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番茄中CBF表达诱导因子1(ICE1)的克隆与转化

Cloning and transformation of INDUCER of CBF EXPRESSION1 (ICE1) in tomato.

作者信息

Yu X H, Juan J X, Gao Z L, Zhang Y, Li W Y, Jiang X M

机构信息

Key Laboratory of Biology and Genetic Improvement of Horticultural Crops, Northeast Region, Ministry of Agriculture, Northeast Agriculture University, Harbin, China.

Heilongjiang Academy of Sciences Natural and Ecological Research Institute, Xiangfang District, Harbin, Heilongjiang, China.

出版信息

Genet Mol Res. 2015 Oct 27;14(4):13131-43. doi: 10.4238/2015.October.26.9.

Abstract

The tomato plant (Solanum lycopersicum Mill.) is sensitive to cold, and low field temperatures can result in shortened growth periods and decreased crop yield. Transcription of CRT/DRE-binding factor (CBF) is regulated by INDUCER of CBF EXPRESSION1 (ICE1). CBF activates many downstream genes that confer cold tolerance on plants. ICE1 has been used in genetic engineering to improve cold-resistance in several plant species. Here, ICE1 in a plant expression vector was used to transform a tissue-cultured rhubarb tomato variety using Agrobacterium tumefaciens. The transgenic and control plants were compared at 4°C for 0, 24, and 72 h. We measured leaf physiological indicators related to cold resistance, including malondialdehyde (MDA) and proline (Pro) contents, and peroxidase (POD) and catalase (CAT) activities. At 72 h, the MDA content in transgenic plants was significantly lower than in control plants, indicating a lower membrane lipid injury. The Pro contents and the CAT and POD activities in the transgenic plants increased significantly compared with those of the control plants. For Pro, the increase continued over the prolonged stress exposure, while CAT and POD activities reached peak levels at 24 h. These results are consistent with the roles of Pro, CAT, and POD in defending the integrity of plant cells. Our study not only improves the cold resistance of tomato, but also provides the foundation for further research on the role of ICE1 as a transcription factor in plant cold resistance.

摘要

番茄植株(Solanum lycopersicum Mill.)对低温敏感,田间低温会导致生长周期缩短和作物产量下降。CRT/DRE结合因子(CBF)的转录受CBF表达诱导因子1(ICE1)调控。CBF激活许多赋予植物耐寒性的下游基因。ICE1已被用于基因工程,以提高多种植物的抗寒性。在此,利用植物表达载体中的ICE1,通过根癌农杆菌转化组织培养的大黄番茄品种。将转基因植株和对照植株在4℃下分别处理0、24和72小时后进行比较。我们测量了与抗寒性相关的叶片生理指标,包括丙二醛(MDA)和脯氨酸(Pro)含量以及过氧化物酶(POD)和过氧化氢酶(CAT)活性。在72小时时,转基因植株中的MDA含量显著低于对照植株,表明膜脂损伤较低。与对照植株相比,转基因植株中的Pro含量以及CAT和POD活性显著增加。对于Pro,在长时间胁迫暴露下其含量持续增加,而CAT和POD活性在24小时时达到峰值水平。这些结果与Pro、CAT和POD在保护植物细胞完整性中的作用一致。我们的研究不仅提高了番茄的抗寒性,还为进一步研究ICE1作为转录因子在植物抗寒中的作用奠定了基础。

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