中国樱桃‘玛瑙红’CpCHS1基因的克隆与表达分析

Cloning and Expression Analysis of CpCHS1 Gene of Cerasus pseudoceresus‘Manaohong’

以‘玛瑙红’樱桃根系组织为试材,采用PCR(RT-PCR)技术对CpCHS1基因进行克隆,并对其序列进行生物信息学分析及表达分析,研究了该基因在植株不同组织中及在逆境处理下的表达情况,以期为后续深入解析该基因在逆境胁迫中功能提供参考依据。结果表明:‘玛瑙红’樱桃CpCHS1全长1227 bp,编码408个氨基酸。相对分子质量为44.6 kD,理论等电点(pI)为6.53;无跨膜结构域,且为稳定的亲水性蛋白;蛋白信号肽预测为非分泌蛋白;二级结构包括α-螺旋及无规则卷曲等;多序列比对表明,CpCHS1与水蜜桃(Prunus persica)亲缘关系最近,相似度为100%。荧光定量PCR结果表明,CpCHS1在根中的表达量最高,其次是果、叶;在干旱、高盐、低温和高温等胁迫处理下CpCHS1表达量出现显著变化。综上所述,‘玛瑙红’樱桃CpCHS1在根系中表达量最高,且在胁迫处理下显著响应逆境胁迫,推测CpCHS1可能与植株抗性有着密切联系。

Taking the root tissue of‘Manaohong’cherry as experimental materials,using PCR(RT-PCR)technology,the CpCHS1 gene was cloned,and its sequence was analyzed by bioinformatics and expression analysis.The expression of this gene in different tissues of the plant and its expression in response to stress were investigated,in order to provide reference for further analysis of the function of this gene in stress.The results showed that,the full length of‘Manaohong’cherry CpCHS1 was 1227 bp,which encoded 408 amino acids.The relative molecular mass was 44.6 kD,the theoretical isoelectric point(pI)was 6.53;there was no transmembrane domain,and it was a stable hydrophilic protein;the protein signal peptide was predicted to be a non-secreted protein;the secondary structure includedα-helix and no regular curling,etc.;multiple sequence alignments showed that CpCHS1 was closely related to Prunus persica,with a similarity of 100%.The results of fluorescence quantitative PCR showed that the expression of CpCHS1 in roots was the highest,followed by fruits and leaves.CpCHS1 expression changed significantly under drought,high salt,low temperature and high temperature.In conclusion‘Manaohong’cherry CpCHS1 had the highest expression level in the root system,and it responded significantly to adversity stress,which might be closely related to plant resistance.