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.

中文翻译：

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鹰嘴豆荚壁的防御反应由于模拟的草食性而展现出差异蛋白质组特征。

已知豆类的豆荚壁可以保护发育中的种子免受害虫和病原体的侵害。但是，还没有破译赋予昆虫防御作用的机制。在这里，我们利用二维凝胶电泳（2D-GE）结合质谱（MS / MS）来鉴定两个不同品种（商品品种：JG 11和耐性品种：ICC 506-棉铃虫施用12小时后的鹰嘴豆EB）口腔分泌物（模拟食草动物）。进行分析的目的是，幼虫是贪食者，会在12小时内对豆荚造成实质性损害。在凝胶上共检测到600个可重现的蛋白质斑点，比较分析有助于在JG 11和11中鉴定35个（上调12个，下调23个）和20个（10个上调，10个下调）差异表达的蛋白质。 ICC 506-EB。MS / MS后每个品种的蛋白质斑点的功能分类表明差异表达的蛋白质与各种代谢活性有关。此外，由于模拟的草食性，与应激相关的蛋白质如甘露醇脱氢酶（MADH），抗病性样蛋白质CSA1，丝氨酸/苏氨酸激酶（D6PKL2），内切葡聚糖酶19等也被上调。使用STRING数据库进一步分析了鉴定出的可能在防御中起作用的蛋白质，以增进我们对其相互作用伴侣的认识。它解码了几种活性氧清除剂和其他参与细胞壁增强的蛋白质的参与。生化分析还证实了在模拟食草过程中ROS清除剂的积极作用。因此，我们的研究为鹰嘴豆提供了有价值的新见解-^ h。棉铃虫在蛋白质水平上的相互作用。