Mahajan Neha S, Mishra Manasi, Tamhane Vaijayanti A, Gupta Vidya S, Giri Ashok P
Division of Biochemical Sciences, CSIR-National Chemical Laboratory, MS, India.
Biochim Biophys Acta. 2013 Jun;1830(6):3414-20. doi: 10.1016/j.bbagen.2013.03.017. Epub 2013 Mar 23.
A multi-domain Pin-II type protease inhibitor from Capsicum annuum (CanPI-7) is known to be effective against the insect pest, Helicoverpa armigera. The present study is an attempt to investigate the optimal dose of recombinant CanPI-7 (rCanPI-7) for effective antibiosis to H. armigera and further to characterize the responses of digestive proteases upon rCanPI-7 ingestion.
The gut protease activity was assessed biochemically and transcript accumulation pattern for selected trypsin and chymotrypsin genes was analyzed by quantitative Real-Time PCR.
The growth retardation upon exposure to rCanPI-7 was more prominent in neonates as compared to third instar larvae. Influence of stage and dosage of rCanPI-7 was conspicuous on the expression and regulation of candidate trypsin and chymotrypsin genes in H. armigera. The transcript accumulation pattern correlated with the protease activity in rCanPI-7 exposed larvae.
We conclude that early exposure and specific dose of protease inhibitor are essential for effective antibiosis despite the large diversity and plasticity in the expression of protease genes in H. armigera. Moreover, it is also evident that the regulation and expression of H. armigera gut proteases are specific to the stage of PI exposure.
These results highlight the requirement of optimal PI concentration for effective growth retardation and for inhibiting the major gut proteases of H. armigera.
已知来自辣椒的一种多结构域Pin-II型蛋白酶抑制剂(CanPI-7)对害虫棉铃虫有效。本研究旨在探究重组CanPI-7(rCanPI-7)对棉铃虫产生有效抗菌作用的最佳剂量,并进一步表征摄入rCanPI-7后消化蛋白酶的反应。
通过生化方法评估肠道蛋白酶活性,并通过定量实时PCR分析所选胰蛋白酶和胰凝乳蛋白酶基因的转录本积累模式。
与三龄幼虫相比,新生幼虫暴露于rCanPI-7后生长迟缓更为明显。rCanPI-7的阶段和剂量对棉铃虫中候选胰蛋白酶和胰凝乳蛋白酶基因的表达和调控有显著影响。转录本积累模式与暴露于rCanPI-7的幼虫中的蛋白酶活性相关。
我们得出结论,尽管棉铃虫蛋白酶基因的表达具有很大的多样性和可塑性,但早期暴露和特定剂量的蛋白酶抑制剂对于有效抗菌至关重要。此外,棉铃虫肠道蛋白酶的调控和表达也明显因PI暴露阶段而异。
这些结果突出了有效抑制棉铃虫生长和抑制其主要肠道蛋白酶所需的最佳PI浓度。