Moulin M M, Rodrigues R, Ribeiro S F F, Gonçalves L S A, Bento C S, Sudré C P, Vasconcelos I M, Gomes V M
Laboratório de Melhoramento Genético Vegetal, Universidade Estadual do Norte Fluminense Darcy Ribeiro, Campos dos Goytacazes, RJ, Brasil
Laboratório de Melhoramento Genético Vegetal, Universidade Estadual do Norte Fluminense Darcy Ribeiro, Campos dos Goytacazes, RJ, Brasil.
Genet Mol Res. 2014 Nov 7;13(4):9229-43. doi: 10.4238/2014.November.7.10.
Several plant organs contain proteinase inhibitors, which are produced during normal plant development or are induced upon pathogen attack to suppress the enzymatic activity of phytopathogenic microorganisms. In this study, we examined the presence of proteinase inhibitors, specifically trypsin inhibitors, in the leaf extract of Capsicum baccatum var. pendulum inoculated with PepYMV (Pepper yellow mosaic virus). Leaf extract from plants with the accession number UENF 1624, which is resistant to PepYMV, was collected at 7 different times (0, 24, 48, 72, 96, 120, and 144 h). Seedlings inoculated with PepYMV and control seedlings were grown in a growth chamber. Protein extract from leaf samples was partially purified by reversed-phase chromatography using a C2/C18 column. Residual trypsin activity was assayed to detect inhibitors followed by Tricine-SDS-PAGE analysis to determine the N-terminal peptide sequence. Based on trypsin inhibitor assays, trypsin inhibitors are likely constitutively synthesized in C. baccatum var. pendulum leaf tissue. These inhibitors are likely a defense mechanism for the C. baccatum var. pendulum- PepYMV pathosystem.
几种植物器官含有蛋白酶抑制剂，这些抑制剂在植物正常发育过程中产生，或在病原体攻击时被诱导产生，以抑制植物病原微生物的酶活性。在本研究中，我们检测了接种辣椒黄化花叶病毒（PepYMV）的灯笼椒叶片提取物中蛋白酶抑制剂，特别是胰蛋白酶抑制剂的存在情况。从对PepYMV具有抗性的保藏编号为UENF 1624的植株上采集叶片提取物，在7个不同时间点（0、24、48、72、96、120和144小时）进行收集。接种PepYMV的幼苗和对照幼苗在生长室中培养。使用C2/C18柱通过反相色谱法对叶片样品的蛋白质提取物进行部分纯化。通过测定残余胰蛋白酶活性来检测抑制剂，随后进行Tricine-SDS-PAGE分析以确定N端肽序列。基于胰蛋白酶抑制剂测定，胰蛋白酶抑制剂可能在灯笼椒叶片组织中组成型合成。这些抑制剂可能是灯笼椒 - PepYMV病理系统的一种防御机制。
