马尾松PmWRKY2和PmWRKY6基因生物信息学及表达分析

黄金龙<sup>1; 2</sup>; 陈 虎<sup>2</sup>; 陈颖豪<sup>2</sup>; 杨章旗<sup>2*</sup>

引用本文:	黄金龙, 陈虎, 陈颖豪, 杨章旗.马尾松PmWRKY2和PmWRKY6基因生物信息学及表达分析[J].广西植物,2025,45(10):1926-1935.[点击复制]
	HUANG Jinlong, CHEN Hu, CHEN Yinghao, YANG Zhangqi.Bioinformatics and expression analysis of PmWRKY2 and PmWRKY6 genes in Pinus massoniana[J].Guihaia,2025,45(10):1926-1935.[点击复制]

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*马尾松PmWRKY2和PmWRKY6基因生物信息学及表达分析*
黄金龙^1,2, 陈虎², 陈颖豪², 杨章旗^2*
1.广西师范大学生命科学学院, 广西桂林 541006;2. 广西壮族自治区林业科学研究院, 国家林业和草原局马尾松工程技术研究中心, 广西马尾松工程技术研究中心, 广西优良用材林资源培育重点实验室, 南宁 530002

摘要:

WRKY家族是高等植物中最大的转录因子家族之一。课题组前期研究发现WRKY家族基因在马尾松响应虫害、生长发育、干旱胁迫等方面发挥重要作用,为进一步研究WRKY家族基因的生物学功能和响应外源激素情况,该研究对PmWRKY2和PmWRKY6基因生物信息学特征进行分析,并利用qPCR技术对其在脱落酸(ABA)、水杨酸(SA)、茉莉酸甲酯(MeJA)和赤霉素(GA)等外源激素和钙离子处理下的表达模式进行研究。结果表明:(1)PmWRKY2和PmWRKY6蛋白分别编码667个和575个氨基酸,亚细胞均定位于细胞核,2个基因N端均含有高度保守的WRKYGQK七肽结构,C端含有锌指结构,属于WRKY转录因子家族。(2)在进化关系上,PmWRKY2和PmWRKY6蛋白与同为裸子植物油松的PtXG20020.1和Pt2G29990.1蛋白关系最近。(3)与CK相比,4种激素处理和相应添加Ca²⁺处理均能显著诱导PmWRKY2和PmWRKY6基因表达,其表达量均在处理后期达到峰值; 在单独Ca²⁺处理下,2个基因均呈先下降后上升的表达趋势,其中Ca²⁺处理前期能显著诱导PmWRKY2基因表达,处理后期与CK无显著性差异,而PmWRKY6基因的表达量在整个处理过程中均显著高于CK。上述结果表明, PmWRKY2和PmWRKY6基因均能响应不同外源信号物质,但表达模式存在差异。

关键词: 马尾松, PmWRKY2, PmWRKY6, 生物信息学, 表达模式

DOI：10.11931/guihaia.gxzw202404036

分类号:Q943

文章编号:1000-3142(2025)10-1926-10

基金项目:国家自然科学基金(32060348); 广西科技基地和人才专项(桂科AD19254004); 八桂学者专项(2019A26); 广西自然科学基金(2019GXNSFDA245033)

Bioinformatics and expression analysis of PmWRKY2 and PmWRKY6 genes in Pinus massoniana

HUANG Jinlong^1,2, CHEN Hu², CHEN Yinghao², YANG Zhangqi^2*

1.College of Life Sciences, Guangxi Normal University, Guilin 541006, Guangxi, China;2. Guangxi Key Laboratory of Superior Timber Trees Resource Cultivation, Engineering Technology Research Center of Masson Pine of State Forestry and Grassland Administration,Engineering Technology Research Center of Masson Pine of Guangxi, Guangxi Institute of Forestry Science, Nanning 530002, China

Abstract:

The WRKY family is one of the largest families of transcription factors in higher plants. Previous studies have found that WRKY family genes play an important role in the response of Pinus massoniana to insect pests, growth and development, and drought stress. In order to further study the biological function of WRKY family genes and the response to exogenous hormones, the bioinformatics characteristics of PmWRKY2 and PmWRKY6 genes were analyzed, and the expression patterns of PmWRKY2 and PmWRKY6 genes under exogenous hormones treatment such as abscisic acid(ABA), salicylic acid(SA), methyl jasmonate(MeJA)and gibberellin acid(GA)and calcium ion were studied by qPCR. The results were as follows:(1)PmWRKY2 and PmWRKY6 proteins encoded 667 and 575 amino acids, respectively, and were subcellularly located in the nucleus, and the N-terminus of the two genes contained a highly conserved WRKYGQK heptapeptide structure, and the C-terminus contained a zinc finger structure, which belonged to the WRKY transcription factor family.(2)In terms of evolutionary relationship, PmWRKY2 protein and PmWRKY6 protein were most closely related to PtXG20020.1 and Pt2G29990.1 proteins from P. tabuliformis, which were also gymnosperm.(3)Compared with CK, the expression of PmWRKY2 and PmWRKY6 genes could be significantly induced by the four hormone treatments and the corresponding Ca²⁺ treatment, and their expression levels reached the peak in the late treatment period. Under Ca²⁺ treatment alone, both genes showed an expression trend of first decreasing and the increasing, in which the expression of PmWRKY2 gene was significantly induced in the early stage, and there was no significant difference between the expression of PmWRKY2 gene in the late treatment and the CK, while the expression level of PmWRKY6 gene was significantly higher than that of the CK during the whole treatment. These results show that both PmWRKY2 and PmWRKY6 genes can respond to different exogenous signaling substances, but the expression patterns are different.

Key words: Pinus massoniana, PmWRKY2, PmWRKY6, bioinformatics, expression pattern