Cloning and subcellular localization analysis of the GmNF-YC4 gene in soybean

doi:10.6048/j.issn.1001-4330.2025.06.008

Xinjiang Agricultural Sciences ›› 2025, Vol. 62 ›› Issue (6): 1365-1370.DOI: 10.6048/j.issn.1001-4330.2025.06.008

• Crop Genetics and Breeding·Cultivation Physiology·Physiology and Biochemistry • Previous Articles Next Articles

Cloning and subcellular localization analysis of the GmNF-YC4 gene in soybean

LIU Leilei¹(), ZHAO Guoqin¹, NIE Junfang², YU Yuehua², NI Zhiyong¹()

1. Key Laboratory of Ecological Adaptation and Evolution of Extreme Environment in Xinjiang, College of Life Sciences, Xinjiang Agricultural University, Urumqi 830052, China
2. College of Agronomy, Xinjiang Agricultural University, Urumqi 830052, China

Received:2024-11-15 Online:2025-06-20 Published:2025-07-29
Correspondence author: NI Zhiyong
Supported by:
National Natural Science Foundation of China(32160446);National Natural Science Foundation of China(32360454);“Tianshan Talents”Training Program Project(2023TSYCCX0012);Natural Science Foundation of Xinjiang Uygur Autonomous Region(2022D01E14);College Students' Innovation and Entrepreneurship Training Program Project of Xinjiang Uygur Autonomous Region(S202210758033)

大豆GmNF-YC4基因的克隆与亚细胞定位分析

刘雷蕾¹(), 赵国勤¹, 聂俊芳², 于月华², 倪志勇¹()

1.新疆极端环境生物生态适应与进化重点实验室/新疆农业大学生命科学学院,乌鲁木齐, 830052
2.新疆农业大学农学院,乌鲁木齐 830052

通讯作者: 倪志勇
作者简介:刘雷蕾(1998-),女,四川人,硕士研究生,研究方向生物学,(E-mail)1908181694@qq.com
基金资助:
国家自然科学基金项目(32160446);国家自然科学基金项目(32360454);“天山英才”培养计划(2023TSYCCX0012);新疆维吾尔自治区自然科学基金(2022D01E14);新疆维吾尔自治区级大学生创新创业训练计划项目(S202210758033)

Abstract

Abstract:

【Objective】 To study the subcellular localization of GmNF-YC4,which lays the foundation for revealing the molecular mechanism of soybean stress resistance. 【Methods】 In this study, GmNF-YC4 was cloned from soybean, bioinformatics for GmNF-YC4 sequence and phylogenetic tree, and the subcellular localization of GmNF-YC4 in tobacco cells was analyzed by transient transformation tobacco. 【Results】 The open reading frame size of GmNF-YC4 (GenBank accession number: NM _ 001362631.1) was 690bp, encoding 229 amino acids, which predicted molecular weight of 24.78 kDa, and isoelectric point of 5.19. The GmNF-YC4 protein contained a HAP 5 conserved domain. The phylogenetic tree analysis showed that GmNF-YC4 and AtNF-YC 1, AtNF-YC 4 were clustered in one clade. The genome of GmNF-YC4 contained 1 exon. The subcellular localization results indicated that GmNF-YC4 protein was localized in the nucleus of tobacco. 【Conclusion】 GmNF-YC4 is a nucleus-localized transcription factor. GmNF-YC4 gene sequence is obtained.

Key words: soybean; nuclear factor subunit C; gene cloning; subcellular localization

摘要：

【目的】为了研究GmNF-YC4的亚细胞定位,为揭示大豆抗逆分子机理奠定基础。【方法】本研究从大豆中克隆基因GmNF-YC4,利用生物信息学对GmNF-YC4的序列进行分析并构建系统发生树,采用瞬时转化烟草方法分析GmNF-YC4在烟草细胞中的亚细胞定位。【结果】GmNF-YC4 (GenBank 登录号:NM_001362631.1)的开放阅读框大小为690 bp,编码229个氨基酸,预测分子量为24.78 kDa,等电点为5.19。GmNF-YC4蛋白含有一个HAP5保守结构域。GmNF-YC4和拟南芥的AtNF-YC1,AtNF-YC4聚在一个分枝中。GmNF-YC4的基因组包含1个外显子。GmNF-YC4蛋白定位于烟草的细胞核中。【结论】 GmNF-YC4是一个细胞核定位的转录因子,获得了GmNF-YC4基因序列。

关键词: 大豆, 核因子亚基C, 基因克隆, 亚细胞定位

CLC Number:

S565.1

LIU Leilei, ZHAO Guoqin, NIE Junfang, YU Yuehua, NI Zhiyong. Cloning and subcellular localization analysis of the GmNF-YC4 gene in soybean[J]. Xinjiang Agricultural Sciences, 2025, 62(6): 1365-1370.

