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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 a key enzyme of phenylpropanoid metabolic pathway, which is closely related to plant disease resistance. Verticillium wilt is the first major disease of cotton. Although the upland cotton genomic sequencing has been completed, but its phenylalanine ammonia lyase family of genes is still lack of systematic and in-depth study. This study identified the phenylalanine ammonia-lyase family genes in upland cotton TM-1 and their chromosomal location, gene structure, evolutionary relationships and tissue expression characteristics using bioinformatics and Realtime RT-PCR. A total of 13 phenylalanine ammonia-lyase family genes were found in the TM-1 genome. The amino acid length of the TM-1 gene ranged from 697 to 723, distributed on 12 chromosomes. There were only two PAL genes of 440 kb apart on A4 and other chromosomes Only 1 PAL gene. In addition, only GhPAL4 has no introns in its genetic structure, and the rest of the genes contain 1 intron. The 13 phenylalanine ammonia lyase family genes are evolutionarily divided into 4 groups, of which the first group contains 8 genes. Using EST database analysis, two genes located on chromosome 6 were not expressed in all the tested tissues. Realtime RT-PCR showed that 10 genes on chromosomes 1, 4, 9, 10 and 11 were expressed, and GhPAL1 and GhPAL8 on chromosome 1 were the highest expressed. The other three were not amplified. The analysis of the ten genes that could be amplified showed that the background expression of GhPAL1 and GhPAL8 in the resistant material was eight times that of the susceptible material, , The gene expression increased 2-fold. These two genes may play an important role in the resistance to Verticillium wilt. Further prove the important role of PAL gene in the resistance to Verticillium wilt and provide references for the follow-up study on the resistance of these genes.