is transient in plants. Less than 1 per cent of the water reaching the leaves is used in photosynthesis and plant growth. Most of it is lost through the stomata in the leaves. This water loss is known as transpiration.

TRANSPIRATION

  • Transpiration is the evaporative loss of water by plants. It occurs mainly through the stomata in the leaves.
  • Besides the loss of water vapour in transpiration, exchange of oxygen and carbon dioxide in the leaf also occurs through pores called stomata (sing.: stoma).
  • Normally stomata are open in the day time and close during the night.
  • The immediate cause of the opening or closing of the stomata is a change in the turgidity of the guard cells.
  • The inner wall of each guard cell, towards the pore or stomatal aperture, is thick and elastic. When turgidity increases within the two guard cells flanking each stomatal aperture or pore, the thin outer walls bulge out and force the inner walls into a crescent shape.
  • The opening of the stoma is also aided due to the orientation of the microfibrils in the cell walls of the guard cells. Cellulose microfibrils are oriented radially rather than longitudinally making it easier for the stoma to open.
  • When the guard cells lose turgor, due to water loss (or water stress) the elastic inner walls regain their original shape, the guard cells become flaccid and the stoma closes.

  • Usually the lower surface of a dorsiventral (often dicotyledonous) leaf has a greater number of stomata while in an isobilateral (often monocotyledonous) leaf they are about equal on both surfaces.
  • Transpiration is affected by several external factors: temperature, light, humidity, wind speed. Plant factors that affect transpiration include number and distribution of stomata, number of stomata open, per cent, water status of the plant, canopy structure etc.
  • The transpiration driven ascent of xylem sap depends mainly on the following physical properties of water:
  • Cohesion – mutual attraction between water molecules.
  • Adhesion – attraction of water molecules to polar surfaces (such as the surface of tracheary elements).
  • Surface Tension – water molecules are attracted to each other in the liquid phase more than to water in the gas phase.
  • These properties give water high tensile strength, i.e., an ability to resist a pulling force, and high capillarity, i.e., the ability to rise in thin tubes. In plants capillarity is aided by the small diameter of the tracheary elements – the tracheids and vessel elements.
  • The process of photosynthesis requires water. The system of xylem vessels from the root to the leaf vein can supply the needed water.
  • As water evaporates through the stomata, since the thin film of water over the cells is continuous, it results in pulling of water, molecule by molecule, into the leaf from the xylem.
  • Also, because of lower concentration of water vapour in the atmosphere as compared to the substomatal cavity and intercellular spaces, water diffuses into the surrounding air. This creates a ‘pull’.
  • Measurements reveal that the forces generated by transpiration can create pressures sufficient to lift a xylem sized column of water over 130 metres high.

Transpiration and Photosynthesis – a Compromise

  • Transpiration has more than one purpose; it
    • createstranspiration pull for absorption and transport of plants
    • supplieswater for photosynthesis
    • transportsminerals from the soil to all parts of the plant
    • cools leaf surfaces, sometimes 10 to 15 degrees, by evaporative cooling
    • maintainsthe shape and structure of the plants by keeping cells turgid
  • An actively photosynthesising plant has an insatiable need for water. Photosynthesis is limited by available water which can be swiftly depleted by transpiration.
  • The humidity of rainforests is largely due to this vast cycling of water from root to leaf to atmosphere and back to the soil.
  • The evolution of the C4 photosynthetic system is probably one of the strategies for maximising the availability of CO2 while minimising water loss.
  • C4 plants are twice as efficient as C3 plants in terms of fixing carbon (making sugar). However, a C4 plant loses only half as much water as a C3 plant for the same amount of CO2 fixed.

Uptake and Transport of Mineral Nutrients

  • Plants obtain their carbon and most of their oxygen from CO2 in the atmosphere. However, their remaining nutritional requirements are obtained from minerals and water for hydrogen in the soil.

Uptake of Mineral Ions

  • Unlike water, all minerals cannot be passively absorbed by the roots.
  • Two factors account for this:

(i) minerals are present in the soil as charged particles (ions) which cannot move across cell membranes and
(ii) the concentration of minerals in the soil is usually lower than the concentration of minerals in the root.

  • Therefore, most minerals must enter the root by active absorption into the cytoplasm of epidermal cells.
  • This needs energy in the form of ATP.
  • The active uptake of ions is partly responsible for the water potential gradient in roots, and therefore for the uptake of water by osmosis. Some ions also move into the epidermal cells passively.
  • Ions are absorbed from the soil by both passive and active transport.
  • Specific proteins in the membranes of root hair cells actively pump ions from the soil into the cytoplasms of the epidermal cells.
  • Like all cells, the endodermal cells have many transport proteins embedded in their plasma membrane; they let some solutes cross the membrane, but not others.
  • Transport proteins of endodermal cells are control points, where a plant adjusts the quantity and types of solutes that reach the xylem.
  • the root endodermis because of the layer of suberin has the ability to actively transport ions in one direction only.

Translocation of Mineral Ions

  • After the ions have reached xylem through active or passive uptake, or a combination of the two, their further transport up the stem to all parts of the plant is through the transpiration stream.
  • The chief sinks for the mineral elements are the growing regions of the plant, such as the apical and lateral meristems, young leaves, developing flowers, fruits and seeds, and the storage organs.
  • Unloading of mineral ions occurs at the fine vein endings through diffusion and active uptake by these cells.
  • Mineral ions are frequently remobilised, particularly from older, senescing parts.
  • Older dying leaves export much of their mineral content to younger leaves.
  • Similarly, before leaf fall in decidous plants, minerals are removed to other parts.
  • Elements most readily mobilised are phosphorus, sulphur, nitrogen and potassium.
  • Some elements that are structural components like calcium are not remobilised.
  • An analysis of the xylem exudates shows that though some of the nitrogen travels as inorganic ions, much of it is carried in the organic form as amino acids and related compounds.
  • Similarly, small amounts of P and S are carried as organic compounds.
  • In addition, small amount of exchange of materials does take place between xylem and phloem.
  • Hence, it is not that we can clearly make a distinction and say categorically that xylem transports only inorganic nutrients while phloem transports only organic materials.

Phloem Transport: Flow from Source to Sink

  • Food, primarily sucrose, is transported by the vascular tissue phloem from a source to a sink.
  • Usually the source is understood to be that part of the plant which synthesises the food, i.e., the leaf, and sink, the part that needs or stores the food.
  • But, the source and sink may be reversed depending on the season, or the plant’s needs.
  • Sugar stored in roots may be mobilised to become a source of food in the early spring when the buds of trees, act as sink; they need energy for growth and development of the photosynthetic apparatus.
  • Since the source-sink relationship is variable, the direction of movement in the phloem can be upwards or downwards, i.e., bi-directional. This contrasts with that of the xylem where the movement is always unidirectional, i.e., upwards.
  • Hence, unlike one-way flow of water in transpiration, food in phloem sap can be transported in any required direction so long as there is a source of sugar and a sink able to use, store or remove the sugar.
  • Phloem sap is mainly water and sucrose, but other sugars, hormones and amino acids are also transported or translocated through phloem.

The Pressure Flow or Mass Flow Hypothesis

  • The accepted mechanism used for the translocation of sugars from source to sink is called the pressure flow hypothesis.
  • As glucose is prepared at the source (by photosynthesis) it is converted to sucrose (a dissacharide). The sugar is then moved in the form of sucrose into the companion cells and then into the living phloem sieve tube cells by active transport.
  • This process of loading at the source produces a hypertonic condition in the phloem.
  • Water in the adjacent xylem moves into the phloem by osmosis. As osmotic pressure builds up the phloem sap will move to areas of lower pressure.
  • At the sink osmotic pressure must be reduced. Again active transport is necessary to move the sucrose out of the phloem sap and into the cells which will use the sugar – converting it into energy, starch, or cellulose.
  • As sugars are removed, the osmotic pressure decreases and water moves out of the phloem.
  • the movement of sugars in the phloem begins at the source, where sugars are loaded (actively transported) into a sieve tube. Loading of the phloem sets up a water potential gradient that facilitates the mass movement in the phloem.
  • Phloem tissue is composed of sieve tube cells, which form long columns with holes in their end walls called sieve plates.
  • Cytoplasmic strands pass through the holes in the sieve plates, so forming continuous filaments.
  • As hydrostatic pressure in the phloem sieve tube increases, pressure flow begins, and the sap moves through the phloem.
  • Meanwhile, at the sink, incoming sugars are actively transported out of the phloem and removed as complex carbohydrates.
  • The loss of solute produces a high water potential in the phloem, and water passes out, returning eventually to xylem.
  • A simple experiment, called girdling, was used to identify the tissues through which food is transported.
  • On the trunk of a tree a ring of bark up to a depth of the phloem layer, can be carefully removed.
  • In the absence of downward movement of food, the portion of the bark above the ring on the stem becomes swollen after a few weeks.
  • This simple experiment shows that phloem is the tissue responsible for translocation of food; and that transport takes place in one direction, i.e., towards the roots.

 

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CHAPTER 11- TRANSPORT IN PLANTS

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CHAPTER 12 : BIOTECHNOLOGY AND ITS APPLICATIONS

CHAPTER 12

BIOTECHNOLOGY AND ITS APPLICATIONS

  • Biotechnology essentially deals with industrial scale production of biopharmaceuticals and biologicals using genetically modified microbes, fungi, plants and animals.
  • The applications of biotechnology include therapeutics, diagnostics, and genetically
    modified crops for agriculture, processed food, bioremediation, waste treatment, and
    energy production.
  • Three critical research areas of biotechnology are:
    (i) Providing the best catalyst in the form of improved organism usually a microbe or pure enzyme.
    (ii) Creating optimal conditions through engineering for a catalyst to act, and
    (iii) Downstream processing technologies to purify the protein/organic compound.

BIOTECHNOLOGICAL APPLICATIONS IN AGRICULTURE

There are three options that can be thought for increasing food production
(i) agro-chemical based agriculture;
(ii) organic agriculture; and
(iii) Genetically engineered crop-based agriculture.

  •  We have succeeded in tripling the food supply by Green Revolution but yet it was not
    enough to feed the growing human population.
  • Increased yields have partly been due to the use of improved crop varieties, but mainly due to the use of better management practices and use of agrochemicals (fertilisers and pesticides).
  • However, for farmers in the developing world, agrochemicals are often too expensive, and further increases in yield with existing varieties are not possible using conventional breeding.
  • So there is a need to find alternative path that our understanding of genetics can show so that farmers may obtain maximum yield from their fields and to minimise the use of fertilisers and chemicals so that their harmful effects on the environment can be reduced. Use of genetically modified crops is a possible solution.
  • Plants, bacteria, fungi and animals whose genes have been altered by manipulation are called Genetically Modified Organisms (GMO).
  • Genetic modification has:
    (i) Made crops more tolerant to abiotic stresses (cold, drought, salt, heat).
    (ii) Reduced reliance on chemical pesticides (pest-resistant crops).
    (iii) Helped to reduce post-harvest losses.
    (iv) Increased efficiency of mineral usage by plants (this prevents early exhaustion of
    fertility of soil).
    (v) Enhanced nutritional value of food, e.g., Vitamin ‘A’ enriched rice.
    In addition to these uses, GM has been used to create tailor-made plants to supply
    alternative resources to industries, in the form of starches, fuels and pharmaceuticals.
  • By applications of biotechnology in agriculture, pest resistant plants are produced,
    which could decrease the amount of pesticide used.
  • Bt toxin is produced by a bacterium called Bacillus thuringiensis (Bt for short).
  • Bt toxin gene has been cloned from the bacteria and been expressed in plants to
    provide resistance to insects without the need for insecticides; in effect created a
    bio-pesticide. Examples are Bt cotton, Bt corn, rice, tomato, potato and soyabean etc.

Bt Cotton:

  • Some strains of Bacillus thuringiensis produce proteins that kill certain insects such as lepidopterans (tobacco budworm, armyworm), coleopterans (beetles) and dipterans (flies, mosquitoes).
  • B. thuringiensis forms protein crystals during a particular phase of their growth. These crystals contain a toxic insecticidal protein.
  • This toxin does not kill the Bacillus because this protein exists as inactive protoxins but once an insect ingest the inactive toxin, it is converted into an active form of toxin due to the alkaline pH of the gut which solubilise the crystals. The activated toxin binds to the surface of midgut epithelial cells and create pores that cause cell swelling and lysis and eventually cause death of the insect.
  • Specific Bt toxin genes were isolated from Bacillus thuringiensis and incorporated into the several crop plants such as cotton. The choice of genes depends upon the crop and the targeted pest, as most Bt toxins are insect-group specific.
  • The toxin is coded by a gene named cry. There are a number of them, for example, the proteins encoded by the genes crylAc and cryllAb control the cotton bollworms, that of crylAb controls corn borer.

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Pest Resistant Plants:

  • Several nematodes parasitise a wide variety of plants and animals including human beings.
  • A nematode Meloidegyne incognitia infects the roots of tobacco plants and causes a
    great reduction in yield.
  • A novel strategy was adopted to prevent this infestation which was based on the
    process of RNA interference (RNAi).
  • RNAi takes place in all eukaryotic organisms as a method of cellular defense.
  • This method involves silencing of a specific mRNA due to a complementary dsRNA
    molecule that binds to and prevents translation of the mRNA (silencing).
  • The source of this complementary RNA could be from an infection by viruses having
    RNA genomes or mobile genetic elements (transposons) that replicate via an RNA
    intermediate.
  • Using Agrobacterium vectors, nematode-specific genes were introduced into the host
    plant.
  • The introduction of DNA was such that it produced both sense and anti-sense RNA in
    the host cells. These two RNA’s being complementary to each other formed a double
    stranded (dsRNA) that initiated RNAi and thus, silenced the specific mRNA of the
    nematode.
  • The consequence was that the parasite could not survive in a transgenic host
    expressing specific interfering RNA. The transgenic plant therefore got itself protected from the parasite.

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BIOTECHNOLOGICAL APPLICATIONS IN MEDICINE

  • By enabling mass production of safe and more effective therapeutic drugs.
  • Further, the recombinant therapeutics do not induce unwanted immunological
    responses as is common in case of similar products isolated from non-human sources.
  • At present, about 30 recombinant therapeutics have been approved for human-use the world over. In India, 12 of these are presently being marketed.

