马铃薯StDWF1基因克隆及表达分析

doi:10.3969/j.issn.1004-1524.2018.06.04

浙江农业学报 ›› 2018, Vol. 30 ›› Issue (6): 909-917.DOI: 10.3969/j.issn.1004-1524.2018.06.04

马铃薯StDWF1基因克隆及表达分析

唐晓¹, 邓孟胜¹, 邹雪², 李立芹^{1, 3}, 祝渊智^{1, 3}, 王西瑶^{1, 3, *}

1.四川农业大学农学院马铃薯研究与开发中心,四川成都 611130;
2.绵阳市农业科学院,四川绵阳 621000;
3.作物科学国家级实验教学示范中心(四川农业大学),四川成都 611130

收稿日期:2018-01-04 出版日期:2018-06-20 发布日期:2018-07-02
通讯作者: 王西瑶,E-mail:1357664714@qq.com
作者简介:唐晓(1992—),男,四川成都人,硕士研究生,研究方向为马铃薯分子生物学。E-mail: 354550329@qq.com
基金资助:
四川省学术和技术带头人培养经费(03120233); 四川农业大学大学生科研兴趣培养计划(ky2016247)

Cloning and expression analysis of StDWF1 in Solanum tuberosum

TANG Xiao¹, DENG Mengsheng¹, ZOU Xue², Li Liqin^{1, 3}, ZHU Yuanzhi^{1, 3}, WANG Xiyao^{1, 3, *}

1. Potato Research and Development Center, College of Agronomy,Sichuan Agricultural University, Chengdu 611130, China;
2. Mianyang Academy of Agricultural Sciences, Mianyang 621000, China;
3. National Demonstration Center for Experimental Crop Science Education (Sichuan Agricultural University), Chengdu 611130, China

Received:2018-01-04 Online:2018-06-20 Published:2018-07-02

摘要/Abstract

摘要： DWF1是油菜素内酯(BR)生物合成途径中的关键基因。为探索StDWF1在马铃薯生长发育中的功能,本实验以马铃薯品种费乌瑞它(Favorita)试管苗为材料,克隆StDWF1基因全长序列,开放阅读框(ORF)为1 704 bp,编码567个氨基酸,蛋白质相对分子质量为66.08 ku,理论等电点为7.96。利用农杆菌介导法在烟草中瞬时表达StDWF1,共聚焦显微镜观察显示StDWF1定位于细胞质。荧光定量分析表明,嫩叶中StDWF1表达量最高,其次是老叶、匍匐茎、主根、茎、块茎。对马铃薯生育期叶片StDWF1表达量进行分析,结果表明在出苗期表达量最高。本实验结果为进一步阐释BR对马铃薯生长发育的调控机制奠定理论基础。

关键词: 马铃薯, StDWF1, 基因克隆, 表达分析, 亚细胞定位

Abstract: The DWF1 is a key gene in brassinosteroid (BR) biosynthesis pathway. To explore the function of StDWF1 in the growth and development of potato, this experiment was based on the potato variety Favorita test-tube plantlets, and we cloned the full-length sequence of StDWF1 gene. The open reading frame (ORF) of StDWF1 gene was 1 704 bp, and encoded 567 amino acids. The molecular weight of protein was 66.08 ku, and the isoelectric point (pI) was 7.96. Agrobacterium tumefaciens-mediated transient expression was carried out in tobacco, and laser scanning confocal microscopy showed that potato StDWF1 was located in cytoplasm. Fluorescence quantitative analysis showed StDWF1 expression quantity was the highest in new leaves, followed by old leaves, stolon, roots, stems and tubers. We analyzed the expression of StDWF1 in potato leaves at the growth period, and the results showed that the highest expression quantity was in potato seedling stage. The results of this experiment laid the theoretical foundation for further interpreting the regulation mechanism of BR in the potato growth and development.

Key words: potato, DWF1, gene cloning, expression analysis, subcellular localization

中图分类号:

S532

唐晓, 邓孟胜, 邹雪, 李立芹, 祝渊智, 王西瑶. 马铃薯StDWF1基因克隆及表达分析[J]. 浙江农业学报, 2018, 30(6): 909-917.

TANG Xiao, DENG Mengsheng, ZOU Xue, Li Liqin, ZHU Yuanzhi, WANG Xiyao. Cloning and expression analysis of StDWF1 in Solanum tuberosum[J]. , 2018, 30(6): 909-917.

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马铃薯StDWF1基因克隆及表达分析

Cloning and expression analysis of StDWF1 in Solanum tuberosum

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