菜用大豆GmDi19-3启动子对盐胁迫和外源ABA、MeJA的响应

doi:10.16420/j.issn.0513-353x.2023-0934

摘要/Abstract

摘要：

为阐明菜用大豆GmDi19-3启动子的调控功能，分析了GmDi19-3的盐胁迫诱导性、外源植物生长调节剂信号诱导性和组织表达特异性，克隆获得GmDi19-3启动子序列并分析其顺式作用元件组成，构建GmDi19-3启动子融合GUS报告基因表达载体并在拟南芥中稳定表达，检测启动子在转基因植株中的活性。GmDi19-3呈现典型的盐胁迫和外源ABA、MeJA信号诱导表达特性，GmDi19-3启动子区域包含逆境胁迫及激素诱导表达元件。GUS组织染色和活性检测显示，GmDi19-3启动子具有活性，在叶片、茎和根中明显表达，盐胁迫及外源ABA和MeJA信号能显著诱导GmDi19-3启动子活性增强。这些研究结果表明GmDi19-3启动子在菜用大豆盐胁迫和外源ABA、MeJA信号应答过程中发挥重要作用。

关键词: 菜用大豆, Di19-3, 启动子, GUS活性, 盐胁迫, ABA, MeJA

Abstract:

To clarify the regulation function of vegetable soybean GmDi19-3 promoter，the expression patterns of GmDi19-3 gene in different tissues and in response to salt stress and exogenous plant growth regulators were analyzed. The promoter of GmDi19-3 was cloned and cis-acting element composition was analyzed. The promoter of GmDi19-3 was fused with the GUS gene in the expression vector which was stably expressed in Arabidopsis. The promoter activity of GmDi19-3 was detected in the transgenic plants. Typical salt stress and exogenous ABA and MeJA signal inducible expression patterns of GmDi19-3 were observed. The promoter of GmDi19-3 harbored abiotic stress and hormone signal responsive elements. Based on the GUS staining and activity detection assays，it was verified that salt stress and exogenous ABA and MeJA signals obviously enhanced the promoter activity of GmDi19-3 in the leaf，stem and root of transgenic plants. These results suggested that GmDi19-3 promoter played key roles in response to salt stress，and exogenous ABA and MeJA signals.

Key words: vegetable soybean, Di19-3, promoter, GUS activity, salt stress, ABA, MeJA

冯志娟, 刘娜, 张古文, 卜远鹏, 王斌, 龚亚明. 菜用大豆GmDi19-3启动子对盐胁迫和外源ABA、MeJA的响应[J]. 园艺学报, 2024, 51(12): 2791-2799.

FENG Zhijuan, LIU Na, ZHANG Guwen, BU Yuanpeng, WANG Bin, GONG Yaming. Promoter of Vegetable Soybean GmDi19-3 Responds to Salt Stress and Exogenous ABA and MeJA[J]. Acta Horticulturae Sinica, 2024, 51(12): 2791-2799.

图/表 7

表1 菜用大豆GmDi19-3启动子顺式作用元件分析

Table 1 Distribution of cis-acting elements in the GmDi19-3 promoter of vegetable soybean

元件名称 Element name	核心序列 Core sequence	数量 Number	位置/bp Location		功能 Function
元件名称 Element name	核心序列 Core sequence	数量 Number	（+）链（+）Strand	（-）链（-）Strand	功能 Function
ABRE	ACGTG	2	452 757		脱落酸响应ABA responsiveness
ARE	AAACCA	2	498 958		厌氧诱导 Anaerobic induction
CGTCA基序 GTCA-motif	CGTCA	1		755	茉莉酸响应MeJA responsiveness
ERE	ATTTTAAA	1		236	乙烯响应ET responsiveness
LTR	CCGAAA	1		947	低温响应Low temperature responsiveness
MYB	CAACCA	2	502	312	激素信号诱导和非生物胁迫响应 Hormone signal induction andabiotic stress responsiveness
MYB识别位点 MYB recognition site	CCGTTG	1	605
MYC	GTTTAC	1		133	植物生长和防御响应Plant growth and defense responsiveness
TCA元件 TCA-element	CCATCTTTTT	1		831	水杨酸响应SA responsiveness
TGA元件 TGA-element	AACGAC	1		485	生长素响应Auxin responsiveness
TGACG基序 TGACG-motif	TGACG	1		191	茉莉酸响应MeJA responsiveness

图1 菜用大豆中GmDi19-3在不同组织中的表达 **：叶与茎、根与茎之间比较有显著差异（t检验，P < 0.01）。

Fig. 1 Expression patterns of GmDi19-3 in different tissues of vegetable soybean **：Significant differences in comparison between the leaf and stem，root and stem（t-test，P < 0.01）.

图2 GmDi19-3启动子在转基因拟南芥幼苗、主根和侧根GUS染色

Fig. 2 GUS staining of GmDi19-3 promoter in young seedling, primary and lateral roots of transgenic Arabidopsis

图3 菜用大豆幼苗在盐、外源ABA和MeJA处理条件下的GmDi19-3表达量 **：处理与对照比较有显著差异（t检验，P < 0.01）。下同。

Fig. 3 Expression patterns of GmDi19-3 in vegetable soybean after salt，exogenous ABA and MeJA treatments **：Significant differences in comparison between the treatment and control seedlings（t-test，P < 0.01）. The same below.

图4 盐胁迫处理下proGmDi19-3::GUS转基因拟南芥幼苗的GUS染色和活性

Fig. 4 GUS staining and activity of proGmDi19-3::GUS promoter in transgenic Arabidopsis seedlings after salt treatment

图5 外源ABA处理下proGmDi19-3::GUS转基因拟南芥幼苗的GUS染色和活性

Fig. 5 GUS staining and activity of proGmDi19-3::GUS promoter in transgenic Arabidopsis seedlings after exogenous ABA treatment

图6 外源MeJA处理下proGmDi19-3::GUS转基因拟南芥幼苗的GUS染色和活性

Fig. 6 GUS staining and activity of proGmDi19-3::GUS promoter in transgenic Arabidopsis seedlings after exogenous MeJA treatment

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