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1
Welcome
to
credit seminar session 2020
Speaker
Bhupendra Sagore
21292
M.Sc. 1st Year
2
“Role of next-generation
elicitors and bio-molecules
for improving
fruit quality”
Topic
Division of Fruits and Horticultural Technology
ICAR-Indian Agricultural Research Institute, New Delhi-110012
3
Seminar outlines
1. Introduction
2. Types of elicitors
3. Bio-molecules
4. Mechanism
5. Factors affecting elicitation
6. Case studies
7. Conclusion
8. Future thrusts
4
Fruit quality
Fruits are mainly consumed for their nutritive value as
well as for the variety of shapes, colours and flavours
that make them attractive for food preparation.
When they are consumed raw or with very little
preparation, the consumer’s main concern is that they
must be free of biotic or non-biotic contaminants that
may affect health.
Types of bio-active compounds
Bioactive
compoundsFlavonoids
Anthocyanins
Tannins
Betalins
Carotenoids
Plant sterols
Walia et al. (2019)
Sources of Carotenoids
Apricot
Mango
Guava
PlumPapaya
Grapefruit Persimmon
IndianAgriculturalResearchInstitute,NewDelhi
Agnieszka Szajdek & E. J. Borowska (2008)
Sources of Anthocyanin
Acai berry
Black currant
Blueberry
CherryRed grapes
IndianAgriculturalResearchInstitute,NewDelhi
Agnieszka Szajdek & E. J. Borowska (2008)
ICMR recommendation
Dietary guidelines for Indians -a manual by ICMR, 2011
WHO, 2014
10
• To improve the phenolic content of fruits, a novel
field of interest is based on results obtained using
elicitors, agrochemicals which were primarily
designed to improve resistance to plant
pathogens.
• Elicitors do not kill pathogens, they trigger plant
defense mechanisms, one of which is to increase
the levels of phenolic compounds.
• Therefore, their application not only allows us to
control plant disease but also to increase the
phenolic content of plant foodstuffs.
Quality improvement tool
11
Elicitors
Elicitors are generally defined as molecules that can
stimulate the defense responses of plants, including the
formation of phytoalexins.
Source: Bioprocessing for Value-Added Products from Renewable Resources, 2007
OR
An elicitor is defined as a compound that, in small
concentrations, can activate different plant responses, such
as endogenous protection responses, including the
production of different secondary metabolites .
(Namdeo, 2007)
The first biotic elicitors were described in the early 1970s.
12
Types of elicitors
Biotic Abiotic
13
Source-Journal of Applied Research on Medicinal and Aromatic
Plants,2013 Review article
14Jesus et al. 2018
15
Effects of the application of different elicitors on polyphenol
content
Source- Ruiz et al. Review 2013
16
17
Source- Molecules 2014, ISSN 1420-3049 (Review)
Elicitation: A Tool for Enriching the Bioactive Composition of Foods
18
Bio-molecules
Bio-molecules are also called biological
molecules, any of numerous substances that are
produced by cells and living organisms. Bio-molecules
have a wide range of sizes and structures and perform
a vast array of functions. The four major types of
biomolecules are
1. Carbohydrate
2. Lipids
3. Proteins
4. Nucleic Acids
Postharvest pathogens/diseases of horticultural
produce and their control by chitosan
19
Source-Sharma and Pongener / Stewart Postharvest review,2010
20
21
22
Elicitations
1. Pre harvest elicitations
Preharvest elicitation could be done as seed
priming, soaking seeds in a water solution with the
elicitor, or after seedling, applying exogenous
spraying treatment over the leaves or in a
hydroponic system.
2. Post harvest elicitations-
The post harvest application of low or high
temperature treatments, ultraviolet (UV) or gas
combinations, phytohormones applied to tissues
will increase phenolic concentration.
23
How
Does
It
Work
24
Mode of action
25
Stress-induced polyphenols synthesis in plants
26
Factors affecting elicitation
Several parameters which decide the
elicitation such as:
• elicitors concentration and selectivity
• duration of elicitation
• age of culture
• cell line
• growth regulation
• nutrient composition
• quality of cell wall materials and
27
Research work related to elicitors & bio-molecules
in ICAR-IARI, New Delhi
S.N. Research work Crop Name of
Division
1.
