大豆转录因子GmDof1.5的克隆与非生物胁迫诱导表达

doi:10.3969/j.issn.1004-1524.2021.01.01

浙江农业学报 ›› 2021, Vol. 33 ›› Issue (1): 1-7.DOI: 10.3969/j.issn.1004-1524.2021.01.01

大豆转录因子GmDof1.5的克隆与非生物胁迫诱导表达

何佳琦¹(), 翟莹¹^,^*(), 张军², 邱爽¹, 李铭杨¹, 赵艳¹, 张梅娟¹, 马天意¹

1.齐齐哈尔大学生命科学与农林学院,抗性基因工程与寒地生物多样性保护黑龙江省重点实验室,黑龙江齐齐哈尔161006
2.黑龙江省农业科学院畜牧兽医分院,黑龙江齐齐哈尔161005

收稿日期:2020-03-18 出版日期:2021-01-25 发布日期:2021-01-25
通讯作者: 翟莹
作者简介:*翟莹,E-mail:fairy39809079@126.com
何佳琦(1995—),女,黑龙江齐齐哈尔人,硕士研究生,研究方向为大豆分子遗传育种。E-mail:he2918350891@163.com
基金资助:
黑龙江省普通本科高等学校青年创新人才培养计划(UNPYSCT-2017153);黑龙江省省属高等学校基本科研业务费科研项目(135209264);黑龙江省省属高等学校基本科研业务费科研项目(植物性食品加工技术特色学科专项)(YSTSXK201878);齐齐哈尔大学研究生创新科研项目(YJSCX2020041)

Cloning and expression analysis of GmDof1.5 in soybean under abiotic stress

HE Jiaqi¹(), ZHAI Ying¹^,^*(), ZHANG Jun², QIU Shuang¹, LI Mingyang¹, ZHAO Yan¹, ZHANG Meijuan¹, MA Tianyi¹

1. Heilongjiang Provincial Key Laboratory of Resistance Gene Engineering and Protection of Biodiversity in Cold Areas, College of Life Science and Agro-Forestry, Qiqihar University, Qiqihar 161006, China
2. Institute of Animal Husbandry, Heilongjiang Institute of Veterinary Science, Qiqihar 161005, China

Received:2020-03-18 Online:2021-01-25 Published:2021-01-25
Contact: ZHAI Ying

摘要/Abstract

摘要：

Dof转录因子在植物的生长发育和非生物胁迫响应中起重要的调控作用。利用RT-PCR技术从大豆中克隆了一个功能未知的Dof转录因子基因GmDof1.5,该基因位于大豆基因组15号染色体上,ORF全长540 bp,编码含有179个氨基酸的蛋白质,相对分子质量为20.15 ku,等电点为9.82。蛋白序列预测发现,GmDof1.5蛋白含有一个典型Zf-Dof结合域,且含有大量磷酸化位点。蛋白系统进化分析表明,大豆GmDof1.5与豆科植物鸡血藤SsDof4的亲缘关系最近。实时荧光定量PCR结果显示,ABA、干旱、高盐、高温和低温胁迫均可不同程度地诱导GmDof1.5的表达,且GmDof1.5对高温胁迫的响应最为显著。

关键词: 大豆, Dof转录因子, 非生物胁迫, 基因克隆, 表达分析

Abstract:

Dof transcription factors play an important regulatory role in plant growth and development and response to abiotic stress. A soybean Dof transcription factor GmDof1.5 with unknown function was cloned through RT-PCR, which was located on chromosome 15 of the soybean genome. The ORF of GmDof1.5 was 540 bp, which encoded 179 amino acid residues with molecular weight of 20.15 ku and isoelectric point of 9.82. Prediction of protein sequence showed that GmDof1.5 contained a typical Zf-Dof binding domain and a large number of phosphorylation sites. Phylogenetic analysis indicated that GmDof1.5 of soybean was highly homologous to SsDof4 of Spatholobus suberectus. Quantitative real-time PCR showed that the expressions of GmDof1.5 were induced by ABA, drought, salt, high temperature and cold stresses to different degrees. Especially, the response of GmDof1.5 to high temperature was the most obvious.

Key words: soybean, Dof transcription factor, abiotic stress, gene clone, expression analysis

中图分类号:

S565.1

何佳琦, 翟莹, 张军, 邱爽, 李铭杨, 赵艳, 张梅娟, 马天意. 大豆转录因子GmDof1.5的克隆与非生物胁迫诱导表达[J]. 浙江农业学报, 2021, 33(1): 1-7.

