Cloning and Functional Analysis of Candidate Gene for SMV Resistance Locus qTsmv-3 in Soybean

doi:10.7668/hbnxb.20195741

Abstract

Abstract:

Soybean mosaic virus(SMV)disease can cause significant yield losses and quality deterioration in soybeans,and breeding resistant cultivars remains the only effective strategy for SMV control.Identifying the functional genes associated with SMV resistance provides essential genetic resources for developing resistant varieties.Six MATE candidate genes involved in SMV resistance were identified using a near-isogenic line(NIL)of the qTsmv-3 locus and a transgenic Arabidopsis thaliana plant.A total of 128 MATE family genes were predicted in the soybean genome,which were classified into five subfamilies.Notably,all six MATE candidate genes located at the qTsmv-3 locus clustered within subfamily Ⅰ,exhibiting significant differences in expression levels and tissue specificity based on public data.Among them,Glyma.03G005600 showed the highest expression in aerial tissues(leaves and stems),while Glyma.03G005800 was predominantly expressed in underground tissues(roots and nodules).Following SMV inoculation,the resistant NIL(#NIL-NC)exhibited a 70% reduction in viral accumulation compared with the susceptible line(#NIL-SMC).Concurrently,the expression levels of GmICS1 and GmPR1,key genes in the salicylic acid(SA)-mediated defense pathway,were upregulated by 2.40,15.16 folds,respectively,in #NIL-NC,indicating that qTsmv-3 confers resistance through SA-dependent signaling.Among the 6 MATE candidate genes,only Glyma.03G005300 and Glyma.03G005600 displayed significant differential expression between NILs,which were down-regulated by 61.0% and 82.1%,respectively.Considering their expression patterns and responses to SMV infection,Glyma.03G005600 was identified as the most promising candidate gene for qTsmv-3.Further,the expression of GmICS1 and GmPR1 in transgenic Arabidopsis thaliana(OE_MATE),which carrying Glyma.03G005600,was significantly up-regulated by 4.22,9.12 folds compared with that of wild type(WT)after UV-B stress.These results strongly indicated that Glyma.03G005600 could significantly enhance or affect the expression of genes in salicylic acid signaling pathway,and preliminarily confirmed that Glyma.03G005600 was a key regulatory gene for qTsmv-3 locus.In all,the results laid a foundation for cloning the key genes regulating SMV resistance and provided gene resources for genetic improvement of SMV resistance in soybean.

Key words: Soybean mosaic virus, Resistance candidate gene, MATE, qTsmv-3, Function analysis

CLC Number:

S435.651

LIN Jing, SHI Xiaolei, XU Junjie, YU Cuihong, CAO Zhimin, TANG Xiaodong, YANG Chunyan, ZHANG Mengchen, YAN Long. Cloning and Functional Analysis of Candidate Gene for SMV Resistance Locus qTsmv-3 in Soybean[J]. Acta Agriculturae Boreali-Sinica, 2025, 40(4): 57-64. doi: 10.7668/hbnxb.20195741.

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Figures/Tables 6

Tab.1 RT-qPCR primers for gene expression analysis

基因编号 Gene ID	前引物(5'-3') Forward primer(5'-3')	后引物(5'-3') Revers primer(5'-3')
Glyma.03G005600	TTTCACTTGCCAGAATTTCCTG	CAGGAATCTCGAGCTTGAATTG
Glyma.03G005200	CAGAAGTTGCTGAGCAGAGAGA	TCCACATCACCTTGCTCTCTTC
Glyma.03G005300	AGCACTGTAGCGTATGTGAACA	CGGGCTATAATCACCTGCTCAT
Glyma.03G005400	CACTGTCATCATACGTTTCGCC	CAGGAAGAAGACAGAGTGCAGT
Glyma.03G005500	TGACTCTCTTAGACCAGGATGA	GTCTGGGTAGTGAATGAGACAA
Glyma.03G005800	CGTTCCTGCAATCTCAAAGCAA	CAAAAGCCAGGAGAGAAACACG
GmActin	CGGTGGTTCTATCTTGGCATC	GTCTTTCGCTTCAATAACCCTA
SMV_CP	AATGGCGTGTGGGTGATGAT	GCCTTCATCTGCGCTATTGC
Glyma.15G062400(GmPR1)	GAGTCACGTTGCCAATGCTC	GCAGACACTCTCCACCAACA
Glyma.03G070600(GmISC1)	CCTTCCTATCCCCTTCACTTTT	TTGGTGTCGTAGAAATGAGTGA
AT1G74710(AtICS1)	TCTCAGGTACGAGCTTTTGTCC	AGAACCCCTTATCCCCCATACA
AT2G14610(AtPR1)	TAAGCGATGTTTACGAACCCCA	GAGTCTGCAGTTGCCTCTTAGT
AT4G20890(AtTub9)	CGGAGCTAAGTTCTGGGAAGTT	AGTTATTCCCAGCACCAGACTG

Fig.1 Phylogenetic and evolutionary analysis of MATE family genes in soybean

Fig.2 Tissue expression analysis of 6 MATE candidate genes

Fig.3 Analysis of SA-related gene expression in near-isogenic lines after SMV inoculation A.Symptoms of near-isogenic lines after SMV inoculation;B,C,D.Relative expression levels of GmICS1,GmPR1,and SMV among near-isogenic lines.Different lowercase letters indicate significant differences (P<0.05),the same as Fig.4-5.

Fig.4 Expression analysis of MATE candidate genes in near-isogenic lines

Fig.5 Functional analysis of Glyma.03G005600 in transgenic plant A.Growth comparison of transgenic Arabidopsis(OE_MATE)and wild-type(WT)under normal conditions;B.Response of OE_MATE and WT to UV-B treatment; C.Root length comparison between OE_MATE and WT;D.Expression analysis of the ICS1 gene in Arabidopsis;E.Expression analysis of the PR1 gene in Arabidopsis.

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