外施水杨酸对白粉菌侵染小麦的影响及白粉菌转录组分析

doi:10.13560/j.cnki.biotech.bull.1985.2024-1201

摘要/Abstract

摘要：

目的白粉病作为危害小麦的重要病害之一，对小麦的产量和品质构成严重威胁，通过外施水杨酸探究其对白粉菌侵染的调控作用，并结合白粉菌转录组分析揭示其侵染机制及水杨酸介导的抗病分子基础。方法以白粉菌侵染感病普通小麦中作9504发病后采集的小麦白粉菌为材料，对小麦叶片感病后1-4 d的小麦白粉菌进行转录组分析，同时通过外施水杨酸以观察植物激素对白粉菌侵染小麦的影响。结果白粉菌转录组结果显示，与1 d相比，4 d时上调了399个基因，下调了1 110个基因。上调基因富集在次级代谢的生物合成、代谢通路、蛋白酶体等通路，下调基因主要富集在缬氨酸、亮氨酸和异亮氨酸的降解等通路。对上述基因进行绘制蛋白互作网络图，发现BGTH12_LOCUS642、BGTH12_LOCUS3045和BGTH12_LOCUS5497为白粉菌核心hub基因，表明这3个基因在侵染小麦过程中发挥重要作用。另外，研究发现外源施加SA显著提高了小麦中ERF109、PP2C30、TIFY6B、HSP70、At4g15970和HERK1这6个hub基因的表达量，进而降低白粉菌的危害。结论白粉菌在侵染小麦的过程中有大量差异基因的表达，BGTH12_LOCUS642、BGTH12_LOCUS3045和BGTH12_LOCUS5497为白粉菌核心hub基因。外施水杨酸可以诱导小麦中抗性基因表达来对抗白粉菌致病基因进而抑制分生孢子的生长发育，从而一定程度上减缓白粉菌的侵染，为小麦白粉病抗性研究增加新的途径。

关键词: 小麦白粉病, 白粉菌侵染, 水杨酸, 转录组分析

Abstract:

Objective Powdery mildew, one of the most devastating diseases affecting wheat, poses a significant threat to wheat yield and quality. This study aims to investigate the regulatory effects of exogenous salicylic acid (SA) on Blumeria graminis f. sp. tritici (abbreviated as Bgt below) infection and to reveal the molecular mechanisms underlying Bgt infection and SA-mediated resistance to disease through transcriptomic analysis. Method In this experiment, we analyzed the transcriptome of wheat powdery mildew fungus under Bgt-infested conditions for 1-4 d using powdery mildew-susceptible common wheat Zhongzuo 9504 as the material. Concurrently, salicylic acid was applied externally to observe the effect of phytohormones on powdery mildew-infected wheat. Result Transcriptomic analysis revealed that, compared to 1 d, 399 genes were upregulated and 1 110 genes were downregulated at 4 d. The upregulated genes were enriched in pathways such as secondary metabolite biosynthesis, metabolic pathways, and proteasome, while the downregulated genes were primarily involved in the degradation of valine, leucine, and isoleucine. Protein-protein interaction network analysis identified three core hub genes of Bgt: BGTH12_LOCUS642, BGTH12_LOCUS3045, and BGTH12_LOCUS5497, indicating their crucial roles during wheat infection. Furthermore, exogenous SA significantly upregulated the expressions of six hub genes in wheat (ERF109, PP2C30, TIFY6B, HSP70, At4g15970, and HERK1), thereby reducing the damage caused by Bgt. Conclusion During the infection process, Bgt demonsttrates extensive differential gene expression, with BGTH12_LOCUS642, BGTH12_LOCUS3045, and BGTH12_LOCUS5497 identified as core hub genes. The external application of salicylic acid may induce the expressions of resistance genes and inhibit the growth and development of conidia under the conditions of powdery mildew infection, thereby slowing down the infection of powdery mildew to a certain extent and increasing the resistance of wheat to powdery mildew.

