杨梅MrSPL4在响应干旱和低温胁迫中的作用研究

doi:10.16420/j.issn.0513-353x.2023-0258

摘要/Abstract

摘要：

通过分析杨梅MrSPL4基因启动子顺式作用元件和对MrSPL4-OE转基因烟草T₂代3个株系进行干旱与低温胁迫处理，观察其种子萌发率和植株表型，测定植株生理指标，以期明确MrSPL4在响应干旱和低温等非生物胁迫中的功能。MrSPL4启动子区包含干旱诱导、低温响应、防御和胁迫响应等8类顺式作用元件。干旱胁迫下，与野生型烟草相比，MrSPL4-OE转基因烟草种子萌发率显著降低，但植株失水和萎蔫程度差异不明显，除H₂O₂含量显著降低外，MDA、脯氨酸和可溶性糖含量以及SOD、POD活性差异不明显；低温胁迫后转移至正常条件下，种子萌发率显著升高，植株失绿和萎蔫程度降低，SOD、POD酶活性显著升高，MDA、H₂O₂、脯氨酸和可溶性糖含量均显著降低。上述结果表明，杨梅MrSPL4正向调控了转基因烟草低温胁迫响应过程，提高了其耐低温能力，但抑制了干旱胁迫下种子的萌发。

关键词: 杨梅, SPL基因, 种子萌发, 干旱胁迫, 低温胁迫

Abstract:

To investigate the function of MrSPL4 in response to drought and low temperature stress in Chinese bayberry，the cis-acting element of MrSPL4 promoter and treating MrSPL4-OE transgenic tobacco T₂ generation strain with drought and low temperature stress were analyzed，observing its seed germination rate and plant phenotype，and measuring plant physiological indexes to clarify the function of MrSPL4. The results showed that the promoter region of MrSPL4 contains cis-acting elements for drought induction，low temperature response，defense and stress response，and the other important cis-elements. Compared with wild-type tobacco（WT），under drought stress，MrSPL4-OE transgenic tobacco seed germination rate was significantly reduced. There was no significant difference in the reduction of plant water loss and wilting between WT and MrSPL4-OE. Except for a significant decrease in H₂O₂ content，there was no significant difference in other indicators. Transfer to normal conditions after low temperature stress，the germination rate of seeds significantly increased. Plant greening and wilting were reduced. SOD and POD enzyme activities increased significantly，while the content of MDA，H₂O₂，proline，and soluble sugar decreased significantly. The results indicated that MrSPL4 positively regulates the response process of tobacco to low temperature stress，improves its ability to tolerate low temperature，but inhibits the germination rate of tobacco seeds under drought stress.

Key words: Chinese bayberry, SPL gene, seed germination, drought stress, low-temperature stress

武祥琪, 孙鹂, 俞浙萍, 俞沁佩, 梁森苗, 郑锡良, 戚行江, 张淑文. 杨梅MrSPL4在响应干旱和低温胁迫中的作用研究[J]. 园艺学报, 2024, 51(5): 927-938.

WU Xiangqi, SUN Li, YU Zheping, YU Qinpei, LIANG Senmiao, ZHENG Xiliang, QI Xingjiang, ZHANG Shuwen. Functional Study of MrSPL4 Gene in Response to Drought and Low Temperature Stress in Chinese Bayberry[J]. Acta Horticulturae Sinica, 2024, 51(5): 927-938.

图/表 7

图1 MrSPL4在烟草转基因株系（MrSPL4-OE）及其野生型（WT）中的相对表达量

Fig. 1 Relative expression levels of MrSPL4 in transgenic tobacco lines（MrSPL4-OE）and wild-type（WT） t-test，*** P < 0.001.

表1 MrSPL4启动子区的相关顺式作用元件

Table 1 Predicted cis-elements in the promoter of the MrSPL4 gene

顺式作用元件	数量	起始位置/bp	终止位置/bp
cis-Element	Number	Starting position	Ending position
赤霉素应答元件 Gibberellin responsiveness	1	193	203
干旱诱导元件 Drought inducibility	3	79	89
		1 450	1 460
		1 491	1 501
光反应元件 Light responsiveness	3	737	747
		1 434	1 444
		1 417	1 427
生长素响应元件 Auxin responsiveness	1	1 373	1 383
低温响应元件 Low-temperature responsiveness	2	251	261
		1 405	1 415
厌氧诱导元件 Anaerobic induction	2	275	285
		1 454	1 464
防御和胁迫响应元件 Defense and stress responsiveness	2	852	862
		1 428	1 438
茉莉酸甲酯响应元件 MeJA responsiveness	4	736	746
		1 462	1 472
		736	746
		1 462	1 472

图2 烟草转基因株系（MrSPL4-OE）及其野生型（WT）种子在甘露醇处理下（14 d）的发芽情况

Fig. 2 Germination of transgenic tobacco lines（MrSPL4-OE）and wild-type（WT）seeds under different concentrations of mannitol treatment（14 days） ** P < 0.01.

图3 干旱胁迫下烟草转基因株系（MrSPL4-OE）及其野生型（WT）植株叶片生理指标的变化

Fig. 3 Leaf physiological indicators of transgenic tobacco lines（MrSPL4-OE）and wild-type（WT）plants under drought stress ** P < 0.01.

图4 低温处理下烟草转基因株系（MrSPL4-OE）及其野生型（WT）种子的萌发率

Fig. 4 Germination rate of transgenic tobacco lines（MrSPL4-OE）and wild-type（WT）seeds under low-temperature treatment ** P < 0.01.

图5 烟草转基因株系（MrSPL4-OE）及其野生型（WT）低温处理前后的表型

Fig. 5 Phenotypes of transgenic tobacco lines（MrSPL4-OE）and wild-type（WT）before and after low-temperature treatment

图6 低温胁迫下烟草转基因株系（MrSPL4-OE）及其野生型（WT）叶片生理指标的变化

Fig. 6 Leaf physiological indicators of transgenic tobacco lines（MrSPL4-OE）and wild-type（WT）under low temperature stress * P < 0.05，** 0.01 < P < 0.05，*** P < 0.001.

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