    Genetically Engineered Insulin

  • Management of adult-onset diabetes is possible by taking insulin at regular time
    intervals.
  • if enough human-insulin was not available, that one would have to isolate and use
    insulin from other animals.
  • Insulin used for diabetes was earlier extracted from pancreas of slaughtered cattle and
    pigs.
  • Insulin from an animal source, though caused some patients to develop allergy or other types of reactions to the foreign protein.
  • Insulin consists of two short polypeptide chains: chain A and chain B, which are linked together by disulphide bridges.
  • In mammals, including humans, insulin is synthesised as a prohormone (like a
    pro-enzyme, the pro-hormone also needs to be processed before it becomes a fully
    mature and functional hormone) which contains an extra stretch called the C peptide.
  • This C peptide is not present in the mature insulin and is removed during maturation
    into insulin.
  • The main challenge for production of insulin using rDNA techniques was getting insulin assembled into a mature form.
  • In 1983, Eli Lilly an American company prepared two DNA sequences corresponding to A and B, chains of human insulin and introduced them in plasmids of E. coli to produce insulin chains. Chains A and B were produced separately, extracted and combined by creating disulfide bonds to form human insulin.

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Gene Therapy

  • Gene therapy is the corrective therapy for hereditary disease.
    Gene therapy is a collection of methods that allows correction of a gene defect that has
    been diagnosed in a child/embryo. Here genes are inserted into a person’s cells and
    tissues to treat a disease.
  • Correction of a genetic defect involves delivery of a normal gene into the individual or
    embryo to take over the function of and compensate for the non-functional gene.
  • The first clinical gene therapy was given in 1990 to a 4-year old girl with adenosine
    deaminase (ADA) deficiency. This enzyme is crucial for the immune system to function.
  • The disorder is caused due to the deletion of the gene for adenosine deaminase.
  • ADA deficiency can be cured by bone marrow transplantation or by enzyme
    replacement therapy, in which functional ADA is given to the patient by injection.
    But the problem with both of these approaches that they are not completely curative.
  • In gene therapy, lymphocytes from the blood of the patient are grown in a culture
    outside the body. A functional ADA cDNA (using a retroviral vector) is then introduced
    into these lymphocytes, which are subsequently returned to the patient. However, as
    these cells are not immortal, the patient requires periodic infusion of such genetically
    engineered lymphocytes. However, if the gene isolate from marrow cells producing
    ADA is introduced into cells at early embryonic stages, it could be a permanent cure.

Molecular Diagnosis

  • For effective treatment of a disease, early diagnosis and understanding its
    pathophysiology is very important but using conventional methods of diagnosis (serum and urine analysis, etc.) early detection is not possible.
  • Recombinant DNA technology, Polymerase Chain Reaction (PCR) and Enzyme Linked
    Immuno-sorbent Assay (ELISA) are some of the techniques that serve the purpose of
    early diagnosis.
  • Presence of a pathogen (bacteria, viruses, etc.) is normally suspected only when the
    pathogen has produced a disease symptom. By this time the concentration of pathogen is already very high in the body. However, very low concentration of a bacteria or virus (at a time when the symptoms of the disease are not yet visible) can be detected by amplification of their nucleic acid by PCR.
  • PCR is now routinely used to detect HIV in suspected AIDS patients. It is being used to
    detect mutations in genes in suspected cancer patients too. It is a powerful techqnique
    to identify many other genetic disorders.
  • PCR –
    A single stranded DNA or RNA, tagged with a radioactive molecule (probe) is allowed to hybridise to its complementary DNA in a clone of cells followed by detection using
    autoradiography. The clone having the mutated gene will hence not appear on the
    photographic film, because the probe will not have complimentarity with the mutated
    gene.
  • ELISA is based on the principle of antigen-antibody interaction. Infection by pathogen can be detected by the presence of antigens (proteins, glycoproteins, etc.) or by detecting the antibodies synthesised against the pathogen.

TRANSGENIC ANIMALS

  • Animals that have had their DNA manipulated to possess and express an extra (foreign) gene are known as transgenic animals.
  • Transgenic rats, rabbits, pigs, sheep, cows and fish have been produced, although over 95 per cent of all existing transgenic animals are mice.
  • common reasons to produce transgenic animals:
    (i) Normal physiology and development:
    Transgenic animals can be specifically designed to allow the study of how genes are
    regulated, and how they affect the normal functions of the body and its
    development, e.g., study of complex factors involved in growth such as insulin-like
    growth factor.
    By introducing genes from other species that alter the formation of this factor and
    studying the biological effects that result, information is obtained about the
    biological role of the factor in the body.
    (ii) Study of disease:
    Many transgenic animals are designed to increase our understanding of how genes
    contribute to the development of disease. These are specially made to serve as
    models for human diseases so that Investigation of new treatments for diseases is
    made possible.
    Today transgenic models exist for many human diseases such as cancer, cystic
    fibrosis, rheumatoid arthritis and Alzheimer’s.
    (iii) Biological products:
    Medicines required to treat certain human diseases can contain biological products,
    but such products are often expensive to make.
    Transgenic animals that produce useful biological products can be created by the
    introduction of the portion of DNA (or genes) which codes for a particular product
    such as human protein (α-1-antitrypsin) used to treat emphysema.
    Similar attempts are being made for treatment of phenylketonuria (PKU) and cystic
    fibrosis.
    In 1997, the first transgenic cow, Rosie, produced human protein-enriched milk (2.4
    grams per litre). The milk contained the human alpha-lactalbumin and was
    nutritionally a more balanced product for human babies than natural cow-milk.
    (iv) Vaccine safety:
    Transgenic mice are being developed for use in testing the safety of vaccines before
    they are used on humans.
    Transgenic mice are being used to test the safety of the polio vaccine. If successful
    and found to be reliable, they could replace the use of monkeys to test the safety of
    batches of the vaccine.
    (v) Chemical safety testing:
    This is known as toxicity/safety testing. The procedure is the same as that used for
    testing toxicity of drugs.
    Transgenic animals are made that carry genes which make them more sensitive to
    toxic substances than non-transgenic animals. They are then exposed to the toxic
    substances and the effects studied. Toxicity testing in such animals will allow us to
    obtain results in less time.

    ETHICAL ISSUES

    The manipulation of living organisms by the human race cannot go on any further, without
    regulation. Some ethical standards are required to evaluate the morality of all human
    activities that might help or harm living organisms.
    Going beyond the morality of such issues, the biological significance of such things is also
    important. Genetic modification of organisms can have unpredicatable results when such
    organisms are introduced into the ecosystem.
    Therefore, the Indian Government has set up organisations such as GEAC (Genetic
    Engineering Approval Committee), which will make decisions regarding the validity of GM
    research and the safety of introducing GM-organisms for public services.

Bio-patent:

  • The modification/usage of living organisms for public services (as food and medicine
    sources, for example) has also created problems with patents granted for the same.
  • There is growing public anger that certain companies are being granted patents for
    products and technologies that make use of the genetic materials, plants and other
    biological resources that have long been identified, developed and used by farmers and
    indigenous people of a specific region/country.
  • Rice is an important food grain, the presence of which goes back thousands of years in
    Asia’s agricultural history. There are an estimated 200,000 varieties of rice in India
    alone. The diversity of rice in India is one of the richest in the world.
  • Basmati rice is distinct for its unique aroma and flavour and 27 documented varieties of Basmati are grown in India. There is reference to Basmati in ancient texts, folklore and poetry, as it has been grown for centuries.
  • In 1997, an American company got patent rights on Basmati rice through the US Patent and Trademark Office. This allowed the company to sell a ‘new’ variety of Basmati, in the US and abroad.
  • This ‘new’ variety of Basmati had actually been derived from Indian farmer’s varieties.
    Indian Basmati was crossed with semi-dwarf varieties and claimed as an invention or a novelty.
  • The patent extends to functional equivalents, implying that other people selling
    Basmati rice could be restricted by the patent.
  • Several attempts have also been made to patent uses, products and processes based
    on Indian traditional herbal medicines, e.g., turmeric neem.
  • If we are not vigilant and we do not immediately counter these patent applications,
    other countries/individuals may encash on our rich legacy and we may not be able to
    do anything about it.

Biopiracy

  • It is the term used to refer to the use of bio-resources by multinational companies and
    other organisations without proper authorisation from the countries and people
    concerned without compensatory payment.
  • Most of the industrialised nations are rich financially but poor in biodiversity and
    traditional knowledge. In contrast the developing and the underdeveloped world is rich in biodiversity and traditional knowledge related to bio-resources. Traditional
    knowledge related to bio-resources can be exploited to develop modern applications
    and can also be used to save time, effort and expenditure during their
    commercialisation.
  • There has been growing realisation of the injustice, inadequate compensation and
    benefit sharing between developed and developing countries. Therefore, some nations
    are developing laws to prevent such unauthorised exploitation of their bio-resources
    and traditional knowledge.
  • The Indian Parliament has recently cleared the second amendment of the Indian
    Patents Bill, that takes such issues into consideration, including patent terms
    emergency provisions and research and development initiative.

 

To download notes in pdf format please click on the following link.

CHAPTER 12 : BIOTECHNOLOGY AND ITS APPLICATIONS

CHAPTER 11 : BIOTECHNOLOGY: PRINCIPLES AND PROCESSES

Chapter 11

Biotechnology : Principles and Processes

[you can download the notes from the link given at the end of theory]

Biotechnology deals with techniques of using live organisms or enzymes from organisms to produce products and processes useful to humans.

  • Traditional form – based on natural capabilities of microorganisms. making curd, bread or wine, which are all microbe-mediated processes, could also be thought as a form of biotechnology. However, it is used in a restricted sense today,
  • Modern form – it uses genetically modified organisms to achieve the same on a larger scale. Further, many other processes/techniques are also included under biotechnology. For example, in vitro fertilisation leading to a ‘test-tube’ baby, synthesising a gene and using it, developing a DNA vaccine or correcting a defective gene, are all part of biotechnology.
  • The European Federation of Biotechnology (EFB) has given a definition of biotechnology that encompasses both traditional view and modern molecular biotechnology. The definition given by EFB is as follows:

‘The integration of natural science and organisms, cells, parts thereof, and molecular analogues for products and services’.

PRINCIPLES OF BIOTECHNOLOGY

  • Among many, the two core techniques that enabled birth of modern biotechnology are :
    • Genetic engineering: Techniques to alter the chemistry of genetic material (DNA and RNA),to introduce these into host organisms and thus change the phenotype of the host organism.
    • Maintenance of sterile (microbial contamination-free) ambience in chemical engineering processes to enable growth of only the desired microbe/eukaryotic cell in large quantities for the manufacture of biotechnological products like antibiotics, vaccines, enzymes, etc.
  • Sexual reproduction has many advantages over asexual reproduction. The former provides opportunities for variations and formulation of unique combinations of genetic setup, some of which may be beneficial to the organism as well as the population. Asexual reproduction preserves the genetic information, while sexual reproduction permits variation.
  • Traditional hybridisation procedures used in plant and animal breeding, very often lead to inclusion and multiplication of undesirable genes along with the desired genes. The techniques of genetic engineering which include creation of recombinant DNA, use of gene cloning and gene transfer, overcome this limitation and allow us to isolate and introduce only one or a set of desirable genes without introducing undesirable genes into the target organism.
  • A piece of DNA, which is somehow transferred into an alien organism, most likely would not be able to multiply itself in the progeny cells of the organism. But, when it gets integrated into the genome of the recipient, it may multiply and be inherited along with the host DNA. This is because the alien piece of DNA has become part of a chromosome, which has the ability to replicate.
  • In a chromosome there is a specific DNA sequence called the origin of replication, which is responsible for initiating replication. Therefore, for the multiplication of any alien piece of DNA in an organism it needs to be a part of a chromosome(s) which has a specific sequence known as ‘origin of replication’. Thus, an alien DNA is linked with the origin of replication, so that, this alien piece of DNA can replicate and multiply itself in the host organism. This can also be called as cloning or making multiple identical copies of any template DNA.
  • The construction of the first recombinant DNA emerged from the possibility of linking a gene encoding antibiotic resistance with a native plasmid (autonomously replicating circular extra-chromosomal DNA) of  Salmonella typhimurium.
  • Stanley Cohen and Herbert Boyer accomplished this in 1972 by isolating the antibiotic resistance gene by cutting out a piece of DNA from a plasmid which was responsible for conferring antibiotic resistance.
  • The cutting of DNA at specific locations became possible with the discovery of the so-called ‘molecular scissors’- restriction enzymes.
  • The cut piece of DNA was then linked with the plasmid DNA. These plasmid DNA act as vectors to transfer the piece of DNA attached to it. A plasmid can be used as vector to deliver an alien piece of DNA into the host organism.
  • The linking of antibiotic resistance gene with the plasmid vector became possible with the enzyme DNA ligase, which acts on cut DNA molecules and joins their ends. This makes a new combination of circular autonomously replicating DNA created in vitro and is known as recombinant DNA.
  • When this DNA is transferred into Escherichia coli, a bacterium closely related to Salmonella, it could replicate using the new host’s DNA polymerase enzyme and make multiple copies. The ability to multiply copies of antibiotic resistance gene in coli was called cloning of antibiotic resistance gene in E. coli.
  • there are three basic steps in genetically modifying an organism
    • identification of DNA with desirable genes;
    • introduction of the identified DNA into the host;
    • maintenance of introduced DNA in the host and transfer of the DNA to its progeny.

TOOLS OF RECOMBINANT DNA TECHNOLOGY

Key tools of Recombinant DNA technology are – restriction enzymes, polymerase enzymes, ligases, vectors and the host organism.

  1. Restriction Enzymes

  • In 1963, the two enzymes responsible for restricting the growth of bacteriophage in Escherichia coli were isolated. One of these added methyl groups to DNA, while the other cut DNA. The later was called restriction endonuclease.
  • The first restriction endonuclease isolated – Hind II.
  • Restriction endonuclease cut DNA molecules at a particular point by recognising a specific sequence of base pairs. This specific base sequence is known as the recognition sequence.(For Hind II – sequence of 6 base pairs).
  • Today we know more than 900 restriction enzymes that have been isolated from over 230 strains of bacteria each of which recognise different recognition sequences.

Naming of enzymes –

  • First letter of the name comes from the genes
  • The second two letters come from the species of the prokaryotic cell from which they were isolated, e.g., EcoRI comes from Escherichia coli RY 13.
  • Next letter derived from the name of strain.
  • Roman numbers following the names indicate the order in which the enzymes were isolated from that strain of bacteria.

Action of enzyme –

  • Restriction enzymes belong to a larger class of enzymes called nucleases. These are of two kinds; exonucleasesand endonucleases.
  • Exonucleases remove nucleotides from the ends of the DNA whereas, endonucleases make cuts at specific positions within the DNA.
  • Each restriction endonuclease functions by ‘inspecting’ the length of a DNA sequence. Once it finds its specific recognition sequence, it will bind to the DNA and cut each of the two strands of the double helix at specific points in their sugar -phosphate backbones.
  • Each restriction endonuclease recognises a specific palindromic nucleotide sequences in the DNA.
  • The palindrome in DNA is a sequence of base pairs that reads same on the two strands when orientation of reading is kept the same. For example, the following sequences reads the same on the two strands in 5→3 This is also true if read in the 3→5direction.

