›› 2007, Vol. 15 ›› Issue (5): 418-422.DOI: 10.11733/j.issn.1007-0435.2007.05.004

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Influencing Factors on Alfalfa Transformation with Atriplex dimorphostegia NHX Gene Mediated by Agrobacterium tumefaciens

ZHAO Hong-juan, ZHANG Bo, LI Pei-ying, CHEN Ai-ping   

  1. Xinjiang Key Laboratory of Grassland Resources and Ecology, College of Grassland Science, Xinjiang Agricultural University, Urumqi, Xinjiang 830052, China
  • Received:2007-06-04 Revised:2007-07-03 Online:2007-10-15 Published:2007-10-15

农杆菌介导犁苞滨藜NHX基因转化苜蓿的影响因素

赵红娟, 张博, 李培英, 陈爱萍   

  1. 新疆农业大学草业工程学院, 新疆草地资源与生态自治区重点实验室, 新疆乌鲁木齐, 830052
  • 通讯作者: 张博,E-mail:xjauzb@126.com
  • 作者简介:赵红娟(1979- ),女,宁夏人,硕士研究生,主要从事牧草生物技术育种研究
  • 基金资助:
    新疆维吾尔自治区教育厅高校重点研究项目(XJEDU2004118)

Abstract: The leaves and cotyledons of Medicago varia Martin cv. Xinmu No.1 and Medicago sativa L. cv. Xinjiang Daye were used as materials and influencing factors on the salt-tolerant gene (NHX) transformation of Atriplex dimorphostegia Kar. et. Kir. by Agrobacterium tumefaciens EHA105 were studied. The results showed that the optimum transformation condition for cotyledon precultured 3 days and bacterial concentration of OD600 0.4~0.5 was immersing for 10~15 minutes, then laying cotyledons in co-culture medium with addition of 20~30 mg/L acetosyringone for 4 days. The PCR analysis showed that the NHX gene had been integrated into the genome of alfalfa.

Key words: Alfalfa, Agrobacterium tumefaciens, NHX gene, Genetic transformation

摘要: 以新牧1号杂花苜蓿(Medicago varia Martin.cv.Xinmu No.1)和新疆大叶苜蓿(Medicago sativa L.cv.Xinjiang Daye)无菌苗叶片和子叶为材料,通过根癌农杆菌EHA105介导转化犁苞滨藜NHX耐盐基因,提高植株耐盐性。本文对农杆菌转化条件进行研究,结果表明:以预培养3d的子叶为转化受体,菌液浓度OD600为0.4~0.5时,浸染10~15min,然后转入附加20~30mg/L乙酰丁香酮的共培养基,培养4d,可以获得较高的转化效率;对抗性芽的PCR检测初步证明,外源基因整合到苜蓿的基因组中。

关键词: 苜蓿, 农杆菌, NHX基因, 遗传转化

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