Cloning and Biology Function Analysis of Soybean GmWRKY44

doi:10.7668/hbnxb.20195201

Abstract

Abstract:

WRKY is a unique class of transcription factors in plants,which plays an important role in plant abiotic stress response,seed dormancy and germination,growth and development,etc.In order to reveal the function and underlying molecular mechanism of GmWRKY44 gene in soybean WRKY transcription factor family,bioinformatics analysis and biology function verification of soybean Williams 82 GmWRKY44 were performed.GmWRKY44 gene was 1 077 bp in length and encoded 358 amino acids;the results of structural prediction and evolutionary analysis showed that,the secondary structure was composed of 23.46% α-helix,4.75% β-fold,58.94% irregular coil and 12.85% extended chain,and the tertiary structure was unified with the secondary structure;It contained a conserved WRKY domain, the zinc finger structure was of the C₂H₂ type, and it belonged to the WRKY IIc subfamily; GmWRKY44 is a homologous gene of Arabidopsis thaliana AtWRKY71 with a similarity of 35.56%, and the two genes had similar gene structures. RT-qPCR analysis showed that GmWRKY44 responded to salt stress and its expression level first decreased and then increased.Under salt stress,the germination rate and root length of wild-type(Col-0)and GmWRKY44 overexpressing Arabidopsis lines were inhibited to a certain extent,but GmWRKY44 overexpressing lines were significantly better than Col-0.In addition,under salt stress,the growth inhibition of GmWRKY44 overexpressing lines was lower than that of Col-0.Physiological index analysis revealed that under salt stress,the overexpression lines of GmWRKY44 exhibited significantly higher activities of superoxide dismutase(SOD),peroxidase(POD),and catalase(CAT)than Col-0,while the content of malondialdehyde(MDA)was significantly lower than Col-0.These data indicated that overexpression of GmWRKY44 could improve salt tolerance in transgenic Arabidopsis.

Key words: Soybean, GmWRKY44, Gene cloning, Bioinformatics analysis, Function verification, Salt stress

HUANG Youju, YU Yongbo, PANG Cuijing, SUN Shixu, LU Chen, YU Yanchong. Cloning and Biology Function Analysis of Soybean GmWRKY44[J]. Acta Agriculturae Boreali-Sinica, 2025, 40(1): 29-35. doi: 10.7668/hbnxb.20195201.

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Figures/Tables 8

Tab.1 The primers used for RT-qPCR

引物名称 Primer name	序列(5'—3') Sequence(5'—3')
GmWRKY44 (Glyma.05G127600)-real-F	CACCCTCACCACCAATATCACA
GmWRKY44 (Glyma.05G127600)-real-R	TCCGGAGGAAGAAACTGGTTG
GmActin (Glyma.02G091900)-real-F	GCGGGAAATTGTAAGGGATGT
GmActin (Glyma.02G091900)-real-R	TCGCCAATAGTGATGACCTG

Fig.1 Physicochemical properties of GmWRKY44 A.Hydrophilic and hydrophobic analysis of GmWRKY44; B.Prediction of the transmembrane structure of GmWRKY44.

Fig.2 Molecular structure prediction of GmWRKY44 A.Secondary structure components of GmWRKY44;B.Prediction of secondary structure of GmWRKY44; C.Prediction of tertiary structure of GmWRKY44;D.GmWRKY44 domain prediction.

Fig.3 Homology analysis of the WRKY IIc family of soybean and Arabidopsis A.Evolutionary tree analysis;B.Conserved motif analysis;C.Domain analysis;D.Gene structure analysis.

Fig.4 Relative expression of GmWRKY44 in soybean seedlings treated with NaCl at 0—24 h Different capital letters represent a extremely significant difference in P<0.01.

Fig.5 Analysis of germination rate and root length of Arabidopsis thaliana under salt stress A.Arabidopsis germination status after 7 days of NaCl treatment;B.Germination rate of Arabidopsis under normal conditions; C.Germination rate of Arabidopsis under NaCl treatment; D.Arabidopsis root growth after 7 days of NaCl treatment; E.Arabidopsis root length after 7 days of NaCl treatment.Different lowercase letters represent a significant difference in P<0.05.The same as Fig.7.

Fig.6 Phenotypic observation of soil-cultured Arabidopsis thaliana under salt stress for 7 and 14 days

Fig.7 Physiological indicators of stress resistance of Arabidopsis thaliana leaves under salt stress

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