Cloning and Expression Pattern Analysis of Brassinosteriod Signal Response Transcription Factors GmBEE1 and GmBEE3 in Soybean

doi:10.13560/j.cnki.biotech.bull.1985.2025-0381

Abstract

Abstract:

Objective This study aims to explore the biological functions of soybean transcription factors GmBEE1a and GmBEE3 involving in brassinosteroid signaling, and provides a scientific basis for the discovery of soybean disease resistance and high-yield gene resources. Method The soybean variety Nandou 12 (Glycine max cv. Nandou 12) was used as the material, bioinformatics methods were employed to analyze GmBEEs' gene structure, physicochemical properties of the encoded proteins, and their spatial conformations, the gene coding sequences (CDS) of GmBEE1a and GmBEE3 were cloned, and the subcellular localizations of the proteins were observed using the tobacco transient expression system. Real-time quantitative PCR was used to analyze the tissue-specific expressions of the two GmBEEs genes and their expression characteristics under different treatments. Result Both GmBEE1a and GmBEE3 in soybean contained six exons and five introns, and identified as the typical bHLH transcription factor. Their encoded proteins were acidic and unstable. Phylogenetic analysis revealed that GmBEE1a and GmBEE3 proteins were closely related to homologous proteins in Arabidopsis thaliana and tobacco (Nicotinan tabacum). Promoter cis-acting element analysis showed that the promoter regions of both genes contained functional elements related to light response, stress response, and hormone regulation. Both Bioinformatics predictions and tobacco transient expression experiments indicated that GmBEE1a and GmBEE3 proteins were localized in the nucleus. Tissue-specific expression analysis revealed that the expressions of GmBEE1a and GmBEE3 in soybean flowers and leaves were higher than those in the seeds, roots, stems, and pods. Treatment with the pathogen strain Fusarium oxysporum B3S1 and the bacterial flagellin peptide flg22 significantly induced the expressions of both GmBEEs, whereas treatment with rhizosphere beneficial bacterial strain Pseudomonas chlororaphis IRHB3 suppressed their expression. Additionally, exogenous treatments with salicylic acid (SA), methyl jasmonate (MeJA), and brassinolide (BL) transiently and significantly induced the expressions of GmBEE1a and GmBEE3, followed by a reduction in expression. Conclusion The soybean bHLH transcription factors GmBEE1a and GmBEE3 not only participate in regulating soybean growth and development but also respond to microbial colonization and exogenous hormone treatments.

Key words: soybean, transcription factor bHLH, GmBEE gene, expression pattern

HE Wen-xin, CHEN Ling, HUANG Ling-lin, XU Yan, SUN Zu-dong, ZENG Wei-ying, CHANG Xiao-li, YANG Feng. Cloning and Expression Pattern Analysis of Brassinosteriod Signal Response Transcription Factors GmBEE1 and GmBEE3 in Soybean[J]. Biotechnology Bulletin, 2025, 41(11): 261-271.

Figures/Tables 11

References 32

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引物名称 Primer name	上游引物 Forward primer （5′‒3′）	下游引物 Reverse primer （5′‒3′）
GmBEE1a-CDS	ATGGCAGAATTCACTGCAGATTT	CAGGGGCCATGACCATGTT
GmBEE3-CDS	ATGGCTGAATTCACTGCAGATTT	CAGAGGCCATGGCCATGTT
qPCR-GmBEE1a	CAGAACATTGTCCCAGGATG	ATTCTTCTCTCACATACCTGTC
qPCR-GmBEE3	CCGGCTCTTGTTCAATGTGTC	TAGCTTGACCACGTTTGGCT
eYFP	GAAGATGCCTCTGCCGACAG	AACTTCAGGGTCAGCTTGCCG
GmActin	GGTGGTTCTATCTTGGCATC	CTTTCGCTTCAATAACCCTA

引物名称 Primer name	上游引物 Forward primer （5′‒3′）	下游引物 Reverse primer （5′‒3′）
GmBEE1a-CDS	ATGGCAGAATTCACTGCAGATTT	CAGGGGCCATGACCATGTT
GmBEE3-CDS	ATGGCTGAATTCACTGCAGATTT	CAGAGGCCATGGCCATGTT
qPCR-GmBEE1a	CAGAACATTGTCCCAGGATG	ATTCTTCTCTCACATACCTGTC
qPCR-GmBEE3	CCGGCTCTTGTTCAATGTGTC	TAGCTTGACCACGTTTGGCT
eYFP	GAAGATGCCTCTGCCGACAG	AACTTCAGGGTCAGCTTGCCG
GmActin	GGTGGTTCTATCTTGGCATC	CTTTCGCTTCAATAACCCTA

基因名 Gene name	基因ID Gene ID	定位标记 Locus tag	外显子个数 Number of exons	内含子个数 Number of introns
GmBEE3	100777160	Glyma08g042000	6	5
GmBEE1a	100810945	Glyma05g234500	6	5
GmBEE1b	100806942	Glyma09g183500	6	5
GmBEE1c	100807773	Glyma14g102200	6	5
GmBEE1d	100809974	Glyma17g223100	6	5
GmBEE1e	100803700	Glyma07g092700	6	5

基因名 Gene name	基因ID Gene ID	定位标记 Locus tag	外显子个数 Number of exons	内含子个数 Number of introns
GmBEE3	100777160	Glyma08g042000	6	5
GmBEE1a	100810945	Glyma05g234500	6	5
GmBEE1b	100806942	Glyma09g183500	6	5
GmBEE1c	100807773	Glyma14g102200	6	5
GmBEE1d	100809974	Glyma17g223100	6	5
GmBEE1e	100803700	Glyma07g092700	6	5

基因名称 Gene name	基因ID Gene ID	氨基酸数 Number of amino acids	分子式 Formula	分子量 Molecular weight （Da）	等电点 pI
GmBEE1a	100810945	246	C₁₂₂₂H₁₉₂₆N₃₄₄O₃₈₁S₁₇	28 077.83	5.40
GmBEE1b	100806942	273	C₁₃₄₆H₂₁₂₆N₃₇₈O₄₁₆S₁₂	30 644.69	6.10
GmBEE1c	100807773	264	C₁₃₃₁H₂₀₇₅N₃₆₃O₄₀₈S₁₁	30 042.96	5.56
GmBEE1d	100809974	268	C₁₃₄₃H₂₀₈₆N₃₇₂O₄₁₉S₁₁	30 500.24	5.43
GmBEE1e	100803700	272	C₁₃₃₉H₂₁₁₅N₃₇₉O₄₁₃S₁₄	30 579.65	6.16
GmBEE3	100777160	252	C₁₂₂₆H₁₉₅₄N₃₄₆O₃₈₇S₁₆	28 246.05	5.53