Preharvest application of methyl jasmonate for
improving post harvest quality of “Pusa Navrang”
Grape Fruits &
Horticultural
Technology
2.
Induction of resistance by synthetic elicitor
molecules against root-knot nematode
Brinjal Nematology
3. Biotic elicitor induced biochemical and molecular
manifestations of drought tolerance in contrasting
rice genotypes
Rice Biochemistry
4. Microbial priming elicits improved plant growth
promotion and nutrient uptake.
Pea Microbiology
5.
Effect of Salicylic Acid and Pseudomonas
fluorescens
Cotton Plant Pathology
6. Postharvest studies in tuberose. Tuberose Floriculture and
landscaping
7. Natural products for postharvest decay control in
horticultural produce.
Horticultur
al crops
Post Harvest
Technology
8. Genome-wide characterization and expression
patterns of chitinase genes.
Pigeon Pea National Institute
for Plant
Biotechnology
28
Case studies
29
CASE STUDY- I Pomegranate (Methyl jasmonate)
The effects of preharvest MeJA treatments on pomegranate ‘Mollar de
Elche’ for-
1.Crop yield,
2.Quality attributes and bioactive compounds content (at harvest or after
long-term storage).
Maria et. al - Oct, 2019
Objectives
30
Experimental details
Particular Details
Name of cultivar “Mollar de Elche”
Plant age 8 years (planted at 6 m × 5 m)
Date of experiment Start from April 2016 to 2017
Place Elche, Spain
Design RBD
Type of research Pre -harvest treatment with post
harvest analysis
Elicitor used Methyl jasmonate ( MeJA)
NPK ratio 160:80:160
Treatment Total 4 (control , 1, 5 and 10 mmol L-1
MeJA)
Application at monthly intervals (94, 64, 34 and 4
days before harvesting)
31
Materials and Methods
Plant material and experimental design
• For each treatment (control, 1, 5 and 10 mmol L-1 MeJA),
three blocks of two trees each one were selected.
• Each block or replicate for the four treatments was set in
a row, leaving an untreated tree between each block and
an untreated row between each treated row in order to
avoid treatment cross effects.
• In addition, at least one tree without treatment was left in
each row to avoid edge effect.
• Treatments were performed by applying 3 L of freshly
prepared MeJA at 1, 5 or 10 mmol L-1, containing 1 mL
L−1 Tween-20, to each tree at monthly intervals (94, 64,
34 and 4 days before harvesting). sprayed with distilled
32
Controlled T-1(1) T-2 (5) T-3 (10)v
Untreated Controlled Treated
33
Result
Fruit Yield
34
Result
Respiration Rate , Firmness, Weight loss & Hue angle
35
Result
Sugar & Organic acids
36
Result
Phenols & anthocyanins content
Total phenols and total anthocyanins content in pomegranate arils in
control and methyl jasmonate (MeJA, 1, 5 and 10 mmol L-1) treated trees
at harvest and during postharvest storage at 10 ∘C.
37
Inference
• Preharvest treatments with MeJA at 1, 5, and 10 mmol L-1 increased
crop yield. In addition, the on-tree fruit ripening process was
accelerated by 1 and 5 mmol L-1 doses.
• Quality parameters after 30 and 60 days of storage at 10 ∘C were
maintained at higher levels in MeJA treated fruit, manifested by
reduced weight loss, respiration rate and losses of firmness and
titratable acidity.
• MeJA treatments improved arils colour and their content in bioactive
compounds (phenolics, anthocyanins and ascorbic acid) and these
effects being maintained during storage.
• Among the assayed doses, the highest effects were found with MeJA at
5 mmol L-1.
• Thus, MeJA has potential application in pre-harvest treatment as a
useful tool for the induction of health benefitting chemicals in the
plant diet.
38
CASE STUDY- II Citrus
( Oligochitosan, Salicylic acid, Pichia membranaefaciens )
1. To study the effects of oligochitosan, salicylic acid (SA), and Pichia
membranaefaciens on inducing disease resistance against Geotrichum
candidum in citrus fruit by using iTRAQ proteomic and physicochemical
analysis.
