Sequence Analysis and Induced Expression Analysis of GmGST Gene Cluster Genes in Soybean

doi:10.13560/j.cnki.biotech.bull.1985.2024-0980

Abstract

Abstract:

Objective To investigate the gene structure and expression characteristics of GmGST, laying the foundation for studying its function in soybean resistance to soybean mosaic virus. Method The gene GmGSTU12 (Glyma.07G139700), GmGSTU13 (Glyma.07G139800), GmGSTU16 (Glyma.07G140100), and GmGSTU47 (Glyma.07G140200) were cloned from the resistant material X149 using RT-PCR. The method of bioinformatics was used to analyze GmGST bioinformatics. RT-qPCR was used to analyze the expressions of GmGST genes induced by SC15, hormones (MeJA, ABA, ETH and SA), H₂O₂and tissue-specific expression in the resistant material X149, and a UV spectrophotometer was used to detect the activities of glutathione S- transferase (GSTs). Result The results of gene amplification showed that the lengths of the open reading frames of GmGSTU12, GmGSTU13, GmGSTU16 and GmGSTU47 were 678, 678, 952, and 669 bp, respectively. Bioinformatics analysis showed that the four genes belonged to the Tau family of GSTs, which were cytoplasmic, cytoplasmic, chloroplast and nuclear proteins without signal peptides, among which GmGSTU12, GmGSTU13 and GmGSTU47 were stable hydrophilic proteins, and GmGSTU16 were hydrophobic proteins, all of which were non-transmembrane structural proteins. RT-qPCR analysis showed that the expression of GmGSTU13 was the highest in the leaves, and the expressions of GmGSTU12,GmGSTU16 and GmGSTU47 in the roots were the highest. All four genes were up-regulated by SC15 and H₂O₂. In addition, GmGSTU12 and GmGSTU13 were mainly expressed in response to MeJA induction. GmGSTU16 was mainly expressed in response to the induction of both MeJA and SA. GmGSTU47 was mainly expressed in response to the induction of both MeJA and ABA. The dectection of GSTs activity showed that GSTs activity increased significantly at 12 h after SC15 induction and reached its maximum at 48 h. Conclusion All four GmGST genes belong to the Tau class of the GST family and can be induced by SC15, exogenous hormones (MeJA, ABA, ETH, SA), and H₂O₂.

Key words: soybean, soybean mosaic virus, GmGST, gene amplification, bioinformatics analysis, expression analysis

ZHAO Jing, GUO Qian, LI Rui-qi, LEI Ying-yang, YUE Ai-qin, ZHAO Jin-zhong, YIN Cong-cong, DU Wei-jun, NIU Jing-ping. Sequence Analysis and Induced Expression Analysis of GmGST Gene Cluster Genes in Soybean[J]. Biotechnology Bulletin, 2025, 41(5): 129-140.

Figures/Tables 17

References 54

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基因 Gene	正向引物 Forward primer （5′‒3′）	反向引物 Reverse primer （5′‒3′）	退火温度 Annealing temperature （℃）
GmGSTU12	TTACCAGCTATACTTGTTCC	ATTCAAACAGCAATAACTCAA	55
GmGSTU13	GGTTAGAAGTGCTACAATACAAA	AATTCAAACAGCAGTAACTCAAC	56
GmGSTU16	ACCTTAGGACCCATCACAGAC	GATTGTGTGAACTACTTGTCG	58
GmGSTU47	AGTCAAGAGAAGAAGTGAAGTG	AGGTTGTTGACATTTTCGCT	55

基因 Gene	正向引物 Forward primer （5′‒3′）	反向引物 Reverse primer （5′‒3′）	退火温度 Annealing temperature （℃）
GmGSTU12	TTACCAGCTATACTTGTTCC	ATTCAAACAGCAATAACTCAA	55
GmGSTU13	GGTTAGAAGTGCTACAATACAAA	AATTCAAACAGCAGTAACTCAAC	56
GmGSTU16	ACCTTAGGACCCATCACAGAC	GATTGTGTGAACTACTTGTCG	58
GmGSTU47	AGTCAAGAGAAGAAGTGAAGTG	AGGTTGTTGACATTTTCGCT	55

软件名称 Software name	软件网址 Software URL
SignalP4.1	http：//www.cbs.dtu.dk/services /SignalP-4.1/
PlantCARE	http：//bioinformatics.psb.ugent.be/webtools/plantcare/html/
ProtParam	http：//web.expasy.org/protparam/
SOPMA	https：//npsaprabi.ibcp. fr/cgibin/npsa_automat.pl？page=/NPSA/npsa_gor4.html
Phyre2	http：//www.sbg.bio.ic.ac.uk/phyre2/html
Evolview	http：//www.omicsclass.com/article/671
MEME	http：//meme suite.org/
GSDS2.0	http：//gsds.cbi.pku.edu.cn/
TMHMM-2.0	http：//www.cbs.dtu.dk/services/TMHMM/
WoLF PSORT	https：//wolfpsort.hgc.jp/
Pfam	https：//www.ebi.ac.uk/interpro/entry/pfam

软件名称 Software name	软件网址 Software URL
SignalP4.1	http：//www.cbs.dtu.dk/services /SignalP-4.1/
PlantCARE	http：//bioinformatics.psb.ugent.be/webtools/plantcare/html/
ProtParam	http：//web.expasy.org/protparam/
SOPMA	https：//npsaprabi.ibcp. fr/cgibin/npsa_automat.pl？page=/NPSA/npsa_gor4.html
Phyre2	http：//www.sbg.bio.ic.ac.uk/phyre2/html
Evolview	http：//www.omicsclass.com/article/671
MEME	http：//meme suite.org/
GSDS2.0	http：//gsds.cbi.pku.edu.cn/
TMHMM-2.0	http：//www.cbs.dtu.dk/services/TMHMM/
WoLF PSORT	https：//wolfpsort.hgc.jp/
Pfam	https：//www.ebi.ac.uk/interpro/entry/pfam

基因 Gene	正向引物 Forward primer （5′‒3′）	反向引物 Reverse primer （5′‒3′）
GmTublin	GGAGTTCACAGAGGCAGAG	CACTTACGCATCACATAGC
GmGSTU12	TGTGTGCAGGGTCCAGATTG	GCTATGGGCTGCTCATTGTG
GmGSTU13	GCGAAGAATCCCTTAACCACC	AGCGGGCTTTGAAGAAGGTA
GmGSTU16	GGGTCCAGATTGCCCTCAAA	ACGGGGTTGGATTTGAGAAGT
GmGSTU47	CAGGGTGCAGATTGCTCTCA	CAAGGGACTCGGATATGGGC