刘雷蕾, 赵国勤, 聂俊芳, 于月华, 倪志勇. 大豆GmNF-YC4基因的克隆与亚细胞定位分析[J]. 新疆农业科学, 2025, 62(6): 1365-1370.

Figures/Tables 3

Fig.1 Sequence of GmNF-YC4 Notes: a: analysis of the phylogenetic tree of GmNF-YC4 and the Arabidopsis NF-YC family; b: prediction of conserved domain of GmNF-YC4 protein and analysis of amino acid sequence alignment with the Arabidopsis NF-YB family; c: prediction of conserved domain of GmNF-YC4 protein; d: schematic diagram of GmNF-YC4 structure

Fig.2 Cloning of GmNF-YC4 gene Notes: M: DL2000 DNA Marker (a). Amplification of the target fragment; 1: the amplified product of the GmNF-YC4 coding region; (b). GmNF-YC4-pEGAD was detected by PCR of the bacterial solutio

Fig.3 GmNF-YC04 subcellular localization

References 21

[1]	宋秋明, 李大勇, 张慧娟, 等. 植物NF-Y转录因子的生物学功能及其研究进展[J]. 植物生理学报, 2015, 51(5): 623-632.
	SONG Qiuming, LI Dayong, ZHANG Huijuan, et al. NF-Y transcription factors and their biological functions in plants[J]. Plant Physiology Journal, 2015, 51(5): 623-632.
[2]	Strader L, Weijers D, Wagner D. Plant transcription factors—being in the right place with the right company[J]. Current Opinion in Plant Biology, 2022, 65: 102136.
[3]	Leyva-González M A, Ibarra-Laclette E, Cruz-Ramírez A, et al. Functional and transcriptome analysis reveals an acclimatization strategy for abiotic stress tolerance mediated by Arabidopsis NF-YA family members[J]. PLoS One, 2012, 7(10): e48138.
[4]	Yamamoto A, Kagaya Y, Toyoshima R, et al. Arabidopsis NF-YB subunits LEC1 and LEC1-LIKE activate transcription by interacting with seed-specific ABRE-binding factors[J]. The Plant Journal, 2009, 58(5): 843-856.
[5]	Laloum T, De Mita S, Gamas P, et al. CCAAT-box binding transcription factors in plants: Y so many?[J]. Trends in Plant Science, 2013, 18(3): 157-166.
[6]	Zhao H, Wu D, Kong F Y, et al. The Arabidopsis thaliana nuclear factor Y transcription factors[J]. Frontiers in Plant Science, 2017, 7: 2045.
[7]	Petroni K, Kumimoto R W, Gnesutta N, et al. The promiscuous life of plant NUCLEAR FACTOR Y transcription factors[J]. The Plant Cell, 2012, 24(12): 4777-4792.
[8]	Baudin M, Laloum T, Lepage A, et al. A phylogenetically conserved group of nuclear factor-Y transcription factors interact to control nodulation in legumes[J]. Plant Physiology, 2015, 169(4): 2761-2773.
[9]	Lee D K, Kim H I, Jang G, et al. The NF-YA transcription factor OsNF-YA7 confers drought stress tolerance of rice in an abscisic acid independent manner[J]. Plant Science, 2015, 241: 199-210.
