| 摘要: |
| 【目的】将具有广谱抗病性的hrpZpsta基因导入大豆，为培育抗灰斑病的转基因大豆新品系奠定基础。【方法】采用农杆菌介导法，以大豆子叶节为受体，将具有广谱抗性的hrpZpsta基因转入大豆品种“吉林30”中，以耐盐基因badh作为筛选标记性基因，经过抗性筛选，对转基因植株进行PCR检测、Southern杂交和RT-PCR检测分析。【结果】确定的NaCl筛选浓度为200 mmol/L。对T1、T2和T3代转基因植株进行PCR检测，得到T1代阳性植株30株，T2代45株，T3代284株，说明外源hrpZpsta基因在转基因后代中能够遗传。Southern杂交结果表明，外源目的基因hrpZpsta已经整合进大豆基因组中，且整合位点不尽相同。RT-PCR结果表明，hrpZpsta基因在受体大豆中获得表达。【结论】获得了hrpZpsta基因遗传表达的T3代转基因大豆株系。 |
| 关键词: hrpZpsta基因 大豆 农杆菌介导法 子叶节 T3代转基因株系 |
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| 基金项目:国家转基因专项资助项目（2011ZX08004-004） |
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| Study on transforming hrpZpsta gene into soybean |
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| Abstract: |
| 【Objective】In order to breed-transgenic soybean which can resist frogeye leaf spot,the present experiment was conducted to transform the resistant gene hrpZpsta into soybean genome.【Method】The hrpZpsta gene was transformed into cotyledonary node of recipient variety Jilin 30 via agrobacterium-mediated.Transgenic plants were screened with salt-tolerance gene badh,and detected by PCR,Southern blot,RT-PCR.【Result】The screening concentration of NaCl was 200 mmol/L in phase of shoots.We got 30 positive transgenic plant lines in T1 generation by PCR detecting,and 45 plant lines in T2 generation,and 284 plant lines in T3 generation,which show that hrpZpsta gene could inherit in transgenic plants.The result of southern blot confirmed that hrpZpsta gene had been integrated into soybean genome,and the integration sites were different.RT-PCR further indicated that hrpZpsta gene had been expressed in soybean recipient.【Conclusion】We obtained transgenic plant lines of T3 generation,and that hrpZpsta gene could inherit in it. |
| Key words: hrpZpsta gene soybean Agrobacterium-mediated cotyledonary node transgenic plant lines of T3 generation |