5—— GAATTC —— 3

3—— CTTAAG —— 5

  • Restriction enzymes cut the strand of DNA a little away from the centre of the palindrome sites, but between the same two bases on the opposite strands. This leaves single stranded portions at the ends. There are overhanging stretches called sticky ends on each strand.
  • These are named so because they form hydrogen bonds with their complementary cut counterparts. This stickiness of the ends facilitates the action of the enzyme DNA ligase.
  • Restriction endonucleases are used in genetic engineering to form ‘recombinant’ molecules of DNA, which are composed of DNA from different sources/genomes.
  • When cut by the same restriction enzyme, the resultant DNA fragments have the same kind of ‘sticky-ends’ and, these can be joined together (end-to-end) using DNA ligases .
  • Normally, unless one cuts the vector and the source DNA with the same restriction enzyme, the recombinant vector molecule cannot be created.

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Fig: Steps in formation of recombinant DNA by action of restriction endonuclease enzyme – EcoRI

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Fig: Diagrammatic representation of recombinant DNA technology

 

Separation and isolation of DNA fragments :

  • The cutting of DNA by restriction endonucleases results in the fragmentes of DNA. These fragments can be separated by a technique known as gel electrophoresis.
  • Since DNA fragments are negatively charged molecules they can be separated by forcing them to move towards the anode under an electric field through a medium/matrix. Nowadays the most commonly used matrix is agarose which is a natural polymer extracted from sea weeds.
  • The DNA fragments separate (resolve) according to their size through sieving effect provided by the agarose gel. Hence, the smaller the fragment size, the farther it moves.
  • The separated DNA fragments can be visualised only after staining the DNA with a compound known as ethidium bromide followed by exposure to UV radiation.
  • We can see bright orange coloured bands of DNA in aethidium bromide stained gel exposed to UV light.
  • The separated bands of DNA are cut out from the agarose gel and extracted from the gel piece. This step is known as elution. The DNA fragments purified in this way are used in constructing recombinant DNA by joining them with cloning vectors.

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Fig: A typical agarose gel electrophoresis showing migration of undigested (lane 1) and digested set of DNA fragments (lane 2 to 4)
  1. Cloning Vectors

  • Plasmids and bacteriophages have the ability to replicate within bacterial cells independent of the control of chromosomal DNA.
  • Bacteriophages because of their high number per cell, have very high copy numbers of their genome within the bacterial cells.
  • If we are able to link an alien piece of DNA with bacteriophage or plasmid DNA, we can multiply its numbers equal to the copy number of the plasmid or bacteriophage.
  • Vectors used at present, are engineered in such way that they help easy linking of foreign DNA and selection of recombinants from non-recombinants.

Features required to facilitate cloning into a vector.

Origin of replication (ori):

  • This is a sequence from where replication starts and any piece of DNA when linked to this sequence can be made to replicate within the host cells.
  • This sequence is also responsible for controlling the copy number of the linked DNA.
  • So, if one wants to recover many copies of the target DNA it should be cloned in a vector whose origin support high copy number.

    Selectable marker :

  • In addition to ‘ori’, the vector requires a selectable marker, which helps in identifying and eliminating nontransformants and selectively permitting the growth of the transformants.
  • Transformation is a procedure through which a piece of DNA is introduced in a host bacterium.
  • Normally, the genes encoding resistance to antibiotics such as ampicillin, chloramphenicol, tetracycline or kanamycin, etc., are considered useful selectable markers for coli. The normal E. coli cells do not carry resistance against any of these antibiotics.

    Cloning sites:

  • In order to link the alien DNA, the vector needs to have very few, preferably single, recognition sites for the commonly used restriction enzymes.
  • Presence of more than one recognition sites within the vector will generate several fragments, which will complicate the gene cloning.
  • The ligation of alien DNA is carried out at a restriction site present in one of the two antibiotic resistance
  • For example, you can ligate a foreign DNA at the Bam H I site of tetracycline resistance gene in the vector pBR322. The recombinant plasmids will lose tetracycline resistance due to insertion of foreign DNA but can still be selected out from non-recombinant ones by plating the transformants on ampicillin containing medium. The transformants growing on ampicillin containing medium are then transferred on a medium containing tetracycline. The recombinants will grow in ampicillin containing medium but not on that containing tetracycline. But, nonrecombinants will grow on the medium containing both the antibiotics. In this case, one antibiotic resistance gene helps in selecting the transformants, whereas the other antibiotic resistance gene gets ‘inactivated due to insertion’ of alien DNA, and helps in selection of recombinants.
  • Selection of recombinants due to inactivation of antibiotics is a cumbersome procedure because it requires simultaneous plating on two plates having different antibiotics. Therefore, alternative selectable markers have been developed which differentiate recombinants from non-recombinants on the basis of their ability to produce colour in the presence of a chromogenic substrate.
  • In this, a recombinant DNA is inserted within the coding sequence of an enzyme, a-galactosidase. This results into inactivation of the enzyme, which is referred to as insertional inactivation. The presence of a chromogenic substrate gives blue coloured colonies if the plasmid in the bacteria does not have an insert. Presence of insert results into insertional inactivation of the a-galactosidase and the colonies do not produce any colour, these are identified as recombinant colonies.

    Vectors for cloning genes in plants and animals :

  • Viruses and bacteria are used to transfer genes into plants and animals which transform eukaryotic cells and force them to do what the bacteria or viruses want.
  • For example, Agrobacterioumtumifaciens, a pathogen of several dicot plants is able to deliver a piece of DNA known as ‘T-DNA’ to transform normal plant cells into a tumor and direct these tumor cells to produce the chemicals required by the pathogen.
  • Similarly, retroviruses in animals have the ability to transform normal cells into cancerous
  • A better understanding of the art of delivering genes by pathogens in their eukaryotic hosts has generated knowledge to transform these tools of pathogens into useful vectors for delivering genes of interest to humans.
  • The tumor inducing (Ti) plasmid of Agrobacterium tumifacienshas now been modified into a cloning vector which is no more pathogenic to the plants but is still able to use the mechanisms to deliver genes of our interest into a variety of plants. Similarly, retroviruses have also been disarmed and are now used to deliver desirable genes into animal cells. So, once a gene or a DNA fragment has been ligated into a suitable vector it is transferred into a bacterial, plant or animal host (where it multiplies).
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Fig: E. coli cloning vector pBR322 showing restriction sites (Hind III, EcoR I, BamH I, Sal I, PvuII, PstI, ClaI), ori and antibiotic resistance genes (ampR and tetR). Rop codes for the proteins involved in the replication of the plasmid.
  1. Competent Host (For Transformation with Recombinant DNA)

  • Since DNA is a hydrophilic molecule, it cannot pass through cell membranes. In order to force bacteria to take up the plasmid, the bacterial cells must first be made ‘competent’ to take up DNA.
  • This is done by treating them with a specific concentration of a divalent cation, such as calcium, which increases the efficiency with which DNA enters the bacterium through pores in its cell wall.
  • Recombinant DNA can then be forced into such cells by incubating the cells with recombinant DNA on ice, followed by placing them briefly at 420oC (heat shock), and then putting them back on ice. This enables the bacteria to take up the recombinant DNA.
  • In micro-injection method, recombinant DNA is directly injected into the nucleus of an animal cell.
  • In another method, suitable for plants, cells are bombarded with high velocity micro-particles of gold or tungsten coated with DNA in a method known as biolisticsor gene gun.
  • And the last method uses ‘disarmed pathogen’ vectors, which when allowed to infect the cell, transfer the recombinant DNA into the host.

PROCESSES OF RECOMBINANT DNA TECHNOLOGY

Recombinant DNA technology involves several steps in specific sequence such as –

  • isolation of DNA,
  • fragmentation of DNA by restriction endonucleases,
  • isolation of a desired DNA fragment,
  • ligation of the DNA fragment into a vector,
  • transferring the recombinant DNA into the host,
  • culturing the host cells in a medium at large scale and
  • extraction of the desired product.
  1. Isolation of the Genetic Material (DNA)

  • Nucleic acid is the genetic material of all organisms without exception. In majority of organisms this is deoxyribonucleic acid or DNA.
  • In order to cut the DNA with restriction enzymes, it needs to be in pure form, free from other macro-molecules. Since the DNA is enclosed within the membranes, we have to break the cell open to release DNA along with other macromolecules such as RNA, proteins, polysaccharides and also lipids. This can be achieved by treating the bacterial cells/plant or animal tissue with enzymes such as lysozyme (bacteria), cellulase(plant cells), chitinase(fungus).
  • genes are located on long molecules of DNA interwined with proteins such as histones.
  • RNA can be removed by treatment with ribonuclease whereas proteins can be removed by treatment with protease. Other molecules can be removed by appropriate treatments and purified DNA ultimately precipitates out after the addition of chilled ethanol. This can be seen as collection of fine threads in the suspension.

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Fig: DNA thatseparates out can beremoved by spooling
  1. Cutting of DNA at Specific Locations

  • Restriction enzyme digestions are performed by incubating purified DNA molecules with the restriction enzyme, at the optimal conditions for that specific enzyme.
  • Agarose gel electrophoresis is employed to check the progression of a restriction enzyme digestion. DNA is a negatively charged molecule, hence it moves towards the positive electrode (anode).
  • The process is repeated with the vector DNA also.
  • The joining of DNA involves several processes. After having cut the source DNA as well as the vector DNA with a specific restriction enzyme, the cut out ‘gene of interest’ from the source DNA and the cut vector with space are mixed and ligase is added. This results in the preparation of recombinant DNA.
  1. Amplification of Gene of Interest using PCR (Polymerase Chain Reaction)

  • In this reaction, multiple copies of the gene (or DNA) of interest is synthesisedin vitro using two sets of primers (small chemically synthesised oligonucleotides that are complementary to the regions of DNA) and the enzyme DNA polymerase.
  • The enzyme extends the primers using the nucleotides provided in the reaction and the genomic DNA as template.
  • If the process of replication of DNA is repeated many times, the segment of DNA can be amplified to approximately billion times.
  • Such repeated amplification is achieved by the use of a thermostable DNA polymerase (isolated from a bacterium, Thermusaquaticus), which remain active during the high temperature induced denaturation of double stranded DNA.
  • The amplified fragment if desired can now be used to ligate with a vector for further cloning.

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Fig: Polymerase chain reaction (PCR) : Each cycle has three steps: (i) Denaturation;

(ii) Primer annealing; and (iii) Extension of primers

  1. Insertion of Recombinant DNA into the Host Cell/Organism

  • There are several methods of introducing the ligated DNA into recipient cells. Recipient cells after making them ‘competent’ to receive, take up DNA present in its surrounding.
  • So, if a recombinant DNA bearing gene for resistance to an antibiotic (e.g., ampicillin) is transferred into coli cells, the host cells become transformed into ampicillin-resistant cells. If we spread the transformed cells on agar plates containing ampicillin, only transformants will grow, untransformed recipient cells will die. Since, due to ampicillin resistance gene, one is able to select a transformed cell in the presence of ampicillin. The ampicillin resistance gene in this case is called a selectable marker.
  1. Obtaining the Foreign Gene Product

  • When you insert a piece of alien DNA into a cloning vector and transfer it into a bacterial, plant or animal cell, the alien DNA gets multiplied.
  • In almost all recominant technologies, the ultimate aim is to produce a desirable protein. Hence, there is a need for the recombinant DNA to be expressed.
  • The foreign gene gets expressed under appropriate conditions. The expression of foreign genes in host cells involve understanding many technical details.
  • After having cloned the gene of interest and having optimised the conditions to induce the expression of the target protein, one has to consider producing it on a large scale.
  • If any protein encoding gene is expressed in a heterologous host, is called a recombinant protein.
  • The cells harbouring cloned genes of interest may be grown on a small scale in the laboratory. The cultures may be used for extracting the desired protein and then purifying it by using different separation techniques.
  • The cells can also be multiplied in a continuous culture system wherein the used medium is drained out from one side while fresh medium is added from the other to maintain the cells in their physiologically most active log/exponential phase. This type of culturing method produces a larger biomass leading to higher yields of desired protein.
  • Small volume cultures cannot yield appreciable quantities of products. To produce in large quantities, the development of bioreactors, where large volumes (100-1000 litres) of culture can be processed, was required. Thus, bioreactors can be thought of as vessels in which raw materials are biologically converted into specific products, individual enzymes, etc., using microbial plant, animal or human cells. A bioreactor provides the optimal conditions for achieving the desired product by providing optimum growth conditions (temperature, pH, substrate, salts, vitamins, oxygen).
  • A stirred-tank reactor is usually cylindrical or with a curved base to facilitate the mixing of the reactor contents. The stirrer facilitates even mixing and oxygen availability throughout the bioreactor. Alternatively air can be bubbled through the reactor.
  • The bioreactor has an agitator system, an oxygen delivery system and a foam control system, a temperature control system, pH control system and sampling ports so that small volumes of the culture can be withdrawn periodically.

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Fig: (a) Simple stirred-tank bioreactor; (b) Sparged stirred-tank bioreactor through whichsterile air bubbles are sparged

 

  1. Downstream Processing
  • After completion of the biosynthetic stage, the product has to be subjected through a series of processes before it is ready for marketing as a finished The processes include separation and purification, which are collectively referred to as downstream processing.
  • The product has to be formulated with suitable preservatives. Such formulation has to undergo thorough clinical trials as in case of drugs. Strict quality control testing for each product is also required. The downstream processing and quality control testing vary from product to product.

To download notes in pdf format please click on the following link.

CHAPTER 11 : BIOTECHNOLOGY: PRINCIPLES AND PROCESSES

CHAPTER 2 : SEXUAL REPRODUCTION IN FLOWERING PLANTS

CHAPTER 2

SEXUAL REPRODUCTION IN FLOWERING PLANTS

[you can download the notes from the link given at the end of theory]

  • All flowering plants show sexual reproduction.

Flower – A Fascinating Organ of Angiosperms

  • Flowers are objects of aesthetic, ornamental, social, religious and cultural value – they have always been used as symbols for conveying important human feelings such as love, affection, happiness, grief, mourning, etc.

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  • Pre-fertilisation: Structures and Events

  • Several hormonal and structural changes are initiated which lead to the differentiation and further development of the floral primordium.
  • Inflorescences are formed which bear the floral buds and then the flowers.
  • In the flower the male and female reproductive structures, the androecium and the gynoecium differentiate and develop.
  • The androecium consists of a whorl of stamens representing the male reproductive organ and the gynoecium represents the female reproductive organ.

Stamen, Microsporangium and Pollen Grain

  • A typical stamen has two parts –

the long and slender stalk called the filament,

and the terminal generally bilobed structure called the anther.

