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草业学报 ›› 2015, Vol. 24 ›› Issue (2): 142-147.DOI: 10.11686/cyxb20150216

• 研究论文 • 上一篇    下一篇

马铃薯ARF基因RNAi载体的遗传转化及对其试管薯生理特性的影响

裴瑞芳1, 2, 刘英1, 2, 刘柏林1, 张宁2, 司怀军1, 2*, *, 王蒂1*, *   

  1. 1.甘肃省作物遗传改良与种质创新重点实验室,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,甘肃 兰州 730070;
    2.甘肃农业大学生命科学技术学院,甘肃 兰州 730070
  • 收稿日期:2014-02-25 出版日期:2015-02-20 发布日期:2015-02-20
  • 作者简介:裴瑞芳(1990-),女,甘肃天水人,在读硕士。E-mail:peirfsmile@163.com
  • 基金资助:
    甘肃省杰出青年基金项目(1308RJDA011)和国家自然科学基金项目(31160298)资助

Genetic transformation using an RNAi vector containing an ARF gene in potato and effects on physiological characteristics of microtuber

PEI Ruifang1, 2, LIU Ying1, 2, LIU Bailin1, ZHANG Ning2, SI Huaijun1, 2, *, WANG Di1, *   

  1. 1.Gansu Key Laboratory of Crop Genetic and Germplasm Enhancement, Gansu Provincial Key Laboratory of Aridland Crop Science, Lanzhou 730070, China;
    2.College of Life Science and Technology, Gansu Agricultural University, Lanzhou 730070, China
  • Received:2014-02-25 Online:2015-02-20 Published:2015-02-20

摘要: 用含有ADP核糖基化因子(ARF)基因的干扰载体的农杆菌转化马铃薯栽培品种“甘农薯2号”,获得转化植株48株,经卡那霉素抗性筛选和PCR检测证明ARF基因已成功整合到马铃薯基因组中。用实时荧光定量PCR检测表明,转化植株中ARF基因的表达较对照均有不同程度的下降,在转基因株系Z-11和Z-12中,ARF基因表达的干扰程度分别高达92.53%和93.22%。对转基因植株诱导的试管薯的淀粉、可溶性糖、蛋白质、蔗糖和葡萄糖含量测定结果表明,与未转基因的对照相比,转基因植株的淀粉含量提高了4.29%~34.27%,蛋白质含量提高了1.47%~7.35%,可溶性糖含量提高了1.44%~17.39%;蔗糖含量提高了11.53%~53.84%,而葡萄糖含量降低了6.06%~21.21%。

Abstract: Using potato (Solanum tuberosum) cultivar ‘Gannongshu 2’ as donor, 48 transgenic plants were obtained through an Agrobacterium-mediated transformation method using an RNAi vector containing an ADP-ribosylation factors (ARF) gene. Testing of the transformed plants on a culture media containing kanmycin and PCR assay showed that the AFR gene had been successfully integrated into the potato genome. The analysis of real time fluorescence quantitative polymerase chain reaction (qRT-PCR) data showed that the ARF gene expression declined to varying degrees in the transgenic plants compared with the controls, and interference degree reached 92.53% and 93.22% in the transgenic lines Z-11 and Z-12, respectively. The starch content of the transgenic plants increased by 4.29%-34.27%, the protein content by 1.47%-7.35%, the soluble sugar content by 1.44%-17.39%, and the sucrose content by 11.53%-53.84% compared with control plants; however, the glucose content of the transgenic plants decreased by 6.06%-21.21% compared with control plants.