[10]	李世贵, 马瑞, 王芳芳, 等. 植物NF-Y转录因子研究进展[J]. 植物生理学报, 2021, 57(2): 248-256.
	LI Shigui, MA Rui, WANG Fangfang, et al. Research progresses on plant NF-Y transcription factors[J]. Plant Physiology Journal, 2021, 57(2): 248-256.
[11]	Zhang H, Liu S J, Ren T M, et al. Crucial abiotic stress regulatory network of NF-Y transcription factor in plants[J]. International Journal of Molecular Sciences, 2023, 24(5): 4426.
[12]	Hwang K, Susila H, Nasim Z, et al. Arabidopsis ABF₃ and ABF4 transcription factors act with the NF-YC complex to regulate SOC1 expression and mediate drought-accelerated flowering[J]. Molecular Plant, 2019, 12(4): 489-505.
[13]	Li S G, Zhang N, Zhu X, et al. Genome-wide analysis of NF-Y genes in potato and functional identification of StNF-YC9 in drought tolerance[J]. Frontiers in Plant Science, 2021, 12: 749688.
[14]	Bi C, Ma Y, Wang X F, et al. Overexpression of the transcription factor NF-YC9 confers abscisic acid hypersensitivity in Arabidopsis[J]. Plant Molecular Biology, 2017, 95(4/5): 425-439.
[15]	Yu T F, Liu Y, Fu J D, et al. The NF-Y-PYR module integrates the abscisic acid signal pathway to regulate plant stress tolerance[J]. Plant Biotechnology Journal, 2021, 19(12): 2589-2605.
[16]	曹岩, 张晓玫, 陈新建, 等. 大豆GmNF-YC₂基因的克隆与功能分析[J]. 作物学报, 2012, 38(9): 1607-1616.
	CAO Yan, ZHANG Xiaomei, CHEN Xinjian, et al. Cloning and functional analysis of GmNF-YC₂ gene in soybean(Glycine max)[J]. Acta Agronomica Sinica, 2012, 38(9): 1607-1616.
[17]	白玉翠, 王萍, 倪志勇, 等. 大豆GmNF-YA11基因克隆及特征分析[J]. 大豆科学, 2019, 38(5): 719-725.
	BAI Yucui, WANG Ping, NI Zhiyong, et al. Cloning and characterization of GmNF-YA11 gene in soybean[J]. Soybean Science, 2019, 38(5): 719-725.
[18]	Hackenberg D, Wu Y F, Voigt A, et al. Studies on differential nuclear translocation mechanism and assembly of the three subunits of the Arabidopsis thaliana transcription factor NF-Y[J]. Molecular Plant, 2012, 5(4): 876-888.
[19]	Shi H T, Ye T T, Zhong B, et al. AtHAP5A modulates freezing stress resistance in Arabidopsis through binding to CCAAT motif of AtXTH21[J]. New Phytologist, 2014, 203(2): 554-567.
[20]	Li L, Zheng W G, Zhu Y B, et al. QQS orphan gene regulates carbon and nitrogen partitioning across species via NF-YC interactions[J]. Proceedings of the National Academy of Sciences of the United States of America, 2015, 112(47): 14734-14739.
[21]	Tong S F, Wang Y B, Chen N N, et al. PtoNF-YC9-SRMT-PtoRD26 module regulates the high saline tolerance of a triploid poplar[J]. Genome Biology, 2022, 23(1): 148.

