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BIOCHEMISTRY OF
SEED
GERMINATION
AND FRUIT
RIPENING
JANANI PALPANDI
II BT
DEPT., OF BIOTECHNOLOGY,
MEPCO SCHLENK ENGG., COLLEGE.
BICHEMISTRY OF
SEED GERMINATION
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BIOCHEMICAL REACTIONS UNDERTHISTOPIC
• Respiration
• Protein and Nucleic Acid Synthesis
• Mobilization of stored reserves
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RESPIRATION
• In imbibed seeds the three metabolic pathways take place:
• Glycolysis
• Pentose Phosphate Pathway
• Citric Acid Cycle
• Through Glycolysis And Pentose Phosphate pathway – ATP, NADPH, FAD
produced.
• Through HMP shunt – NADP
• NADPH , NADP – Readily available reducing agents – utilized in bio-reductive
reactions
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RESPIRATION
• Dry seeds - 10-15 per cent water.
• Dry seeds - low level of respiration.
• Notable metabolic event during imbibition occurring in less than 15 min is the
reformation of keto acids from amino acids by deamination and
transamination.
• Divided into four phases.
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RESPIRATION
PHASE I
• Characterized by a sharp rise in
respiration for about 10 hours.
• Due to the activation and hydration
of mitochondrial enzymes.
• Enzymes - belonging to the cycle
and electron transport chain.
• There is rapid oxygen uptake into
seeds with intact testas during
phase I .
• Known as Early imbibition
PHASE II
• Involves a lag in respiration between
10 and 25 hours - after the start of
imbibition.
• Hydration of the cotyledons-
completed and all pre-existing
enzymes- activated.
• Testa impedes oxygen uptake in
phase II. (slows down).
• Between phase II and phase III, the
radicle penetrates the testa.
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RESPIRATION
PHASE III
• A second respiratory surge.
• Due to increased oxygen supply
through pierced testa.
• Another reason - newly synthesized
mitochondria and respiratory
enzymes in the dividing cells of the
growing axis.
PHASE IV
• Marked fall in respiration.
• disintegration of the cotyledons .
• exhaustion of the stored food.
• Early stages of germination,
respiration is cyanide-resistant and
the alternative oxidase instead of
cytochrome oxidase plays a role in
germination.
• . At later stages of germination,
however, respiration becomes
sensitive to cyanide.
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ROLE OF CYANIDE IN GERMINATION
• Cyanide is a germination regulatory agent.
• In some plants like Amaranthus at lower concentration it promotes seed
germination.
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PROTEIN AND NUCLEIC ACID SYNTHESIS
• Pre-requisite for radicle emergence .
• Protein synthesis does not occur in the dry seeds.
• Starts when these are hydrated and cytoplasmic ribosomes (eukaryotic SOS)
get associated with messenger RNA (mRNA).
• Radicle is the embryonic part from which root system develops.
• It is the first developed system.
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PROTEIN AND NUCLEIC ACID SYNTHESIS
• Initiation of protein synthesis - attachment of the
• small (40S) ribosomal subunit.
• tRNA molecule .
• Attachment to the initiation site on the mRNA.
• After formation of the initiation complex - the large (60S) ribosomal subunit
becomes attached .
• Protein synthesis begins.
• Formation of the initiation complex requires CTP and perhaps ATP.
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PROTEIN AND NUCLEIC ACID SYNTHESIS
• Other soluble protein factors - required for the transfer of aminoacyl-tRNAs to
their appropriate codons on the ribosome-mRNA complex.
• To move the message through the ribosome.
• These are the elongation factors requiring CTP for activity.
• New mRNA synthesis - not essential for resumption of protein synthesis
during the first hour of imbibition.
• mRNA conserved in the dry embryo - utilized for early protein synthesis.
• Even if new mRNA is synthesized as an early event, it is not involved in early
protein synthesis.
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PROTEIN AND NUCLEIC ACID SYNTHESIS
• Ribosomal RNA (rRNA) synthesis commences as early as mRNA
• Increases during and after imbibition - after the 6th hour coinciding with the
time of radicle elongation and
• At 16-18h it reaches a rate about 12-fold greater than the earlier stage.
