• Green plants carry out ‘photosynthesis’, a physico-chemical process by which they use light energy to drive the synthesis of organic compounds.
  • Ultimately, all living forms on earth depend on sunlight for energy.
  • The use of energy from sunlight by plants doing photosynthesis is the basis of life on earth. Photosynthesis is important due to two reasons:

(a) it is the primary source of all food on earth.

(b) It is also responsible for the release of oxygen into the atmosphere by green plants.

  • chlorophyll (green pigment of the leaf), light and CO2 are required for photosynthesis to occur.


  • Two leaves experiment for importance of chlorophyll for starch formation – a variegated leaf or a leaf that was partially covered with black paper, and one that was exposed to light. On testing these leaves for starch it was clear that photosynthesis occurred only in the green parts of the leaves in the presence of light.
  • Half-leaf experiment for importance of CO2 for starch formation – a part of a leaf is enclosed in a test tube containing some KOH soaked cotton (which absorbs CO2), while the other half is exposed to air. The setup is then placed in light for some time. On testing for starch later in the two halves of the leaf the exposed part of the leaf tested positive for starch while the portion that was in the tube, tested negative.


Early Experiments

Joseph Priestley (1770)               – revealed the essential role of air in the growth of green plants

– discovered oxygen in 1774.

– Bell jar experiment

– hypothesised that Plants restore to the air whatever breathing animals and burning candles remove.



Jan Ingenhousz (1730-1799)      – showed that sunlight is essential to the photosynthesis.

Julius von Sachs (1854)              – provided evidence for production of glucose when plants grow.

                                                      – Glucose is usually stored as starch.

– showed that the green substance in plants is located in special bodies (later called chloroplasts) within plant cells.

T.W Engelmann (1843 – 1909)   – Used a prism to split light into its spectral components and then illuminated a green alga, Cladophora, placed in a suspension of aerobic bacteria. The bacteria were used to detect the sites of O2 evolution.

observed that the bacteria accumulated mainly in the region of blue and red light of the split spectrum.

described first action spectrum of photosynthesis, which resembles roughly the absorption spectra of chlorophyll a and b.

The empirical equation representing the total process of photosynthesis for oxygen evolving organisms was then understood as:

CO2 + H2O —Light –> [CH2O] + O2

where [CH2O] represented a carbohydrate (e.g., glucose, a six-carbon sugar).

Cornelius van Niel (1897-1985)  – a microbiologist

– studies of purple and green bacteria,

– demonstrated that photosynthesis is essentially a light-dependent   reaction in which hydrogen from a suitable oxidisable compound reduces carbon dioxide to carbohydrates.

2H2A + CO2Light –> 2A + CH2O + H2O

– In green plants H2O is the hydrogen donor and is oxidised to O2.

– When H2S is the hydrogen donor for purple and green sulphur bacteria, the ‘oxidation’ product is sulphur or sulphate and not O2.

– Hence, he inferred that the O2 evolved by the green plant comes from H2O, not from carbon dioxide.

– This was later proved by using radioisotopic techniques.

The correct equation, that would represent the overall process of photosynthesis is therefore:

6CO2 +12H2O —Light –> C6H12O6 + 6H2O + 6O2



  • Photosynthesis occurs in green leaf in the chloroplasts.
  • mesophyll cells in the leaves, have a large number of chloroplasts. Usually the chloroplasts align themselves along the walls of the mesophyll cells, such that they get the optimum quantity of the incident light.
  • Within the chloroplast there is the membranous system consisting of grana, the stroma lamellae, and the fluid stroma. There is a clear division of labour within the chloroplast. The membrane system is responsible for trapping the light energy and also for the synthesis of ATP and NADPH. In stroma, enzymatic reactions incorporate CO2 into the plant leading to the synthesis of sugar, which in turn forms starch.
  • The former set of reactions, since they are directly light driven are called light reactions. The latter are not directly light driven but are dependent on the products of light reactions (ATP and NADPH). Hence, to distinguish the latter they are called, by convention, as dark reactions. However, this should not be construed to mean that they occur in darkness or that they are not light- dependent.


