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作物学报 ›› 2024, Vol. 50 ›› Issue (10): 2503-2514.doi: 10.3724/SP.J.1006.2024.31075

• 作物遗传育种·种质资源·分子遗传学 • 上一篇    下一篇

大麦HvMBF1c耐盐机制分析

湛潇潇1,2(), 冯举伶1,2, 张震欢1,3, 张宏1,2, 汪军成1,2, 李葆春1,3, 杨轲1,2, 司二静1,2, 孟亚雄1,2, 马小乐1,2, 王化俊1,2, 姚立蓉1,2,*()   

  1. 1省部共建干旱生境作物学国家重点实验室 / 甘肃省作物遗传改良与种质创新重点实验室, 甘肃兰州 730070
    2甘肃农业大学农学院, 甘肃兰州 730070
    3甘肃农业大学生命科学技术学院, 甘肃兰州 730070
  • 收稿日期:2023-11-30 接受日期:2024-06-20 出版日期:2024-10-12 网络出版日期:2024-07-11
  • 通讯作者: *姚立蓉, E-mail: ylr0384@163.com
  • 作者简介:E-mail: 1363472842@qq.com
  • 基金资助:
    财政部和农业农村部国家现代农业产业技术体系建设专项(大麦青稞, CARS-05-03B-03);财政部和农业农村部国家现代农业产业技术体系建设专项(CARS-05-03B-03);国家自然科学基金项目(30771331);国家自然科学基金项目(32160496);甘肃农业大学学生科研训练计划(202301047);甘肃农业大学学生科研训练计划(202210733008);甘肃农业大学学生科研训练计划(S202210733003);甘肃省青年科技基金计划(20JR5RA010);甘肃省教育厅创新基金(2021A-055);甘肃省教育厅产业支撑计划项目(2021CYZC-12)

Salt tolerance analysis of HvMBF1c in barley

ZHAN Xiao-Xiao1,2(), FENG Ju-Ling1,2, ZHANG Zhen-Huan1,3, ZHANG Hong1,2, WANG Jun-Cheng1,2, LI Bao-Chun1,3, YANG Ke1,2, SI Er-Jing1,2, MENG Ya-Xiong1,2, MA Xiao-Le1,2, WANG Hua-Jun1,2, YAO Li-Rong1,2,*()   

  1. 1State Key Laboratory of Crop Science in Arid Habitat Jointly Established by the Ministry of Province and Ministry of Finance / Key Laboratory of Crop Genetic Improvement and Germplasm Innovation in Gansu Province, Lanzhou 730070, Gansu, China
    2Agronomy College, Gansu Agricultural University, Lanzhou 730070, Gansu, China
    3College of Life Science and Technology, Gansu Agricultural University, Lanzhou 730070, Gansu, China
  • Received:2023-11-30 Accepted:2024-06-20 Published:2024-10-12 Published online:2024-07-11
  • Contact: *E-mail: ylr0384@163.com
  • Supported by:
    China Agriculture Research System of MOF and MARA(Barley, CARS-05-03B-03);China Agriculture Research System of MOF and MARA(CARS-05-03B-03);National Natural Science Foundation of China(30771331);National Natural Science Foundation of China(32160496);Gansu Agricultural University Student Research Training Program(202301047);Gansu Agricultural University Student Research Training Program(202210733008);Gansu Agricultural University Student Research Training Program(S202210733003);Gansu Youth Science and Technology Fund Program(20JR5RA010);Innovation Fund of Education Department of Gansu Province(2021A-055);Industry Support Plan of Education Department of Gansu Province(2021CYZC-12)