Key words: wheat powdery mildew, Blumeria graminis f. sp. tritici, exogenous salicylic acid, transcriptome analysis

李成花, 豆飞飞, 任毓昭, 刘彩霞, 刘凤楼, 王掌军, 李清峰. 外施水杨酸对白粉菌侵染小麦的影响及白粉菌转录组分析[J]. 生物技术通报, 2025, 41(7): 272-280.

LI Cheng-hua, DOU Fei-fei, REN Yu-zhao, LIU Cai-xia, LIU Feng-lou, WANG Zhang-jun, LI Qing-feng. Effect of Exogenous Salicylic Acid on Wheat Infested with Blumeria graminis f. sp. tritici and Its Transcriptome Analysis[J]. Biotechnology Bulletin, 2025, 41(7): 272-280.

图/表 8

表1 实时荧光定量PCR引物

Table 1 Real-time fluorescence quantitative PCR Primers

基因名称 Gene name	基因ID Gene ID	正向引物 Forward primer （5′-3′）	反向引物 Reverse primer （5′-3′）
ERF109	TraesCS1A02G370700.1	AGGGGCACGAGTACATGATC	AGCTTCATCAGGTCCTGCAG
T7H20-70	TraesCS1D02G280200.2	TGGACGCCAAGAAGAGGTC	CTTGCCACCAATCACCTCTG
CIGR2	TraesCS2A02G189600.1	ATCCACCCCTTCAGCAACAT	GAGGGGCTTGCTCTTCCAG
HSP70	TraesCS4B02G205700.1	TCGGCACCACATACTCCTG	CAGTCCACTTCTTCGATCTTGG
PP2C30	TraesCS4B02G210100.1	ATGTCAGAGATCCGCCGC	GTTGTCCGAGCTCTGCCG
TIFY6B	TraesCS5A02G204900.1	GGAGTCAGCTTACTTTGGGG	CTAAAGCTGATGTTCCTGGGC
DUF4228	TraesCS5D02G414000.1	ACGGCGAGGGTGGTTCTC	GAGATGGCGGTTAGGTCGG
At4g15970	TraesCS5D02G536500.1	CAGCAGCAACATTAGCCCC	CACGTCCAACACATTCCTCA
tmem53	TraesCS7B02G502200.1	AAACTCTCCCACCACCACAG	CGCAGTCCTCCAAGAAGTTG
HERK1	TraesCS7D02G338800.3	CTCTACTCCCCTTTCCGCTG	TCACCTCCCCTCAGCTTTG
FLA17	TraesCS2B02G263900.1	TCCTCCTCTGCCTGGTACT	CCTTCCAACAACTTCCCGC
Actin	-	TACTCCCTCACAACAACCG	AACAAGAGTCACCTCCAAGA

图1 1-4 d的白粉菌转录组分析A：样本主成分分析；B：韦恩图；C：差异基因数目

Fig. 1 Transcriptome analysis of powdery mildew from 1-4 dA: Principal component analysis for samples. B: Venn plot. C: Histogram of number of DEGs

图2 1-4 d时白粉菌中DEGS的GO富集分析

Fig. 2 GO enrichment analysis of DEGS inBgt at 1-4 d

图3 1-4 d时白粉菌中DEGS的KEGG富集分析

Fig. 3 KEGG enrichment analysis of DEGS in Bgtat 1-4 d

图4 上调差异基因（A）和下调差异基因（B）的KEGG富集气泡图

Fig. 4 Bubble plot of KEGG enrichment for up-regulated DEGs (A) and down-regulated DEGs (B)

图5 蛋白质-蛋白质互作网络图

Fig. 5 Protein-protein interaction network diagram

图6 外源水杨酸诱导后接种白粉菌时关键激素基因表达情况不同小写字母代表在P<0.05水平上差异显著

Fig. 6 Expressions of key hormone genes during inoculation of Bgt after induction by exogenous salicylic acidDifferent letters indicate significant difference at P<0.05 level

图7 显微镜下外源水杨酸对白粉菌生长状态的影响A：白粉菌正常生长状态；B：施加外源水杨酸后白粉菌生长状态

Fig. 7 Effect of exogenous salicylic acid on the growth state of Bgt under microscopeA: The normal growth state of Bgt. B: The growth state of Bgt after the application of exogenous salicylic acid

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