  • A chromatographic separation of the leaf pigments shows that the colour that we see in leaves is not due to a single pigment but due to four pigments:

Chlorophyll a (bright or blue green in the chromatogram),

chlorophyll b (yellow green),

xanthophylls (yellow) and

carotenoids (yellow to yellow-orange).

  • Pigments are substances that have an ability to absorb light, at specific wavelengths.
  • wavelengths at which there is maximum absorption by chlorophyll a, is in the blue and the red regions, also shows higher rate of photosynthesis. Hence, we can conclude that chlorophyll a is the chief pigment associated with photosynthesis.
  • These graphs, together, show that most of the photosynthesis takes place in the blue and red regions of the spectrum; some photosynthesis does take place at the other wavelengths of the visible spectrum.
  • Though chlorophyll is the major pigment responsible for trapping light, other thylakoid pigments like chlorophyll b, xanthophylls and carotenoids, which are called accessory pigments, also absorb light and transfer the energy to chlorophyll a. Indeed, they not only enable a wider range of wavelength of incoming light to be utilised for photosyntesis but also protect chlorophyll a from photo-oxidation.

Screenshot (43)


  • Light reactions or the ‘Photochemical’ phase include light absorption, water splitting, oxygen release, and the formation of high-energy chemical intermediates, ATP and NADPH.
  • several complexes are involved in the process.
  • The pigments are organised into two discrete photochemical light harvesting complexes (LHC) within the Photosystem I (PS I) and Photosystem II (PS II). These are named in the sequence of their discovery, and not in the sequence in which they function during the light reaction.
  • The LHC are made up of hundreds of pigment molecules bound to proteins.
  • Each photosystem has all the pigments (except one molecule of chlorophyll a) forming a light harvesting system also called antennae.
  • These pigments help to make photosynthesis more efficient by absorbing different wavelengths of light. The single chlorophyll a molecule forms the reaction centre.
  • The reaction centre is different in both the photosystems.
  • In PS I the reaction centre chlorophyll a has an absorption peak at 700 nm, hence is called P700, while in PS II it has absorption maxima at 680 nm, and is called P680


  • In photosystem II the reaction centre chlorophyll a absorbs 680 nm wavelength of red light causing electrons to become excited and jump into an orbit farther from the atomic nucleus. These electrons are picked up by an electron acceptor which passes them to an electrons transport system consisting of cytochromes.
  • This movement of electrons is downhill, in terms of an oxidation-reduction or redox potential scale.
  • The electrons are not used up as they pass through the electron transport chain, but are passed on to the pigments of photosystem PS I.
  • Simultaneously, electrons in the reaction centre of PS I are also excited when they receive red light of wavelength 700 nm and are transferred to another accepter molecule that has a greater redox potential.
  • These electrons then are moved downhill again, this time to a molecule of energy-rich NADP+.
  • The addition of these electrons reduces NADP+ to NADPH + H+.
  • This whole scheme of transfer of electrons, starting from the PS II, uphill to the accepter, down the electron transport chain to PS I, excitation of electrons,transfer to another accepter, and finally down hill to NADP+ causing it to be reduced to NADPH + H+ is called the Z scheme, due to its characterstic shape. This shape is formed when all the carriers are placed in a sequence on a redox potential scale.

Screenshot (45)  Splitting of Water

  • The electrons that were moved from photosystem II must be replaced.
  • This is achieved by electrons available due to splitting of water.
  • The splitting of water is associated with the PS II; water is split into H+, [O] and electrons.
  • This creates oxygen, one of the net products of photosynthesis.
  • The electrons needed to replace those removed from photosystem I are provided by photosystem II.
  • 2H2O ——-> 4H+ + O2 + 4e
  • water splitting complex is associated with the PS II, which itself is physically located on the inner side of the membrane of the thylakoid.