  • The proximal end of the filament is attached to the thalamus or the petal of the flower.
  • The number and length of stamens are variable in flowers of different species.
  • A typical angiosperm anther is bilobed with each lobe having two theca, i.e., they are
  • The anther is a four-sided (tetragonal) structure consisting of four microsporangia located at the corners, two in each lobe. tetrasporangiate
  • The microsporangia develop further and become pollen sacs. They extend longitudinally all through the length of an anther and are packed with pollen grains.

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Structure of microsporangium:

  • In a transverse of an anther section, a typical microsporangium appears nearcircular in outline.
  • It is generally surrounded by four wall layers :- the epidermis, endothecium, middle layers and the tapetum.
  • The outer three wall layers perform the function of protection and help in dehiscence of anther to release the pollen.
  • The innermost wall layer is the It nourishes the developing pollen grains. Cells of the tapetum possess dense cytoplasm and generally have more than one nucleus.
  • When the anther is young, a group of compactly arranged homogenous cells called the sporogenous tissue occupies the centre of each microsporangium.

Microsporogenesis :

  • As the anther develops, the cells of the sporogenous tissue undergo meiotic divisions to form microspore tetrads, which is haploid.
  • As each cell of the sporogenous tissue is capable of giving rise to a microspore tetrad. Each one is a potential pollen or microspore mother cell (PMC).
  • The process of formation of microspores from a pollen mother cell through meiosis is called
  • The microspores, as they are formed, are arranged in a cluster of four cells-the microspore tetrad.
  • As the anthers mature and dehydrate, the microspores dissociate from each other and develop into pollen grains.
  • Inside each microsporangium several thousands of microspores or pollen grains are formed that are released with the dehiscence of anther.

 

Pollen grain:

  • The pollen grains represent the male gametophytes.
  • Pollen grains are generally spherical measuring about 25-50 micrometers in diameter.
  • It has a prominent two-layered wall.
    • The hard outer layer called the exine is made up of sporopollenin which is one of the most resistant organic material known. It can withstand high temperatures and strong acids and alkali. No enzyme that degrades sporopollenin is so far known. Pollen grain exine has prominent apertures called germ pores where sporopollenin is absent. Pollen grains are well- preserved as fossils because of the presence of sporopollenin. The exine exhibits a fascinating array of patterns and designs.
    • The inner wall of the pollen grain is called the It is a thin and continuous layer made up of cellulose and pectin.
  • The cytoplasm of pollen grain is surrounded by a plasma membrane.
  • When the pollen grain is mature it contains two cells, the vegetative cell and generative cell.
    • The vegetative cell is bigger, has abundant food reserve and a large irregularly shaped nucleus.
    • The generative cell is small and floats in the cytoplasm of the vegetative cell. It is spindle shaped with dense cytoplasm and a nucleus.
  • In over 60 per cent of angiosperms, pollen grains are shed at this 2-celled stage. In the remaining species, the generative cell divides mitotically to give rise to the two male gametes before pollen grains are shed (3-celled stage).

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  • Pollen grains of many species cause severe allergies and bronchial afflictions in some people often leading to chronic respiratory disorders – asthma, bronchitis, etc.
  • It may be mentioned that Parthenium or carrot grass that came into India as a contaminant with imported wheat, has become ubiquitous in occurrence and causes pollen allergy.
  • Pollen grains are rich in nutrients. It has become a fashion in recent years to use pollen tablets as food supplements.
  • In western countries, a large number of pollen products in the form of tablets and syrups are available in the market.
  • Pollen consumption has been claimed to increase the performance of athletes and race horses.
  • When once they are shed, pollen grains have to land on the stigma before they lose viability if they have to bring about fertilisation.
  • The period for which pollen grains remain viable is highly variable and to some extent depends on the prevailing temperature and humidity.
  • In some cereals such as rice and wheat, pollen grains lose viability within 30 minutes of their release, and in some members of Rosaceae, Leguminoseae and Solanaceae, they maintain viability for months.
  • It is possible to store pollen grains of a large number of species for years in liquid nitrogen (-1960C). Such stored pollen can be used as pollen banks, similar to seed banks, in crop breeding programmes.

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The Pistil, Megasporangium (ovule) and Embryo sac (Female gametophyte):

  • The gynoecium represents the female reproductive part of the flower.
  • It may consist of a single pistil (monocarpellary) or may have more than one pistil (multicarpellary). When there are more than one, the pistils may be fused together (syncarpous) or may be free (apocarpous).
  • Each pistil has three parts – the stigma, style and ovary.

The stigma serves as a landing platform for pollen grains.

The style is the elongated slender part beneath the stigma.

The basal bulged part of the pistil is the ovary. Inside the ovary is the ovarian cavity (locule). The placenta is located inside the ovarian cavity.

  • Arising from the placenta are the megasporangia, commonly calledovules.
  • The number of ovules in an ovary may be one (wheat, paddy, mango) to many (papaya, water melon, orchids).

The Megasporangium (Ovule):

  • The ovule is a small structure attached to the placenta by means of a stalk calledfunicle.
  • The body of the ovule fuses with funicle in the region calledhilum. Thus, hilum represents the junction between ovule and funicle.
  • Each ovule has one or two protective envelopes calledinteguments. Integuments encircle the ovule except at the tip where a small opening called themicropyle is organised.
  • Opposite the micropylar end, is thechalaza, representing the basal part of the ovule.
  • Enclosed within the integuments is a mass of cells called thenucellus. Cells of the nucellus have abundant reserve food materials.
  • Located in the nucellus is theembryo sac or female gametophyte.
  • An ovule generally has a single embryo sac formed from a megaspore through reduction division.

Megasporogenesis:

  • The process of formation of megaspores from the megaspore mother cell is called
  • Ovules generally differentiate a single megaspore mother cell (MMC) in the micropylar region of the nucellus. It is a large cell containing dense cytoplasm and a prominent nucleus.
  • The MMC undergoes meiotic division.Meiosis results in the production of four megaspores.

Female gametophyte:

  • In a majority of flowering plants, one of the megaspores is functional while the other three degenerate.
  • Only the functional megaspore develops into the female gametophyte (embryo sac). This method of embryo sac formation from a single megaspore is termed monosporic
  • Ploidy of the cells of the

nucellus – 2n,

MMC – 2n,

the functional megaspore – n,

female gametophyte – n.

  • Process of development –

    The nucleus of the functional megaspore divides mitotically to form two nuclei which move to the opposite poles, forming the 2-nucleate embryo sac. Two more sequential mitotic nuclear divisions result in the formation of the 4-nucleate and later the 8-nucleate stages of the embryo sac.

these mitotic divisions are strictly free nuclear. (nuclear divisions are not followed immediately by cell wall formation.)

After the 8-nucleate stage, cell walls are laid down leading to the organisation of the typical female gametophyte or embryo sac.

  • Structure –

Six of the eight nuclei are surrounded by cell walls and organised into cells; the remaining two nuclei, called polar nuclei are situated below the egg apparatus in the large central cell.

Three cells are grouped together at the micropylar end and constitute the egg apparatus. The egg apparatus, in turn, consists of two synergids and one egg cell.

The synergids have special cellular thickenings at the micropylar tip called filiform apparatus, which play an important role in guiding the pollen tubes into the synergid.

Three cells are at the chalazal end and are called the antipodals.

The large central cell, as mentioned earlier, has two polar nuclei.

Thus, a typical angiosperm embryo sac, at maturity, though 8-nucleate is 7-celled.

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Pollination

  • both male and female gametes are non-motile, so they have to be brought together for fertilisation to occur.
  • The transfer of pollen grains (shed from the anther) to the stigma of a pistil is termedpollination.
  • Kinds of Pollination : Depending on the source of pollen, pollination can be divided into three types.
    • Autogamy : Transfer of pollen grains from the anther to the stigma of the same flower. In a normal flower which opens and exposes the anthers and the stigma, complete autogamy is rather rare. Autogamy in such flowers requires synchrony in pollen release and stigma receptivity and also, the anthers and the stigma should lie close to each other so that self-pollination can occur.

Some plants such as Viola (common pansy), Oxalis, and Commelina produce two types of flowers –

chasmogamous flowers which are similar to flowers of other species with exposed anthers and stigma, and

cleistogamous flowers which do not open at all. In such flowers, the anthers and stigma lie close to each other. When anthers dehisce in the flower buds, pollen grains come in contact with the stigma to effect pollination. Thus, cleistogamous flowers are invariably autogamous as there is no chance of cross-pollen landing on the stigma. Cleistogamous flowers produce assured seed-set even in the absence of pollinators.

  • Geitonogamy – Transfer of pollen grains from the anther to the stigma of another flower of the same plant. Although geitonogamy is functionally cross-pollination involving a pollinating agent, genetically it is similar to autogamy since the pollen grains come from the same plant.
  • Xenogamy – Transfer of pollen grains from anther to the stigma of a different plant. This is the only type of pollination which during pollination brings genetically different types of pollen grains to the stigma.

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 Agents of Pollination:

  • Plants use two abiotic (wind and water) and one biotic (animals) agents to achieve pollination.
  • Majority of plants use biotic agents for pollination. Only a small proportion of plants use abiotic agents.
  • Pollen grains coming in contact with the stigma is a chance factor in both wind and water pollination. To compensate for this uncertainties and associated loss of pollen grains, the flowers produce enormous amount of pollen when compared to the number of ovules available for pollination.
  • Wind poliination –
    • Pollination by wind is more common amongst abiotic pollinations.
    • Wind pollination requires that the pollen grains are light and non-sticky so that they can be transported in wind currents.
    • These plants often possess well-exposed stamens (so that the pollens are easily dispersed into wind currents) and large often-feathery stigma to easily trap air-borne pollen grains.
    • Wind- pollinated flowers often have a single ovule in each ovary and numerous flowers packed into an inflorescence.
    • g., corn cob, grasses.
  • Water pollination –
    • Pollination by water is quite rare in flowering plants, present in mostly monocotyledons.
    • water is a regular mode of transport for the male gametes among the lower plant groups such as algae, bryophytes and pteridophytes.
    • g., Vallisneria and Hydrilla (fresh water) and Zostera (marine sea-grasses).
    • Not all aquatic plants use water for pollination.
    • In a majority of aquatic plants such as water hyacinth and water lily, the flowers emerge above the level of water and are pollinated by insects or wind as in most of the land plants.
    • In Vallisneria, the female flower reach the surface of water by the long stalk and the male flowers or pollen grains are released on to the surface of water. They are carried passively by water currents; some of them eventually reach the female flowers and the stigma.
    • In seagrasses, female flowers remain submerged in water and the pollen grains are released inside the water. Pollen grains in many such species are long, ribbon like and they are carried passively inside the water; some of them reach the stigma and achieve pollination.
    • In most of the water-pollinated species, pollen grains are protected from wetting by a mucilaginous covering.
    • Both wind and water pollinated flowers are not very colourful and do not produce nectar.
  • Animal pollination –
    • Mode of pollination in majority of flowering plants.
    • Bees, butterflies, flies, beetles, wasps, ants, moths, birds (sunbirds and humming birds) and bats are the common pollinating agents.
    • Among the animals, insects, particularly bees are the dominant biotic pollinating agents.
    • Even larger animals such as some primates (lemurs), arboreal (tree-dwelling) rodents, or even reptiles (gecko lizard and garden lizard) are also pollinators in some species.
    • Often flowers of animal- pollinated plants are specifically adapted for a particular species of animal.
    • Majority of insect-pollinated flowers are large, colourful, fragrant and rich in nectar.
    • When the flowers are small, a number of flowers are clustered into an inflorescence to make them conspicuous.
    • Animals are attracted to flowers by colour and/or fragrance.
    • The flowers pollinated by flies and beetles secrete foul odours to attract these animals.
    • To sustain animal visits, the flowers have to provide rewards to the animals. Nectar and pollen grains are the usual floral rewards.
    • For harvesting the reward(s) from the flower the animal visitor comes in contact with the anthers and the stigma. The body of the animal gets a coating of pollen grains, which are generally sticky in animal pollinated flowers. When the animal carrying pollen on its body comes in contact with the stigma, it brings about pollination.
    • In some species floral rewards are in providing safe places to lay eggs; e.g., tallest flower of Amorphophallus (6 feet in height).
    • A similar relationship exists between a species of moth and the plant Yucca where both species – moth and the plant – cannot complete their life cycles without each other. The moth deposits its eggs in the locule of the ovary and the flower, in turn, gets pollinated by the moth. The larvae of the moth come out of the eggs as the seeds start developing.
    • Other examples of insect pollinated plants – Cucumber, Mango, PeepaL, Coriander, Papaya, Onion, Lobia, Cotton, Tobacco, Rose, Lemon, Eucalyptus, Banana.
    • Pollen/ Nectar robbers – Many insects may consume pollen or the nectar without bringing about pollination. Such floral visitors are referred to as pollen/nectar robbers.

Outbreeding Devices:

  • Majority of flowering plants produce hermaphrodite flowers and pollen grains are likely to come in contact with the stigma of the same flower.
  • Continued self-pollination result in inbreeding depression.
  • Flowering plants have developed many devices to discourage self- pollination and to encourage cross-pollination.
  • In some species, pollen release and stigma receptivity are not synchronised. Either the pollen is released before the stigma becomes receptive or stigma becomes receptive much before the release of pollen.
  • In some other species, the anther and stigma are placed at different positions so that the pollen cannot come in contact with the stigma of the same flower. Both these devices prevent autogamy.
  • self-incompatibility – This is a genetic mechanism and prevents self-pollen (from the same flower or other flowers of the same plant) from fertilising the ovules by inhibiting pollen germination or pollen tube growth in the pistil.
  • production of unisexual flowers.

If both male and female flowers are present on the same plant such as castor and maize (monoecious), it prevents autogamy but not geitonogamy.

In several species such as papaya, male and female flowers are present on different plants, that is each plant is either male or female (dioecy). This condition prevents both autogamy and geitonogamy.