Wang et. al 2020
Objective
39
Genesis of the research
• During transporation of citrus fruits severe quality losses
are commonly occurred mainly due to the fungal disease
like Sour rot (Geotrichum candidum)
• The most effective way to control these fungi is the
application of synthetic fungicides, such as 2,4-D, imazalil,
thiabendazole, pyrimethanil, prochloraz, fludioxonil, etc.
• Salicylic acid (SA), Pichia membranaefaciens and
oligochitosan have been reported to be able to induce
multiple defensive reactions against particular biotic and
abiotic stresses in some postharvest fruits, including citrus,
apple, peach, pear, sweet cherry, etc.
40
Materials and methods
• Harvested Citrus sinensis (L.) Osbeck cv.
Jincheng fruits were selected based on
their uniformity in color, size and
lack of physical injury or infection.
The fruits were superficially
disinfected for 2 min via dipping in
2% (v/v) sodium hypochlorite,
followed by washing with water, and
air-dried at room temperature (25
°C).
• The antagonistic yeast maintained on
yeast extract peptone dextrose
medium at 4 °C in lab.
Citrus Sinesis cv.
Jincheng
41
• The solution of SA and oligochitosan was prepared
with SDW to the final concentrations of 2.5 mmol
L−1 and 15 g L−1 , respectively.
• The pathogen G. candidum maintained in the
laboratory was isolated from decayed citrus fruit
showing symptoms of the disease and identified on
the basis of morphology and internal transcribed
spacer (ITS) sequence region of the rDNA analysis
(Zhao et al., 2017).
42
Induction of resistance treatment and sample preparation
Two wounds (3 mm diameter × 3 mm deep) per fruit were
created on the opposite sides of fruit equator using a sterile
needle. Each wound was then inoculated with 30 μL of:
(1) SDW as the control
(2) SA (2.5 mmol L−1 )
(3) P. membranaefaciens (1 × 108 cells mL−1 )
(4) Oligochitosan (15 g L−1 ).
43
Result
Effects of SA, P. membranaefaciens and oligochitosan on disease incidence
and lesion diameter of citrus fruit caused by G. candidum when treatment
solutions and pathogens were inoculated on the same (A and B) and
different wounds (C and D).
44
Effects of SA, P. membranaefaciens and oligochitosan on the relative contents of sugars:
glucose (A), fructose (B), sucrose (C), inositol (D), glucopyranose (E), arabinose (F),
galactose (G), mannose (H), ribose (I) and xylose (J) contents in citrus peel
45
Effects of SA, P. membranaefaciens and oligochitosan on the relative contents of organic
acids: citrate acid (A), α-ketoglutarate acid (B), succinic acid (C), malic acid (D), fumaric (E),
2-keto-D-glucosaccharic acid (F), gulonic acid (G) and oxalic acid (H) contents in citrus peel .
46
Inference
1.These three elicitors significantly improved disease resistance against
G. candidum in citrus, by regulating the DEPs involved in pathways of
starch and sucrose metabolism, carbon metabolism and amino acid
metabolism.
2. Besides, the three elicitors induced accumulation of soluble sugars,
key organic acids in TCA circle and amino acids, which resulted in
reprogramming the energy and resources relevant to the disease
resistance system, via different modes of action directly or indirectly
enhanced the resistance reaction of the citrus fruit.
3. The primary metabolism played a vital role in the disease resistance of
citrus induced by oligochitosan, SA and P. membranaefaciens.
47
1. The use of elicitors may be regarded as a simple and useful
technique to increase the phenolic content of fruit, protecting,
at the same time, both plants and fruits from biotic and abiotic
stresses, without the disadvantage of the environment.
e
2. Major advantage of the post harvest elicitations is cell cultures
includes synthesis of bioactive secondary metabolites,
independently of environmental and soil conditions.
3. However, more research is needed to better understand
the effect of these elicitors in the different phytochemical
synthesis pathways in order to be able to increase the health
related properties of fruit products without decreasing the
sensory properties of these products..
Conclusion
48
Future thrusts
 Utilization of next-generation elicitors and bio-molecules
for improving fruit quality and disease management in fruit
crops.
 Elicitor treatments could be an alternative to genetically
modified (GM) plants for better attraction of natural enemies
of pest organisms on cultivated plants.
 Elicitor-treated plants bear lower ecological risks than GM
plants.
