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为了探讨14-3-3基因在小麦逆境胁迫应答中的调控作用,利用RACE技术克隆了两个包含完整编码框的14-3-3基因(命名为Ta14R1和Ta14R2),其中Ta14R1 cDNA长999 bp,编码262个氨基酸,而Ta14R2 cDNA长897 bp,编码261个氨基酸。Ta14R1/Ta14R2-GFP融合载体瞬时表达结果显示,Ta14R1和Ta14R2蛋白均定位于细胞质和细胞膜,但不在叶绿体中。荧光定量PCR分析表明,Ta14R1和Ta14R2均在萌发1 d的胚芽鞘中表达量最高;在高温、低温、模拟干旱和ABA处理下,两个基因在小麦的根和叶中都受胁迫诱导而且显著上调表达,推测这两个14-3-3基因通过依赖ABA的非生物胁迫响应途径发挥作用,可能参与了小麦中高温、低温和干旱胁迫的耐受调节过程。
In order to investigate the role of 14-3-3 in the regulation of stress response in wheat, two 14-3-3 genes (named Ta14R1 and Ta14R2) were cloned by RACE technique. The Ta14R1 cDNA was 999 bp in length , Encoding 262 amino acids, while Ta14R2 cDNA is 897 bp in length and encodes 261 amino acids. The transient expression results of Ta14R1 / Ta14R2-GFP fusion vector showed that both Ta14R1 and Ta14R2 proteins localized in the cytoplasm and cell membrane, but not in the chloroplast. Quantitative real-time PCR analysis showed that both Ta14R1 and Ta14R2 expressed the highest level in the coleoptile on the first day of germination. Under the conditions of high temperature, low temperature, simulated drought and ABA treatment, both genes were induced by stress and significant in wheat roots and leaves Upregulated expression of these two 14-3-3 genes presumably through abiotic ABA-dependent stress response pathway, which may be involved in the regulation of tolerance to high temperature, low temperature and drought stress in wheat.