花花柴蜡质合成相关基因的克隆及分析

Cloning and Analysis of Genes Related to Waxy Synthesis in Karelinia caspia

植物表皮蜡质是由脂肪酸、烷烃、醇类等物质组成的一类有机混合物,在植物抵御逆境胁迫中起着至关重要的作用。为深入研究花花柴(Karelinia caspica)蜡质合成相关基因P450-77A,HHT和FAD2的结构与功能,运用分子克隆技术,获得花花柴蜡质合成相关基因P450-77A和HHT和FAD2的CDS全长,命名为KcP450-77A,KcHHT和KcFAD2,Genebank登录号分别为MW528399,MW528400和MW528401。并对以上基因的编码蛋白进行生物信息学分析,利用RT-PCR技术对其在高温处理下不同时间点进行表达模式分析。结果发现KcP450-77A全长1521bp,编码507个氨基酸;KcHHT全长1347 bp,编码449个氨基酸;KcFAD2全长1152 bp,编码384个氨基酸。通过对上述3个基因在高温处理下不同时间点的表达模式分析发现,3个基因的表达均表现为高-低-高的趋势,其中,KcP450-77A基因在40℃处理8 h时表达量最高,KcHHT基因在处理4 h时表达量最高,KcFAD2基因在处理16 h时表达最为显著。本研究结果表明,花花柴表皮蜡质合成相关基因的表达受高温影响,表现出胁迫-诱导-适应的特性,这种应激反应与植物受到环境胁迫时的生理反应一致。

Plant epicuticular wax is a kind of organic mixture composed of fatty acids,alkanes and alcohols,which plays an important role in plant resistance to adversity stress.In order to study the structure and function of the genes P450-77A,HHT and FAD2 related to paraffin wax synthesis in Karelinia caspia,the full-length coding sequence of the genes P450-77A,HHT and FAD2 were obtained by molecular cloning technique.These genes were named as KcP450-77A,KcHHT and KcFAD2,and submitted to GenBank,the entry numbers were MW-528399,MW528400 and MW528401,respectively.Bioinformatics analysis was performed on those genes,and RT-PCR technology was employed to analyze their expression patterns at different time points under the condition of high temperature.The full lengths of KcP450-77A,KcHHT and KcFAD2 were 1521 bp,1347 bp and 1152 bp,encoding 507,449 and 384 amino acids,respectively.The expression patterns of above three genes were analyzed at different time points under the condition of high temperature,and it was found that the expression of the three genes showed a trend of high-low-high.Under 40℃comdition,KcP450-77A,KcHHT and KcFAD2 genes had the highest expression at 8 h,4 h and 16 h,respectively.The results showed that the expression of above genes related to epidermal wax synthesis of K.caspia was affected by high temperature and showed a character of stress-induction-adaptation,which was consistent with the physiological res-ponse of plants under environmental stress.