 Improved quality fruit with high phenolic and anti-oxidant is
a basic need to fight against several disease, COVID-19 is one
of them.
49
‘Quality
improvement never
ends…’
Thank You _/_

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Elicitors

  • 1. 1 Welcome to credit seminar session 2020 Speaker Bhupendra Sagore 21292 M.Sc. 1st Year
  • 2. 2 “Role of next-generation elicitors and bio-molecules for improving fruit quality” Topic Division of Fruits and Horticultural Technology ICAR-Indian Agricultural Research Institute, New Delhi-110012
  • 3. 3 Seminar outlines 1. Introduction 2. Types of elicitors 3. Bio-molecules 4. Mechanism 5. Factors affecting elicitation 6. Case studies 7. Conclusion 8. Future thrusts
  • 4. 4 Fruit quality Fruits are mainly consumed for their nutritive value as well as for the variety of shapes, colours and flavours that make them attractive for food preparation. When they are consumed raw or with very little preparation, the consumer’s main concern is that they must be free of biotic or non-biotic contaminants that may affect health.
  • 5. Types of bio-active compounds Bioactive compoundsFlavonoids Anthocyanins Tannins Betalins Carotenoids Plant sterols Walia et al. (2019)
  • 6. Sources of Carotenoids Apricot Mango Guava PlumPapaya Grapefruit Persimmon IndianAgriculturalResearchInstitute,NewDelhi Agnieszka Szajdek & E. J. Borowska (2008)
  • 7. Sources of Anthocyanin Acai berry Black currant Blueberry CherryRed grapes IndianAgriculturalResearchInstitute,NewDelhi Agnieszka Szajdek & E. J. Borowska (2008)
  • 8. ICMR recommendation Dietary guidelines for Indians -a manual by ICMR, 2011
  • 10. 10 • To improve the phenolic content of fruits, a novel field of interest is based on results obtained using elicitors, agrochemicals which were primarily designed to improve resistance to plant pathogens. • Elicitors do not kill pathogens, they trigger plant defense mechanisms, one of which is to increase the levels of phenolic compounds. • Therefore, their application not only allows us to control plant disease but also to increase the phenolic content of plant foodstuffs. Quality improvement tool
  • 11. 11 Elicitors Elicitors are generally defined as molecules that can stimulate the defense responses of plants, including the formation of phytoalexins. Source: Bioprocessing for Value-Added Products from Renewable Resources, 2007 OR An elicitor is defined as a compound that, in small concentrations, can activate different plant responses, such as endogenous protection responses, including the production of different secondary metabolites . (Namdeo, 2007) The first biotic elicitors were described in the early 1970s.
  • 13. 13 Source-Journal of Applied Research on Medicinal and Aromatic Plants,2013 Review article
  • 15. 15 Effects of the application of different elicitors on polyphenol content Source- Ruiz et al. Review 2013
  • 16. 16
  • 17. 17 Source- Molecules 2014, ISSN 1420-3049 (Review) Elicitation: A Tool for Enriching the Bioactive Composition of Foods
  • 18. 18 Bio-molecules Bio-molecules are also called biological molecules, any of numerous substances that are produced by cells and living organisms. Bio-molecules have a wide range of sizes and structures and perform a vast array of functions. The four major types of biomolecules are 1. Carbohydrate 2. Lipids 3. Proteins 4. Nucleic Acids
  • 19. Postharvest pathogens/diseases of horticultural produce and their control by chitosan 19 Source-Sharma and Pongener / Stewart Postharvest review,2010
  • 20. 20
  • 21. 21
  • 22. 22 Elicitations 1. Pre harvest elicitations Preharvest elicitation could be done as seed priming, soaking seeds in a water solution with the elicitor, or after seedling, applying exogenous spraying treatment over the leaves or in a hydroponic system. 2. Post harvest elicitations- The post harvest application of low or high temperature treatments, ultraviolet (UV) or gas combinations, phytohormones applied to tissues will increase phenolic concentration.
