Trasfering Soybean Gene into Hedysarum mongolicum Mature Seeds by Low Energy N+ Implantation

doi:10.11733/j.issn.1007-0435.2011.06.025

›› 2011, Vol. 19 ›› Issue (6): 1036-1041.DOI: 10.11733/j.issn.1007-0435.2011.06.025

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Trasfering Soybean Gene into Hedysarum mongolicum Mature Seeds by Low Energy N⁺ Implantation

CHEN Ling-ling², NA Ri¹, WU Yan-hong², LUAN Shou-quan², ZHENG Yue²

1. School of Physical Science and Technology/Key Laboratory of Ion Beam Bioengineering, Inner Mongolia University, Hohhot, Inner Mongolia 010021, China;
2. Chifeng Academy of Agricultural and Animal Husbandry Sciences, Chifeng, Inner Mongolia 024031, China

Received:2011-03-02 Revised:2011-07-20 Online:2011-12-15 Published:2012-07-09

应用低能N⁺介导法将大豆外源基因转入羊柴成熟种子的初步研究

陈玲玲², 那日¹, 乌艳红², 栾守泉², 正月²

1. 内蒙古大学物理科学与技术学院/内蒙古大学离子束生物工程自治区重点实验室, 内蒙古呼和浩特010021;
2. 赤峰市农牧科学研究院, 内蒙古赤峰024031

通讯作者: 乌艳红,E-mail:xuyanwuyanhong@sina.com
作者简介:陈玲玲(1982- ),女,蒙古族,内蒙古赤峰人,硕士研究生,助理研究员,研究方向为牧草栽培与良种繁育,E-mail:chenlingling001@tom.com
基金资助:
国家自然科学基金(50267001);内蒙古自治区自然科学基金重大项目(200711020101);内蒙古教育厅基金项目(NJ05025)资助

Abstract

Abstract: Effects of low energy N⁺ implantation on Hedysarum mongolicum mature seeds are studied.The germination rates and survival rates of Hedysarum mongolicum dry seeds with different N⁺ implantation dosages are investigated.Results show that the feasible transformation dosages are 900·2.6·10¹³ N⁺·cm^-2.The blue cells expressing β-glucuronidase are observed at Hedysarum mongolicum callus with 13% transient expression.PCR analysis of transformed plants shows that the soybean gene had been integrated into Hedysarum mongolicum genome.

Key words: Ion beam, Hedysarum mongolicum, Mature seeds, Genetic transformation

摘要： 为研究低能N⁺介导大豆(Glycine max)基因遗传转化的影响,以羊柴(Hedysarum mongolicum)成熟种子为材料,根据不同注入剂量下羊柴种子的发芽率,计算出羊柴种子发芽率的差异性并绘制出剂量-存活率曲线,找出最适宜的转化剂量。结果表明:经离子束(25keV,900·2.6·10¹³ N⁺·cm^-2)注入后的羊柴成熟种子发芽率的差异性和剂量-存活率曲线可以确定900·2.6·10¹³ N⁺·cm^-2是最适宜的转化剂量。将处理后的羊柴种子浸入含有大豆质粒DNA的SSC导入介质,24h后观察到受离子束直接作用的胚部有外源基因的导入和表达,检测到的GUS基因瞬间表达率为13%,转化体的PCR检测结果初步证实了大豆基因已经整合到羊柴基因组中。

关键词: 离子束, 羊柴, 成熟种子, 遗传转化

CLC Number:

Q785

CHEN Ling-ling, NA Ri, WU Yan-hong, LUAN Shou-quan, ZHENG Yue. Trasfering Soybean Gene into Hedysarum mongolicum Mature Seeds by Low Energy N⁺ Implantation[J]. , 2011, 19(6): 1036-1041.

陈玲玲, 那日, 乌艳红, 栾守泉, 正月. 应用低能N⁺介导法将大豆外源基因转入羊柴成熟种子的初步研究[J]. , 2011, 19(6): 1036-1041.

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Trasfering Soybean Gene into Hedysarum mongolicum Mature Seeds by Low Energy N⁺ Implantation

应用低能N⁺介导法将大豆外源基因转入羊柴成熟种子的初步研究

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Trasfering Soybean Gene into Hedysarum mongolicum Mature Seeds by Low Energy N+ Implantation

应用低能N+介导法将大豆外源基因转入羊柴成熟种子的初步研究

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Trasfering Soybean Gene into Hedysarum mongolicum Mature Seeds by Low Energy N⁺ Implantation

应用低能N⁺介导法将大豆外源基因转入羊柴成熟种子的初步研究