黄瓜苯丙氨酸解氨酶基因CsPAL的克隆及响应白粉菌侵染的表达分析  

Cloning of CsPAL Gene and Its Expression Analysis in response to Powdery Mildew Infection

作　　者：高红胜[1] 张仁英[1] 许学文[1] 齐晓花[1] 徐强[1] 陈学好[1] Gao Hongsheng;Zhang Renying;Xu Xuewen;Qi Xiaohua;Xu Qiang;Chen Xuehao(School of Horticulture and Plant Protection,Yangzhou University,Yangzhou,225009)

机构地区：[1]扬州大学园艺与植物保护学院,扬州225009

出　　处：《分子植物育种》2019年第6期1757-1762,共6页Molecular Plant Breeding

基　　金：国家自然科学基金(31672176);江苏省农业科技自主创新资金(CX(17)2004);江苏省农业三新工程项目(SXGC[2017]303)共同资助

摘　　要：为了分析苯丙氨酸解氨酶基因(CsPAL)在黄瓜侵染白粉菌中的转录应答响应,对CsPAL基因(GenBank No.JN 675927)及其启动子进行了克隆与序列分析,再利用qRT-PCR技术分析了该基因在黄瓜高抗白粉病品种‘Jin5-508’接种白粉菌后不同时间的相对表达量。结果表明:CsPAL基因全长2 142 bp,编码713个氨基酸;推测CsPAL蛋白存在于细胞质中;系统发育进化树分析发现与拟南芥基因进化距离较近;启动子克隆及序列分析显示该基因能够响应真菌的侵染;组织特异性表达发现CsPAL在在叶片中的表达量最高;qRTPCR结果表明‘:Jin5-508’叶片在接菌16 h内,CsPAL基因的相对表达量于对照间无显著差异;接菌16 h后,CsPAL基因表达量迅速上升,显著高于对照,并在24 h达到最大值;接菌48 h后,CsPAL基因的表达量逐渐下降,但在96 h内仍显著高于相应对照的表达量。综上所述,黄瓜CsPAL基因为白粉菌侵染响应基因,可能与黄瓜对白粉病的抗性具有一定的相关性。To analyze the role of Cucumis sativus Phenylalanine ammonia-lyase(CsPAL) gene in responding of powdery mildew infection, we cloned and sequenced the CsPAL gene(GenBank No.JN675927) and its promoter, then analyzed the relative expression levels of the gene in the breed of high resistance to powdery mildew ’Jin5-508’at different time-points after inoculation of powdery mildew(PM) pathogen. The results showed that the full length of CsPAL was 2 142 bp, encodes 713 amino acids and located in cytoplasm. Phylogenetic tree analysis showed that the gene was evolutionary close to Arabidopsis PAL gene. Cloning and sequences analysis of the promoter showed that CsPAL gene was able to respond to fungal infection. Tissue-special expression showed that the gene was highly expressed in leaves;qRT-PCR results showed that: there was no significant different in CsPAL expression within 16 h of PM inoculation and control. The relative expression of CsPAL was significantly higher in PM inoculated leaves than those in the control 16 h after treatment and reached the maximum at 24 h. The relative expression of CsPAL was down-regulated gradually after 48 h of treatment, but still significantly higher than control within 96 h. In conclusion, the cloned cucumber CsPAL might be a PM inoculation responsive gene, and might correlate with cucumber PM resistance.

关 键 词：黄瓜 白粉病 CsPAL 克隆 QRT-PCR 

分 类 号：S436.421.12[农业科学—农业昆虫与害虫防治]