HE Jiaqi, ZHAI Ying, ZHANG Jun, QIU Shuang, LI Mingyang, ZHAO Yan, ZHANG Meijuan, MA Tianyi. Cloning and expression analysis of GmDof1.5 in soybean under abiotic stress[J]. Acta Agriculturae Zhejiangensis, 2021, 33(1): 1-7.

图/表 4

图1 大豆GmDof1.5 PCR扩增 M,DL 2000相对分子质量标记物;1-2,GmDof1.5 PCR 扩增产物。

Fig.1 PCR amplification of GmDof1.5 from soybean M, DL 2000 marker; 1-2, PCR product of GmDof1.5.

图2 GmDof1.5的核苷酸与氨基酸序列下划线代表Zf-Dof结构域;黑体代表磷酸化位点;*代表终止密码子。

Fig.2 Nucleotide and deduced amino acid sequence of GmDof1.5 Zf-Dof domain was underlined; Phosphorylation sites were indicated by boldface type; Stop codon was shown in *.

图3 Dof蛋白系统进化树

Fig.3 Phylogenetic tree of Dof proteins

图4 干旱(A)、高盐(B)、高温(C)、低温(D)和ABA(E)处理下GmDof1.5在大豆幼苗中的表达 **和*分别代表在1%和5%概率水平上差异显著。

Fig.4 Expression of GmDof1.5 under drought(A), salt(B), high temperature(C), low temperature(D) and ABA(E) treatments in soybean seedlings ** and * indicate significant difference at P<0.01 and P<0.05, respectively.