  • 26. 26 Factors affecting elicitation Several parameters which decide the elicitation such as: • elicitors concentration and selectivity • duration of elicitation • age of culture • cell line • growth regulation • nutrient composition • quality of cell wall materials and
  • 27. 27 Research work related to elicitors & bio-molecules in ICAR-IARI, New Delhi S.N. Research work Crop Name of Division 1. Preharvest application of methyl jasmonate for improving post harvest quality of “Pusa Navrang” Grape Fruits & Horticultural Technology 2. Induction of resistance by synthetic elicitor molecules against root-knot nematode Brinjal Nematology 3. Biotic elicitor induced biochemical and molecular manifestations of drought tolerance in contrasting rice genotypes Rice Biochemistry 4. Microbial priming elicits improved plant growth promotion and nutrient uptake. Pea Microbiology 5. Effect of Salicylic Acid and Pseudomonas fluorescens Cotton Plant Pathology 6. Postharvest studies in tuberose. Tuberose Floriculture and landscaping 7. Natural products for postharvest decay control in horticultural produce. Horticultur al crops Post Harvest Technology 8. Genome-wide characterization and expression patterns of chitinase genes. Pigeon Pea National Institute for Plant Biotechnology
  • 29. 29 CASE STUDY- I Pomegranate (Methyl jasmonate) The effects of preharvest MeJA treatments on pomegranate ‘Mollar de Elche’ for- 1.Crop yield, 2.Quality attributes and bioactive compounds content (at harvest or after long-term storage). Maria et. al - Oct, 2019 Objectives
  • 30. 30 Experimental details Particular Details Name of cultivar “Mollar de Elche” Plant age 8 years (planted at 6 m × 5 m) Date of experiment Start from April 2016 to 2017 Place Elche, Spain Design RBD Type of research Pre -harvest treatment with post harvest analysis Elicitor used Methyl jasmonate ( MeJA) NPK ratio 160:80:160 Treatment Total 4 (control , 1, 5 and 10 mmol L-1 MeJA) Application at monthly intervals (94, 64, 34 and 4 days before harvesting)
  • 31. 31 Materials and Methods Plant material and experimental design • For each treatment (control, 1, 5 and 10 mmol L-1 MeJA), three blocks of two trees each one were selected. • Each block or replicate for the four treatments was set in a row, leaving an untreated tree between each block and an untreated row between each treated row in order to avoid treatment cross effects. • In addition, at least one tree without treatment was left in each row to avoid edge effect. • Treatments were performed by applying 3 L of freshly prepared MeJA at 1, 5 or 10 mmol L-1, containing 1 mL L−1 Tween-20, to each tree at monthly intervals (94, 64, 34 and 4 days before harvesting). sprayed with distilled
  • 32. 32 Controlled T-1(1) T-2 (5) T-3 (10)v Untreated Controlled Treated
  • 34. 34 Result Respiration Rate , Firmness, Weight loss & Hue angle
  • 36. 36 Result Phenols & anthocyanins content Total phenols and total anthocyanins content in pomegranate arils in control and methyl jasmonate (MeJA, 1, 5 and 10 mmol L-1) treated trees at harvest and during postharvest storage at 10 ∘C.
  • 37. 37 Inference • Preharvest treatments with MeJA at 1, 5, and 10 mmol L-1 increased crop yield. In addition, the on-tree fruit ripening process was accelerated by 1 and 5 mmol L-1 doses. • Quality parameters after 30 and 60 days of storage at 10 ∘C were maintained at higher levels in MeJA treated fruit, manifested by reduced weight loss, respiration rate and losses of firmness and titratable acidity. • MeJA treatments improved arils colour and their content in bioactive compounds (phenolics, anthocyanins and ascorbic acid) and these effects being maintained during storage. • Among the assayed doses, the highest effects were found with MeJA at 5 mmol L-1. • Thus, MeJA has potential application in pre-harvest treatment as a useful tool for the induction of health benefitting chemicals in the plant diet.
  • 38. 38 CASE STUDY- II Citrus ( Oligochitosan, Salicylic acid, Pichia membranaefaciens ) 1. To study the effects of oligochitosan, salicylic acid (SA), and Pichia membranaefaciens on inducing disease resistance against Geotrichum candidum in citrus fruit by using iTRAQ proteomic and physicochemical analysis. Wang et. al 2020 Objective
  • 39. 39 Genesis of the research • During transporation of citrus fruits severe quality losses are commonly occurred mainly due to the fungal disease like Sour rot (Geotrichum candidum) • The most effective way to control these fungi is the application of synthetic fungicides, such as 2,4-D, imazalil, thiabendazole, pyrimethanil, prochloraz, fludioxonil, etc. • Salicylic acid (SA), Pichia membranaefaciens and oligochitosan have been reported to be able to induce multiple defensive reactions against particular biotic and abiotic stresses in some postharvest fruits, including citrus, apple, peach, pear, sweet cherry, etc.
