Cloning and Functional Analysis of the Cotton GhMYB113 Gene in Verticillium Wilt Resistance

doi:10.7668/hbnxb.20196180

Abstract

Abstract:

This study investigated the function of the cotton gene GhMYB113 in resistance to Verticillium wilt.GhMYB113 encoded a 249 amino acid protein with a molecular weight of 29.07 ku and an isoelectric point of 6.46.Phylogenetic and subcellular localization analyses revealed that GhMYB113 shared the highest homology with soybean GmMYB113 and was localized to the nucleus.Expression profiling by qPCR showed that GhMYB113 was predominantly expressed in roots and was upregulated upon infection by Verticillium dahliae as well as by treatment with jasmonic acid(JA)and salicylic acid(SA),suggesting a role in disease resistance signaling.Silencing of GhMYB113 via virus-induced gene silencing(VIGS)led to a significant decrease in leaf anthocyanin accumulation and downregulated the expression of key anthocyanin biosynthesis genes,indicating that GhMYB113 positively regulates anthocyanin synthesis in cotton.This was further confirmed using a GUS reporter system,which demonstrated that GhMYB113 activates the promoter of GhDFR.Inoculation assays performed on GhMYB113-silenced plants resulted in more severe disease symptoms,higher disease index(DI)values,increased disease incidence,enhanced stem browning,and significantly greater fungal biomass in stems compared with control plants,demonstrating that GhMYB113 positively regulates cotton resistance to Verticillium wilt.In summary,GhMYB113 plays a critical role in both anthocyanin biosynthesis and Verticillium wilt resistance in cotton,providing a theoretical foundation for breeding disease-resistant cotton varieties.

Key words: Cotton, MYB113, Verticillium dahliae, Verticillium wilt, Defense, Anthocyanin

CLC Number:

ZHONG Guimai, LIAO Chanjuan, LI Dan, WU Qi, HU Guang. Cloning and Functional Analysis of the Cotton GhMYB113 Gene in Verticillium Wilt Resistance[J]. Acta Agriculturae Boreali-Sinica, 2026, 41(4): 215-221. doi: 10.7668/hbnxb.20196180.

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http://www.hbnxb.net/EN/Y2026/V41/I4/215

Figures/Tables 6

Tab.1 The primer sequences used in this study

引物 Primer	序列(5'—3') Sequence (5'—3')	用途 Employ
MYB113-GFP-F	CGAGATCTGATGGAGGGCTCATCTTTAAGAG	PCR
MYB113-GFP-R	CGCACTAGTTGGGTTAAACACATTCCACAG	PCR
qMYB113-F	TTGAGAAATATGGTGAAGGGAA	qPCR
qMYB113-R	GGAGGTTATGGAGGCGAATA	qPCR
GhUBI3-F	GAAGGCATTCCACCTGACCAAC	qPCR
GhUBI3-R	CTTGACCTTCTTCTTCTTGTGCTTG	qPCR
GhCHI-F	TGATGCAGAAGCAAAAGCCAT	qPCR
GhCHI-R	AGCTCAGTACCAGAAGGTGGGA	qPCR
GhF3H-F	GGGCCTAGCTTGCAAGCTTCTT	qPCR
GhF3H-R	AAGCAAGAGTGTGATGGTGCCTG	qPCR
GhDFR-F	GACCCTGGCAACTCGAAGAA	qPCR
GhDFR-R	GGACTCGAAGTCCATAGGCG	qPCR
GhANS-F	CCGCAGCTGTGTCTATCAACAC	qPCR
GhANS-R	AGCAAATTTCCAAGCTGCCTT	qPCR
GhANR-F	GAGGCCTAGCGAGCAAAATAC	qPCR
GhANR-R	GTGAGCTTCGACACCGAGAG	qPCR
GhUFGT-F	TCTTCTCCGCATCCCAACTA	qPCR
GhUFGT-R	TCCATTGCCTTCCTCAGATT	qPCR
vMYB113-F	CGGAATTCTGGAGGGCTCATCTTTAAG	VIGS
vMYB113-R	CGGGATCCTGAACTGCCACTTGCACTGATAC	VIGS
VdTUB1-F	AACAACAGTCCGATGGATAATTC	qPCR
VdTUB1-R	GTACCGGGCTCGAGATCG	qPCR
GhACT7-F	TTGCAGACCGTATGAGCAAG	qPCR
GhACT7-R	ATCCTCCGATCCAGACACTG	qPCR
DFRpro-F	CGAAGCTTGATTGTTCATACAGGGACATGACCGAC	PCR
DFRpro-R	CCGGGATCCTGGGAAATAACTTGAACACTCTGCTC	PCR

Fig.1 Phylogenetic analysis and subcellular localization of GhMYB113 A.Phylogenetic analysis of cotton MYB transcription factors;B.Phylogenetic analysis of GhMYB113 alongside MYB113 from various other plant species;C.Construction of the vector for subcellular localization;D.Fluorescence observation of GhMYB113-GFP fusion.H2B-RFP transgenic N.benthamiana plants were used as a nucleus marker.Scale bars.30 μm.

Fig.2 GhMYB113 expression profiles Different lowercase letters denoted significant differences(P<0.05),based on the Tukey HSD test.The same as Fig.3—5.

Fig.3 GhMYB113 regulates anthocyanin synthesis A.Construction of the GhMYB113-TRV silencing vector;B.Representative diagram of TRV:PDS plants;C.Relative expression of GhMYB113 in silenced plants;D,E.Anthocyanin accumulation in GhMYB113-silenced and control plants;F.The expression levels of anthocyanin synthesis pathway genes.

Fig.4 GhMYB113 promotes GhDFR transcriptional expression A.Diagram illustrating the construction of effector and reporter vectors;B.GUS staining in tobacco leaves;C.Enzyme activity assays in tobacco leaves.

Fig.5 Resistance analysis of GhMYB113-silenced plants to Verticillium wilt A.Representative images of disease symptoms on GhMYB113-silenced and control plants inoculated with V.dahliae;B,C.Disease index(DI)and the rate of diseased plants of GhMYB113-silencing and control plants;D.Representative images of the longitudinal sections of cotton stems infested with V.dahliae;E.Fungal recovery assay;F.Fungal biomass was quantified as the ratio of V.dahlia TUB1 gene to cotton ACT7 gene DNA levels.

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