Pollen-pistil Interaction:

  • Pollination does not guarantee the transfer of the right type of pollen (compatible pollen of the same species as the stigma). Often, pollen of the wrong type, either from other species or from the same plant (if it is self-incompatible), also land on the stigma.
  • The pistil has the ability to recognise the pollen, whether it is of the right type (compatible) or of the wrong type (incompatible). If it is of the right type, the pistil accepts the pollen and promotes post-pollination events that leads to fertilisation. If the pollen is of the wrong type, the pistil rejects the pollen by preventing pollen germination on the stigma or the pollen tube growth in the style.
  • The ability of the pistil to recognise the pollen followed by its acceptance or rejection is the result of a continuous dialogue between pollen grain and the pistil. This dialogue is mediated by chemical components of the pollen interacting with those of the pistil.
  • As mentioned earlier, following compatible pollination, the pollen grain germinates on the stigma to produce a pollen tube through one of the germ pores. The contents of the pollen grain move into the pollen tube. Pollen tube grows through the tissues of the stigma and style and reaches the ovary.
  • In some plants, pollen grains are shed at two-celled condition (a vegetative cell and a generate cell). In such plants, the generative cell divides and forms the two male gametes during the growth of pollen tube in the stigma. In plants which shed pollen in the three-celled condition, pollen tubes carry the two male gametes from the beginning.
  • Pollen tube, after reaching the ovary, enters the ovule through the micropyle and then enters one of the synergids through the filiform apparatus. Filiform apparatus present at the micropylar part of the synergids guides the entry of pollen tube.
  • All these events-from pollen deposition on the stigma until pollen tubes enter the ovule-are together referred to as pollen-pistil interaction.
  • pollen-pistil interaction is a dynamic process involving pollen recognition followed by promotion or inhibition of the pollen.

Artificial hybridisation

  • it is one of the major approaches of crop improvement programme. In such crossing experiments it is important to make sure that only the desired pollen grains are used for pollination and the stigma is protected from contamination (from unwanted pollen). This is achieved by emasculation and bagging techniques.
  • If the female parent bears bisexual flowers, removal of anthers from the flower bud before the anther dehisces using a pair of forceps is necessary. This step is referred to as
  • Emasculated flowers have to be covered with a bag of suitable size, generally made up of butter paper, to prevent contamination of its stigma with unwanted pollen. This process is called
  • When the stigma of bagged flower attains receptivity, mature pollen grains collected from anthers of the male parent are dusted on the stigma, and the flowers are rebagged, and the fruits allowed to develop.
  • If the female parent produces unisexual flowers, there is no need for emasculation. The female flower buds are bagged before the flowers open. When the stigma becomes receptive, pollination is carried out using the desired pollen and the flower rebagged.

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Double Fertilisation

  • After entering one of the synergids, the pollen tube releases the two male gametes into the cytoplasm of the synergid.
  • One of the male gametes moves towards the egg cell and fuses with its nucleus thus completing the This results in the formation of a diploid cell, the zygote.
  • The other male gamete moves towards the two polar nuclei located in the central cell and fuses with them to produce a triploid primary endosperm nucleus (PEN). As this involves the fusion of three haploid nuclei it is termed triple fusion.
  • Since two types of fusions, syngamy and triple fusion take place in an embryo sac the phenomenon is termed double fertilisation, an event unique to flowering plants.
  • The central cell after triple fusion becomes the primary endosperm cell (PEC) and develops into the endosperm while the zygote develops into an

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  • Post-Fertilisation : Structures And Events
  • Following double fertilisation, events of endosperm and embryo development, maturation of ovule(s) into seed(s) and ovary into fruit, are collectively termed post-fertilisation events.

Endosperm

  • Endosperm development precedes embryo development.
  • The primary endosperm cell divides repeatedly and forms a triploid endosperm tissue. The cells of this tissue are filled with reserve food materials and are used for the nutrition of the developing embryo.
  • In the most common type of endosperm development, the PEN undergoes successive nuclear divisions to give rise to free nuclei. This stage of endosperm development is called free-nuclear endosperm.
  • Subsequently cell wall formation occurs and the endosperm becomes cellular. The number of free nuclei formed before cellularisation varies greatly.
  • The coconut water from tender coconut that you are familiar with, is nothing but free-nuclear endosperm (made up of thousands of nuclei) and the surrounding white kernel is the cellular endosperm.
  • Endosperm may either be completely consumed by the developing embryo (e.g., pea, groundnut, beans) before seed maturation or it may persist in the mature seed (e.g. castor and coconut) and be used up during seed germination.

Embryo

  • Embryo develops at the micropylar end of the embryo sac where the zygote is situated.
  • Most zygotes divide only after certain amount of endosperm is formed. This is an adaptation to provide assured nutrition to the developing embryo.
  • Though the seeds differ greatly, the early stages of embryo development (embryogeny) are similar in both monocotyledons and dicotyledons.
  • The zygote gives rise to the proembryo and subsequently to the globular, heart-shaped and mature embryo.
  • A typical dicotyledonous embryo, consists of an embryonal axis and two
  • The portion of embryonal axis above the level of cotyledons is the epicotyl, which terminates with the plumule or stem tip.
  • The cylindrical portion below the level of cotyledons is hypocotyl that terminates at its lower end in the radical or root tip. The root tip is covered with a root cap.
  • Embryos of monocotyledons possess only one cotyledon. In the grass family the cotyledon is called scutellum that is situated towards one side (lateral) of the embryonal axis.
  • At its lower end, the embryonal axis has the radical and root cap enclosed in an undifferentiated sheath called
  • The portion of the embryonal axis above the level of attachment of scutellum is the epicotyl. Epicotyl has a shoot apex and a few leaf primordia enclosed in a hollow foliar structure, the

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Seed

  • In angiosperms, the seed is the final product of sexual reproduction. It is often described as a fertilised ovule. Seeds are formed inside fruits.
  • A seed typically consists of seed coat(s), cotyledon(s) and an embryo axis.
  • The cotyledons of the embryo are simple structures, generally thick and swollen due to storage of food reserves (as in legumes).
  • Mature seeds may be non-albuminous or
  • Non-albuminous seeds have no residual endosperm as it is completely consumed during embryo development (e.g., pea, groundnut). Albuminous seeds retain a part of endosperm as it is not completely used up during embryo development (e.g., wheat, maize, barley, castor, sunflower).
  • Occasionally, in some seeds such as black pepper and beet, remnants of nucellus are also persistent. This residual, persistent nucellus is the
  • Integuments of ovules harden as tough protective seed
  • The micropyle remains as a small pore in the seed coat. This facilitates entry of oxygen and water into the seed during germination.
  • As the seed matures, its water content is reduced and seeds become relatively dry (10-15 per cent moisture by mass).
  • The general metabolic activity of the embryo slows down.
  • The embryo may enter a state of inactivity called dormancy, or if favourable conditions are available (adequate moisture, oxygen and suitable temperature), they germinate.
  • As ovules mature into seeds, the ovary develops into a fruit, i.e., the transformation of ovules into seeds and ovary into fruit proceeds simultaneously.
  • The wall of the ovary develops into the wall of fruit called
  • The fruits may be fleshy as in guava, orange, mango, etc., or may be dry, as in groundnut, and mustard, etc.
  • Many fruits have evolved mechanisms for dispersal of seeds.
  • In most plants, by the time the fruit develops from the ovary, other floral parts degenerate and fall off.
  • However, in a few species such as apple, strawberry, cashew, etc., the thalamus also contributes to fruit formation. Such fruits are called false fruits. Most fruits however develop only from the ovary and are called true fruits.
  • Although in most of the species, fruits are the results of fertilisation, there are a few species in which fruits develop without fertilisation. Such fruits are called parthenocarpic fruits.g., Banana. Parthenocarpy can be induced through the application of growth hormones and such fruits are seedless.

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Advantages offered by seeds –

  • since reproductive processes such as pollination and fertilisation are independent of water, seed formation is more dependable.
  • Seeds have better adaptive strategies for dispersal to new habitats and help the species to colonise in other areas.
  • As Seeds have sufficient food reserves, young seedlings are nourished until they are capable of photosynthesis on their own.
  • The hard seed coat provides protection to the young embryo.
  • Being products of sexual reproduction, they generate new genetic combinations leading to variations.
  • Seed is the basis of our agriculture.
  • Dehydration and dormancy of mature seeds are crucial for storage of seeds which can be used as food throughout the year and also to raise crop in the next season.
  • Oldest recorded viable seeds – a lupine, Lupinus arcticus excavated from Arctic Tundra. (10,000 years), and date palm, Phoenix dactylfera (2000 years)excavated from King Herod’s palace near the Dead Sea.
  • In Orchid each fruit contain thousands of tiny seeds. Similar is the case in fruits of some parasitic species such as Orobanche and Striga.

Apomixis and Polyembryony

  • Although seeds, in general are the products of fertilisation, a few flowering plants such as some species of Asteraceae and grasses, have evolved a special mechanism, to produce seeds without fertilisation, called
  • apomixis is a form of asexual reproduction that mimics sexual reproduction.
  • There are several ways of development of apomictic seeds.
  • In some species, the diploid egg cell is formed without reduction division and develops into the embryo without fertilisation.
  • in many Citrus and Mangovarieties some of the nucellar cells surrounding the embryo sac start dividing, protrude into the embryo sac and develop into the embryos. In such species each ovule contains many embryos. Occurrence of more than one embryo in a seed is referred as
  • Hybrid varieties of several of our food and vegetable crops are being extensively cultivated. Cultivation of hybrids has tremendously increased productivity.
  • One of the problems of hybrids is that hybrid seeds have to be produced every year. If the seeds collected from hybrids are sown, the plants in the progeny will segregate and do not maintain hybrid characters. Production of hybrid seeds is costly and hence the cost of hybrid seeds become too expensive for the farmers.
  • If these hybrids are made into apomicts, there is no segregation of characters in the hybrid progeny. Then the farmers can keep on using the hybrid seeds to raise new crop year after year and he does not have to buy hybrid seeds every year. Because of the importance of apomixis in hybrid seed industry, active research is going on in many laboratories around the world to understand the genetics of apomixis and to transfer apomictic genes into hybrid varieties.

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CHAPTER 2 : SEXUAL REPRODUCTION IN FLOWERING PLANTS

 

NEET Phase-II (24-07-2016) Biology Solution {Code – XX}

NEET Phase-II Biology solution

Code – XX

Date – 24-07-2016

 

  1. A foreign DNA and plasmid cut by the samerestriction endonuclease can be joined toform a recombinant plasmid using
    1. ligase
    2. Eco RI
    3. Taq polymerase
    4. polymerase III

Ans.        (1) Ligase                                                                              [NCERT class 12, page 197]      

 

  1. Which of the following is not a component ofdownstream processing?
    1. Expression
    2. Separation
    3. Purification
    4. Preservation

Ans.        (1) Expression                                                  [NCERT class 12, page 205]

 

  1. Which of the following restriction enzymesproduces blunt ends?
    1. Hind III
    2. Sal I
    3. Eco RV
    4. Xho I

Ans.        (3) Eco RV

 

  1. Which kind of therapy was given in 1990 to afour-year-old girl with adenosine deaminase(ADA) deficiency?
    1. Radiation therapy
    2. Gene therapy
    3. Chemotherapy
    4. Immunotherapy

Ans.        (2) Gene therapy                                                                      [NCERT class 12, page 211]

 

  1. How many hot spots of biodiversity in theworld have been identified till date byNorman Myers?
    1. 43
    2. 17
    3. 25
    4. 34

Ans.        (4) 34                                                                            [NCERT class 12, page 266]

 

  1. The primary producers of the deep-seahydrothermal vent ecosystem are
    1. coral reefs
    2. green algae
    3. chemosynthetic bacteria
    4. blue-green algae

Ans.        (3) chemosynthetic bacteria                                             [NCERT class 12, page 226]

  

  1. Which of the following is correct forr-selected species?
    1. Small number of progeny with large size
    2. Large number of progeny with small size
    3. Large number of progeny with large size
    4. Small number of progeny with small size

Ans.        (2) Large number of progeny with small size

 

  1. If’+’ sign is assigned to beneficial interaction, ‘-’ sign to detrimental and ‘0’ sign to neutralinteraction, then the population interactionrepresented by ‘+’ refers to
    1. parasitism
    2. mutualism
    3. amensalism
    4. commensalism

Ans.        (1) parasitism                                                [NCERT class 12, page 232]

 

  1. Which of the following is correctly matched?
    1. Stratification—Population
    2. Aerenchyma—Opuntia
    3. Age pyramid—Biome
    4. Parthenium hysterophorus—Threat to biodiversity

Ans.        (4) Parthenium hysterophorus—Threat to biodiversity                                        [NCERT class 12, page 265]

 

  1. Red List contains data or information on
    1. marine vertebrates only
    2. all economically important plant
    3. plants whose products are ininternational trade
    4. threatened species

Ans.        (4) threatened species                                                           [NCERT class 12, page 263]

 

  1. Which one of the following is wrongfor fungi?
    1. They are both unicellular and
    2. They are eukaryotic.
    3. All fungi possess a purely cellulosic cell wall.
    4. They are heterotrophic

Ans.        (3) All fungi possess a purely cellulosic cell wall                                                        [NCERT class 11, page 22]

 

  1. Methanogens belong to
    1. Slime moulds
    2. Eubacteria
    3. Archaebacteria
    4. Dinoflagellates

Ans.        (3) Archaebacteria                                                             [NCERT class 11, page 19]

 

  1. Select thewrong
    1. Diatoms are microscopic and floatpassively in water.
    2. The walls of diatoms are easily
    3. ‘Diatomaceous earth’ is formed by thecell walls of diatoms.
    4. Diatoms are chief producers in the

Ans.        (2) The walls of diatoms are easily destructible.            [NCERT class 11, page 20]

 

 

  1. The label of a herbarium sheetdoes not carry information on
    1. height of the plant
    2. date of collections
    3. name of collector
    4. local names

Ans.        (1) height of the plant                                                           [NCERT class 11, page 12]

 

  1. Conifers are adapted to tolerate extremeenvironmental conditions because of
    1. presence of vessels
    2. broad hardy leaves
    3. superficial stomata
    4. thick cuticle

Ans.        (4) thick cuticle                                                              [NCERT class 11, page 38]

 

  1. which one of the following statements iswrong?
    1. Laminaria and Sargassum are used as
    2. Algae increase the level of dissolved oxygen in the immediate environment.
    3. Algin is obtained from red algae, andcarrageenan from brown algae.
    4. Agar-agar is obtained from Gelidium and

Ans.        (3) Algin is obtained from red algae, and carrageenan from brown algae   [NCERT class 11, page 32]

 

  1. The term ‘polyadelphous’ is related to
    1. calyx
    2. gynoecium
    3. androecium
    4. corolla

Ans.        (3) androecium                                                                   [NCERT class 11, page 75]

 

  1. How many plants among, indigophera, Sesbania, Salvia, Allium, Aloe, Mustard, Groundnut, Radish, Gram, and Turnip have stamens with different lengths in their flowers?
    1. six
    2. Three
    3. Four
    4. Five

Ans.        (3) Four (Salvia, Mustard, Radish, Turnip)              [NCERT class 11, page 75]

 

  1. Radial symmetry is found in the flowers of
    1. Cassia
    2. Brassica
    3. Trifolium
    4. Pisum

Ans.        (2) Brassica                                                              [NCERT class 11, page 72,79]

 

  1. Free-central placentation is found in
    1. Citrus
    2. Dianthus
    3. Argemone
    4. Brassica

Ans.        (2) Dianthus                                                                      [NCERT class 11, page 75]

 

 

  1. Cortex is the region found between
    1. endodermis and vascular bundle
    2. epidermis and stele
    3. pericycle and endodermis
    4. endodermis and pith

Ans.        (2) epidermis and stele                                              [NCERT class 11, page 91]

 

  1. The balloon-shaped structures called tyloses
    1. are linked to the ascent of sap through xylem vessels
    2. originate in the lumen of vessels
    3. Characterize the sapwood
    4. are extensions of xylem parenchyma cells into vessels

Ans.        (4) are extensions of xylem parenchyma cells into vessels

 

  1. A non-proteinaceous enzyme is
    1. deoxyribonuclease
    2. lysozyme
    3. ribozyme
    4. ligase

Ans.        (3) ribozyme                                                                    [NCERT class 11, page 154]

 

  1. Select the
    1. Methanogens—Prokaryote
    2. Gas vacuoles—Green bacteria
    3. Large central vacuoles—Animal cells
    4. Protists—Eukaryotes

Ans.        (3) Large central vacuoles—Animal cells            [NCERT class 11, page 129]

 

  1. Select the wrong
    1. Mycoplasma is a wall-less
    2. Bacterial cell wall is made up of
    3. Pili and fimbriae are mainly involved inmotility of bacterial cells.
    4. Cyanobacteria lack flagellated cells.

