›› 2014, Vol. 16 ›› Issue (5): 147-153.DOI: 10.13304/j.nykjdb.2014.170

• 方法与技术创新 • 上一篇    下一篇

玉米生长条件对玉米原生质体转化的影响研究

谢礼,宗娜,赵军*   

  1. (中国农业科学院生物技术研究所作物基因组与遗传改良研究室玉米功能基因组创新团队|农作物基因资源与基因改良国家重大科学工程, 北京 100081)
  • 出版日期:2014-10-15 发布日期:2014-10-14
  • 通讯作者: 赵军,研究员,博士,主要从事植物抗逆性和籽粒发育分子生物学和分子育种研究。E\|mail: zhaojun01@caas.cn
  • 作者简介:谢礼,硕士研究生|主要从事植物非生物逆境传导研究。E\|mail: xieli200709@163.com。
  • 基金资助:

    国家自然基金项目(31170235)资助。

Effects of Plant Growth Conditions on Maize Protoplast Transformation

XIE Li, ZONG Na, ZHAO Jun*   

  1. (Faculty of Maize functional Genomics, Department of Crop Genome Research and Genetic Improvement, Biotechnology Research Institute,Chinese Academy of Agricultural Sciences|National Key Facility for Crop Gene Resources and Genetic Improvement, CAAS, Beijing 100081, China)
  • Online:2014-10-15 Published:2014-10-14

摘要:

植物原生质体瞬时转化体系是一种高效、快捷的基因表达分析工具。为研究玉米生长条件对玉米原生质体活力和转化效率的影响,以玉米自交系Q319幼苗为研究对象,利用正交试验L9(34)探讨了玉米的生长条件(光照强度、光照周期、生长温度和龄期)对玉米叶肉细胞原生质体转化的影响。实验结果显示上述四个因素都能够影响玉米叶肉细胞原生质体活力与转化效率,而光照强度的影响最大;进一步研究发现光照强度与玉米叶肉细胞原生质体的活力及转化效率呈负相关关系,玉米黄化苗原生质体转化效果最好,其原生质体活力及转化效率分别达到了100%和85%。另外,龄期为13 d的玉米黄化苗原生质体转化效率最高,龄期过小或过大,黄化苗的原生质体转化效率都会下降。

关键词: 玉米Q319;原生质体转化;生长条件;转化效率

Abstract:

Plant protoplast transient transformation is a versatile tool for transient gene expression analysis. To study the effects of plant growth conditions on maize protoplast transformation, maize inbred line Q319 was used as material to discuss the effect of maize growth condition(light intensity, photoperiod, growth temperature and seedlings age)on maize protoplast transformation by orthogonal experiment L9(34) design. The results showed that the 4 growth conditions all affected maize protoplast transformation, among which light intensity played the most important role on protoplast viability and transformation efficiency. Further study showed that there was a notable negative correlation between light intensity and protoplast viability and transformation efficiency. The maize etiolated seedlings were the best material for maize protoplast transformation. The protoplast viability and transformation efficiency of etiolated seedlings could reach 100% and 85%, respectively. In addition, the highest transformation efficiency was obtained using the protoplasts from etiolated seedlings 13 d after sowing.

Key words: maize Q319, protoplast transformation, growth condition, transformation efficiency

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