Cyclic and Non-cyclic Photo-phosphorylation

  • Living organisms have the capability of extracting energy from oxidisable substances and store this in the form of bond energy.
  • Special substances like ATP, carry this energy in their chemical bonds.
  • The process of whichATP is synthesised by cells (in mitochondria and chloroplasts) is named phosphorylation.
  • Photo- phosphorylation is the synthesis of ATP from ADP and inorganic phosphate in the presence of light.
  • When the two photosystems work in a series, first PS II and then the PS I, a process called non-cyclic photo-phosphorylation occurs.
  • The two photosystems are connected through an electron transport chain, as seen earlier – in the Z scheme.
  • Both ATP and NADPH + H+ are synthesised by this kind of electron flow.
  • When only PS I is functional, the electron is circulated within the photosystem and the phosphorylation occurs due to cyclic flow of electrons.
  • A possible location where this could be happening is in the stroma lamellae.
  • While the membrane or lamellae of the grana have both PS I and PS II the stroma lamellae membranes lack PS II as well as NADP reductase enzyme.
  • The excited electron does not pass on to NADP+ but is cycled back to the PS I complex through the electron transport chain. The cyclic flow hence, results only in the synthesis of ATP, but not of NADPH + H+.
  • Cyclic photophosphorylation also occurs when only light of wavelengths beyond 680 nm are available for excitation.

Chemiosmotic Hypothesis

  • The chemiosmotic hypothesis has been put forward to explain the mechanism of synthesis of ATP.
  • Like in respiration, in photosynthesis too, ATP synthesis is linked to development of a proton gradient across a membrane.
  • This time these are membranes of the thylakoid.
  • There is one difference though, here the proton accumulation is towards the inside of the membrane, i.e., in the lumen.
  • In respiration, protons accumulate in the intermembrane space of the mitochondria when electrons move through the ETS.
  • Processes that take place during the activation of electrons and their transport to determine the steps that cause a proton gradient to develop are –

(a) Since splitting of the water molecule takes place on the inner side of the membrane, the protons or hydrogen ions that are produced by the splitting of water accumulate within the lumen of the thylakoids.

(b) As electrons move through the photosystems, protons are transported across the membrane. This happens because the primary accepter of electron which is located towards the outer side of the membrane transfers its electron not to an electron carrier but to an H carrier. Hence, this molecule removes a proton from the stroma while transporting an electron. When this molecule passes on its electron to the electron carrier on the inner side of the membrane, the proton is released into the inner side or the lumen side of the membrane.

(c) The NADP reductase enzyme is located on the stroma side of the membrane. Along with electrons that come from the accepter of electrons of PS I, protons are necessary for the reduction of NADP+ to NADPH+ H+. These protons are also removed from the stroma.

  • Hence, within the chloroplast, protons in the stroma decrease in number, while in the lumen there is accumulation of protons.
  • This creates a proton gradient across the thylakoid membrane as well as a measurable decrease in pH in the lumen.
  • This proton gradient is important because it is the breakdown of this gradient that leads to release of energy.
  • The gradient is broken down due to the movement of protons across the membrane to the stroma through the transmembrane channel of the F0 of the ATPase.
  • The ATPase enzyme consists of two parts:

one called the F0 is embedded in the membrane and forms a transmembrane channel that carries out facilitated diffusion of protons across the membrane.

The other portion is called F1 and protrudes on the outer surface of the thylakoid membrane on the side that faces the stroma. The breakdown of the gradient provides enough energy to cause a conformational change in the F1 particle of the ATPase, which makes the enzyme synthesise several molecules of energy-packed ATP.

  • Chemiosmosis requires a membrane, a proton pump, a proton gradient and ATPase.
  • Energy is used to pump protons across a membrane, to create a gradient or a high concentration of protons within the thylakoid lumen.
  • ATPase has a channel that allows diffusion of protons back across the membrane; this releases enough energy to activate ATPase enzyme that catalyses the formation of ATP.
  • Along with the NADPH produced by the movement of electrons, the ATP will be used immediately in the biosynthetic reaction taking place in the stroma, responsible for fixing CO2, and synthesis of sugars.