Ans.        (3) Pili and fimbriae are mainly involved in motility of bacterial cells           [NCERT class 11, page 129]

 

  1. A cell organelle containing hydrolyticenzymes is
    1. mesosome
    2. lysosome
    3. microsome
    4. ribosome

Ans.        (2) lysosomes                                                                  [NCERT class 11, page 134]

 

  1. During cell growth, DNA synthesis takesplace in
    1. M phase
    2. S phase
    3. G1 phase
    4. G2 phase

Ans.        (2) S phase                                                                        [NCERT class 11, page 163]

 

 

  1. Which of the following biomolecules iscommon to respiration-mediated breakdownof fats, carbohydrates and proteins?
    1. Acetyl CoA
    2. Glucose-6-phosphate
    3. Fructose 1,6-bisphosphate
    4. Pyruvic acid

Ans.        (1) Acetyl CoA                                                                 [NCERT class 11, page 236]

 

  1. A few drops of sap were collected by cuttingacross a plant stem by a suitable method.The sap was tested chemically. Which one ofthe following test results indicates that it isphloem sap?
    1. Absence of sugar
    2. Acidic
    3. Alkaline
    4. Low refractive index

Ans.        (3) Alkaline

 

  1. You are given a tissue with its potential fordifferentiation in an artificial culture. Whichof the following pairs of hormones would youadd to the medium to secure shoots as wellas roots?
    1. Gibberellin and abscisic acid
    2. IAA and gibberellins
    3. Auxin and cytokinin
    4. Auxin and abscisic acid

Ans.        (3) Auxin and cytokinin                                              [NCERT class 12, page 177]

 

  1. Phytochrome is a
    1. Chromoprotein
    2. Flavoprotein
    3. Glycoprotein
    4. Lipoprotein

Ans.        (1) Chromoprotein

 

  1. Which is essential for the growth of root tip?
    1. Mn
    2. Zn
    3. Fe
    4. Ca

Ans.        (2) Zn                                                                    [NCERT class 11, page 198,248]

 

  1. The process which makes major differencebetween C3 and C4 plants is
    1. respiration
    2. glycolysis
    3. Calvin cycle
    4. Photorespiration

Ans.        (4) Photorespiration                                                 [NCERT class 11, page 220]

 

  1. Which one of the following statements is not correct?
    1. Water hyacinth, growing in the standingwater, drains oxygen from water thatleads to the death of fishes.
    2. Offspring produced by the asexualreproduction are called clone.
    3. Microscopic, motile asexual reproductivestructures are called zoospores.
    4. In potato, banana and ginger, theplantlets arise from the internodespresent in the modified stem.

Ans.        (4) In potato, banana and ginger, the plantlets arise from the internodes present in the modified stem.                                                                                                                                                                        [NCERT class 12, page 8]

 

  1. Which one of the following generates newgenetic combinations leading to variation?
    1. Nucellar polyembryony
    2. Vegetative reproduction
    3. Parthenogenesis
    4. Sexual reproduction

Ans.        (4) Sexual reproduction                                             [NCERT class 12, page 38]

 

  1. Match Column—I with Column—II andselect the correct option using the codesgiven below:
  Column—I   Column—II
a. Pistils fused together (i) Gametogenesis
b. Formation of gametes (ii) Pistillate
c. Hyphae of higher Ascomycetes (iii) Syncarpous
d. Unisexual female flower (iv) Dikaryotic

Codes :

a             b              c                 d 

  1. (iii)            (i)            (iv)          (ii)
  2. (iv)             (iii)          (i)            (ii)
  3. (ii)            (i)            (iv)          (iii)
  4. (i)               (ii)           (iv)          (iii)

Ans.        (1) a-(iii), b-(i), c-(iv), d-(ii)                                        [NCERT class 11, page 23,75]

 

  1. In majority of angiosperms
    1. a small central cell is present in theembryo sac
    2. egg has a filiform apparatus
    3. there are numerous antipodal cells
    4. reduction division occurs in themegaspore mother cells

Ans.        (4) reduction division occurs in the megaspore mother cells                               [NCERT class 12, page 26,27]

 

  1. Pollination in water hyacinth and water lily isbrought about by the agency of
    1. Bats
    2. Water
    3. insects or wind
    4. birds

Ans.     (3) insects or wind                                                                                                [NCERT class 12, page 29]

 

  1. The ovule of an angiosperm is technicallyequivalent to
    1. megaspore
    2. megasporangium
    3. megasporophyll
    4. megaspore mother cell

Ans.     (2) megasporangium                                               [NCERT class 12, page 25]

 

  1. Taylor conducted the experiments to provesemiconservative mode of chromosomereplication on
    1. coli
    2. Vinca rosea
    3. Vicia faba
    4. Drosophila melanogaster

Ans.     (3) Vicia faba                                                              [NCERT class 12, page 106]

 

  1. The mechanism that causes a gene to movefrom one linkage group to another is called
    1. crossing-over
    2. inversion
    3. duplication
    4. translocation

Ans.     (4) translocation

 

  1. The equivalent of a structural gene is
    1. recon
    2. muton
    3. cistron
    4. operon

Ans.     (3) Cistron                                                                    [NCERT class 12, page 109]

 

  1. A true breeding plant is
    1. always homozygous recessive in itsgenetic constitution
    2. one that is able to breed on its own
    3. produced due to cross-pollination amongunrelated plant
    4. near homozygous and produces offspringof its own kind

Ans.     (4) near homozygous and produces offspring of its own kind                   [NCERT class 12, page 70]

 

  1. Which of the following rRNAs acts asstructural RNA as well as ribozyme inbacteria?
    1. 8 S rRNA
    2. 5 S rRNA
    3. 18 S rRNA
    4. 23 S rRNA

Ans.     (4) 23 S rRNA                                                                              [NCERT class 12, page 115]

 

  1. Stirred-tank bioreactors have been designedfor
    1. ensuring anaerobic conditions in theculture vessel
    2. purification of product
    3. addition of preservatives to the product
    4. availability of oxygen throughout theprocess

Ans.     (4) availability of oxygen throughout the process                                      [NCERT class 12, page 204]

 

  1. A molecule that can act as a genetic materialmust fulfill the traits given below, except
    1. it should provide the scope for slow changes that are required for evolution
    2. it should be able to express itself in theform of ‘Mendelian characters’
    3. it should be able to generate its replica
    4. it should be unstable structurally andchemically

Ans.     (4) it should be unstable structurally and chemically                         [NCERT class 12, page 103]

 

  1. DNA-dependent RNA polymerase catalyzestranscription on one strand of the DNAwhich is called the
    1. antistrand
    2. template strand
    3. coding strand
    4. alpha strand

Ans.     (2) template strand                                                  [NCERT class 12, page 108]

 

  1. Interspecific hybridization is the mating of
    1. more closely related individuals withinsame breed for 4-6 generations
    2. animals within same breed withouthaving common ancestors
    3. two different related species
    4. superior males and females of differentbreeds

Ans.     (3) two different related species                                         [NCERT class 12, page 168]

 

  1. which of the following is correctregardingAIDS causative agent HIV?
    1. HIV does not escape but attacks theacquired immune response.
    2. HIV is enveloped virus containing onemolecule of single-stranded RNA and onemolecule of reverse transcriptase.
    3. HIV is enveloped virus that contains twoidentical molecules of single-stranded RNA and two molecules of reverse
    4. HIV is unenveloped retrovirus.

Ans.     (3) HIV is enveloped virus that contains two identical molecules of single-stranded RNA and two molecules of reverse transcriptase                                                                

 

  1. Among the following edible fishes, whichone is a marine fish having rich source ofomega-3 fatty acids?
    1. Mackerel
    2. Mystus
    3. Mangur
    4. Mrigala

Ans.     (1) Mackerel                                                                  [NCERT class 12, page 169]

 

  1. Match Column—I with Column—II and select the correct option using the codesgiven below:

Column—I                              Column—II

  1. Citric acid                                (i)    Trichoderma
  2. Cyclosporin A                         (ii)   Clostridium
  3. Statins                                    (iii) Aspergillus
  4. Butyric acid                           (iv) Monascus

Codes :

a          b         c          d

  1. (iii)      (iv)     (i)        (ii)
  2. (iii)        (i)        (ii)       (iv)
  3. (iii)        (i)        (iv)     (ii)
  4. (i)          (iv)     (ii)       (iii)

Ans.     (3) a-(iii), b-(i), c-(iv), d-(ii)                                              [NCERT class 12, page 183]

 

  1. Biochemical Oxygen Demand (BOD) may not be a good index for pollution for water bodiesreceiving effluents from.
    1. sugar industry
    2. domestic sewage
    3. dairy industry
    4. petroleum industry

Ans.     (4) petroleum industry                                                          [NCERT class 12, page 276]

 

  1. The principle of competitive exclusion wasstated by
    1. Verhulst and Pearl
    2. Darwin
    3. F. Gause
    4. MacArthur

Ans.     (3) G. F. Gause                                                           [NCERT class 12, page 234]

 

  1. Which of the following National Parks is home to the famous musk deer or hangul?
    1. Dachigam National Park, Jammu &Kashmir
    2. Keibul Lamjao National Park, Manipur
    3. Bandhavgarh National Park, MadhyaPradesh
    4. Eaglenest Wildlife Sanctuary, ArunachalPradesh

Ans.     (1) Dachigam National Park, Jammu & Kashmir                                                  

 

  1. A lake which is rich in organic waste mayresult in
    1. mortality of fish due to lack of oxygen
    2. increased population of aquaticorganisms due to minerals
    3. drying of the lake due to algal bloon
    4. increased population of fish due to lots ofnutrients

Ans.     (1) mortality of fish due to lack of oxygen                        [NCERT class 12, page 275]

 

  1. The highest DDT concentration in aquaticfood chain shall occur in
    1. eel
    2. phytoplankton
    3. seagull
    4. crab

Ans.     (3) seagull                                                                [NCERT class 12, page 276]

 

  1. Which-of the following sets of diseases iscaused by bacteria?
    1. Herpes and influenza
    2. Cholera and tetanus
    3. Typhoid and smallpox
    4. Tetanus and mumps

Ans.     (2) Cholera and tetanus                                               [NCERT class 11, page 26]

 

 

  1. Match Column—I with Column—II forhousefly classification and select the correctoption using the codes given below :

Column—I                      Column—II

  1. Family                   (i)   Diptera
  2. order                      (ii) Arthropoda
  3. Class                 (iii)  Muscidae
  4. Phylum                   (iv) Insecta

Codes :

a      b    c  d

  1.          (iv)    (ii)       (i)        (iii)
  2.         (iii)    (i)        (iv)      (ii)
  3.        (iii)     (ii)       (iv)      (i)
  4.       (iv)      (iii)      (ii)       (i)

Ans.     (2) a-(iii), b-(i), c-(iv), d-(ii).                                       [NCERT class 11, page 11]

 

  1. Choose the correct
    1. All Pisces have gills covered by an
    2. All mammals are viviparous.
    3. All cyclostomes do not possess jaws and paired fins.
    4. All reptiles have a three-chambered

Ans.     (3) All cyclostomes do not possess jaws and paired fins                      [NCERT class 11, page 56-59]

 

  1. Study the four statements (A-D) given belowand select the two correct ones out of them:
    1. Definition of biological species was givenby Ernst Mayr.
    2. Photoperiod does not affect reproductionin plants.
    3. Binomial nomenclature system wasgiven by R. H.
    4. In unicellular organisms, reproduction issynonymous with growth.

