陆地棉苯丙氨酸解氨酶家族基因的鉴定及分析

发布时间：2018-02-03 03:23

本文关键词： 棉花苯丙氨酸解氨酶基因结构表达分析　出处：《分子植物育种》2017年04期 　论文类型：期刊论文

【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。
[Abstract]:Phenylalanine ammonia-lyase is a key enzyme in phenylpropane metabolic pathway and is closely related to plant disease resistance. Verticillium wilt is the first major disease of cotton although genome sequencing of upland cotton has been completed. However, the gene of phenylalanine ammonia-lyase family is still lack of systematic and in-depth study. Bioinformatics and Realtime RT-PCR methods are used in this study. The genes of phenylalanine ammonia-lyase family in TM-1 of Upland cotton (Gossypium hirsutum) were identified and their chromosomal location and gene structure were identified. There are 13 phenylalanine ammonia-lyase family genes in TM-1 genome, their amino acid length is between 697 and 723, distributed on 12 chromosomes. There were only two PAL genes in A4 and only one PAL gene on other chromosomes. In addition, only GhPAL4 had no intron in gene structure. All the other genes contained one intron. 13 phenylalanine ammonia-lyase family genes were divided into 4 groups. The first group contained 8 genes. The results were analyzed by EST database. The 2 genes on chromosome 6 were not expressed in all the tissues examined. Realtime RT-PCR indicated that the gene was located at 1 / 4 / 9 / 10. Ten genes on chromosome 11 can be expressed. GhPAL1 and GhPAL8, which were located on chromosome 1, were the most expressed. The other three genes were not amplified. The expression of 10 genes was analyzed in the resistant materials before and after the induction of Fusarium oxysporum. The results showed that the expression of GhPAL1 and GhPAL8 in the resistant materials was 8 times higher than that in the susceptible materials, and it was induced by Fusarium wilt. These two genes may play an important role in the resistance to Verticillium wilt. It is further proved that PAL gene plays an important role in the resistance to Verticillium wilt and provides a reference for the further study of resistance to Verticillium wilt.
【作者单位】：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;
【基金】：江苏省自主创新项目(CX(14)5057)资助
【分类号】：Q943.2;S562
【正文快照】： 苯丙氨酸解氨酶(phenylalanine ammonia-lyase,PAL,EC4.3.1.5)催化L-苯丙氨酸形成反肉桂酸和游离铵离子,是苯丙烷类代谢途径的第一步酶,也是关键酶和限速酶,该酶在1961年被首次报道(Koukol and Conn,1961)。苯丙烷类代谢途径是植物最重要的次生代谢途径之一,产物包括多种抗病

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