Screenshot (46)


  • The products of light reaction are ATP, NADPH and O2.
  • Of these O2 diffuses out of the chloroplast while ATP and NADPH are used to drive the processes leading to the synthesis of food, more accurately, sugars.
  • This is the biosynthetic phase of photosynthesis.
  • This process does not directly depend on the presence of light but is dependent on the products of the light reaction, i.e., ATP and NADPH, besides CO2 and H2
  • Immediately after light becomes unavailable, the biosynthetic process continues for some time, and then stops. If then, light is made available, the synthesis starts again.
  • CO2 is combined with H2O to produce (CH2O)n or sugars.
  • Melvin Calvin used radioactive 14C in algal photosynthesis studies which led to the discovery that the first CO2 fixation product was a 3-carbon organic acid or 3-phosphoglyceric acid (PGA).
  • He also contributed to working out the complete biosynthetic pathway; hence it was called Calvin cycle after him.
  • In another group of plants, first stable product of CO2 fixation is 4 carbon organic acid oxaloacetic acid or OAA.
  • The Primary Acceptor of CO2 is a 5-carbon ketose sugar – ribulose bisphosphate (RuBP).


  • Calvin and his co-workers then worked out the whole pathway and showed that the pathway operated in a cyclic manner; the RuBP was regenerated.
  • The Calvin pathway occurs in all photosynthetic plants; it does not matter whether they have C3 or C4 (or any other) pathways.
  • The Calvin cycle can be described under three stages: carboxylation, reduction and regeneration.
  1. Carboxylation –

Carboxylation is the fixation of CO2 into a stable organic intermediate.

Carboxylation is the most crucial step of the Calvin cycle where CO2 is utilised for the carboxylation of RuBP.

This reaction is catalysed by the enzyme RuBP carboxylase which results in the formation of two molecules of 3-PGA.

Since this enzyme also has an oxygenation activity it would be more correct to call it RuBP carboxylase-oxygenase or RuBisCO.

  1. Reduction –

These are a series of reactions that lead to the formation of glucose.

The steps involve utilisation of 2 molecules of ATP for phosphorylation and two of NADPH for reduction per CO2 molecule fixed.

The fixation of six molecules of CO2 and 6 turns of the cycle are required for the removal of one molecule of glucose from the pathway.

  1. Regeneration –

Regeneration of the CO2 acceptor molecule RuBP is crucial if the cycle is to continue uninterrupted.

The regeneration steps require one ATP for phosphorylation to form RuBP.

  • Hence for every CO2 molecule entering the Calvin cycle, 3 molecules of ATP and 2 of NADPH are required.
  • It is probably to meet this difference in number of ATP and NADPH used in the dark reaction that the cyclic phosphorylation takes place.
  • To make one molecule of glucose 6 turns of the cycle are required.
  • Total 18 ATP, 12 NADPH, 12 CO2 are used synthesis of for 1 molecule of glucose.

 Screenshot (47)


