Response Mechanism of Cotton GhDMT7 Gene to 5-azacytidine

doi:10.13304/j.nykjdb.2024.0522

Journal of Agricultural Science and Technology ›› 2025, Vol. 27 ›› Issue (8): 28-35.DOI: 10.13304/j.nykjdb.2024.0522

• BIOTECHNOLOGY & LIFE SCIENCE • Previous Articles Next Articles

Response Mechanism of Cotton GhDMT7 Gene to 5-azacytidine

Zhining YANG¹^,²(), Xuke LU²^,³, Yapeng FAN², Yuping SUN², Xin YU², Liang WANG⁴, Yongjian GAO⁴, Gulijiayinashen Habuli⁴, Gulishaxi Nasiyi⁴, Wumuer Abudu⁴, Hui HUANG², Menghao ZHANG², Lidong WANG², Xiao CHEN², Lei XIAO², Xinrui ZHANG², Shuai WANG²^,³, Xiugui CHEN²^,³, Junjuan WANG², Lixue GUO², Wenwei GAO¹(), Wuwei YE¹^,²^,³()

^1.Cotton Engineering Research Center of Ministry of Education，College of Agriculture，Xinjiang Agricultural University，Urumqi 830052，China
^2.State Key Laboratory of Cotton Bio-breeding and Integrated Utilization，Institute of Cotton Research，Chinese Academy of Agricultural Sciences，Henan Anyang 455000，China
^3.National Center of Technology Innovation for Comprehensive Utilization of Saline-alkali Land，Shandong Dongying 257000，China
^4.Agricultural Technology Extension Center of Shawan City，Xinjiang Shawan 832100，China

Received:2024-06-27 Accepted:2024-09-24 Online:2025-08-15 Published:2025-08-26
Contact: Wenwei GAO,Wuwei YE

棉花GhDMT7基因对5-氮杂胞嘧啶核苷的响应机制

杨志宁¹^,²(), 陆许可²^,³, 范亚朋², 孙玉平², 喻欣², 王亮⁴, 高永健⁴, 古丽加依那什·哈不力⁴, 古丽沙西·拿斯依⁴, 吾木尔·阿不都⁴, 黄慧², 张梦豪², 王立东², 陈筱², 肖磊², 张鑫蕊², 王帅²^,³, 陈修贵²^,³, 王俊娟², 郭丽雪², 高文伟¹(), 叶武威¹^,²^,³()

^1.新疆农业大学农学院，棉花教育部工程研究中心，乌鲁木齐 830052
^2.中国农业科学院棉花研究所，棉花生物育种与综合利用全国重点实验室，河南安阳 455000
^3.国家盐碱地综合利用技术创新中心，山东东营 257000
^4.沙湾市农业技术推广中心，新疆沙湾 832100

通讯作者: 高文伟,叶武威
作者简介:杨志宁 E-mail：18399191165@163.com
基金资助:
国家棉花产业技术体系建设项目(CARS-15-02);中国农业科学院科技创新工程项目

Abstract

Abstract:

To investigate the effects of 5-azacytidine （5-azaC） on the expression of the GhDMT7 gene and seed germination of cotton （Gossypium hirsutum L.）， this paper analyzed the expression level of GhDMT7 and the growth traits of cotton seed. The GhDMT7 gene was cloned and performed comprehensive bioinformatics analysis. The results showed that GhDMT7 protein was a stable hydrophilic protein with no signal peptide and transmembrane domains. Under salt stress conditions， the malondialdehyde content in cotton leaves increased， indicating exacerbated membrane damage. 5-azaC exhibited a dosage effect on GhDMT7 expression with slightly up-regulated expression at low dosage and down-regulated expression at high dosage， and decreased 5-methylcytosine （^5mC） content. Furthermore， the treatment with 50 μmol·L^-1 5-azaC significantly enhanced seed germination potential and growth parameters， which promoted seed germination. Above results laid a foundation for further studying the function of GhDMT7 on cotton growth and development， and provided genetic resources for breeding new varieties of saline-resistant cotton.

