棉花黄萎病菌β-葡萄糖苷酶基因的鉴定及其在致病中的功能

doi:10.3864/j.issn.0578-1752.2026.07.002

Abstract

Abstract:

【Objective】Verticillium dahliae is a major soil-borne pathogenic fungus that can infect more than 200 plant species, causing Verticillium wilt and leading to severe economic losses in global agricultural production. Cell wall-degrading enzymes (CWDEs) play a critical role in fungal pathogenesis. This study aimed to identify β-glucosidase genes in V. dahliae and explore their functions, thereby providing new molecular targets for cotton disease-resistant breeding.【Method】β-glucosidase genes were identified from the whole genome of V. dahliae through bioinformatic analysis, and their evolutionary relationships, conserved domains, and expression patterns were systematically analyzed. Host-induced gene silencing (HIGS) technology was used to silence Vdbg4 and Vdbg6 in cotton to evaluate the effects of this silencing on cotton disease resistance. Artificial small interfering RNAs (asiRNAs) targeting Vdbg4 (asiR1364) and Vdbg6 (asiR1444) were designed and co-cultured with V. dahliae, respectively. The growth and development, carbon source utilization and stress response abilities, and pathogenicity of V. dahliae in co-culture were investigated. The secretory activity of Vdbg6 was verified using a yeast signal peptide trapping assay, and whether the gene could trigger plant immune responses was detected using an Agrobacterium-mediated tobacco transient expression assay.【Result】A total of 18 β-glucosidase genes were identified from V. dahliae. Among them, the expression levels of Vdbg4 and Vdbg6 were significantly up-regulated after induction by root exudates of a susceptible cotton cultivar. Silencing Vdbg4, Vdbg6 alone, or silencing both genes simultaneously via HIGS, significantly alleviated the disease symptoms of cotton, reduced the disease index, and decreased the fungal biomass. The asiRNAs targeting Vdbg4 and Vdbg6 (asiR1364, asiR1444, and asiR1364+1444) could inhibit the fungal colony and mycelial growth, reduce the sporulation and spore germination rates, and impair the carbon source utilization and stress response abilities, and pathogenicity of V. dahliae. Vdbg6 exhibited secretory activity, but it could neither induce programmed cell death (PCD) nor suppress BAX-induced PCD in tobacco cells.【Conclusion】Both Vdbg4 and Vdbg6 are involved in the growth and development, carbon source utilization, stress response, and pathogenic processes of V. dahliae.

Key words: cotton, Verticillium dahliae, cell wall-degrading enzymes, β-glucosidase, host-induced gene silencing (HIGS), artificial small interfering RNA (asiRNA)

LI YuanJing, YUAN RuiXiang, LI YongTai, SUN TianGe, LIU Feng, LI YanJun, ZHANG XinYu. Identification and Functional Characterization of β-Glucosidase Genes in Verticillium dahliae for Pathogenicity on Cotton[J].Scientia Agricultura Sinica, 2026, 59(7): 1380-1399.

Figures/Tables 10

Table 1

Table 2

Identification of beta-glucosidase genes in V. dahliae"

基因ID ID gene	基因名称 Gene name	CDS长度 CDS length (bp)	蛋白长度 Protein length (aa)	染色体Chromosome	分子量 Molecular weight (kDa)	理论等电点Isoelectric point	跨膜结构域数量Transmembrane domain number	亚细胞定位预测Subcellular localization prediction	基因描述 Gene description
VDAG_00134	Vdbg1	2878	813	2	87.54	6.34	1	质膜 Plasma membrane	β-葡萄糖苷酶Beta-glucosidase
VDAG_01062	Vdbg2	3076	897	1	98.58	5.41	0	细胞质 Cytosol	耐热β-葡萄糖苷酶Thermostable Beta-glucosidase
VDAG_01263	Vdbg3	1886	611	1	66.56	6.28	0	细胞质 Cytosol	β-葡萄糖苷酶Beta-glucosidase
VDAG_02542	Vdbg4	2796	798	3	86.19	5.67	0	线粒体Mitochondrion	耐热β-葡萄糖苷酶Thermostable Beta-glucosidase
VDAG_05580	Vdbg5	2489	774	2	85.27	4.98	0	细胞质 Cytosol	β-葡萄糖苷酶Beta-glucosidase
VDAG_06279	Vdbg6	2758	683	8	74.29	5.54	0	细胞外 Extracellular	β-葡萄糖苷酶Beta-glucosidase
VDAG_08139	Vdbg7	2500	816	6	87.91	4.69	0	细胞外 Extracellular	β-葡萄糖苷酶Beta-glucosidase
VDAG_09280	Vdbg8	2605	823	4	89.29	5.93	0	细胞质 Cytosol	耐热β-葡萄糖苷酶Thermostable Beta-glucosidase
VDAG_09750	Vdbg9	3169	876	3	94.85	4.78	0	细胞外 Extracellular	β-葡萄糖苷酶Beta-glucosidase
VDAG_09393	Vdbg10	1943	609	4	67.13	4.88	0	细胞外 Extracellular	周质β-葡萄糖苷酶Periplasmic Beta-glucosidase
VDAG_05611	Vdbg11	2676	833	2	91.40	5.61	0	细胞质 Cytosol	β-葡萄糖苷酶Beta-glucosidase
VDAG_03973	Vdbg12	2598	803	3	104.64	5.51	0	细胞外 Extracellular	β-葡萄糖苷酶Beta-glucosidase
VDAG_02717	Vdbg13	2942	955	3	86.67	5.24	1	细胞质 Cytosol	β-葡萄糖苷酶Beta-glucosidase
VDAG_08263	Vdbg14	2313	746	6	78.45	4.77	0	细胞外 Extracellular	耐热β-葡萄糖苷酶Thermostable Beta-glucosidase
VDAG_08240	Vdbg15	2632	836	6	89.78	5.41	0	细胞质 Cytosol	耐热β-葡萄糖苷酶Thermostable Beta-glucosidase
VDAG_06333	Vdbg16	1995	476	8	53.78	4.87	0	细胞质 Cytosol	β-葡萄糖苷酶Beta-glucosidase
VDAG_02175	Vdbg17	2420	781	7	84.91	5.39	0	细胞外 Extracellular	β-葡萄糖苷酶Beta-glucosidase
VDAG_03143	Vdbg18	648	215	6	23.10	4.88	0	细胞质 Cytosol	β-葡萄糖苷酶Beta-glucosidase

Table 2

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基因 Gene	asiRNA	正义链 Sense strand (5′-3′)	反义链 Antisense strand (5′-3′)
Vdbg4	asiR1364	GGCUGAAGAUCACAACCAATT	UUGGUUGUGAUCUUCAGCCTT
Vdbg6	asiR1444	CGAUUGUGGUCUUCCAUAATT	UUAUGGAAGACCACAAUCGTT

Identification and Functional Characterization of β-Glucosidase Genes in Verticillium dahliae for Pathogenicity on Cotton

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