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中国农学通报

中国农学通报 ›› 2017, Vol. 33 ›› Issue (15): 36-42.doi: 10.11924/j.issn.1000-6850.casb16060067

所属专题： 园艺

• 生物技术科学 • 上一篇    下一篇

‘晶瑶’草莓高效再生及遗传转化体系的建立

向发云,韩永超,曾祥国,张庆华,陈丰滢,顾玉成   

  1. (湖北省农业科学院经济作物研究所,武汉 430064）
  • 收稿日期:2016-06-13 修回日期:2017-05-16 接受日期:2016-12-14 出版日期:2017-05-26 发布日期:2017-05-26
  • 通讯作者: 顾玉成
  • 基金资助:
    国家农业支撑计划“草莓新品种选育”(2013BAD02B04-02-04)；湖北省科技支撑计划项目“果树新种质创制与品种培育应用技术研究”(2015BBA209)；湖北省农业科技创新中心资助项目“草莓新品种选育及设施栽培技术创新”(2016-620-000-001-014)。

Regeneration and Transformation of ‘Jingyao’ Strawberry: Some Influencing Factors

Xiang Fayun, Han Yongchao, Zeng Xiangguo, Zhang Qinghua, Chen Fengying, Gu Yucheng   

  1. (Industrial Crops Institute of Hubei Academy of Agricultural Sciences, Wuhan 430064)
  • Received:2016-06-13 Revised:2017-05-16 Accepted:2016-12-14 Online:2017-05-26 Published:2017-05-26

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摘要： 以草莓‘晶瑶’为试材，研究不同激素种类和浓度配比对叶片诱导愈伤、愈伤分化出芽的影响，建立‘晶瑶’草莓叶片的高效离体再生体系。结果表明，‘晶瑶’叶片在MS 2,4-D 0.05 mg/L TDZ 2.0 mg/L的诱导培养基上不定芽再生率达到79.18%。在‘晶瑶’草莓遗传转化试验中，叶盘对卡那霉素的敏感性最适筛选浓度为30 mg/L，羧苄青霉素的最适抑菌浓度为400~500 mg/L，150 μmol/L乙酰丁香酮的加入可提高GUS的瞬时表达率；10 min是最适合农杆菌侵染‘晶瑶’叶片的时间；共培养48 h对叶片转化较适宜，得到抗卡那霉素并呈GUS阳性的抗性芽频率为2.38%，获得了转化阳性植株。

关键词: 叶用莴苣, 叶用莴苣, 热激蛋白基因, LsHsp70-2711, LsHsp70-3711, 抽薹, 相关性

Abstract: In this paper, strawberry investigated for factors affecting the regeneration of adventitious bud. We established the plant regeneration of “Jingyao” strawberry with leaf. The results showed that the optimum callus inducing medium of the petioles from Jingyao was MS TDZ 2.0 mg / L 2,4-D 0.05 mg / L and the regeneration of adventitious buds rate was 79.17%. Factors on Agrobacterium tumifaciens-mediated gene transfer was discussed in detail.The optional concentration of kanamycin to strain fungus was 30 mg/L and of carbenicillin was 450 mg/L.Transient expression radio of GUS improved by adding of 150 μmol/L AS to medium.The ideal time for leaves dipping in Agrobacterium was 10-15min and the optional co-cultivation was 3 days. Kanamycin -resistant shoots regenerated with the radio of 1.1were preliminarily identified as transgenin shoots.