亚麻LuWRI1a在旱盐胁迫响应中的功能分析

doi:10.3724/SP.J.1006.2024.34171

作物学报 ›› 2024, Vol. 50 ›› Issue (7): 1750-1761.doi: 10.3724/SP.J.1006.2024.34171

• 作物遗传育种·种质资源·分子遗传学 • 上一篇下一篇

亚麻LuWRI1a在旱盐胁迫响应中的功能分析

李闻娟(), 王利民, 齐燕妮, 赵玮, 谢亚萍, 党照, 赵丽蓉, 李雯, 徐晨梦, 王琰, 张建平^*()

甘肃省农业科学院作物研究所, 甘肃兰州 730070

收稿日期:2023-10-19 接受日期:2024-01-30 出版日期:2024-07-12 网络出版日期:2024-02-20
通讯作者: *张建平, E-mail: zhangjpzw3@gsagr.ac.cn
作者简介:E-mail: liwenjuan@gsagr.ac.cn
基金资助:
国家自然科学基金项目(31460388);国家自然科学基金项目(32360502);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-14-1-05);甘肃省农业科学院现代生物育种项目(2022GAAS04);兰州市科技计划项目(2023-3-37)

Functional analysis of flax LuWRI1a in response to drought and salt stresses

LI Wen-Juan(), WANG Li-Min, QI Yan-Ni, ZHAO Wei, XIE Ya-Ping, DANG Zhao, ZHAO Li-Rong, LI Wen, XU Chen-Meng, WANG Yan, ZHANG Jian-Ping^*()

Institute of Crop Research, Gansu Academy of Agricultural Sciences, Lanzhou 730070, Gansu, China

Received:2023-10-19 Accepted:2024-01-30 Published:2024-07-12 Published online:2024-02-20
Contact: *E-mail: zhangjpzw3@gsagr.ac.cn
Supported by:
National Natural Science Foundation of China(31460388);National Natural Science Foundation of China(32360502);China Agriculture Research System(CARS-14-1-05);China Agriculture Research System of MOF and MARA(2022GAAS04);Science and Technology Program of Lanzhou(2023-3-37)

摘要/Abstract

摘要：

AP2/ERF转录因子家族参与植物对生物和非生物胁迫响应的调控。前期我们从亚麻中克隆了一个WRINKLED1的同源基因LuWRI1a, 蛋白序列分析发现, LuWRI1a包含2个AP2的DNA结合域, 属于AP2/ERF转录因子家族。对LuWRI1a的顺式作用元件进行分析发现, pLuWRI1a包含响应光、干旱、低温和激素等多个非生物胁迫应激元件。本研究以亚麻栽培品种陇亚10号和LuWRI1a过表达转基因纯合株系为试验材料, 用200 mmol L^-1NaCl营养液和25% PEG营养液模拟盐胁迫和干旱胁迫处理。结果表明, 在盐胁迫和干旱胁迫处理后, 转基因植株的相对株高、主根长度、侧根数目及叶片数均升高; 3种抗氧化酶的活性均显著高于对照, 而MDA含量低于对照; 非生物胁迫响应基因LuAREB、LuDREB、LuLEA和LuNCED的表达水平均上调。通过探究LuWRI1a在逆境胁迫下的生物学功能发现, LuWRI1a通过抵抗盐胁迫和干旱胁迫对亚麻生长的抑制, 增强活性氧清除能力、减轻膜脂的氧化损伤, 激活逆境胁迫响应基因的表达等途径, 增强了亚麻的耐逆性。综上所述, LuWRI1a可能是一个多功能基因, 它不仅参与脂肪酸合成代谢途径, 还有可能参与植物非生物胁迫信号途径。本研究为亚麻耐逆品种改良提供了新的基因资源。

关键词: 亚麻, LuWRI1a, 盐胁迫, 干旱胁迫, 功能分析

Abstract:

