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中国农学通报

中国农学通报 ›› 2007, Vol. 23 ›› Issue (4): 64-64.

所属专题： 生物技术； 园艺

• 目次 • 上一篇    下一篇

DNA浓度及注射时间对苹果花粉管通道法基因转化率的影响

冯莎莎,杜国强,师校欣,周瑞金,王晓蔓,王燕霞   

  • 出版日期:2007-04-05 发布日期:2007-04-05

Concentration of DNA and Time of Injection on Gene Transformation Efficiency via Pollen-tube Pathway in Apple

Feng Shasha, Du Guoqiang, Shi Xiaoxin, Zhou Ruijin, Wang Xiaoman, Wang Yanxia   

  • Online:2007-04-05 Published:2007-04-05

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摘要： 通过花粉管通道法，将携带CBF3和GUS基因的pWBVec10a质粒DNA导入苹果。坐果后70d左右采收种子，将收获的种子进行离体培养，培养基为MS+BA0.2mg/L+GA2.0mg/L，附加蔗糖50g /L，琼脂6 g/L，培养40d后对转化子叶进行GUS染色检测。结果表明，最佳注射时间为授粉后11~24h之间。随着注入外源DNA浓度的增加，坐果率逐渐降低，DNA浓度为500μg/ml时，坐果率可达3.1%；DNA浓度为1000μg/ml时GUS基因阳性表达率可达12.5%。

关键词: 刺五加, 刺五加, 外植体, 培养基, 组织培养, 继代培养, 激素

Abstract: Using pollen tube pathway transformation, plasmid pWBVec10a containing gene CBF3 and report gene GUS was introduced into apple. The seeds were harvested and the embryos were cultured at 70d after fruit setting. The medium was MS+BA0.2mg/L+GA2.0mg/L supplying with 50g/L sucrose and 6 g /L agar. The results showed that the best time to inoculate was at 11 hours to 24 hours after pollination. With the increase of the concentration of DNA, the rate of fruit setting decreased gradually, when the concentration of DNA was 500μg/ml, it could reach the rate of fruit setting of 3.1%. The expression of GUS gene showed that the rate of expression of GUS was gained with 12.5% when 1000μg/ml plasmid pWBVec10a DNA was inoculated into cotyledon.

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