  • 40. 40 Materials and methods • Harvested Citrus sinensis (L.) Osbeck cv. Jincheng fruits were selected based on their uniformity in color, size and lack of physical injury or infection. The fruits were superficially disinfected for 2 min via dipping in 2% (v/v) sodium hypochlorite, followed by washing with water, and air-dried at room temperature (25 °C). • The antagonistic yeast maintained on yeast extract peptone dextrose medium at 4 °C in lab. Citrus Sinesis cv. Jincheng
  • 41. 41 • The solution of SA and oligochitosan was prepared with SDW to the final concentrations of 2.5 mmol L−1 and 15 g L−1 , respectively. • The pathogen G. candidum maintained in the laboratory was isolated from decayed citrus fruit showing symptoms of the disease and identified on the basis of morphology and internal transcribed spacer (ITS) sequence region of the rDNA analysis (Zhao et al., 2017).
  • 42. 42 Induction of resistance treatment and sample preparation Two wounds (3 mm diameter × 3 mm deep) per fruit were created on the opposite sides of fruit equator using a sterile needle. Each wound was then inoculated with 30 μL of: (1) SDW as the control (2) SA (2.5 mmol L−1 ) (3) P. membranaefaciens (1 × 108 cells mL−1 ) (4) Oligochitosan (15 g L−1 ).
  • 43. 43 Result Effects of SA, P. membranaefaciens and oligochitosan on disease incidence and lesion diameter of citrus fruit caused by G. candidum when treatment solutions and pathogens were inoculated on the same (A and B) and different wounds (C and D).
  • 44. 44 Effects of SA, P. membranaefaciens and oligochitosan on the relative contents of sugars: glucose (A), fructose (B), sucrose (C), inositol (D), glucopyranose (E), arabinose (F), galactose (G), mannose (H), ribose (I) and xylose (J) contents in citrus peel
  • 45. 45 Effects of SA, P. membranaefaciens and oligochitosan on the relative contents of organic acids: citrate acid (A), α-ketoglutarate acid (B), succinic acid (C), malic acid (D), fumaric (E), 2-keto-D-glucosaccharic acid (F), gulonic acid (G) and oxalic acid (H) contents in citrus peel .
  • 46. 46 Inference 1.These three elicitors significantly improved disease resistance against G. candidum in citrus, by regulating the DEPs involved in pathways of starch and sucrose metabolism, carbon metabolism and amino acid metabolism. 2. Besides, the three elicitors induced accumulation of soluble sugars, key organic acids in TCA circle and amino acids, which resulted in reprogramming the energy and resources relevant to the disease resistance system, via different modes of action directly or indirectly enhanced the resistance reaction of the citrus fruit. 3. The primary metabolism played a vital role in the disease resistance of citrus induced by oligochitosan, SA and P. membranaefaciens.
  • 47. 47 1. The use of elicitors may be regarded as a simple and useful technique to increase the phenolic content of fruit, protecting, at the same time, both plants and fruits from biotic and abiotic stresses, without the disadvantage of the environment. e 2. Major advantage of the post harvest elicitations is cell cultures includes synthesis of bioactive secondary metabolites, independently of environmental and soil conditions. 3. However, more research is needed to better understand the effect of these elicitors in the different phytochemical synthesis pathways in order to be able to increase the health related properties of fruit products without decreasing the sensory properties of these products.. Conclusion
  • 48. 48 Future thrusts  Utilization of next-generation elicitors and bio-molecules for improving fruit quality and disease management in fruit crops.  Elicitor treatments could be an alternative to genetically modified (GM) plants for better attraction of natural enemies of pest organisms on cultivated plants.  Elicitor-treated plants bear lower ecological risks than GM plants.  Improved quality fruit with high phenolic and anti-oxidant is a basic need to fight against several disease, COVID-19 is one of them.