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摘要：

大麦HvMBF1c基因参与调控植物盐胁迫表达, 为探究其对盐胁迫的响应模式, 本文对大麦HvMBF1基因进行生物信息学分析, 并以中川大麦(耐盐型)、GN18 (盐敏感型)及WT与转HvMBF1c基因拟南芥为材料, 分别取200 mmol L-1 NaCl胁迫0 h、6 h、12 h、24 h、48 h及72 h样品, 对其进行qRT-PCR分析和生理指标的测定。 结果显示, 大麦HvMBF1c基因与小麦TaMBF1c亲缘关系最近, 位于7H染色体上。HvMBF1c的外显子比HvMBF1a和HvMBF1b的大, 且HvMBF1a和HvMBF1b两者具有典型的motif 4结构域, 而HvMBF1c基因具有独特的motif 5结构域。HvMBF1c基因的启动子序列含有光响应、根特定调节元件和缺氧特异性诱导增强元件, 与小麦2号、3号和7号染色体存在同源性。qRT-PCR分析表明, 随着盐处理时间的延长, 大麦HvMBF1c基因在2份种质材料苗期、拔节期、抽穗期和灌浆期的表达量呈上升趋势, 且中川大麦中该基因的表达量高于GN18中的表达量; 转HvMBF1c拟南芥中其表达量随着盐处理时间的延长而升高。盐胁迫下, 中川大麦苗期、拔节期、抽穗期及灌浆期的SOD、POD、CAT、MDA和可溶性蛋白低于GN18, 转基因拟南芥中各生理指标的变化量低于WT。本研究结果为进一步探索HvMBF1c的功能提供参考。

关键词: 大麦, HvMBF1c, 生理指标, 共线性分析, 表达量

Abstract:

The HvMBF1c gene in barley is implicated in the plant’s response to salt stress. To investigate the HvMBF1c response mechanism to salt stress, we conducted bioinformatics analysis of the HvMBF1 gene, qRT-PCR analysis, and physiological index determination in Nakagawa barley (salt-tolerant type), GN18 (salt-sensitive type), wild-type (WT) and transgenic HvMBF1c Arabidopsis thaliana treated with 200 mmol L-1 NaCl for 0, 6, 12, 24, 48, and 72 hours. The results showed that the barley HvMBF1c gene is closely related to the wheat TaMBF1c gene, and is located on chromosome 7H. The exon of HvMBF1c is larger than those of HvMBF1a and HvMBF1b, with HvMBF1a and HvMBF1b containing a typical motif 4 domain, while the HvMBF1c gene has a unique motif 5 domain. The promoter sequence of the HvMBF1c gene includes photoreactive, root-specific regulatory elements, and hypoxia-specific inducible enhancing elements, homologous to wheat chromosomes 2, 3, and 7. qRT-PCR analysis revealed significant upregulation of HvMBF1c expression in two barley genotypes at the seedling, jointing, heading, and filling stages, with higher expression in Nakagawa barley compared to GN18 as salt treatment duration increased. Similarly, the expression of HvMBF1c in Arabidopsis thaliana also increased with prolonged salt treatment time. Physiological indices of Nakagawa barley at each growth stage were lower than those in GN18, and transgenic Arabidopsis exhibited lower indices compared to WT under different salt stress conditions. These findings provide a foundation for further exploration of the HvMBF1c gene function.

Key words: barley, HvMBF1c, physiological indicators, collinearity analysis, the extension

表1

引物序列及用途"

引物名称
Primer name		 引物序列
Primer sequence (5′-3′)		 用途
Purpose
HvMBF1c-qF
HvMBF1c-qR
HvActin-F
HvActin-R		 CATCCAGAAGGCTCGCGT
CTCTCGTACTCCTGCACCAC
AAGCAGCCAGAATGTACAGCGAGAAC
GGTACAGACCAGCAAAGCCAGAAATG		 荧光定量PCR
Fluorescence quantitative PCR
内参基因引物
Internal reference gene primer

图1

大麦、小麦、水稻、玉米和拟南芥中MBF1基因家族系统发育进化树 Hv: 大麦; Ta: 小麦; Os: 水稻; Zm: 玉米; At: 拟南芥。"

图2

大麦HvMBF1基因家族成员在大麦染色体上的分布"

图3

大麦HvMBF1基因家族成员基因结构 UTR: 非翻译区; CDS: 编码区序列。"

图4

大麦HvMBF1基因家族成员保守基序 Motif: 保守基序。"

图5

大麦HvMBF1基因保守基序序列Logo"

图6

大麦HvMBF1基因启动子顺式作用元件"

图7

大麦与小麦HvMBF1基因共线性分析"

图8

盐胁迫下大麦各个时期和拟南芥HvMBF1c表达量 不同字母表示差异水平显著(P < 0.05)。"

图9

盐胁迫下大麦SOD、POD、MDA、CAT和可溶性蛋白活性变化 不同字母表示差异水平显著(P < 0.05)。"

图10

盐胁迫下拟南芥SOD、POD、MDA、CAT和可溶性蛋白活性变化 不同字母表示差异水平显著(P < 0.05)。"

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