Cloning and subcellular localization analysis of the GmNF-YC4 gene in soybean

大豆GmNF-YC4基因的克隆与亚细胞定位分析

RichHTML

PDF

Knowledge

Abstract

Cite this article

share this article

Figures/Tables 3

References 21

Related Articles 15

Recommended Articles

Metrics

[1]	LI Ming, ZHENG Huanbin, WANG Mingjing, ZHAO Beifang, ZHANG Sitong, YANG Suxin. Effect of salt stress on soybean roots and GWAS analysis in seedling stage [J]. Xinjiang Agricultural Sciences, 2025, 62(3): 627-643.
[2]	CHENG Guanfu, LU Guoqiang, CUI Yongxiang, HOU Aocheng, ZHANG Guoshuai, LIANG Chunyan, LEI Jianfeng, LU Wei, DAI Peihong, LI yue. Functional analysis of a C2H2 zinc protein GhZFP8 gene in Gossypium hirsutum resistance to Verticillium wilt [J]. Xinjiang Agricultural Sciences, 2025, 62(1): 137-145.
[3]	MA Shangjie, LI Shengmei, YANG Tao, WANG Honggang, ZHAO Kang, PANG Bo, GAO Wenwei. Cloning and subcellular localization of the GHWAT1-35 gene in Gossypium hirsutum [J]. Xinjiang Agricultural Sciences, 2024, 61(6): 1310-1317.
[4]	YANG Xiangbo, CHEN Liangyu, YANG Songnan, CHEN Xifeng, XING Weiming, LI Xueying, CONG Weixuan, ZANG Zhenyuan, ZANG Yuanbo, ZHANG Jun. Phenotype analysis and comprehensive evaluation of spring soybean germplasm resources from northeast China [J]. Xinjiang Agricultural Sciences, 2024, 61(12): 2921-2933.
[5]	JIN Juan, Subina Xiaokelaiti, Abudoukayoumu Ayimaiti, YANG Lei, HAO Qing, FAN Dingyu. Cloning and sequence analysis of expansin gene ZjEXPA8 in Huizao [J]. Xinjiang Agricultural Sciences, 2023, 60(9): 2223-2230.
[6]	SHI Xiaolei, DING Sunlei, CONG Hua, ZHANG Jinbo, QU Kejia, WANG Xingzhou, HAN dai, YAN Yongliang. Gray correlation analysis of reast soybean yield correlation traits [J]. Xinjiang Agricultural Sciences, 2023, 60(7): 1641-1652.
[7]	YAN Yongliang, ZHANG Heng, QU Kejia, SHI Xiaolei, WANG Xingzhou, ZHANG Jinbo, CONG Hua. Comparative study on the agronomic traits and yields of different soybean varieties [J]. Xinjiang Agricultural Sciences, 2023, 60(7): 1653-1662.
[8]	ZHAO Lianjia, LI Gan, XU Lin, YAN Guorong, LIU Ning, WANG Fan, DENG Chaohong, Abudukeyoumu Abudurezike, WANG Cong, WANG Wei. Analysis of the main characters of soybean varieties in Xinjiang and their correlation with yield [J]. Xinjiang Agricultural Sciences, 2023, 60(7): 1663-1670.
[9]	GENG Feifei, MENG Chaomin, QING Guixia, ZHOU Jiamin, ZHANG Fuhou, LIU Fengju. Cloning and expression analysis of phosphorus efficient gene GhMYB4 in Gossypium hirsutum L. [J]. Xinjiang Agricultural Sciences, 2023, 60(6): 1406-1412.
[10]	ZHU Baoguo, KUANG Enjun, TENG Zhanglin, MENG Qingying, WANG Nannan, FENG Haoyuan, QIU Lei, GAO Xuedong, ZHANG Chunfeng. Effects of different bio-organic fertilizers application combined with conventional fertilization on growth, disease resistance and yield of soybean [J]. Xinjiang Agricultural Sciences, 2023, 60(5): 1127-1133.
[11]	ZHANG Jianxun, REN Jintao, WU Haijun, WANG Cong, ZHANG Jianxin. Effect of Densities on Flower and Pod Formation and Yield of Spring Soybean Varieties with Different Plant Types [J]. Xinjiang Agricultural Sciences, 2023, 60(3): 539-546.
[12]	Alibieligen Hazitai, Hanipa Hazaisi, Zulehumaer Wusimanjiang, YAO Qing, Shouhereti Yodika, HOU Xanfei, LI Qiang, ZHANG Zheng. Adaptability Analysis of Introduced Soybean Varieties in Yili Valley of Xinjiang [J]. Xinjiang Agricultural Sciences, 2023, 60(2): 326-335.
[13]	YI Yuanyang, XIE Fang, TIAN Shiying, ZHANG Zhidong, GU Meiying, PENG Xiaowu. Effect of Organic Fertilizer of Thioerythromycin Residue on Soybean Soil Drug-resistant Bacteria and Their Resistance Genes [J]. Xinjiang Agricultural Sciences, 2023, 60(2): 424-431.
[14]	Nasibai Abuduwahapu, GAO Xiaojuan, Kunduziayi Abudushalamu, LI Jiangwei. Cloning and Sequence Analysis of cDNA of C3d Gene of Bactrian Camel from Xinjiang [J]. Xinjiang Agricultural Sciences, 2023, 60(2): 493-500.
[15]	LIU Chenxi, ZHU Yuting, ZHOU Qiang, CHEN Jin, ZHAO Wenjie, ZHENG Kai. Cloning and bioinformatics analysis of β-carotene isomerase GbD27-6 gene in sea island cotton [J]. Xinjiang Agricultural Sciences, 2023, 60(12): 2869-2877.