陆地棉苯丙氨酸解氨酶家族基因的鉴定及分析  

Identification and Analysis of the Gene Family of Phenylalanine Ammonialyase in Upland Cotton

作　　者：杨郁文[1] 李双[1] 黄俊宇[1] 张保龙[1] 

机构地区：[1]江苏省农业科学院农业生物技术研究所,江苏省农业生物学重点实验室,南京210014

出　　处：《分子植物育种》2017年第4期1184-1191,共8页Molecular Plant Breeding

基　　金：江苏省自主创新项目(CX(14)5057)资助

摘　　要：苯丙氨酸解氨酶是苯丙烷类代谢途径的关键酶,与植物抗病性密切相关。黄萎病是棉花的第一大病害。虽然陆地棉的基因组测序已经完成,但是对其苯丙氨酸解氨酶家族基因还缺乏系统深入的研究。本研究利用生物信息学和Realtime RT-PCR方法,鉴定了陆地棉TM-1中苯丙氨酸解氨酶家族基因以及它们的染色体定位,基因结构,进化关系和组织表达特征。TM-1基因组中共有13个苯丙氨酸解氨酶家族基因,其氨基酸长度介于697~723,分布在12条染色体上,仅A4上有2个相距440 kb的PAL基因,其他染色体上仅有1个PAL基因。另外,在基因结构上仅GhPAL4没有内含子,其余基因都含有1个内含子。13个苯丙氨酸解氨酶家族基因在进化上分为4组,其中第一组含有8个基因。利用EST数据库分析发现,位于6号染色体上的2个基因在所有检测的组织中不表达。Realtime RT-PCR表明位于1、4、9、10、11号染色体上的10个基因能表达,表达量最高的是位于1号染色体上的GhPAL1和GhPAL8。而其他3个未扩增出。分析能扩增出的10个基因在黄萎菌诱导前后抗感材料中的表达,表明GhPAL1和GhPAL8在抗病材料中的本底表达量是感病材料的8倍,并且黄萎菌诱导后,基因的表达量增加2倍。这2个基因可能在抗黄萎病中起重要作用。进一步证明PAL基因在抗黄萎病中的重要作用,并为后续该类基因的抗性研究提供借鉴。Phenylalanine ammonia-lyase is the key enzyme of the metabolic pathway of phenylpropanoid, which is closely related to plant disease resistance. Verticillium wilt is a major threat to cotton production. Although the sequencing project of upland cotton genome has been completed, but there isn＇t systematic research of the phenylalanine ammonia-lyase family genes of cotton yet. Combining the bioinformatics and Realtime RT-PCR method, we have identified the PAL gene members and analyzed their chromosomal location, gene structure, evolutionary relationships and the expressional characteristics in TM-1. The results indicated there were 13 members of PAL genes in the genome of cotton, the length of their amino acids sequence rangesd from 697 to 723. The genes were distributed on 12 chromosomes, and only one gene located on each chromosome except the chromosome A4 possess 2 PAL genes apart away from 440 kb. In addition, only the GhPA L4 had no intron and the other genes contained one intron. The 13 PAL genes can be divided into 4 groups evolutionarily, and among which 8 genes were clustered into the first group. By scanning the cotton EST database, 2 genes located on chromosome 6 were identified not expressed in all the analyzed organizations. The results of Realtime RT-PCR indicated 10 PAL genes located on the chromosome 1, 4, 9, 10 and 11 can expressed and GhPAL1 and GhPAL8 on the chromosome 1 had the highest expression, while the other 3 genes were not be detected. The inductive expression pattern by verticillium inoculation of the 8 PAL genes in the resistant and susceptible plants were also analyzed, and the results showed the background expression of GhPAL1 and GhPAL8 was about 8 fold in the resistant varieties than that of the susceptible ones and their expression increased 2 fold by verticillium treatment. GhPA L1 and GhPA L8 may played vital role in the verticillium resistantce. These results proved the important role of PAL genes in verticillium wilt, and provided reference for further study of these genes in

关 键 词：棉花 苯丙氨酸解氨酶 基因结构 表达分析 

分 类 号：Q943.2[生物学—植物学] S562[农业科学—作物学]