| 花粉介导法获得油菜转基因植株研究 |
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| 引用本文： | 杜春芳,刘惠民,李朋波,孙毅,李润植.花粉介导法获得油菜转基因植株研究[J].作物学报,2006,32(5):749-754. |
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| 作者姓名： | 杜春芳 刘惠民 李朋波 孙毅 李润植 |
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| 作者单位： | 山西省农业科学院棉花研究所, 山西运城044000 |
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| 摘 要： | 以甘蓝型冬油菜品种晋油7号为受体，载有GUS基因的质粒pBI121为供体，在7.5％的蔗糖等渗溶液中，通过花粉介导转化方法将GUS基因导入油菜花粉，随着花粉管的萌发进入胚囊，参与有丝分裂，将外源质粒转入受体材料。经田间植株性状比较、GUS组织化学定位检测、PCR扩增检测及PCR-Southern杂交检测，证明外源GUS基因已整合到油菜基因组中。
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| 关 键 词： | 甘蓝型油菜 花粉介导转化法 GUS基因 |
| 收稿时间： | 2005-04-22 |
| 修稿时间： | 2005-10-06 |
Pollen-mediated Transformation of Rape |
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| DU Chun-Fang,LIU Hui-Min,LI Peng-Bo,SUN Yi,LI Run-Zhi.Pollen-mediated Transformation of Rape[J].Acta Agronomica Sinica,2006,32(5):749-754. |
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| Authors: | DU Chun-Fang LIU Hui-Min LI Peng-Bo SUN Yi LI Run-Zhi |
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| Institution: | Cotton Research Institute, Shanxi Academy of Agricultural Sciences, Yuncheng 044000, Shanxi |
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| Abstract: | Plasmid DNA of pBI121 containing GUS gene and fresh pollen of rape Jinyou 7 were mixed in 7.5% sucrose solution and treated with ultrasonic.After that,pollens were collected,pollinated on stigmas of rape.With pollen tube growth through the style tissue and entering the ovaries,the target gene was integrated in rape genome.Transformed plants were confirmed by field character identification(Fig.2),histochemical assay of GUS expression(Fig.3),DNA dot blot hybridization(Fig.4),PCR amplification(Fig.5) and PCR-Southern blot hybridization(Fig.6).In this study,some regeneration conditions,such as the concentration of sucrose solution,the intensity of ultrasonic and the effective of auxiliary agent for transformation system were discussed.The highly efficient transformation system on rape was established with 7.5% of sucrose solution(Fig.7) or 5% of manna sugar solution(Fig.8),300 W of ultrasonic(Fig.9).Compared with other approaches,the pollen-mediated transformation approach can avoid the tedious tissue culture procedures like in Agrobacterium infection and particle bombardment,etc.This approach is simple,easy to operate,and can be widely used in breeding practice. |
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| Keywords: | Rapeseed Pollen-mediated transformation GUS gene |
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