  • Plants that are adapted to dry tropical regions have the C4 pathway
  • Though these plants have the C4 oxaloacetic acid as the first CO2 fixation product they use the C3 pathway or the Calvin cycle as the main biosynthetic pathway.
  • C4 plants are special: They have a special type of leaf anatomy, they tolerate higher temperatures, they show a response to highlight intensities, they lack a process called photorespiration and have greater productivity of biomass.
  • The particularly large cells around the vascular bundles of the C4 pathway plants are called bundle sheath cells, and the leaves which have such anatomy are said to have ‘Kranz’ anatomy.
  • ‘Kranz’ means ‘wreath’ and is a reflection of the arrangement of cells.
  • The bundle sheath cells may form several layers around the vascular bundles; they are characterised by having a large number of chloroplasts, thick walls impervious to gaseous exchange and no intercellular spaces.
  • This pathway also known as Hatch and Slack Pathway, is a cyclic process.
  • The primary CO2 acceptor is a 3-carbon molecule phosphoenol pyruvate (PEP) and is present in the mesophyll cells. The enzyme responsible for this fixation is PEP carboxylase or PEPcase.
  • The mesophyll cells lack RuBisCO enzyme.
  • The C4 acid OAA is formed in the mesophyll cells.
  • It then forms other 4-carbon compounds like malic acid or aspartic acid in the mesophyll cells itself, which are transported to the bundle sheath cells.
  • In the bundle sheath cells these C4 acids are broken down to release CO2 and a 3-carbon molecule.
  • The 3-carbon molecule is transported back to the mesophyll where it is converted to PEP again, thus, completing the cycle.
  • The CO2 released in the bundle sheath cells enters the C3 or the Calvin pathway, a pathway common to all plants. The bundle sheath cells are rich in an enzyme Ribulose bisphosphate carboxylase-oxygenase (RuBisCO), but lack PEPcase.
  • Thus, the basic pathway that results in the formation of the sugars, the Calvin pathway, is common to the C3 and C4
  • the Calvin pathway occurs in all the mesophyll cells of the C3 plants while in the C4 plants it does not take place in the mesophyll cells but does so only in the bundle sheath cells.

Screenshot (48)


  • RuBisCO is the most abundant enzyme in the world.
  • It is characterised by the fact that its active site can bind to both CO2 and O2 – hence the name.
  • RuBisCO has a much greater affinity for CO2 than for O2.
  • This binding is competitive. It is the relative concentration of O2 and CO2 that determines which of the two will bind to the enzyme.
  • In C3 plants some O2 does bind to RuBisCO, and hence CO2 fixation is decreased. Here the RuBP instead of being converted to 2 molecules of PGA binds with O2 to form one molecule and phosphoglycolate in a pathway called photorespiration.
  • In the photorespiratory pathway, there is neither synthesis of sugars, nor of ATP. Rather it results in the release of CO2 with the utilisation of ATP.
  • In the photorespiratory pathway there is no synthesis of ATP or NADPH. Therefore, photorespiration is a wasteful process.
  • In C4 plants photorespiration does not occur. This is because they have a mechanism that increases the concentration of CO2 at the enzyme site.
  • This takes place when the C4 acid from the mesophyll is broken down in the bundle cells to release CO2 – this results in increasing the intracellular concentration of CO2.
  • In turn, this ensures that the RuBisCO functions as a carboxylase minimising the oxygenase activity.
  • Because of absence of photorespiration in C4 plants, productivity and yields are better in these plants.
  • These plants show tolerance to higher temperatures.
Characteristics C3 Plants C4 Plants
Cell type in which the Calvin cycle takes place Both Bundle sheath
Cell type in which the initial carboxylation reaction occurs Both Mesophyll
How many cell types does the leaf have that fix Co2. Three: Bundle sheath, palisade, spongy mesophyll Two: Bundle sheath and


Which is the primary Co2 acceptor RuBP PEP
Number of carbons in the primary Co2 acceptor 5 3
Which is the primary Co2 fixation product PGA OAA
No. of carbons in the primary Co2 fixation product 3 4
Does the plant have RuBisCo? Yes Yes
Does the plant have PEP Case? No Yes
Which cells in the plant have Rubisco? Mesophyll/Bundle sheath/none Mesophyll/Bundle sheath/none
Co2 fixation rate under high light conditions Medium High
Whether photorespiration is present at low light intensities Sometimes Negligible
Whether photorespiration is present at high light intensities Sometimes Negligible
Whether photorespiration would be present at low CO2 concentrations High Negligible
Whether photorespiration would be present at high CO2 concentrations Sometimes Negligible
Temperature optimum 20-25C 30-40 C