The two correctstatements are

(1) A and B                              (2) B and C                              (3) C and D                      (4) A and D

Ans.     (4) A and D                                                            [NCERT class 11, page 2-7]

 

  1. In male cockroaches, sperms are stored in which part of the reproductive system?
    1. Vas deferens
    2. Seminal vesicles
    3. Mushroom glands
    4. Testes

Ans.     (2) Seminal vesicles                                                                 [NCERT class 11, page 114]

 

  1. Smooth muscles are
    1. voluntary, spindle-shaped, uninucleate
    2. involuntary, fusiform, non-striated
    3. voluntry, multinucleate, cylindrical
    4. involuntary, cylindrical, striated

Ans.     (2) involuntary, fusiform, non-striated                    [NCERT class 11, page 105,303]

 

 

  1. Oxidative phosphorylation is
    1. formation of ATP by energy released from electrons removed during substrateoxidation
    2. formation of ATP by transfer ofphosphate group from a substrateto ADP
    3. oxidation of phosphate group in ATP
    4. addition of phosphate group to ATP

Ans.     (1) formation of ATP by energy released from electrons removed during substrate oxidation                                                                        [NCERT class 11, page 233]

 

  1. Which of the following is the least likely to beinvolved in stabilizing the three-dimensionalfolding of most proteins?
    1. Ester bonds
    2. Hydrogen bonds
    3. Electrostatic interaction
    4. Hydrophobic interaction

Ans.     (1) Ester bonds                                                 [NCERT class 11, page 150]

 

  1. Which of the following describes the given graph correctly?

1

  1. Exothermic reaction with energy A inabsence of enzyme and B in presence ofenzyme
  2. Endothermic reaction with energy A inpresence of enzyme and B in absence ofenzyme
  3. Exothermic reaction with energy A inpresence of enzyme and B in absence ofenzyme
  4. Endothermic reaction with energy A inabsence of enzyme and B in presence ofenzyme

Ans.     (3) Exothermic reaction with energy A in presence of enzyme and B in absence of enzyme                                                                                       [NCERT class 11, page 156]

 

  1. When cell has stalled DNA replication fork,which checkpoint should be predominantlyactivated?
    1. Both G2/M and M
    2. G1/S
    3. G2/M
    4. M

Ans.     (2) G1/S                                                                         [NCERT class 11, page 164]

 

  1. Match the stages of meiosis in Column—I to their characteristic features in Column—II and select the correct option using the codes given below:
  Column—I   Column—II
a. Pachytene (i) Pairing of homologous chromosomes
b. Metaphase-I (ii) Terminalization of chiasmata
c. Diakinesis (iii) Crossing-over takes place
d. Zygotene (iv) Chromosomes align at equatorial plate

a          b         c          d

  1. (iv) (iii)      (ii)       (i)
  2. (iii) (iv)     (ii)       (i)
  3. (i) (iv)     (ii)       (iii)
  4. (ii) (iv)     (iii)      (i)

Ans.     (2) a-(iii), b-(iv), c-(ii), d-(i)                               [NCERT class 11, page 168]

 

  1. Which hormones do stimulate theproduction of pancreatic juice andbicarbonate?
    1. Insulin and glucagon
    2. Angiotensin and epinephrine
    3. Gastrin and insulin
    4. Cholecystokinin and secretin

Ans.     (4) Cholecystokinin and secretin                             [NCERT class 11, page 338]

 

  1. The partial pressure of oxygen in the alveoliof the lungs is
    1. less than that of carbon dioxide
    2. equal to that in the blood
    3. more than that in the blood
    4. less than that in the blood

Ans.     (3) more than that in the blood                                        [NCERT class 11, page 272]

 

  1. Choose the correct
    1. Receptors do not produce graded
    2. Nociceptors respond to changes in
    3. Meissner’s corpuscles are thermo­receptors.
    4. Photoreceptors in the human eye aredepolarized during darkness and becomehyperpolarized in response to the light

Ans.     (4) Photoreceptors in the human eye are depolarized during darkness and become hyperpolarized in response to the light stimulus.                          

 

  1. Graves’ disease is caused due to
    1. hypersecretion of adrenal gland
    2. hyposecretion of thyroid gland
    3. hypersecretion of thyroid gland
    4. hyposecretion of adrenal gland

Ans.     (3) hypersecretion of thyroid gland                                

 

 

  1. Name the ion responsible for unmasking ofactive sites for myosin for cross-bridgeactivity during muscle contraction.
    1. Potassium
    2. Calcium
    3. Magnesium
    4. Sodium

Ans.     (2) Calcium                                                     [NCERT class 11, page 307]

 

  1. Name the blood cells, whose reduction innumber can cause clotting disorder, leadingto excessive loss of blood from the
    1. Thrombocytes
    2. Erythrocytes
    3. Leucocytes
    4. Neutrophils

Ans.     (1) Thrombocyte                                                                   [NCERT class 11, page 280]

 

  1. Name a peptide hormone which acts mainlyon hepatocytes, adipocytes and enhancescellular glucose uptake and utilization.
    1. Gastrin
    2. Insulin
    3. Glucagon
    4. Secretin

Ans.     (2) Insulin                                                                   [NCERT class 11, page 336]

 

  1. Osteoporosis, an age-related disease ofskeletal system, may occur due to
    1. accumulation of uric acid leading toinflammation of joints
    2. immune disorder affecting neuro­muscular junction leading to fatigue
    3. high concentration of Ca++ and Na+
    4. decreased level of estrogen

Ans.     (4) decreased level of estrogen                                [NCERT class 11, page 312]

 

  1. Serum differs from blood in.
    1. lacking antibodies
    2. lacking globulins
    3. lacking albumins
    4. lacking clotting factors

Ans.     (4) lacking clotting factors                                          [NCERT class 11, page 279]

 

  1. Lungs do not collapse between breaths andsome air always remains in the lungs whichcan never be expelled because
    1. pressure in the lungs is higher than theatmospheric pressure.
    2. there is a negative pressure in the lungs
    3. there is a negative intrapleural pressure pulling at the lung walls
    4. there is a positive intrapleural pressure

Ans.     (3) there is a negative intrapleural pressure pulling at the lung walls       

 

  1. The posterior pituitary gland is nota trueendocrine gland because
    1. it secretes enzymes
    2. it is provided with a duct
    3. only stores and releases hormones
    4. it is under the regulation of hypo­thalamus

Ans.     (3) only stores and releases hormones                           [NCERT class 11, page 332]

 

  1. The part of nephron involved in activereabsorption of sodium is
    1. descending limb of Henle’s loop
    2. distal convoluted tubule
    3. proximal convoluted tubule
    4. Bowman’s capsule

Ans.     (3) proximal convoluted tubule                     [NCERT class 11, page 294]

 

  1. Which of the following is hormone-releasing IUD?
    1. Cu7
    2. LNG-20
    3. Multiload 375
    4. Lippes loop

Ans.     (2) LNG-20                                                                [NCERT class 12, page 60]

 

  1. Which of the following is incorrectregarding vasectomy?
    1. Irreversible sterility
    2. No sperm occurs in seminal fluid
    3. No sperm occurs in epididymis
    4. Vasa deferentia is cut and tied.

Ans.     (3) No sperm occurs in epididymis                                   [NCERT class 12, page 62]

 

  1. Embryo with more than 16 blastomeresformed due to in vitro fertilization istransferred into
    1. cervix
    2. uterus
    3. fallopian tube
    4. fimbriae

Ans.     (2) Uterus                                                                 [NCERT class 12, page 64]

 

  1. Which of the following depicts the correct pathway of transport of sperms?
    1. Efferent ductules →Rete testis → Vasdeferens → Epididymis
    2. Rete testis →Efferent ductules→ Epididymis → Vas deferens
    3. Rete testis → Epididymis → Efferentductules → Vas deferens
    4. Rete testis → Vas deferens → Efferentductules → Epididymis

Ans.     (2) Rete testis → Efferent ductules → Epididymis → Vas deferens             [NCERT class 12, page 43]

 

  1. Match Column—I with Column—II andselect the correct option using the codesgiven below :

ColumnI                              ColumnII

  1. Mons pubis (i)    Embryo formation
  2. Antrum (ii)   Sperm
  3. Trophectoderm (iii)  Female external genitalia
  4. Nebenkern (iv) Graafian follicle

Codes :

a          b        c          d

  1. (i) (iv)     (iii)      (ii)
  2. (iii) (iv)     (ii)       (i)
  3. (iii) (iv)     (i)        (ii)
  4. (iii) (i)        (iv)     (ii)

Ans.     (3) a-(iii), b-(iv), c-(i), d-(ii)                                 [NCERT class 12, page 46,48]

 

  1. Several hormones like hCG, hPL, estrogenprogesterone are produced by
    1. pituitary
    2. ovary
    3. placenta
    4. fallopian tube

Ans.     (3) placenta                                           [NCERT class 12, page 53]

 

  1. If a colour-blind man marries a woman whois homozygous for normal colour vision, the probability of their son being colour-blind is

(1) 1 (2)  0                                          (3)  5                              (4)  0.75

Ans.     (2) 0                                                                                   

 

  1. Genetic drift operates in
    1. slow reproductive population
    2. small isolated population
    3. large isolated population
    4. non-reproductive population

Ans.     (2) small isolated population                                               [NCERT class 12, page 137]

 

  1. In Hardy-Weinberg equation, the frequentof heterozygous individual is represented by-

(1) q2 (2) p2                                                              (3) 2pq                              (4) pq

Ans.     (3) 2pq                                                                               [NCERT class 12, page 137]

 

  1. The chronological order of human evolution/ from early to the recent is
    1. Australopithecus → Homo habilis → Ramapithecus → Homo erectus
    2. Australopithecus → Ramapithecus → Homo habilis → Homo erectus
    3. Ramapithecus → Australopithecus → Homo habilis → Homo erectus
    4. Ramapithecus → Homo habilis → Australopithecus → Homo erectus

Ans.     (3) Ramapithecus → Australopithecus → Homo habilis → Homo erectus  [NCERT class 12, page 140]

 

  1. Which of the following is the correct sequence of events in the origin of life?

I- Formation of protobionts

II-Synthesis of organic monomers

III-Synthesis of organic polymers

IV-Formation of DNA-based genetic system

  1. II, III, IV, I
  2. I, II, III, IV
  3. I, III, II, IV
  4. ll, III, I, IV

Ans.     (4) ll, III, I, IV                                                                             [NCERT class 12, page 127]

 

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NEET Phase 2 2016 Solution Set XX

 

CHAPTER 1 : REPRODUCTION IN ORGANISMS

CHAPTER 1

REPRODUCTION IN ORGANISMS

  • Life span –The period from birth to the natural death of an organism represents its life span.
  • life spans of organisms are not necessarily correlated with their sizes.
  • Life span of various organisms –

Name of organism

Life-span
Elephant 60–90 years
Dog 20–30 years
Butterfly 1-2 weeks
Crow 15 years
Parrot 140 years
Cow 20–25 years
Horse 60 years
Crocodile 60 years
Fruit fly 30 days
Tortoise 100-150 years
Rose 5–7 years
Banana tree 25 years
Rice plant 3–4 months
Banyan tree 200 years

Whatever be the life span, death of every individual organism is a certainty, i.e., no individual is immortal, except single-celled organisms.

  • There is no natural death in single-celled organisms as they divide and form 2 new cells.
  • Reproduction–
    • it is defined as a biological process in which an organism gives rise to young ones (offspring) similar to itself.
    • The offspring grow, mature and in turn produce new offspring. Thus, there is a cycle of birth, growth and death.
    • Reproduction enables the continuity of the species, generation after generation.
    • genetic variation is created and inherited during reproduction.
    • There is a large diversity in the mechanism of reproduction of organisms. The organism’s habitat, its internal physiology and several other factors are collectively responsible for how it reproduces.
  • Type of reproduction –

Reproduction is of two types–

When offspring is produced by a single parent with or without the involvement of gamete formation, the reproduction is Asexual.

When two parents (opposite sex) participate in the reproductive process and also involve fusion of male and female gametes, it is called sexual reproduction.

  • Asexual Reproduction
    • In this method, a single individual (parent) is capable of producing offspring.
    • The offspring that are produced are not only identical to one another but are also exact copies of their parent.These offspring are also genetically identical to each other. The term clone is used to describe such morphologically and genetically similar individuals.
    • Asexual reproduction is common among single-celled organisms, and in plants and animals with relatively simple organisations.
        • Binary Fission – In many single-celled organisms cell divides into two halves and each rapidly grows into an adult (e.g., Amoeba, Paramecium).
        • Budding – In yeast, the division is unequal and small buds are produced that remain attached initially to the parent cell which, eventually gets separated and mature into new yeast organisms (cells).
        • Special reproductive structures –Members of the Kingdom Fungi and simple plants such as algae reproduce through special asexual reproductive structures. The most common of these structures are zoospores that usually are microscopic motile structures. Other common asexual reproductive structures are conidia (Penicillium), buds (Hydra) and gemmules (sponge).
        • Vegetative propagation –vegetative reproduction is also asexual process as only one parent is involved. in plants, the term vegetative reproduction is frequently used. e.g., the units of vegetative propagation in plants –runner, rhizome, sucker, tuber, offset, bulb. These structures are called vegetative propagules.In Protists and Monerans, (All unicellular) the organism or the parent cell divides into two to give rise to new individuals. Thus, in these organisms cell division is itself a mode of reproduction.

Water hyacinth, an aquatic weed, also known as ‘terror of Bengal’ propagate vegetatively. Earlier this plant was introduced in India because of its beautiful flowers and shape of leaves. Since it can propagate vegetatively at a phenomenal rate and spread all over the water body in a short period of time, it drain oxygen from water body and cause death of fishes. (Eutrophication)

Bryophyllumshow vegetative propagation from the notches present at margins of leaves.

    • A sexual reproduction is the common method of reproduction in organisms that have a relatively simple organisation, like algae and fungi.
    • These organisms shift to sexual method of reproduction just before the onset of adverse conditions.
    • In higher plants both Asexual (vegetative) as well as sexual modes of reproduction are exhibited.
    • In most of the animals only sexual mode of reproduction is present.

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  • Sexual Reproduction

    • Sexual reproduction involves formation of the male and female gametes, either by the same individual or by different individuals of the opposite sex. These gametes fuse to form the zygote which develops to form the new organism.
    • It is an elaborate, complex and slow process as compared to asexual reproduction.
    • Because of the fusion of male and female gametes, sexual reproduction results in offspring that are not identical to the parents or amongst themselves.
    • Plants, animals, fungishow great diversity in external morphology, internal structure and physiology, but in sexual reproduction they share a similar pattern.
    • Juvenile / vegetative phase – All organisms have to reach a certain stage of growth and maturity in their life, before they can reproduce sexually. That period of growth is called the juvenile phase. It is known as vegetative phase in plants.
    • Reproductive phase –the beginning of the reproductive phase can be seen easily in the higher plants when they come to flower.
    • In some plants, where flowering occurs more than once, inter-flowering period is also known as juvenile period.
    • Plants-the annual and biennial types, show clear cut vegetative, reproductive and senescent phases, but in the perennial species it is very difficult to clearly define these phases.
    • Bamboo species flower only once in their life time, generally after 50-100 years, produce large number of fruits and die.
    • Strobilanthus kunthiana (neelakuranji), flowers once in 12 years. It is found in hilly areas in Kerala, Karnataka and Tamil Nadu.
    • In animals, the juvenile phase is followed by morphological and physiological changes prior to active reproductive behaviour.
    • birds living in nature lay eggs only seasonally. However, birds in captivity (as in poultry farms) can be made to lay eggs throughout the year. In this case, laying eggs is not related to reproduction but is a commercial exploitation for human welfare.
    • The females of placental mammals exhibit cyclical changes in the activities of ovaries and accessory ducts as well as hormones during the reproductive phase.
    • In non-primate mammals like cows, sheep, rats, deers, dogs, tiger, etc., such cyclical changes during reproduction are called oestrus cycle where as in primates (monkeys, apes, and humans) it is called menstrual cycle.
    • Many mammals, especially those living in natural, wild conditions exhibit such cycles only during favourable seasons in their reproductive phase and are therefore called seasonal breeders. Many other mammals are reproductively active throughout their reproductive phase and hence are called continuous breeders.
    • Senescent phase – The end of reproductive phase can be considered as one of the parameters of senescence or old age. There are concomitant changes in the body (like slowing of metabolism, etc.) during this last phase of life span. Old age ultimately leads to death.
    • In both plants and animals, hormones are responsible for the transitions between the three phases. Interaction between hormones and certain environmental factors regulate the reproductive processes and the associated behavioural expressions of organisms.
  • Events in sexual reproduction
    • Sexual reproduction is characterised by the fusion (or fertilisation) of the male and female gametes, the formation of zygote and embryo
    • These sequential events may be grouped into three distinct stages namely, the pre-fertilisation, fertilisation and the post-fertilisation events.
  • Pre-fertilisation Events
    • These include all the events of sexual reproduction prior to the fusion of gametes.
    • The two main pre-fertilisation events aregametogenesisandgamete transfer.
    • Gametogenesis
      • It refers to the process of formation of the two types of gametes – male and female.
      • Gametes are haploid cells.
      • In some algae the two gametes are so similar in appearance that it is not possible to categorise them into male and female gametes.They are hence, are calledhomogametes (isogametes).
      • However, in a majority of sexually reproducing organisms the gametes produced are of two morphologically distinct types (heterogametes). In such organisms the male gamete is called theantherozoid or sperm and the female gamete is called the egg or

