参薯WRKY基因家族鉴定及其在线虫胁迫响应及激素信号调控中的作用

doi:10.13560/j.cnki.biotech.bull.1985.2025-1287

摘要/Abstract

摘要：

目的系统鉴定参薯（Dioscorea alata）WRKY基因家族成员并解析其基本特征，为研究该家族成员在参薯响应根结线虫胁迫中的功能奠定基础。方法基于参薯全基因组数据，利用生物信息学方法对DaWRKY基因家族进行系统鉴定，并分析其蛋白理化性质、系统进化关系、基因结构、染色体定位及启动子顺式作用元件。进一步结合转录组数据与RT-qPCR实验，分析DaWRKY基因在不同组织、南方根结线虫胁迫以及植物激素（ABA、JA、SA）处理下的表达模式。结果 DaWRKY基因家族包含70个成员，编码蛋白氨基酸数量为127-651，均为亲水性蛋白，预测定位均在细胞核。系统进化分析将其划分为I、Ⅱ、Ⅲ三个亚族，其中II亚族占比最高（65.71%）。染色体定位结果显示，该家族成员不均匀分布在15条染色体上。表达谱分析表明，DaWRKY基因在根和微型块茎中显著富集。在南方根结线虫胁迫下，8个DaWRKY基因在7 d均上调表达，其中DaWRKY2上调表达倍数最高；在ABA处理中，仅DaWRKY33在1-5 d下调，而DaWRKY26上调最显著；JA处理诱导DaWRKY33、DaWRKY69表达呈现先降后升的趋势，且8个基因在12 h均表现为上调；SA处理中，除DaWRKY16外，其他基因均在12 h表达上调。结论鉴定出参薯70个DaWRKY转录因子，其中DaWRKY2、DaWRKY33、DaWRKY46等成员在根结线虫侵染与ABA、JA、SA等防御激素处理下显著响应，表明DaWRKY基因家族在参薯应对生物与非生物胁迫中发挥重要作用。

关键词: 参薯, WRKY基因家族, 转录因子, 线虫胁迫, 激素诱导, 基因表达

Abstract:

Objective Systematic identification and characterization of WRKY gene family members in greater yam (Dioscorea alata) were performed to establish a foundation for subsequent research on their functions in response to root-knot nematode stress. Method Based on the whole-genome data of Dioscorea alata, we conducted a genome-wide identification of the DaWRKY gene family using bioinformatics methods. The analyses included physicochemical properties of the encoded proteins, phylogenetic relationships, gene structures, chromosomal locations, and promoter cis-acting elements. The expression patterns of the DaWRKY gene were analyzed using a combined approach of transcriptome data and RT-qPCR experiments across various tissues, under Meloidogyne incognita stress, and in response to treatments with plant hormones (ABA, MeJA, and SA). Result A total of 70 DaWRKY genes were identified in D. alata. The encoded proteins, ranging from 127 to 651 amino acids in length, were all predicted to be hydrophilic and localized to the nucleus. Phylogenetic analysis classified them into three groups (I, II, and III), with group II being the largest proportion (65.71%). Chromosomal location analysis revealed that these genes were unevenly distributed across 15 chromosomes. Expression profile analysis indicated that DaWRKY genes were significantly enriched in roots and microtubers. Under M. incognita stress, eight DaWRKY genes were up-regulated at 7 d post-infection, among which DaWRKY2 exhibited the highest fold change in up-regulation. During ABA treatment, only DaWRKY33 was down-regulated from 1 to 5 d, while DaWRKY26 showed the most significant up-regulation. JA treatment induced an initial decrease followed by an increase in the expression of DaWRKY33 and DaWRKY69, and eight genes were all up-regulated at 12 h post-treatment. Under SA treatment, all genes except DaWRKY16 were up-regulated at 12 h. Conclusion A total of 70 DaWRKY transcription factors were identified in D. alata. Among them, members such as DaWRKY2, DaWRKY33, and DaWRKY46 showed significant responses to root-knot nematode infection as well as treatment with defense-related hormones including ABA, JA, and SA. These results indicate that the DaWRKY gene family plays crucial roles in the response of D. alata to biotic and abiotic stresses, providing a foundation for elucidating the stress resistance mechanisms of this crop.

