Construction of Blue Light Receptor CRY2B over Expression Vector and Genetic Transformation in Medicago Sativa

doi:10.11733/j.issn.1007-0435.2017.05.017

Acta Agrestia Sinica ›› 2017, Vol. 25 ›› Issue (5): 1029-1036.DOI: 10.11733/j.issn.1007-0435.2017.05.017

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Construction of Blue Light Receptor CRY2B over Expression Vector and Genetic Transformation in Medicago Sativa

FAN Wen-na¹, LI Run-lin¹, WANG Zhan-bin¹, DU Hong-qi², ZHAO Ling-ping¹, WANG Cheng-zhang²

1. College of Animal Science and Technology, Henan University of Science and Technology, Luoyang, Henan Province 471003, China;
2. College of Animal Science and Veterinary Medicine, Henan Agricultural University, Zhengzhou, Henan Province 450002, China

Received:2016-04-26 Revised:2017-09-14 Online:2017-10-15 Published:2018-01-25

紫花苜蓿蓝光受体CRY2B过表达载体构建及转化

樊文娜¹, 李润林¹, 王占彬¹, 杜红旗², 赵凌平¹, 王成章²

1. 河南科技大学动物科技学院, 河南洛阳 471003;
2. 河南农业大学牧医工程学院, 河南郑州 450002

通讯作者: 王成章,E-mail:wangchengzhang@263.net
作者简介:樊文娜(1981-),女,河南许昌人,博士,讲师,E-mail:chou0516@163.com
基金资助:
河南科技大学博士科研启动基金（13480078）；国家牧草产业技术体系（CARS-35）资助

Abstract

Abstract:

The short-day is one of the main factors affected alfalfa fall dormancy, which is called as the photoperiodic effect. Based on the results of alfalfa transcriptome sequencing, the relationship between the main photoreceptors-CRY2B gene and alfalfa fall dormancy may reveal the regulation mechanism of alfalfa fall dormancy. In this experiment, cryptochrome gene——CRY2B of Reindeer (the variety of fall dormancy class NO.1) with complete open reading frame (696~2553 bp) was cloned. For the design of specific primers with the restriction sites (CRY2B-F and CRY2B-R), the cloned fragment was connected to the plant expression vector pCAMBIA3301 to get a recombinant vector pCAMBIA3301-CRY2B after enzymes digestion, clean up and ligation. Agrobacterium-mediated genetic transformation using alfalfa cotyledonary node was explored, it laid the foundation for the further research of CRY2B regulating fall dormancy molecular mechanism.

Key words: Medicago sativa, CRY2B, Fall dormancy, Vector construct, Transformation

摘要：

短日照是影响苜蓿秋眠性的主要因素，即光周期效应。根据转录组测序的结果，对主要的蓝光受体隐花色素CYR2B基因和苜蓿秋眠性之间关系的研究可以从光受体途径来揭示苜蓿秋眠性调控机制，本试验首先克隆紫花苜蓿1级品种‘驯鹿’隐花色素CRY2B基因的完整开放阅读框（696~2553 bp），设计1对含有酶切位点的特异性引物CRY2B-F，CRY2B-R，通过多次酶切和连接，将克隆目的片段连接到植物表达载体pCAMBIA3301上得到重组载体pCAMBIA3301-CRY2B，成功地构建了紫花苜蓿CRY2B的过表达载体，并利用农杆菌介导pCAMBIA3301-CRY2B成功转化苜蓿植株，为进一步研究CRY2B调控苜蓿秋眠性分子机制奠定基础。

关键词: 苜蓿, CRY2B, 秋眠性, 载体构建, 转化

CLC Number:

S541

FAN Wen-na, LI Run-lin, WANG Zhan-bin, DU Hong-qi, ZHAO Ling-ping, WANG Cheng-zhang. Construction of Blue Light Receptor CRY2B over Expression Vector and Genetic Transformation in Medicago Sativa[J]. Acta Agrestia Sinica, 2017, 25(5): 1029-1036.

樊文娜, 李润林, 王占彬, 杜红旗, 赵凌平, 王成章. 紫花苜蓿蓝光受体CRY2B过表达载体构建及转化[J]. 草地学报, 2017, 25(5): 1029-1036.

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Construction of Blue Light Receptor CRY2B over Expression Vector and Genetic Transformation in Medicago Sativa

紫花苜蓿蓝光受体CRY2B过表达载体构建及转化

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