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园艺学报 ›› 2021, Vol. 48 ›› Issue (3): 531-538.doi: 10.16420/j.issn.0513-353x.2020-0230

• 研究论文 • 上一篇    下一篇

STTM技术沉默马铃薯Stu-miR156对其侧根发育的影响

罗红玉1, 杨江伟1, 冯亚1, 张欢欢1, 刘升燕1, 张宁1,2,*(), 司怀军1,2,*   

  1. 1甘肃农业大学生命科学技术学院,兰州 730070
    2甘肃省干旱生境作物学省部共建国家重点实验室培育基地,兰州 730070
  • 收稿日期:2020-05-10 出版日期:2021-03-25 发布日期:2021-04-02
  • 通讯作者: 张宁,司怀军 E-mail:ningzh@gsau.edu.cn
  • 基金资助:
    国家自然科学基金项目(31660416);国家自然科学基金项目(31960444);国家自然科学基金项目(31860400)

The Effect of Stu-miR156 Silencing by STTM Technology on Potato Lateral Root Development

LUO Hongyu1, YANG Jiangwei1, FENG Ya1, ZHANG Huanhuan1, LIU Shengyan1, ZHANG Ning1,2,*(), SI Huaijun1,2,*   

  1. 1College of Life Science and Technology,Gansu Agricultural University,Lanzhou 730070,China
    2Gansu Provincial Key Laboratory of Aridland Crop Science,Lanzhou 730070,China
  • Received:2020-05-10 Online:2021-03-25 Published:2021-04-02
  • Contact: ZHANG Ning,SI Huaijun E-mail:ningzh@gsau.edu.cn

摘要:

为了阐明Stu-miR156在马铃薯侧根发育中的生物学功能,以马铃薯栽培品种‘Desiree’为试验材料,利用短串联靶标模拟物(Short tandem target mimic,STTM)技术,成功构建了Stu-miR156沉默表达载体,通过农杆菌介导法转化马铃薯获得了转化植株,qRT-PCR技术检测Stu-miR156及其靶基因StSPL9在转基因植株中的表达量。并通过构建pCAM-GFP-StSPL9融合蛋白表达载体转化烟草,发现靶基因StSPL9位于细胞质和细胞核中。qRT-PCR分析结果表明:在马铃薯转基因株系L1和L2中,Stu-miR156表达量受到严重抑制,在根、茎和叶中均不同程度地下调,其靶基因StSPL9均上调表达。表型分析表明:与野生型对照相比,转基因植株的侧根数量显著减少,生长受到抑制。

关键词: 马铃薯, Stu-miR156, STTM, StSPL9, 遗传转化, 侧根

Abstract:

To elucidate the biological function of Stu-miR156 in potato lateral root development,plantlets in vitro of potato cultivar‘Desiree’were used as dornors. Using STTM(Short tandem target mimic,STTM)technology,the Stu-miR156 silencing expression vector was successfully constructed and further the transgenic plants were obtained viaAgrobacterium-mediated method. qRT-PCR was used to determine the differential expression of Stu-miR156 and its target gene StSPL9in the transgenic potato plants. We constructed a pCAM-GFP-StSPL9 fusion protein expression vector and transformed it into tobacco,thereby confirming that StSPL9 was localized in the cytoplasm and nucleus. The result of qRT-PCR showed that the expression levels of Stu-miR156 in roots,stems and leaves were severely inhibited,while the target gene StSPL9 was up-regulated in the transgenic potato lines L1 and L2. The number of lateral roots of the transgenic plants was significantly reduced and the growth is inhibited compared with the wild type plants.

Key words: potato, Stu-miR156, STTM, StSPL9, genetic transformation, lateral root

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