  • The rate of photosynthesis is very important in determining the yield of plants including crop plants.
  • Photosynthesis is under the influence of several factors, both internal (plant) and external.
  • The plant factors include the number, size, age and orientation of leaves, mesophyll cells and chloroplasts, internal CO2 concentration and the amount of chlorophyll. The plant or internal factors are dependent on the genetic predisposition and the growth of the plant.
  • The external factors include the availability of sunlight, temperature, CO2 concentration and water.
  • As a plant photosynthesises, all these factors will simultaneously affect its rate. Hence, though several factors interact and simultaneously affect photosynthesis or CO2 fixation, usually one factor is the major cause or is the one that limits the rate. Hence, at any point the rate will be determined by the factor available at sub-optimal levels.
  • Blackman’s (1905) Law of Limiting Factors – If a chemical process is affected by more than one factor, then its rate will be determined by the factor which is nearest to its minimal value: it is the factor which directly affects the process if its quantity is changed
  • For example, despite the presence of a green leaf and optimal light and CO2 conditions, the plant may not photosynthesise if the temperature is very low. This leaf, if given the optimal temperature, will start photosynthesising.


  • It includes light quality, light intensity and the duration of exposure to light.
  • There is a linear relationship between incident light and CO2 fixation rates at low light intensities.
  • At higher light intensities, gradually the rate does not show further increase as other factors become limiting.
  • Light saturation occurs at 10 per cent of the full sunlight.
  • Hence, except for plants in shade or in dense forests, light is rarely a limiting factor in nature.
  • Increase in incident light beyond a point causes the breakdown of chlorophyll and a decrease in photosynthesis.

Screenshot (50)

Carbon dioxide Concentration

  • Carbon dioxide is the major limiting factor for photosynthesis.
  • The concentration of CO2 is very low in the atmosphere (between 0.03 and 0.04 per cent).
  • Increase in concentration upto 0.05 per cent can cause an increase in CO2 fixation rates; beyond this the levels can become damaging over longer periods.
  • The C3 and C4 plants respond differently to CO2 At low light conditions neither group responds to high Co2 conditions. At high light intensities, both C3 and C4 plants show increase in the rates of photosynthesis.
  • C4 plants show saturation at about 360 µlL-1 while in C3 plants saturation is seen only beyond 450 µlL-1. Thus, current availability of CO2 levels is limiting to the C3
  • The fact that C3 plants respond to higher CO2 concentration by showing increased rates of photosynthesis leading to higher productivity has been used for some greenhouse crops such as tomatoes and bell pepper. They are allowed to grow in carbon dioxide enriched atmosphere that leads to higher yields.


  • The dark reactions being enzymatic are temperature controlled.
  • Though the light reactions are also temperature sensitive they are affected to a much lesser extent. The C4 plants respond to higher temperatures and show higher rate of photosynthesis while C3 plants have a much lower temperature optimum.
  • The temperature optimum for photosynthesis of different plants also depends on the habitat that they are adapted to.
  • Tropical plants have a higher temperature optimum than the plants adapted to temperate climates.


  • Even though water is one of the reactants in the light reaction, the effect of water as a factor is more through its effect on the plant, rather than directly on photosynthesis.
  • Water stress causes the stomata to close hence reducing the CO2 Besides, water stress also makes leaves wilt, reducing the surface area of the leaves and their metabolic activity as well.

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  • 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.


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.


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
  • Using Agrobacterium vectors, nematode-specific genes were introduced into the host
  • 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
  • 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.



  • 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
  • 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
  • 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.


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
  • 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.


  • 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
    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.


    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.


  • 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.


  • 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
  • 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.

biotechnology and its applications


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’.


  • 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.


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.


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


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.


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).
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.


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.


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.


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.


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.

principles and processes in biotechnology

NEET-2016 allotment list (after 1st Counseling)

Neet2016 1st Counseling results are out..
Cutoff rank
MBBS (UR) 4733
(OBC) 4781
(SC) 30020
(ST) 53031
MBBS (UR-PH) 160789
(SC-PH) 362248
(ST-PH) 366239
BDS (UR) 7579
(OBC) 7589
(SC) 38204
(ST) 67190
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(ST-PH) 329022

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Round 1 Allotment


Congratulations all for your performance in NEET-2016..

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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. 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





  • 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

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.




  • 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

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


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.


  • 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…