转内质网小分子量热激蛋白CaHSP22.5基因甜椒植株的获得

Obtain of Transgenic Sweet Pepper Plants with Gene from Endoplasmic Reticulum Small Heat Shock Protein CaHSP22.5

低温胁迫是喜温蔬菜和作物生产中的主要障碍因素。内质网是细胞内合成蛋白质和膜脂的重要细胞器,内质网小分子量热激蛋白与植物的耐冷性密切相关。我们克隆甜椒内质网小分子量热激蛋白(Ca HSP22.5)基因,构建真核表达载体,用农杆菌LBA4404介导初步获得10个T0代具有卡那抗性的转基因株系,开花结实获得T1代种子94粒。对获得T1代种子进行卡那抗性培养基的萌发和生长筛选,然后对T1代抗性强的转基因株系进行RT-PCR检测,鉴定获得6个转基因株系。通过叶绿素荧光分析证实,组成性表达Ca HSP22.5的转基因株系与野生型相比,显著减轻低温引起的光抑制,表明获得了抗冷性强的种质新材料。可进一步研究Ca HSP22.5的作用机制,从而指导喜温农作物的反季节栽培和育种。

Low temperature s tress is the main obstacle in the development of warm vegetables and crops. Endoplasmic reticulum is an important organelle for intracellular synthesis of proteins and membrane lipids. The endoplasmic reticulum small heat shock protein was closely related to the cold resistance of plants. We cloned the endoplasmic reficulum small heat shock protein of the sweet pepper （CaHSP22.5） gene and constructed the euka- ryotic expression vector. Ten preliminary transgenic strains of To generation were obtained by mediation of Agrobacterium LBA4404.94 seeds ofT1 generation were obtained. The gemination ofT1 seeds were determined on resistant culture medium. RT-PCR was performed to detect T1 generation transgenic strains, and six transgenic strains were obtained. Chlorophyll fluorescence analysis showed that the transgenic strain with constituent expression of CaHSP22.5 reduced the damage caused by low temperature than the wild type, indicating that new materials of germplasm with strong cold tolerance were obtained. The mechanism of CaHSP22.5 could be further studied to guide the anti-seasonal cultivation and breeding of crops.