Abstract
The pod wall of legumes is known to protect the developing seeds from pests and pathogens. However, the mechanism of conferring defense against insects has not yet been deciphered. Here, we have utilized 2-dimensional gel electrophoresis (2D-GE) coupled with mass spectrometry (MS/MS) to identify over expressed proteins in the pod wall of two different cultivars (commercial cultivar: JG 11 and tolerant cultivar: ICC 506-EB) of chickpea after 12 h of application of Helicoverpa armigera oral secretions (simulated herbivory). The assays were performed with a view that larvae are a voracious feeder and cause substantial damage to the pod within 12 h. A total of 600 reproducible protein spots were detected on gels, and the comparative analysis helped identify 35 (12 up-regulated, 23 down-regulated) and 20 (10 up-regulated, 10 down-regulated) differentially expressed proteins in JG 11 and ICC 506-EB, respectively. Functional classification of protein spots of each cultivar after MS/MS indicated that the differentially expressed proteins were associated with various metabolic activities. Also, stress-related proteins such as mannitol dehydrogenase (MADH), disease resistance-like protein-CSA1, serine/threonine kinase (D6PKL2), endoglucanase-19 etc. were up-regulated due to simulated herbivory. The proteins identified with a possible role in defense were further analyzed using the STRING database to advance our knowledge on their interacting partners. It decoded the involvement of several reactive oxygen species (ROS) scavengers and other proteins involved in cell wall reinforcement. The biochemical analysis also confirmed the active role of ROS scavengers during simulated herbivory. Thus, our study provides valuable new insights on chickpea-H.armigera interactions at the protein level.
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Acknowledgements
We are grateful to the International Crop Research Institute for the Semi-Arid Tropics (ICRISAT), Hyderabad for providing chickpea seeds for analyses. MB and SD would like to thank the ICAR-National Professor (Norman Borlaug Chair) Fund and Department of Biotechnology (DBT)-Assam Agricultural University (AAU) Centre for providing funds and the necessary facility to pursue this work. MB and SD would also like to express their sincere gratitude towards Mr Jitendra Maharana from Assam Agricultural University for helping us create well defined images to support our work. All the authors cordially acknowledge the Dr. Rukmini Govekar form Mumbai allowing us to avail the proteomic infrastructure at the Advanced Centre for Treatment, Research and Education in Cancer (ACTREC), Mumbai.
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DBT, Government of India and ICAR-National Professor (Norman Borlaug Chair) Fund grant.
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MB, SA, and BKS designed the experiments. MB and SD performed the experiments. MB and SA analyzed the proteomics data and co-wrote the manuscript. PJH and BKS reviewed the article. All authors read and approved the final manuscript.
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Bhattacharjee, M., Dhar, S., Handique, P.J. et al. Defense Response in Chickpea Pod Wall due to Simulated Herbivory Unfolds Differential Proteome Profile. Protein J 39, 240–257 (2020). https://doi.org/10.1007/s10930-020-09899-9
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DOI: https://doi.org/10.1007/s10930-020-09899-9