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Fatty Acid Accumulation in High Oleic Groundnut Genotypes during Seed Developmental Stages

This research investigates the fatty acid accumulation in high oleic groundnut genotypes during different seed developmental stages. The study explores the benefits of high oleic groundnuts, such as improved shelf-life, lower oxidation, better flavor, and suppression of tumorigenesis. The experimental plot at ICRISAT, Patancheru, during the rainy season of 2017, was used to collect seed samples for fatty acid analysis. The study also examines the expression of key genes involved in fatty acid biosynthesis and identifies factors responsible for the accumulation of high oleic acid. The findings provide valuable insights for the development of high oleic groundnut lines.

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Fatty Acid Accumulation in High Oleic Groundnut Genotypes during Seed Developmental Stages

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  1. Fatty acid accumulation in high oleic groundnut genotypes during seed developmental stages Dnyaneshwar B Deshmukh Research Scholar, ICRISAT, Hyderabad Advisor: Dr P Janila, Principal Scientist, ICRISAT, Hyderabad

  2. High oleic groundnut and its importance X Vulnerable to oxidative rancidity X Thermodynamically unstable FAE X Increases the risk of cardiovascular diseases KAS FAT B FAT B FAD2 Acetyl co-A Palmitic acid (16:0) Oleic acid (18:1) Linoleic acid (18:2) Steric acid (18:0) Legend: FAT B- palmitoyl-ACP thioesterase KAS- ketoacyl ACP synthetase FAD2- Fatty acid desaturase FAE- Fatty acid elongase • Benefits of high oleic in groundnut • Improved shelf-life • Lower oxidation and avoids rancidity • Have much better flavor • Suppression of tumorigenesis • Reduction of inflammatory diseases • Maintains the levels of high-density lipoproteins (HDL) Happy farmer Happy Consumer Happy Trader Fatty acid biosynthesis pathway (Heppard et al. 1996; Singh et al. 2002)

  3. Development of high oleic lines • The oleic: linoleic acid ratio (O/L) in normal peanut genotypes-1.0-4.0 whereas it can reach 35 to 40 in high O/L mutant lines Janila et al. Plant Science, (2016)

  4. Investigation of fatty acids accumulation at different seed development stages Experimental plot at ICRISAT, Patancheru during Rainy, 2017 Ref: AICRP-G Annual Report 2017) • Plant Material: 13 genotypes (8 high oleic lines, 4 non oleic cultivars and SunOleic95R) • Molecular markers:

  5. Workflow of experiment: Rainy 2017 • Experimental plots of Alfisols (Alfisol-Patancheru Soil Series; UdicRhodustolf) at ICRISAT, Patancheru, India (at 17.53◦ N latitude and 78.27◦ E) • Field design: Randomized Complete Block Design • Recommended agronomic management practices were adopted to raise a healthy crop In-situ collection of seed samples for fatty acid analysis 60 DAF 30 Days after fertilization (DAF) 90 DAF Tagging of individual flower (~500 flowers/genotype) Fresh kernels collected for RNA isolation Fresh kernels collected for RNA isolation ~ 150 seeds were collected and dried for fatty acid evaluation Fresh kernels collected for RNA isolation ~ 150 seeds were collected and dried for fatty acid evaluation ~ 150 seeds were collected and dried for fatty acid evaluation

  6. Workflow of lab experiments Target genes in fatty acid bio-synthesis pathway: 9 key genes • Δ12 fatty acid desaturase (FAD2A) • FAD2B • Microsomal omega-6 FA desaturase (FAD2-1) • FAD2-2 • FAD6 (Plastidial D12 desaturase) • KAS-II (Ketoacyl ACP synthetase) • Stearoyl-ACP desaturase (SAD) • FAT-A and FAT-B • Reference genes: Alcohol Dehydrogenase (ADH3), Elongation Factor 1B (ELF1B), and Glucose-6-Phospahate 1-Dehydrogenase (G6PD) A quantitative Polymerase Chain Reaction (qPCR) analysis based on SYBR green dye chemistry Validation of reference genes for qRT-PCR Reddy et al. (2013)

  7. Expression analysis of fatty acid genes in developing seeds SunOleic 95R ICGV 06420 ICGV 15006

  8. Near Infrared Reflectance Spectroscopy showed increase in Oleic acid over time Near Infrared Reflectance Spectroscopy (NIRS) for seed quality analysis • Oleic acid accumulates simultaneously in the seed along with protein and oil

  9. Gas chromatography result revealed a clear picture of fatty acid accumulation • Oleic acid accumulates with increase in developmental stage • High oleate lines were reduced in unfavorable fatty acid like palmitic acid and linoleic acid

  10. Salient finding of the current study • The expression of FAD2 alleles was found to gradually decrease with the developmental stage in high oleic lines compared to the normal types • Oleic acid accumulates simultaneously in the seed along with protein and oil without fluctuating the oil and protein content of the seeds • Downregulation of Plastidial D12 desaturase (FAD6) found to one of the factor responsible of accumulation of high oleic (Up to 80%) • The line, ICGV 15006 showed 71.26%, 77.92% and 78.63% of oleic acid concentration at 30th, 60th and 90th days of seed development and had reduced amounts of palmitic acid (up to 8%)and linoleic acid (up to 33%)

  11. Team Groundnut I will be free to take your questions Photo caption: Vidyasagar Rao, ICRISAT Thank you…. Acknowledgements: Dr P Janila, ICRISAT, Patancheru Dr M Balram, PJTSAU, Hyderabad Dr T V Murali, ICRISAT, Patancheru Mr V Papaiah, ICRISAT, Patancheru Dr P Bhatanagar, ICRISAT, Patancheru Dr S Reddy, ICRISAT, Patancheru RK Laboratories, Junagadh

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