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组成型过表达水稻糜蛋白酶抑制剂基因 OCPI2 导致转基因拟南芥植物生长增强、耐盐和耐渗透胁迫。

Constitutive over-expression of rice chymotrypsin protease inhibitor gene OCPI2 results in enhanced growth, salinity and osmotic stress tolerance of the transgenic Arabidopsis plants.

机构信息

Department of Plant Molecular Biology, University of Delhi South Campus, New Delhi 110021, India.

Department of Plant Molecular Biology, University of Delhi South Campus, New Delhi 110021, India.

出版信息

Plant Physiol Biochem. 2015 Jul;92:48-55. doi: 10.1016/j.plaphy.2015.03.012. Epub 2015 Apr 16.

DOI:10.1016/j.plaphy.2015.03.012
PMID:25910649
Abstract

Protease inhibitors are involved primarily in defense against pathogens. In recent years, these proteins have also been widely implicated in response of plants to diverse abiotic stresses. Rice chymotrypsin protease inhibitor gene OCPI2 is highly induced under salt and osmotic stresses. The construct containing the complete coding sequence of OCPI2 cloned downstream to CaMV35S promoter was transformed in Arabidopsis and single copy, homozygous transgenic lines were produced. The transgenic plants exhibited significantly enhanced tolerance to NaCl, PEG and mannitol stress as compared to wild type plants. Importantly, the vegetative and reproductive growth of transgenic plants under unstressed, control conditions was also enhanced: transgenic plants were more vigorous than wild type, resulting into higher yield in terms of silique number. The RWC values and membrane stability index of transgenic in comparison to wild type plants was higher. Higher proline content was observed in the AtOCPI2 lines, which was associated with higher transcript expression of pyrroline-5-carboxylate synthase and lowered levels of proline dehydrogenase genes. The chymotrypsin protease activities were lower in the transgenic as against wild type plants, under both unstressed, control as well as stressed conditions. It thus appears that rice chymotrypsin protease inhibitor gene OCPI2 is a useful candidate gene for genetic improvement of plants against salt and osmotic stress.

摘要

蛋白酶抑制剂主要参与防御病原体。近年来,这些蛋白质也被广泛认为与植物对各种非生物胁迫的反应有关。水稻糜蛋白酶抑制剂基因 OCPI2 在盐和渗透胁迫下高度诱导。含有 OCPI2 完整编码序列的构建体被克隆到 CaMV35S 启动子的下游,并转化到拟南芥中,产生了单拷贝、纯合的转基因系。与野生型植物相比,转基因植物对 NaCl、PEG 和甘露醇胁迫表现出显著增强的耐受性。重要的是,在非胁迫、对照条件下,转基因植物的营养和生殖生长也得到了增强:转基因植物比野生型植物更健壮,导致在蒴果数量方面的产量更高。与野生型植物相比,转基因植物的 RWC 值和膜稳定性指数更高。在 AtOCPI2 系中观察到较高的脯氨酸含量,这与吡咯啉-5-羧酸合酶的转录表达升高和脯氨酸脱氢酶基因水平降低有关。在非胁迫、对照和胁迫条件下,转基因植物的糜蛋白酶活性均低于野生型植物。因此,水稻糜蛋白酶抑制剂基因 OCPI2 是一种用于遗传改良植物耐盐和渗透胁迫的有用候选基因。

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