• Although transfer RNA (tRNA) synthesis begins within 20 minutes in imbibing
embryos - not certain whether - involved in early protein synthesis.
• Its new synthesis is necessary during later stages to sustain-growth.
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PROTEIN AND NUCLEIC ACID SYNTHESIS
• The radicle within the seed initially grows by cell elongation.
• Emergence through the seed coat may be associated with cell division.
• DNA synthesis which is closely linked to mitotic cell division is mainly
concerned with the post-germination period of seedling development.
• During 9th hour – Inhibition of protein synthesis inhibits DNA synthesis.
• After 9th hour – Inhibition of protein synthesis – no action – in DNA synthesis.
• DNA polymerase synthesized de novo during germination during the first 9 h.
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MOBILIZATION OF STORED RESERVES
• II types of catabolic reactions:
• Hydrolytic involving Amylases
• Phosphorolytic
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HYDROLYTIC CATABOLISM
• In dicot seeds:
• starch degradation yields more glucose.
• more maltose is produced.
• This is related partly to the relative activity of cc-amylase in the two classes of
seeds.
• Two classes of seed – angiosperms and gymnosperms.
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• Storage proteins of seeds are hydrolyzed into their constituent amino acids by
proteinases or proteases.
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PHOSPOROLYTIC CATABOLISM
• Sucrose is the major form in which the products of carbohydrate catabolism
are transported into the developing seedling.
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BIOCHEMISTRY OF
FRUIT RIPENING
TYPES OF RIPENING
• Climacteric fruits – Able to ripen even after they a picked from the plants.
• Non-Climacteric Fruits – Able to ripen only if they are in plants.
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FRUIT RIPENING – PHYSIOLOGICAL CHANGES
• Softening – Both the skin and pulp.
• Flavor and Aroma – Increases.
• Chlorophyll loss – Development of new pigments ie., Carotenoids.
• Carotenoid accumulation.
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BIOCHEMICAL REACTIONS UNDERTHISTOPIC
• Hormonal changes – Ethylene.
• Change in chlorophyll content.
• Synthesis of pigments like carotenoids, anthocyanins and Xanthophylls.
• Increased activity of enzymes.
• Volatile components get synthesized.
• Simple Sugars get synthesized (gluconeogenesis).
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ETHYLENE
• Fruit ripening agent.
• Ripens the fruit even within minutes.
• Ethylene biosynthesis in plants:
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ETHYLENE
• Hastens the climacteric effects in the fruits.
• Accompanied by increased oxygen uptake.
• This process is not reversed.
• In the case of no-climacteric fruits:
• Once when the ethylene synthesis terminated
• Oxygen uptake and respiratory activity comes to control.
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DEGRADATION OF CHLOROPHYLL
• Ethylene promotes chlorophyll degradation.
• Change occurs in 3 steps:
• Replacement of Mg+ ion by hydrogen atom.
• Hydrloysis of chlorophyll.
• Bleaching of chlorophyll.
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Replacement of Mg+ ion
• Mg+ ion is replaced by hydrogen atom.
• Under acidic conditions.
• Associated with formation of Pheophytin.
• Pheophytin formation leads to color change – bright green to dull olive green.
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Hydrolysis of chlorophyll
• Chlorophyll hydrolysed to chlorophyllide and phytol.
• Catalysed by chlorophyllase.
• Results in formation of Pheophorbide.
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Bleaching of chlorophyll
• Through the action of:
• Lipoxygenase
• Peroxidase
• Catalase
• Degradation of fatty acids hydro peroxidases.
• Redox reaction in the presence of oxygen.
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SYNTHESIS OF OTHER PIGMENTS -
CAROTENOIDS
• Color change from green to red or orange.
• Isoprenoid compounds.
• Composed of isoprene units joined from head to tail by conjugated double
bonds.
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ANTHOCYANINS
• Responsible for red, pink, purple and blue color.
• Water soluble pigments.
• Flavonoid pigments.
• Composed of phenyl proponoid nucleus.