参考文献 25

[1]	蔡晓锋, 张余洋, 张俊红, 等. 植物Dof基因家族功能研究进展[J]. 植物生理学报, 2013,49(1):1-12.
	CAI X F, ZHANG Y Y, ZHANG J H, et al. Advances in research on function of the dof gene family in plant[J]. Plant Physiology Journal, 2013,49(1):1-12.(in Chinese with English abstract)
[2]	YANAGISAWA S. Dof domain proteins: plant-specific transcription factors associated with diverse phenomena unique to plants[J]. Plant and Cell Physiology, 2004,45(4):386-391. DOI URL PMID
[3]	YANAGISAWA S, IZUI K. Molecular cloning of two DNA-binding proteins of maize that are structurally different but interact with the same sequence motif[J]. The Journal of Biological Chemistry, 1993,268(21):16028-16036. URL PMID
[4]	郭彦秀, 陈静, 王艳芳, 等. Dof转录因子在植物中的调控作用[J]. 生物技术通报, 2019(5):146-156.
	GUO Y X, CHEN J, WANG Y F, et al. Roles of dof transcription factors in the regulation of plant[J]. Biotechnology Bulletin, 2019(5):146-156.(in Chinese with English abstract)
[5]	NOGUERO M, ATIF R M, OCHATT S, et al. The role of the DNA-binding One Zinc Finger (DOF) transcription factor family in plants[J]. Plant Science, 2013,209:32-45. URL PMID
[6]	SHINOZAKI K, YAMAGUCHI-SHINOZAKI K, SEKI M. Regulatory network of gene expression in the drought and cold stress responses[J]. Current Opinion in Plant Biology, 2003,6(5):410-417. URL PMID
[7]	HE L, SU C, WANG Y C, et al. ATDOF5.8 protein is the upstream regulator of ANAC069 and is responsive to abiotic stress[J]. Biochimie, 2015,110:17-24. DOI URL PMID
[8]	周淑芬, 颜静宛, 刘华清, 等. 水稻Dof基因家族的组织表达谱及胁迫诱导表达特征分析[J]. 分子植物育种, 2012,10(6):635-643.
	ZHOU S F, YAN J W, LIU H Q, et al. Transcriptional profiling analysis of OsDof gene family in various rice tissues and their expression characteristics under different stresses[J]. Molecular Plant Breeding, 2012,10(6):635-643.(in Chinese with English abstract)
[9]	MA J, LI M Y, WANG F, et al. Genome-wide analysis of Dof family transcription factors and their responses to abiotic stresses in Chinese cabbage[J]. BMC Genomics, 2015,16(1):33.
[10]	WU Z M, CHENG J W, CUI J J, et al. Genome-wide identification and expression profile of dof transcription factor gene family in pepper (Capsicum annuum L.)[J]. Frontiers in Plant Science, 2016,7:574. DOI URL PMID
[11]	唐跃辉, 包欣欣, 王健, 等. 小桐子Dof基因家族生物信息学与表达分析[J]. 江苏农业学报, 2019,35(1):15-25.
	TANG Y H, BAO X X, WANG J, et al. Bioinformatics and expression analysis of the Dof gene family in physic nut[J]. Jiangsu Journal of Agricultural Sciences, 2019,35(1):15-25.(in Chinese with English abstract)
[12]	SHAW L M, MCINTYRE C L, GRESSHOFF P M, et al. Members of the Dof transcription factor family in Triticum aestivum are associated with light-mediated gene regulation[J]. Functional Integrative Genomics, 2009,9(4):485. DOI URL PMID
[13]	GUO Y, QIU L J. Genome-wide analysis of the dof transcription factor gene family reveals soybean-specific duplicable and functional characteristics[J]. PLoS One, 2013,8(9):e76809. DOI URL PMID
[14]	WANG H W, ZHANG B, HAO Y J, et al. The soybean Dof-type transcription factor genes, GmDof4 and GmDof11, enhance lipid content in the seeds of transgenic Arabidopsis plants[J]. The Plant Journal, 2007,52(4):716-729. DOI URL PMID
[15]	ZHAI Y, SHAO S L, SHA W, et al. Overexpression of soybean GmERF9 enhances the tolerance to drought and cold in the transgenic tobacco[J]. Plant Cell, Tissue and Organ Culture, 2017,128(3):607-618.
[16]	SKIRYCZ A, JOZEFCZUK S, STOBIECKI M, et al. Transcription factor AtDOF4;2 affects phenylpropanoid metabolism in Arabidopsis thaliana[J]. New Phytologist, 2007,175(3):425-438.
[17]	KANG H G, SINGH K B. Characterization of salicylic acid-responsive, Arabidopsis Dof domain proteins: overexpression of OBP3 leads to growth defects[J]. The Plant Journal, 2000,21(4):329-339. DOI URL PMID
[18]	KANG H G, FOLEY R C, OÑATE-SÁNCHEZ L, et al. Target genes for OBP3, a Dof transcription factor, include novel basic helix-loop-helix domain proteins inducible by salicylic acid[J]. The Plant Journal, 2003,35(3):362-372. URL PMID
[19]	VENKATESH J, PARK S W. Genome-wide analysis and expression profiling of DNA-binding with one zinc finger (Dof) transcription factor family in potato[J]. Plant Physiology and Biochemistry, 2015,94:73-85. URL PMID
[20]	李辉, 黄蔚, 刘志薇, 等. 茶树两个Dof转录因子的分离及其在温度胁迫中的响应分析[J]. 茶叶科学, 2016,36(3):312-322.
	LI H, HUANG W, LIU Z W, et al. Isolation and expression analysis of two temperature responsive dof genes from Camellia sinensis[J]. Journal of Tea Science, 2016,36(3):312-322.(in Chinese with English abstract)
[21]	王海波, 唐利洲. 基于基因组鉴定小桐子Dof转录因子家族及其表达分析[J]. 分子植物育种, 2018,16(3):764-771.
	WANG H B, TANG L Z. Genome-wide identification and expression analysis of Dof transcription factor family in Jatropha curcas[J]. Molecular Plant Breeding, 2018,16(3):764-771.(in Chinese with English abstract)
[22]	YANAGISAWA S, SCHMIDT R J. Diversity and similarity among recognition sequences of Dof transcription factors[J]. The Plant Journal, 1999,17(2):209-214. DOI URL PMID
[23]	丁恺, 麻鹏达, 贾彦彦, 等. 丹参3个R2R3-MYB转录因子的亚细胞定位和自激活活性分析[J]. 西北农业学报, 2018,27(4):586-594.
	DING K, MA P D, JIA Y Y, et al. Subcellular localization and transactivation analysis of three R2R3-MYB in Salvia miltiorrhiza bunge[J]. Acta Agriculturae Boreali-Occidentalis Sinica, 2018,27(4):586-594.(in Chinese with English abstract)
[24]	ZHAO T J, SUN S, LIU Y, et al. Regulating the drought-responsive element (DRE)-mediated signaling pathway by synergic functions of trans-active and trans-inactive DRE binding factors in Brassica napus[J]. Journal of Biological Chemistry, 2006,281(16):10752-10759.
[25]	吴智明, 张圣旭, 梁关生. 马铃薯基因组中Dof转录因子家族的鉴定与表达特征分析[J]. 核农学报, 2015,29(7):1260-1270.
	WU Z M, ZHANG S X, LIANG G S. Genome-wide identification and expression analysis of the dof transcription factor family in potato(Solanum tuberosum L.)[J]. Journal of Nuclear Agricultural Sciences, 2015,29(7):1260-1270.(in Chinese with English abstract)

大豆转录因子GmDof1.5的克隆与非生物胁迫诱导表达

Cloning and expression analysis of GmDof1.5 in soybean under abiotic stress

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