The AP2/ERF family of transcription factors is involved in the regulation of plant responses to biotic and abiotic stresses. Previously, we cloned LuWRI1a, a WRINKLED1 homologous gene from flax. Protein sequence analysis showed that LuWRI1a contained two AP2 DNA-binding domains and belonged to the AP2/ERF transcription factor family. The cis-acting elements of LuWRI1a were analyzed that pLuWRI1a was found containing multiple abiotic stress elements in response to light, drought, low temperature and hormones. In this study, the flax cultivar Longya 10 and LuWRI1a overexpression transgenic pure lines were used as the experimental materials, and salt stress and drought stress treatments were simulated with 200 mmol L^-1NaCl nutrient solution and 25% PEG nutrient solution. The results showed that the relative plant height, primary root length, lateral root number, and leaf number of transgenic plants were elevated after salt and drought stress treatments. The activities of three antioxidant enzymes were significantly higher than the control, while MDA content was lower. The relative expression levels of the abiotic stress-responsive genes, LuAREB, LuDREB, LuLEA, and LuNCED, were up-regulated. By exploring the biological function of LuWRI1a under adversity stress, it was found that LuWRI1a enhanced flax tolerance by resisting the inhibition of flax growth by salt stress and drought stress, enhancing the scavenging ability of reactive oxygen species, reducing the oxidative damage of membrane lipids, and activating the expression of adversity stress response genes. In summary, LuWRI1a may be a multifunctional gene, which was not only involved in fatty acid synthesis and metabolism pathway, but also may be involved in plant abiotic stress signaling pathway. This study provides a new genetic resource for the improvement of stress-tolerant varieties of flax.

Key words: flax, LuWRI1a, salt stress, drought stress, functional analysis

李闻娟, 王利民, 齐燕妮, 赵玮, 谢亚萍, 党照, 赵丽蓉, 李雯, 徐晨梦, 王琰, 张建平. 亚麻LuWRI1a在旱盐胁迫响应中的功能分析[J]. 作物学报, 2024, 50(7): 1750-1761.

LI Wen-Juan, WANG Li-Min, QI Yan-Ni, ZHAO Wei, XIE Ya-Ping, DANG Zhao, ZHAO Li-Rong, LI Wen, XU Chen-Meng, WANG Yan, ZHANG Jian-Ping. Functional analysis of flax LuWRI1a in response to drought and salt stresses[J]. Acta Agronomica Sinica, 2024, 50(7): 1750-1761.

图/表 9

表1

图1

表2

LuWRI1a启动子顺式作用元件的推测功能"

调控元件 Cis-element	核心序列 Sequence (5′-3′)	位置 Position	功能 Function
ACE	CTAACGTATT	-1239	光响应顺式作用元件 Cis-acting element involved in light responsiveness
AE-box	AGAAACAA	+144	光响应模块的组成部分 Part of a module for light response
ARE	AAACCA	-1057, -1801, +2406	厌氧诱导必需的顺式作用调控元件 Cis-acting regulatory element essential for the anaerobic induction
Box 4	ATTAAT	+579, -2178	光响应保守模块的一部分 Part of a conserved DNA module involved in light responsiveness
CGTCA-motif	CGTCA	+963, +1169	MeJA响应顺式作用元件 Cis-acting regulatory element involved in the MeJA-responsiveness
GT1-motif	GGTTAA(T)	+17, -679, -680	光响应元件 Light responsive element
Gap-box	CAAATGAA(A/G)A	+524, +1038	光响应元件的一部分 Part of a light responsive element
LTR	CCGAAA	+1983	低温响应顺式作用元件 Cis-acting element involved in low-temperature responsiveness
MBS	CAACTG	+2127	参与干旱诱导的MYB结合位点 MYB binding site involved in drought-inducibility
MBSI	aaaAaaC(G/C)GTTA	-2215	参与干旱诱导的MYB结合位点 MYB binding site involved in flavonoid biosynthetic genes regulation
MYB	CAACCA, TAACCA	-16, +391, +681, -759, +1853, +2105, +2123	干旱、盐、ABA 和低温响应 Drought, salt, ABA, and low-temperature responsive
MYC	CATTTG	+73, +267, -524, -569, -1038, -1363	干旱和ABA响应 Drought and ABA responsive
O2-site	GATGATGTGG	+350	参与玉米醇溶蛋白新陈代谢调节的顺式调控元件 Cis-acting regulatory element involved in zein metabolism regulation
STRE	AGGGG	+1024, -1548, -1580, +1604, +2307	由热激、渗透应激、低pH值和营养缺乏激活的元件 Activated by heat shock, osmotic stress, low pH, and nutrient starvation
ERE	ATTTTAAA, ATTTCATA	+1439, -1905	乙烯响应元件 Ethylene-responsive element
TCA-element	CCATCTTTTT	+2343	水杨酸响应的顺式作用元件 Cis-acting element involved in salicylic acid responsiveness
TGACG-motif	TGACG	-963, -1169	MeJA 响应顺式作用元件 Cis-acting regulatory element involved in the MeJA-responsiveness

表2

图2

图3

图4

图5

图6

图7

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亚麻LuWRI1a在旱盐胁迫响应中的功能分析

Functional analysis of flax LuWRI1a in response to drought and salt stresses

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