东南景天耐镉相关基因SaFer的克隆与功能初步分析

赵婷; 韩小娇; 刘明英; 乔桂荣; 蒋晶; 姜彦成; 卓仁英

doi:10.11833/j.issn.2095-0756.2015.01.004

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东南景天耐镉相关基因SaFer的克隆与功能初步分析

赵婷 , 韩小娇 , 刘明英 , 乔桂荣 , 蒋晶 , 姜彦成 , 卓仁英

文章导航 > 浙江农林大学学报 > 2015 > 32(1): 25-32

赵婷, 韩小娇, 刘明英, 乔桂荣, 蒋晶, 姜彦成, 卓仁英. 东南景天耐镉相关基因SaFer的克隆与功能初步分析[J]. 浙江农林大学学报, 2015, 32(1): 25-32. doi: 10.11833/j.issn.2095-0756.2015.01.004

引用本文:

ZHAO Ting, HAN Xiaojiao, LIU Mingying, QIAO Guirong, JIANG Jing, JIANG Yancheng, ZHUO Renying. Isolation and expression of a cadmium-resistant gene （SaFer） from Sedum alfredii[J]. Journal of Zhejiang A&F University, 2015, 32(1): 25-32. doi: 10.11833/j.issn.2095-0756.2015.01.004

Citation:

ZHAO Ting, HAN Xiaojiao, LIU Mingying, QIAO Guirong, JIANG Jing, JIANG Yancheng, ZHUO Renying. Isolation and expression of a cadmium-resistant gene （SaFer） from Sedum alfredii[J]. Journal of Zhejiang A&F University, 2015, 32(1): 25-32. doi: 10.11833/j.issn.2095-0756.2015.01.004

东南景天耐镉相关基因SaFer的克隆与功能初步分析

doi: 10.11833/j.issn.2095-0756.2015.01.004

赵婷^1,2,3,
韩小娇^2,3,
刘明英^2,3,
乔桂荣^2,3,
蒋晶^2,3,
姜彦成¹,
卓仁英^2,3, ,

1.
新疆大学生命科学与技术学院，新疆乌鲁木齐 830046;
2.
中国林业科学研究院亚热带林业研究所，浙江富阳 311400;
3.
中国林业科学研究院林木遗传育种国家重点实验室，北京 100091

基金项目:

国家自然科学基金资助项目（31200465）;国家高技术研究发展计划（863计划）项目（2013AA102701-3）

详细信息

作者简介: 赵婷，从事植物分子生物学研究.

通信作者: 卓仁英，研究员，博士，从事植物抗逆分子生物学研究

中图分类号: S718.3；Q943.2

Isolation and expression of a cadmium-resistant gene （SaFer） from Sedum alfredii

1.
Department of Biology and Technology, Xinjiang University, Urumqi 830046, Xinjiang, China;
2.
The Research Institute of Subtropical Forestry, Chinese Academy of Forestry, Fuyang 311400, Zhejiang, China;
3.
State Key Laboratory of Tree Genetics and Breeding, Chinese Academy of Forestry, Beijing 100091, China

摘要: 铁蛋白（ferritin）是一种专门存储铁的重要胁迫相关蛋白，参与镉离子吸收的调控。从构建的东南景天Sedum alfredii cDNA文库中筛选出东南景天Ferritin基因cDNA全长，命名为SaFer，编码序列为1 117 bp，开放阅读框为759 bp，编码252个氨基酸。以东南景天基因组DNA为模板分离到SaFer基因组序列，长度为1 702 bp，含有7个内含子。氨基酸序列同源性分析表明：SaFer与苹果Malus domestica Ferritin亲缘关系最近，同源性高达76%。实时荧光定量聚合酶链式反应（qRT-PCR）分析发现：根系中SaFer 基因在镉胁迫12 h后表达显著上调。镉胁迫实验显示，转基因拟南芥Arabidopsis thaliana比野生型有更高的耐镉胁迫能力。SaFer 基因的分离及其初步功能验证为研究东南景天耐镉机制和林木耐镉转基因育种提供了理论依据。图9表1参23
- 植物学 /
- 东南景天 /
- 铁蛋白（ferritin） /
- 镉胁迫 /
- 表达分析
Abstract: Abiotic stress was a serious problem that affect plants growth and products. In plants, ferritin is a special iron storage protein closely related to stress. A cDNA from the Sedum alfredii cDNA library, designated as SaFer, was isolated and analyzed by homologous analysis and by Real-Time Reverse Transcription-PCR （qRT-PCR）. Then, cadmium-stress experiments were conducted to compare transgenic Arabidopsis thaliana overexpressed SaFer to a wild type. Results showed that SaFer cDNA was 1 117 bp long with an opening reading frame （ORF） of 759 bp. The ORF of SaFer encoded a polypeptide of 252 amino acids with a calculated molecular weight of about 27.8 kDa, and the homologous analysis showed that it was most closely related to ferritin of Malus domestica with 76% identities. The length of the genomic sequence of SaFer was 1 702 bp and contained 7 introns. Expression profiles in roots were analyzed by qRT-PCR and the results showed that the transcription level of SaFer was enhanced after 12 h cadmium stress. Transgenic Arabidopsis thaliana overexpressing SaFer displayed much higher Cd tolerance than the wild type which was further supported by physiological indexes such as SOD activity and electric conductivity. The results showed that SaFer could provide a glimpse into S. alfredii cadmium-tolerance and could contribute to the breeding of cadmium-tolerant plants. ［Ch，9 fig. 1 tab. 23 ref. ］
- botany /
- Sedum alfredii /
- ferritin /
- cadmium stress /
- expression change

