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从君迁子中克隆DlERF10基因及该基因在转基因烟草植株中的耐寒性分析

Cloning of the DlERF10 gene from Diospyros lotus L. and cold tolerance analysis of the DlERF10 gene in transgenic tobacco plants.

作者信息

Zhou Ruijin, Li Shuda, Zhang Xiaona, Wang Yingying, Hu Huiling

机构信息

School of Horticulture Landscape Architecture, Henan Institute of Science and Technology, Xinxiang, Henan, China.

Henan Province Engineering Research Centers of Horticultural Plant Resource Utilization and Germplasm Enhancement, Xinxiang, Henan, China.

出版信息

PLoS One. 2025 Mar 3;20(3):e0314135. doi: 10.1371/journal.pone.0314135. eCollection 2025.

Abstract

In the north of China, Diospyros plants are vulnerable to low-temperature damage in winter and is considered as a major factor restricting the development of the persimmon industry in Northern China. Diospyros lotus L. is featured by high survival potential of seedlings, cold tolerance, and grafting affinity with D. kaki Thunb. D. lotus has been frequently used as rootstocks for Diospyros spp. ERF transcriptional factors are a subfamily of the AP2/ERF gene family and play an important role in plant growth and stress tolerance. To explore the structure and function of the ERF transcription factors in D. lotus, we performed RT-PCR to clone DlERF10 from the leaves. The DlERF10 gene was 1104 bp long, encoding 367 amino acids. In order to deeply study the cold tolerance of DlERF10 gene, the pBI121-DlERF10 overexpression vector was constructed, and agrobacterium-mediated transformation was carried out to transfer the gene into tobacco plants. The wild-type and transgenic tobacco plants were subjected to low-temperature stress. The results showed that the transgenic plants were less severely damaged by low-temperature stress than the wild-type plants. Besides, the SOD, POD and CAT activities of leaves enhanced, and PRO contents of leaves increased, while the MDA content decreased. It was concluded that the DlERF10 gene increased the activity of protective enzymes in tobacco plants, thereby strengthening the tolerance to low-temperature stress. The present study proposes a candidate gene for engineering cold stress tolerance in Diospyros spp.

摘要

在中国北方,柿属植物冬季易受低温伤害,这被认为是限制中国北方柿产业发展的主要因素。君迁子具有幼苗成活率高、耐寒以及与柿嫁接亲和力强的特点。君迁子常被用作柿属植物的砧木。ERF转录因子是AP2/ERF基因家族的一个亚家族,在植物生长和胁迫耐受性中发挥重要作用。为了探究君迁子中ERF转录因子的结构与功能,我们通过RT-PCR从叶片中克隆了DlERF10。DlERF10基因长1104 bp,编码367个氨基酸。为深入研究DlERF10基因的耐寒性,构建了pBI121-DlERF10过表达载体,并通过农杆菌介导的转化将该基因导入烟草植株。对野生型和转基因烟草植株进行低温胁迫处理。结果表明,转基因植株受低温胁迫的损伤程度低于野生型植株。此外,叶片的超氧化物歧化酶(SOD)、过氧化物酶(POD)和过氧化氢酶(CAT)活性增强,脯氨酸(PRO)含量增加,而丙二醛(MDA)含量降低。得出结论:DlERF10基因提高了烟草植株中保护酶的活性,从而增强了对低温胁迫的耐受性。本研究为柿属植物抗寒基因工程提供了一个候选基因。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/936d/11875332/18a6cfeb3b1d/pone.0314135.g001.jpg

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