Key words: Gossypium hirsutum, DNA methyl transferase, gene cloning, bioinformatics analysis, qRT-PCR

摘要：

为探究5-氮杂胞嘧啶（5-azacytidine，5-azaC）对陆地棉（Gossypium hirsutum L.）GhDMT7基因表达及种子萌发的影响，分析了5-azac处理下GhDMT7的表达水平和棉花种子生长性状。克隆了GhDMT7基因，通过生物信息学分析发现，GhDMT7蛋白为稳定的亲水性蛋白，无信号肽和跨膜结构。在盐胁迫条件下，棉花叶片中的丙二醛含量增加，表明膜损伤加剧。5-azaC对GhDMT7的表达呈剂量效应，低水平下轻微上调基因表达，高水平下则抑制表达，并降低5-甲基胞嘧啶（^5mC）含量。此外，50 μmol·L^-1的5-azaC处理显著提高了种子的发芽势和生长指标，对种子萌发起促进作用。以上研究结果为进一步研究GhDMT7在棉花生长发育中的功能奠定了基础，也为棉花耐盐新品种的培育提供了基因资源。

关键词: 陆地棉, DNA甲基转移酶, 基因克隆, 生物信息学分析, qRT-PCR

CLC Number:

S562

Zhining YANG, Xuke LU, Yapeng FAN, Yuping SUN, Xin YU, Liang WANG, Yongjian GAO, Gulijiayinashen Habuli, Gulishaxi Nasiyi, Wumuer Abudu, Hui HUANG, Menghao ZHANG, Lidong WANG, Xiao CHEN, Lei XIAO, Xinrui ZHANG, Shuai WANG, Xiugui CHEN, Junjuan WANG, Lixue GUO, Wenwei GAO, Wuwei YE. Response Mechanism of Cotton GhDMT7 Gene to 5-azacytidine[J]. Journal of Agricultural Science and Technology, 2025, 27(8): 28-35.

杨志宁, 陆许可, 范亚朋, 孙玉平, 喻欣, 王亮, 高永健, 古丽加依那什·哈不力, 古丽沙西·拿斯依, 吾木尔·阿不都, 黄慧, 张梦豪, 王立东, 陈筱, 肖磊, 张鑫蕊, 王帅, 陈修贵, 王俊娟, 郭丽雪, 高文伟, 叶武威. 棉花GhDMT7基因对5-氮杂胞嘧啶核苷的响应机制[J]. 中国农业科技导报, 2025, 27(8): 28-35.

Figures/Tables 9

Table 1 Proportion of amino acid

氨基酸 Amino acid	数量 Number	占比 Percentage/%
丙氨酸Ala	43	6.8
精氨酸Arg	33	5.2
天冬酰胺Asn	38	6.0
天冬氨酸Asp	46	7.2
半胱氨酸Cys	11	1.7
谷氨酰胺Gln	18	2.8
谷氨酸Glu	51	8.0
甘氨酸Gly	47	7.4
组氨酸His	11	1.7
异亮氨酸Ile	28	4.4
亮氨酸Leu	65	10.2
赖氨酸Lys	30	4.7
甲硫氨酸Met	14	2.2
苯丙氨酸Phe	28	4.4
脯氨酸Pro	30	4.7
丝氨酸Ser	52	8.2
色氨酸Trp	10	1.6
酪氨酸Tyr	17	2.7
缬氨酸Val	36	5.7
吡咯赖氨酸Pyl	0	0.0
硒代半胱氨酸Sec	0	0.0

Fig. 1 Prediction of phosphorylation sites of GhDMT7 protein

Fig. 2 Prediction of secondary structure of GhDMT7 protein

Fig. 3 Prediction of tertiary structure of GhDMT7 protein

Fig. 4 Phenotype of seedling under salt stress

Fig. 5 MDA content in leaves of seedling under salt stressNote：Different lowercase letters indicate significant differences between different treatments at P<0.05 level．

Fig. 6 Expression level of GhDMT7 gene and 5mC content under different 5-azaC treatmentsNote：Different lowercase letters indicate significant differences between different treatments at P<0.05 level．

Fig. 7 Germination phenotype of cotton seed under different 5-azaC treatments

Fig. 8 Germination potential， root length and fresh weight of cotton seed under different 5-azaC treatmentsNote：Different lowercase letters indicate significant differences between different treatments at P<0.05 level．

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Response Mechanism of Cotton GhDMT7 Gene to 5-azacytidine

棉花GhDMT7基因对5-氮杂胞嘧啶核苷的响应机制

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