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XANTHOPHYLLS
• Isoprenoid pigment.
• Isoprene subunits joined form head to tail by conjugated double bonds.
• Substituted by oxy, hydroxyl, epoxy compounds.
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INCRESED ENZYME ACTIVITY
• Softening due to enzyme activity.
• Softening due to degradation of pectins.
• Catalysed by pectinases.
• Types of pectinases:
• PMG - Polymethyl galacturonases.
• PG – polygalaturonases.
• PE – Pectin esterases.
• PL – Pectin lyases.
• Degradation of Cellulose by cellulose.
• Hydrolysis of galactans to galactoses by B-galactosidase.
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SYTHESIS OFVOLATILE COMPOUNDS
• Aroma is produced by volatiles synthesized during ripening.
• Includes aldehydes, esters, lactones, terpenes, and sulfur compounds.
• Volatiles originate from proteins, carbohydrates, lipids, and vitamins.
• Taste is provided by many nonvolatile components, including sugars and acids
present in fruits.
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SYNTHESIS OF ORGANIC ACIDS AND SIMPLE
SUGARS
• Typical taste of fruits is determined by the content of sugars and organic
acids.
• Also phenolic compounds and tannins may affect the taste.
• During ripening starch is converted into sucrose, glucose and fructose.
• Starch hydrolysis is a major change during ripening of climacteric fruits.
• The concentration of organic acids also reach to a maximum during growth
and development of fruit on tree.
• Citric and malic acids-intermediates of Krebs cycle.
• Other acids: ascorbic acid (reduced form), oxalic acid.
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Biochemistry of seed germination and fruit ripening

  • 1. BIOCHEMISTRY OF SEED GERMINATION AND FRUIT RIPENING JANANI PALPANDI II BT DEPT., OF BIOTECHNOLOGY, MEPCO SCHLENK ENGG., COLLEGE.
  • 3. 3 8/15/2019 Add a footer
  • 4. BIOCHEMICAL REACTIONS UNDERTHISTOPIC • Respiration • Protein and Nucleic Acid Synthesis • Mobilization of stored reserves 4 8/15/2019 Add a footer
  • 5. RESPIRATION • In imbibed seeds the three metabolic pathways take place: • Glycolysis • Pentose Phosphate Pathway • Citric Acid Cycle • Through Glycolysis And Pentose Phosphate pathway – ATP, NADPH, FAD produced. • Through HMP shunt – NADP • NADPH , NADP – Readily available reducing agents – utilized in bio-reductive reactions 5 8/15/2019 Add a footer
  • 6. RESPIRATION • Dry seeds - 10-15 per cent water. • Dry seeds - low level of respiration. • Notable metabolic event during imbibition occurring in less than 15 min is the reformation of keto acids from amino acids by deamination and transamination. • Divided into four phases. 6 8/15/2019 Add a footer
  • 7. RESPIRATION PHASE I • Characterized by a sharp rise in respiration for about 10 hours. • Due to the activation and hydration of mitochondrial enzymes. • Enzymes - belonging to the cycle and electron transport chain. • There is rapid oxygen uptake into seeds with intact testas during phase I . • Known as Early imbibition PHASE II • Involves a lag in respiration between 10 and 25 hours - after the start of imbibition. • Hydration of the cotyledons- completed and all pre-existing enzymes- activated. • Testa impedes oxygen uptake in phase II. (slows down). • Between phase II and phase III, the radicle penetrates the testa. 7 8/15/2019 Add a footer
  • 8. RESPIRATION PHASE III • A second respiratory surge. • Due to increased oxygen supply through pierced testa. • Another reason - newly synthesized mitochondria and respiratory enzymes in the dividing cells of the growing axis. PHASE IV • Marked fall in respiration. • disintegration of the cotyledons . • exhaustion of the stored food. • Early stages of germination, respiration is cyanide-resistant and the alternative oxidase instead of cytochrome oxidase plays a role in germination. • . At later stages of germination, however, respiration becomes sensitive to cyanide. 8 8/15/2019 Add a footer
