Elicitors

Role of next generation elicitors and bio-molecule for improving fruit quality

1. 1 Welcome to credit seminar session 2020 Speaker Bhupendra Sagore 21292 M.Sc. 1 st 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 Introduction Types of elicitors Bio-molecules Mechanism Factors affecting elicitation Case studies Conclusion 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 Walia et al . (2019)

6. Sources of Carotenoids Apricot Mango Guava Plum Papaya Grapefruit Persimmon Indian Agricultural Research Institute, New Delhi Agnieszka Szajdek & E. J. Borowska (2008)

7. Sources of Anthocyanin Acai berry Black currant Blueberry Cherry Red grapes Indian Agricultural Research Institute, New Delhi Agnieszka Szajdek & E. J. Borowska (2008)

8. ICMR recommendation Dietary guidelines for Indians -a manual by ICMR, 2011

9. WHO, 2014

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 E licitors 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. 12 Types of elicitors Biotic Abiotic

13. 13 Source- Journal of Applied Research on Medicinal and Aromatic Plants,2013 Review article

14. 14 Jesus et al. 2018

15. 15 Effects of the application of different elicitors on polyphenol content Source- Ruiz et al. Review 2013

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17. 17 Source- M olecules 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

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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.

23. 23 How Does It Work

24. 24 Mode of action

25. 25 Stress-induced polyphenols synthesis in plants

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. Horticultural crops Post Harvest Technology 8. Genome-wide characterization and expression patterns of chitinase genes. Pigeon Pea National Institute for Plant Biotechnology 

28. 28 Case studies

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 water containing 1 mL L−1 Tween-20 .

32. 32 Controlled T-1(1) T-2 (5) T-3 (10) v Untreated Controlled Treated

33. 33 Result Fruit Yield

34. 34 Result Respiration Rate , Firmness, Weight loss & Hue angle

35. 35 Result Sugar & Organic acids

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 m embranaefaciens ) 1. To study the ef fects 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. E licitor 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. 49 ‘Quality improvement never ends…’ Thank You _ /\_