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通常在逆境胁迫下,植物细胞会诱导产生一系列的逆境蛋白来保护细胞免受伤害,其中胚胎发育晚期丰富(LEA)蛋白是目前研究最为普遍的一种。为了研究辣椒LEA蛋白的耐寒性,以豫椒101为材料,根据已经报道的LEA基因序列设计引物,通过PCR技术从辣椒基因组中获得LEA基因,命名为CaLEA5。经生物信息学分析,该基因具有一个完整的ORF为297 bp,编码98个氨基酸。辣椒CaLEA5蛋白质含有LEA基因家族保守的结构域。对其亚细胞定位、跨膜结构进行分析,预测其定位在叶绿体,存在跨膜结构。荧光定量PCR检测结果表明,低温胁迫可诱导CaLEA5基因的表达,且其表达量在迅速达到峰值后又降低,说明CaLEA5是一个快速响应基因,推测其在辣椒抗逆机制中起着重要的作用。在低温胁迫的条件下,硅处理可延缓CaLEA5基因的上调表达。
Usually under stress, plant cells can induce a series of adversity proteins to protect cells from injury. Among them, late embryo abundance (LEA) protein is the most common one. In order to study the chilling tolerance of pepper LEA protein, primers were designed according to the reported LEA gene sequence, and the LEA gene was named as CaLEA5 from pepper genome by PCR. Bioinformatics analysis showed that this gene has a complete ORF of 297 bp encoding a protein of 98 amino acids. Pepper CaLEA5 protein contains a conserved domain of the LEA gene family. Its subcellular localization, transmembrane structure analysis, prediction of its positioning in the chloroplast, there is a transmembrane structure. The results of real-time quantitative PCR showed that low temperature stress induced the expression of CaLEA5 gene, and the expression of CaLEA5 decreased rapidly after reaching the peak value, indicating that CaLEA5 is a rapid response gene and it is presumed that CaLEA5 plays an important role in pepper stress resistance mechanism. Silicon treatment delayed the up-regulation of CaLEA5 gene under low temperature stress.