Harsulkar AM, Giri AP, Patankar AG, Gupta VS, Sainani MN, Ranjekar PK, Deshpande VV
Plant Molecular Biology Unit, Division of Biochemical Sciences, National Chemical Laboratory, Pune 411 008, India.
Plant Physiol. 1999 Oct;121(2):497-506. doi: 10.1104/pp.121.2.497.
We report on the efficacy of proteinase inhibitors (PIs) from three host plants (chickpea [Cicer arietinum], pigeonpea [Cajanus cajan], and cotton [Gossypium arboreum]) and three non-host (groundnut [Arachis hypogea], winged bean [Psophocarpus tetragonolobus], and potato [Solanum tuberosum]) in retarding the growth of Helicoverpa armigera larvae, a devastating pest of important crop plants. Enzyme assays and electrophoretic analysis of interaction of H. armigera gut proteinases (HGPs) with PIs revealed that non-host PIs inhibited HGP activity efficiently whereas host PIs were ineffective. In the electrophoretic assay, trypsin inhibitor activity bands were detected in all of the host and non-host plants, but HGP inhibitor activity bands were present only in non-host plants (except cotton in the host plant group). H. armigera larvae reared on a diet containing non-host PIs showed growth retardation, a reduction in total and trypsin-like proteinase activity, and the production of inhibitor-insensitive proteinases. Electrophoretic analysis of PI-induced HGP showed differential regulation of proteinase isoforms. Interestingly, HGP activity induced in response to dietary potato PI-II was inhibited by winged bean PIs. The optimized combination of potato PI-II and winged bean PIs identified in the present study and their proposed successive use has potential in developing H. armigera-resistant transgenic plants.
我们报告了三种寄主植物(鹰嘴豆[Cicer arietinum]、木豆[Cajanus cajan]和棉花[Gossypium arboreum])以及三种非寄主植物(花生[Arachis hypogea]、四棱豆[Psophocarpus tetragonolobus]和马铃薯[Solanum tuberosum])中的蛋白酶抑制剂(PIs)对棉铃虫幼虫生长的抑制效果,棉铃虫是重要农作物的毁灭性害虫。对棉铃虫肠道蛋白酶(HGPs)与PIs相互作用的酶活性测定和电泳分析表明,非寄主PIs能有效抑制HGP活性,而寄主PIs则无效。在电泳分析中,在所有寄主和非寄主植物中均检测到胰蛋白酶抑制剂活性条带,但HGP抑制剂活性条带仅存在于非寄主植物中(寄主植物组中的棉花除外)。以含有非寄主PIs的饲料饲养的棉铃虫幼虫表现出生长迟缓、总蛋白酶活性和类胰蛋白酶活性降低,以及产生对抑制剂不敏感的蛋白酶。对PI诱导的HGP进行电泳分析显示蛋白酶同工型受到差异调节。有趣的是,由膳食马铃薯PI-II诱导的HGP活性受到四棱豆PIs的抑制。本研究中确定的马铃薯PI-II和四棱豆PIs的优化组合及其建议的连续使用在培育抗棉铃虫转基因植物方面具有潜力。