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东南景天耐镉相关基因SaFer的克隆与功能初步分析

doi: 10.11833/j.issn.2095-0756.2015.01.004

1. 新疆大学生命科学与技术学院，新疆乌鲁木齐 830046;

2. 中国林业科学研究院亚热带林业研究所，浙江富阳 311400;

3. 中国林业科学研究院林木遗传育种国家重点实验室，北京 100091

基金项目:

国家自然科学基金资助项目（31200465）;国家高技术研究发展计划（863计划）项目（2013AA102701-3）

作者简介:
赵婷，从事植物分子生物学研究.

通信作者: 卓仁英，研究员，博士，从事植物抗逆分子生物学研究

收稿日期: 2014-03-01
修回日期: 2014-04-14
刊出日期: 2015-02-20

中图分类号: S718.3；Q943.2

关键词:

摘要: 铁蛋白（ferritin）是一种专门存储铁的重要胁迫相关蛋白，参与镉离子吸收的调控。从构建的东南景天Sedum alfredii cDNA文库中筛选出东南景天Ferritin基因cDNA全长，命名为SaFer，编码序列为1 117 bp，开放阅读框为759 bp，编码252个氨基酸。以东南景天基因组DNA为模板分离到SaFer基因组序列，长度为1 702 bp，含有7个内含子。氨基酸序列同源性分析表明：SaFer与苹果Malus domestica Ferritin亲缘关系最近，同源性高达76%。实时荧光定量聚合酶链式反应（qRT-PCR）分析发现：根系中SaFer 基因在镉胁迫12 h后表达显著上调。镉胁迫实验显示，转基因拟南芥Arabidopsis thaliana比野生型有更高的耐镉胁迫能力。SaFer 基因的分离及其初步功能验证为研究东南景天耐镉机制和林木耐镉转基因育种提供了理论依据。图9表1参23

English Abstract

Isolation and expression of a cadmium-resistant gene （SaFer） from Sedum alfredii

1. Department of Biology and Technology, Xinjiang University, Urumqi 830046, Xinjiang, China;

2. The Research Institute of Subtropical Forestry, Chinese Academy of Forestry, Fuyang 311400, Zhejiang, China;

3. State Key Laboratory of Tree Genetics and Breeding, Chinese Academy of Forestry, Beijing 100091, China

Received Date: 2014-03-01
Rev Recd Date: 2014-04-14
Publish Date: 2015-02-20

Keywords:

Abstract: Abiotic stress was a serious problem that affect plants growth and products. In plants, ferritin is a special iron storage protein closely related to stress. A cDNA from the Sedum alfredii cDNA library, designated as SaFer, was isolated and analyzed by homologous analysis and by Real-Time Reverse Transcription-PCR （qRT-PCR）. Then, cadmium-stress experiments were conducted to compare transgenic Arabidopsis thaliana overexpressed SaFer to a wild type. Results showed that SaFer cDNA was 1 117 bp long with an opening reading frame （ORF） of 759 bp. The ORF of SaFer encoded a polypeptide of 252 amino acids with a calculated molecular weight of about 27.8 kDa, and the homologous analysis showed that it was most closely related to ferritin of Malus domestica with 76% identities. The length of the genomic sequence of SaFer was 1 702 bp and contained 7 introns. Expression profiles in roots were analyzed by qRT-PCR and the results showed that the transcription level of SaFer was enhanced after 12 h cadmium stress. Transgenic Arabidopsis thaliana overexpressing SaFer displayed much higher Cd tolerance than the wild type which was further supported by physiological indexes such as SOD activity and electric conductivity. The results showed that SaFer could provide a glimpse into S. alfredii cadmium-tolerance and could contribute to the breeding of cadmium-tolerant plants. ［Ch，9 fig. 1 tab. 23 ref. ］

引用本文: