HARNESSING METABOLOMICS TO COMBAT FUSARIUM WILT IN PIGEONPEA: A LC-MS AND GC-MS PERSPECTIVE
Authors: Megha Ujinwal, Nisha Singh and Sapna Langyan
Megha Ujinwal: Gujarat Biotechnology University, Gujarat-382355, India.
Nisha Singh: Gujarat Biotechnology University, Gujarat-382355, India.
Sapna Langyan: ICAR-National Bureau of Plant genetic Resources, New Delhi, India.
ABSTRACT
Introduction
Pigeonpea has been a traditional staple in India, valued for its nutritional and therapeutic benefits since ancient times. Renowned for its diverse pharmacological properties, it exhibits antibacterial, anti-inflammatory, antitubercular, antioxidant, neuroprotective, antihypertensive, antihyperglycemic, and anticancer activities. However, its cultivation is challenged by various biotic and abiotic stress factors. Among them, Fusarium wilt (Fusarium udum Butler), a devastating soil-borne disease, causes significant yield losses across major pigeonpea production areas.
Objectives
This study aims to identify key metabolites involved in defense pathways and their interactions with the host plant by analyzing and comparing the metabolite profiles of healthy and diseased (fusarium wilt-infected) pigeonpea genotype UP 9-I using GC-MS and LC-MS/MS.
Methods
LC-MS and GC-MS untargeted metabolomics were used to explore and analyze pigeonpea genotype (UP 9-I). For LC-MS/MS, 0.25g of fresh leaves (healthy & diseased) were mixed with 30 mL water-methanol, filtered (0.2 μm), and 5 μL was injected for analysis. For GC-MS, 50 mg of leaves were crushed with 2 mL 80% methanol, concentrated (100 μL), derivatized with 100 μL MSTFA, incubated (65°C, 1 hour), filtered (0.2 μm), and injected. The extract was filtered (0.2 μm filter) and transferred for GC-MS analysis.
Results
Metabolomic analysis identified 152 metabolites in diseased plants, 159 metabolites in healthy plants, and 65 metabolites common to both healthy and diseased plants, including flavonoids, terpenoids, fatty acids, alkaloids, and amino acids. In addition, seven differential metabolic pathways (biosynthesis of unsaturated fatty acids, fatty acid biosynthesis, steroid biosynthesis, cutin, suberine, and wax biosynthesis, fatty acid elongation, brassinosteroid biosynthesis, and fatty acid degradation) were elucidated. The LC-MS analysis identified the presence of Sakuranetin (an antimycobacterial agent), Apigetrin (a non-steroidal anti-inflammatory and antibacterial compound), Pinocembrin (an antineoplastic, vasodilatory, and neuroprotective agent), and Vitexin (a platelet aggregation inhibitor) in healthy pigeonpea leaves.
Conclusion These compounds highlight the potential of pigeonpea extracts in addressing fungal pathogen-related disorders, providing insights into the plant’s response to wilting stress. This study establishes a foundation for further research into the mechanisms of fusarium wilt resistance in pigeonpea, offering valuable information for plant breeding efforts.
Keywords: Enrichment, Fusarium wilt, Metabolites, Pathways, secondary metabolites.