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Sexuality in organisms:

  • Plants may have both male and female reproductive structures in the same plant (bisexual) or on different plants (unisexual).
  • In several fungi and plants, terms such as homothallic and monoecious are used to denote the bisexual condition and heterothallic and dioecious are the terms used to describe unisexual condition.
  • In flowering plants, the unisexual male flower is staminate, e., bearing stamens, while the female ispistillate or bearing pistils.
  • e.g., examples of monoecious plants – cucurbitsand coconuts
  • dioecious plants – Papayaand date palm.
  • Earthworms, sponge, tapeworm and leech are examples of bisexual animals (hermaphrodite). Cockroach is an example of a unisexual species.
  • Cell division during gamete formation:
  • Gametes in all heterogametic species are of two types namely, male and Gametes are haploid though the parent plant body from which they arise may be either haploid or diploid.
  • A haploid parent produces gametes by mitotic division like in monera, fungi, algae and bryophytes
  • In pteridophytes, gymnosperms, angiosperms and most of the animals including human beings, the parental body isIn these, specialised cells calledmeiocytes (gamete mother cell) undergo meiosis.
  • At the end of meiosis, only one set of chromosomesgets incorporated into each

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Name of organism Chromosome number in meiocyte (2n) Chromosome number in gamete (n)
Human beings 46 23
House fly 12 6
Rat 42 21
Dog 78 39
Cat 38 19
Fruit fly 8 4
Ophioglossum (a fern) 1260 630
Apple 34 17
Rice 24 12
Maize 20 10
Potato 48 24
Butterfly 380 190
Onion 32 16
  • Gamete Transfer:
  • After formation, male and female gametes must be physically brought together to facilitate fusion (fertilisation).
  • In most of organisms, male gamete is motile and the female gamete is stationary.
  • Exceptions – few fungi and algae in which both types of gametes are motile.
  • For transfer of male gametes, a medium is needed. In several simple plants like algae, bryophytes and pteridophytes, water is the medium for gamete transfer.
  • A large number of the male gametes, however, fail to reach the female gametes. To compensate this loss of male gametes during transport, the number of male gametes produced is very high.
  • In seed plants, pollen grains are the carriers of male gametes and ovule have the egg. Pollen grains produced in anthers therefore, have tobe transferred to the stigma before it can lead to fertilization.
  • In bisexual, self-fertilising plants, e.g., peas, transfer of pollen grains to the stigma is relatively easy as anthers and stigma are located close to each other; pollen grains soon after they are shed, come in contact with the stigma.
  • in cross pollinating plants (including dioecious plants), a specialised event called pollination facilitates transfer of pollen grains to the stigma.
  • Pollen grains germinate on the stigma and the pollen tubes carrying the male gametes reach the ovule and discharge male gametes near the egg.
  • In dioecious animals, since male and female gametes are formed in different individuals, the organism must evolve a special mechanism for gamete transfer. Successful transfer and coming together of gametes is essential for the most critical event in sexual reproduction, the fertilisation.

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  • Fertilisation
  • The most vital event of sexual reproduction is perhaps the fusion of gametes. This process is also calledsyngamyresults in the formation of a diploid
  • in some organisms like rotifers, honeybees and even some lizards and birds (turkey), the female gamete undergoes development to form new organisms without fertilisation. This phenomenon is called
  • In most aquatic organisms, such as a majority of algae and fishes as well as amphibians, syngamy occurs in the external medium (water), i.e., outside the body of the organism. This type of gametic fusion is called external fertilisation.

Organisms exhibiting external fertilisation show great synchrony between the sexes and release a large number of gametes into the surrounding medium (water) in order to enhance the chances of syngamy. This happens in the bony fishes and frogs where a large number of offspring are produced. A major disadvantage is that the offspring are extremely vulnerable to predators threatening their survival up to adulthood.

  • In many terrestrial organisms, belonging to fungi, higher animals such as reptiles birds, mammals and in a majority of plants (bryophytes, pteridophytes, gymnosperms and angiosperms), syngamy occurs insidethe body of the organism, hence the process is called internal fertilisation.

In all these organisms, egg is formed inside the female body where they fuse with the male gamete. In organisms exhibiting internal fertilisation, the male gamete is motile and has to reach the egg in order to fuse with it. In these even though the number of sperms produced is very large, there is a significant reduction in the number of eggs produced. In seed plants, however, the non-motile male gametes are carried to female gamete by pollen tubes.

  • Post-fertilisation Events
  • Events in sexual reproduction after the formation of zygote are called post-fertilisation events.
  • Zygote :
    • Formation of the diploid zygote is universal in all sexually reproducing organisms.
    • In organisms with external fertilisation, zygote is formed in the external medium (usually water), whereas in those exhibiting internal fertilisation, zygote is formed inside the body of the organism.
    • Further development of the zygote depends on the type of life cycle the organism has and the environment it is exposed to.
    • In organisms belonging to fungi and algae, zygote develops a thick wall that is resistant to dessication and damage. It undergoes a period of rest before germination.
    • In organisms with haplontic life cycle, zygote divides by meiosis to form haploid spores that grow into haploid individuals.
    • Zygote is the vital link that ensures continuity of species between organisms of one generation and the next.
    • Every sexually reproducing organism, including human beings begin life as a single cell-the zygote.
  • Embryogenesis :
    • It refers to the process of development ofembryo from the zygote.
    • During embryogenesis, zygote undergoes cell division (mitosis) and cell differentiation. While cell divisions increase the number of cells in the developing embryo; cell differentiation helps groups of cells to undergo certain modifications to form specialised tissues and organs to form an organism.
    • Animals are categorised into oviparous and viviparous based on whether the development of the zygote take place outside the body of the female parent or inside, i.e., whether they lay fertilised/unfertilised eggs or give birth to young ones.
    • In oviparous animals like reptiles and birds,the fertilised eggs covered by hard calcareous shell are laid in a safe place in the environment; after a period of incubation young ones hatch out.
    • in viviparous animals (majority of mammals including human beings), the zygote develops into a young one inside the body of the female organism. After attaining a certain stage of growth, the young ones are delivered out of the body of the female organism. Because of proper embryonic care and protection, the chances of survival of young ones is greater in viviparous organisms.
    • In flowering plants, the zygote is formed inside the ovule. After fertilisation the sepals, petals and stamens of the flower wither and fall off.
    • The pistil however, remains attached to the plant. The zygote develops into the embryo and the ovules develop into the seed. The ovary develops into the fruit which develops a thick wall called pericarp that is protective in function. After dispersal, seeds germinate under favourable conditions to produce new plants.1downloadble pdf file is available…please click on the link below…

CHAPTER 1 – REPRODUCTION IN ORGANISMS

CHAPTER 10 – CELL CYCLE AND CELL DIVISION

CELL CYCLE AND CELL DIVISION

  • Growth and reproduction are characteristics of living cells and organisms.

Cell Cycle –

  • The sequence of events by which a cell duplicates its genome, synthesizes the other constituents of the cell and eventually divides into two daughter cells is termed cell cycle.
  • Cell cycle includes three processes cell division, DNA replication and cell growth in coordinated way.
  • Duration of cell cycle can vary from organism to organism and also from cell type to cell type. (e.g., in Yeast cell cycle is of 90 minutes, in human 24 hrs.)

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Interphase

  • It is divided into 3 further phases G1, S, and G2.

G1 phase (Gap 1 Phase)

  • Corresponds to the interval between mitosis and initiation of DNA replication.
  • During G1 phase the cell is metabolically active and continuously grows but does not replicate its DNA.

S phase (synthesis phase)

  • period during which DNA synthesis or replication takes place.
  • During this time the amount of DNA per cell doubles. (only amount of DNA is doubled, no of chromosomes remain same.)
  • In animal cells, during the S phase, DNA replication begins in the nucleus, and the centriole duplicates in the cytoplasm.

G2 phase (Gap 2 Phase)

  • Proteins are synthesised in preparation for mitosis while cell growth continues.

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  • Some cells do not exhibit division like heart cells, nerve cells etc. these cells enter in an inactive phase called G0 or quiescent phase from G1 phase.
  • Cells in this phase are metabolically active but they do not divide unless they are called on to do so.

Mitosis or M phase

  • In animals, mitotic cell division is only seen in the diploid somatic cells while in the plants mitotic divisions can be seen in both haploid and diploid cells.
  • it is also called as equational division as the number of chromosomes in the parent and progeny cells are the same.
  • Mitosis is divided into the following four stages:
    • Prophase
    • Metaphase
    • Anaphase
    • Telophase

Prophase

  • It follows the S and G2 phases of interphase.
  • The centrioles now begin to move towards opposite poles of the cell.
  • In prophase Chromosomal material condenses to form compact mitotic chromosomes.
  • Initiation of the assembly of mitotic spindle with the help of the microtubules.
  • Cell organelles like Golgi complexes, endoplasmic reticulum, nucleolus and the nuclear envelope disappear.

Metaphase

  • Start of metaphase is marked by the complete disintegration of the nuclear envelope.
  • The chromosomes are spread through the cytoplasm of the cell.
  • condensation of chromosomes is completed and they can be observed clearly under the microscope.
  • This is the stage at which morphology of chromosomes is most easily studied.
  • At this stage, metaphase chromosome is made up of two sister chromatids, which are held together by the centromere.
  • centromere serve as the sites of attachment of spindle fibres to the chromosomes.
  • chromosomes are moved into position at the centre of the cell.
  • the metaphase is characterised by all the chromosomes coming to lie at the equator with one chromatid of each chromosome connected by its kinetochore to spindle fibres from one pole and its sister chromatid connected by its kinetochore to spindle fibres from the opposite pole.
  • The plane of alignment of the chromosomes at metaphase is referred to as the metaphase plate or equatorial plate.

Anaphase

  • At the onset of anaphase, each chromosome arranged at the metaphase plate is split simultaneously and the two daughter chromatids begin to move towards the two opposite poles.
  • As each chromosome moves away from the equatorial plate, the centromere of each chromosome is towards the pole and hence at the leading edge, with the arms of the chromosome trailing behind

Telophase

  • At the beginning of telophase, the chromosomes at their respective poles decondense and form chromatin network.
  • Nuclear envelope assembles around the chromatin network.
  • Nucleolus, Golgi complex and ER etc cell organelles reform.

Cytokinesis

  • After karyokinesis the cell itself is divided into two daughter cells by a separate process called cytokinesis.
  • In an animal cell, this is achieved by the appearance of a furrow in the plasma membrane.
  • The furrow gradually deepens and ultimately joins in the centre dividing the cell cytoplasm into two.
  • Plant cells undergo cytokinesis by cell plate method. In cell plate method wall formation starts in the centre of the cell and grows outward to meet the existing lateral walls.
  • The formation of the new cell wall begins with the formation of a simple precursor, called the cell-plate that represents the middle lamella between the walls of two adjacent cells.
  • At the time of cytoplasmic division, organelles like mitochondria and plastids get distributed between the two daughter cells.
  • In some organisms karyokinesis is not followed by cytokinesis as a result of which multinucleate condition arises leading to the formation of syncytium (e.g., liquid endosperm in coconut). (should be coenocytic)

Significance of mitosis

  • Mitosis results in the production of diploid daughter cells with identical genetic complement usually.
  • The growth of multicellular organisms is due to mitosis.
  • Cell growth results in disturbing the ratio between the nucleus and the cytoplasm. Therefore, cell divide to restore the nucleo-cytoplasmic ratio.
  • mitosis is important in cell repair. The cells of the upper layer of the epidermis, cells of the lining of the gut, and blood cells are being constantly replaced.
  • Mitotic divisions in the meristematic tissues – the apical and the lateral cambium, result in a continuous growth of plants throughout their life.

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Meiosis

  • The specialised kind of cell division that reduces the chromosome number by half results in the production of haploid daughter cells called
  • It is responsible for formation of haploid gametes, which during sexual reproduction form diploid zygote by fusion.
  • Meiosis involves two sequential cycles of nuclear and cell division called meiosis I and meiosis II but only a single cycle of DNA replication.
  • Interphase of meiosis is similar to interphase of mitosis.

 

Meiosis I

Prophase I

  • Prophase of the meiosis I division is typically longer and more complex than prophase of mitosis.
  • It has been further subdivided into the following five phases based on chromosomal behavior.

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 Metaphase I:

  • The bivalent chromosomes align on the equatorial plate.
  • The microtubules from the opposite poles of the spindle attach to the pair of homologous chromosomes.

Anaphase I:

  • The homologous chromosomes separate, while sister chromatids remain associated at their centromeres.

Telophase I

  • The nuclear membrane and nucleolus reappear.
  • cytokinesis follows telophase I.
  • Although in many cases the chromosomes do undergo some dispersion, they do not reach the extremely extended state of the interphase nucleus. The stage between the two meiotic divisions is called interkinesis and is generally short lived.
  • Interkinesis is followed by prophase II, a much simpler prophase than prophase I.

 

Meiosis II

Meiosis II resembles a normal mitosis.

Prophase II:

  • Meiosis II is initiated immediately after cytokinesis.
  • The nuclear membrane disappears by the end of prophase II.
  • The chromosomes again become compact.

Metaphase II:

  • At this stage the chromosomes align at the equator and the microtubules from opposite poles of the spindle get attached to the kinetochores of sister chromatids.

Anaphase II:

  • splitting of the centromere of each chromosome.
  • Chromosomes move toward opposite poles of the cell.

Telophase II:

  • the two groups of chromosomes once again get enclosed by a nuclear envelope.
  • cytokinesis follows resulting in the formation of four haploid daughter cells).

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SIGNIFICANCE OF MEIOSIS

  • by meiosis conservation of specific chromosome number of each species is achieved across generations in sexually reproducing organisms.
  • It also increases the genetic variability in the population of organisms from one generation to the next. Variations are very important for the process of evolution.

 

 

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CHAPTER 10 – CELL CYCLE AND CELL DIVISION