  • 9. ROLE OF CYANIDE IN GERMINATION • Cyanide is a germination regulatory agent. • In some plants like Amaranthus at lower concentration it promotes seed germination. 9 8/15/2019 Add a footer
  • 10. PROTEIN AND NUCLEIC ACID SYNTHESIS • Pre-requisite for radicle emergence . • Protein synthesis does not occur in the dry seeds. • Starts when these are hydrated and cytoplasmic ribosomes (eukaryotic SOS) get associated with messenger RNA (mRNA). • Radicle is the embryonic part from which root system develops. • It is the first developed system. 10 8/15/2019 Add a footer
  • 11. PROTEIN AND NUCLEIC ACID SYNTHESIS • Initiation of protein synthesis - attachment of the • small (40S) ribosomal subunit. • tRNA molecule . • Attachment to the initiation site on the mRNA. • After formation of the initiation complex - the large (60S) ribosomal subunit becomes attached . • Protein synthesis begins. • Formation of the initiation complex requires CTP and perhaps ATP. 11 8/15/2019 Add a footer
  • 12. PROTEIN AND NUCLEIC ACID SYNTHESIS • Other soluble protein factors - required for the transfer of aminoacyl-tRNAs to their appropriate codons on the ribosome-mRNA complex. • To move the message through the ribosome. • These are the elongation factors requiring CTP for activity. • New mRNA synthesis - not essential for resumption of protein synthesis during the first hour of imbibition. • mRNA conserved in the dry embryo - utilized for early protein synthesis. • Even if new mRNA is synthesized as an early event, it is not involved in early protein synthesis. 12 8/15/2019 Add a footer
  • 13. PROTEIN AND NUCLEIC ACID SYNTHESIS • Ribosomal RNA (rRNA) synthesis commences as early as mRNA • Increases during and after imbibition - after the 6th hour coinciding with the time of radicle elongation and • At 16-18h it reaches a rate about 12-fold greater than the earlier stage. • Although transfer RNA (tRNA) synthesis begins within 20 minutes in imbibing embryos - not certain whether - involved in early protein synthesis. • Its new synthesis is necessary during later stages to sustain-growth. 13 8/15/2019 Add a footer
  • 14. PROTEIN AND NUCLEIC ACID SYNTHESIS • The radicle within the seed initially grows by cell elongation. • Emergence through the seed coat may be associated with cell division. • DNA synthesis which is closely linked to mitotic cell division is mainly concerned with the post-germination period of seedling development. • During 9th hour – Inhibition of protein synthesis inhibits DNA synthesis. • After 9th hour – Inhibition of protein synthesis – no action – in DNA synthesis. • DNA polymerase synthesized de novo during germination during the first 9 h. 14 8/15/2019 Add a footer
  • 15. MOBILIZATION OF STORED RESERVES • II types of catabolic reactions: • Hydrolytic involving Amylases • Phosphorolytic 15 8/15/2019 Add a footer
  • 16. HYDROLYTIC CATABOLISM • In dicot seeds: • starch degradation yields more glucose. • more maltose is produced. • This is related partly to the relative activity of cc-amylase in the two classes of seeds. • Two classes of seed – angiosperms and gymnosperms. 16 8/15/2019 Add a footer
  • 17. • Storage proteins of seeds are hydrolyzed into their constituent amino acids by proteinases or proteases. 17 8/15/2019 Add a footer
  • 18. PHOSPOROLYTIC CATABOLISM • Sucrose is the major form in which the products of carbohydrate catabolism are transported into the developing seedling. 18 8/15/2019 Add a footer
  • 20. TYPES OF RIPENING • Climacteric fruits – Able to ripen even after they a picked from the plants. • Non-Climacteric Fruits – Able to ripen only if they are in plants. 20 8/15/2019 Add a footer