Key words: Dioscorea alata, WRKY gene family, transcription factor, nematode stress, hormone induction, gene expression

王明珠, 孙林涛, 常英英, 张江利, 高锦, 王瑞飞, 张昊, 李明军, 杨清香. 参薯WRKY基因家族鉴定及其在线虫胁迫响应及激素信号调控中的作用[J]. 生物技术通报, 2026, 42(10): 108-121.

WANG Ming-zhu, SUN Lin-tao, CHANG Ying-ying, ZHANG Jiang-li, GAO Jin, WANG Rui-fei, ZHANG Hao, LI Ming-jun, YANG Qing-xiang. Role of the WRKY Gene Family in Dioscorea alata L. in Response to Nematode Stress Response and Hormonal Signal Regulation[J]. Biotechnology Bulletin, 2026, 42(10): 108-121.

图/表 8

图1 参薯与拟南芥WRKY基因家族系统进化树

Fig. 1 Phylogenetic tree of WRKY gene family in D. alata and A. thaliana

图2 DaWRKY的基序分布（A）、保守结构域（B）及基因结构分析（C）为便于排版，对较长的基因进行了部分截断（用\表示），示意图中只展示基序、结构域、CDS区的相对位置，不代表它们在基因或蛋白上所处的绝对位置

Fig. 2 Motif distribution (A), conserved domain (B), and gene structure (C) analysis of DaWRKYFor typesetting purposes, longer genes have been partially truncated (indicated by \). The schematic diagram only shows the relative positions of motifs, domains, and CDS regions, and does not indicate their absolute locations in the gene or protein

图3 DaWRKY基因顺式作用元件分析中间面板是显示顺式作用元件富集水平的热图。右边的堆积图显示了顺式作用元件的丰度：蓝色矩形表示光响应顺式作用元件；绿色矩形表示激素响应顺式元件；黄色矩形表示胁迫响应的顺式元件；青色矩形表示与生长和发育相关的顺式元件

Fig. 3 Analysis of the cis-acting elements of the DaWRKY geneThe central panel is a heatmap displaying the enrichment levels of cis-acting elements. The stacked bar chart on the right illustrates the abundance of cis-acting elements: blue rectangles represent light-responsive cis-acting elements; green rectangles, hormone-responsive cis-elements; yellow rectangles, stress-responsive cis-elements; while cyan rectangles denote cis-elements associated with growth and development

图4 参薯DaWRKY基因家族的染色体位置（A）和共线性分析（B）A：彩色矩形条表示D. alata的染色体；Chr1-20表示对应的染色体；红色粗轮廓表示基因串联重复，0-40 Mb的刻度表示染色体长度。B：红色线条表示片段重复，通过红色线条连接的两个基因是同源的。基因密度由热图（内圈）表示，外圈显示染色体的长度，最外圈的黑色线段表示基因在染色体上的定位

Fig. 4 Chromosomal location (A) and collinearity analysis (B) of DaWRKY gene family in D. alataA: Colored rectangular bars represent the chromosomes of D. alata; Chr1-20 indicate the corresponding chromosomes; the red thick outlines indicate gene tandem duplications, and the scale bar of 0-40 Mb represents the chromosome length. B: Red lines indicate segmental duplications, and the two genes connected by red lines are homologous. Gene density is represented by heatmap (inner circle), the outer circle shows the chromosome length, and the black line segments on the outermost circle indicate the gene locations on the chromosomes

图5 参薯（D. alata）、几内亚山药（D. rotundata）、水稻（O. sativa）和拟南芥（A. thaliana）WRKY基因的共线性分析

Fig. 5 The collinearity analysis of WRKY genes in D. alata, D. rotundata, O. sativa, and A. thaliana

图6 DaWRKY基因在不同组织中的相对表达量不同小写字母表示组间存在显著差异（P<0.05），下同

Fig. 6 Relative expression of DaWRKY in different tissuesDifferent lowercase letters indicate significant differences among groups (P<0.05). The same below

图7 DaWRKY基因在接种南方根结线虫不同时期的相对表达量

Fig. 7 Relative expression of DaWRKY genes at different stages after M. incognita inoculation

图8 DaWRKY基因在不同激素处理下不同时间的相对表达量A：ABA处理；B：MeJA处理；C：SA处理

Fig. 8 Relative expression of the DaWRKY gene at different time under various hormonal treatmentsA: Treatment with ABA. B: Treatment with MeJA. C: Treatment with SA

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