  • 21. FRUIT RIPENING – PHYSIOLOGICAL CHANGES • Softening – Both the skin and pulp. • Flavor and Aroma – Increases. • Chlorophyll loss – Development of new pigments ie., Carotenoids. • Carotenoid accumulation. 21 8/15/2019 Add a footer
  • 22. BIOCHEMICAL REACTIONS UNDERTHISTOPIC • Hormonal changes – Ethylene. • Change in chlorophyll content. • Synthesis of pigments like carotenoids, anthocyanins and Xanthophylls. • Increased activity of enzymes. • Volatile components get synthesized. • Simple Sugars get synthesized (gluconeogenesis). 22 8/15/2019 Add a footer
  • 23. ETHYLENE • Fruit ripening agent. • Ripens the fruit even within minutes. • Ethylene biosynthesis in plants: 23 8/15/2019 Add a footer
  • 24. ETHYLENE • Hastens the climacteric effects in the fruits. • Accompanied by increased oxygen uptake. • This process is not reversed. • In the case of no-climacteric fruits: • Once when the ethylene synthesis terminated • Oxygen uptake and respiratory activity comes to control. 24 8/15/2019 Add a footer
  • 25. DEGRADATION OF CHLOROPHYLL • Ethylene promotes chlorophyll degradation. • Change occurs in 3 steps: • Replacement of Mg+ ion by hydrogen atom. • Hydrloysis of chlorophyll. • Bleaching of chlorophyll. 25 8/15/2019 Add a footer
  • 26. Replacement of Mg+ ion • Mg+ ion is replaced by hydrogen atom. • Under acidic conditions. • Associated with formation of Pheophytin. • Pheophytin formation leads to color change – bright green to dull olive green. 26 8/15/2019 Add a footer
  • 27. Hydrolysis of chlorophyll • Chlorophyll hydrolysed to chlorophyllide and phytol. • Catalysed by chlorophyllase. • Results in formation of Pheophorbide. 27 8/15/2019 Add a footer
  • 28. Bleaching of chlorophyll • Through the action of: • Lipoxygenase • Peroxidase • Catalase • Degradation of fatty acids hydro peroxidases. • Redox reaction in the presence of oxygen. 28 8/15/2019 Add a footer
  • 29. SYNTHESIS OF OTHER PIGMENTS - CAROTENOIDS • Color change from green to red or orange. • Isoprenoid compounds. • Composed of isoprene units joined from head to tail by conjugated double bonds. 29 8/15/2019 Add a footer
  • 30. ANTHOCYANINS • Responsible for red, pink, purple and blue color. • Water soluble pigments. • Flavonoid pigments. • Composed of phenyl proponoid nucleus. 30 8/15/2019 Add a footer
  • 31. XANTHOPHYLLS • Isoprenoid pigment. • Isoprene subunits joined form head to tail by conjugated double bonds. • Substituted by oxy, hydroxyl, epoxy compounds. 31 8/15/2019 Add a footer
  • 32. INCRESED ENZYME ACTIVITY • Softening due to enzyme activity. • Softening due to degradation of pectins. • Catalysed by pectinases. • Types of pectinases: • PMG - Polymethyl galacturonases. • PG – polygalaturonases. • PE – Pectin esterases. • PL – Pectin lyases. • Degradation of Cellulose by cellulose. • Hydrolysis of galactans to galactoses by B-galactosidase. 32 8/15/2019 Add a footer
  • 33. SYTHESIS OFVOLATILE COMPOUNDS • Aroma is produced by volatiles synthesized during ripening. • Includes aldehydes, esters, lactones, terpenes, and sulfur compounds. • Volatiles originate from proteins, carbohydrates, lipids, and vitamins. • Taste is provided by many nonvolatile components, including sugars and acids present in fruits. 33 8/15/2019 Add a footer
  • 34. SYNTHESIS OF ORGANIC ACIDS AND SIMPLE SUGARS • Typical taste of fruits is determined by the content of sugars and organic acids. • Also phenolic compounds and tannins may affect the taste. • During ripening starch is converted into sucrose, glucose and fructose. • Starch hydrolysis is a major change during ripening of climacteric fruits. • The concentration of organic acids also reach to a maximum during growth and development of fruit on tree. • Citric and malic acids-intermediates of Krebs cycle. • Other acids: ascorbic acid (reduced form), oxalic acid. 34 8/15/2019 Add a footer