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目的:建立当归试管苗ISSR-PCR反应体系,在此基础上对当归试管苗进行遗传稳定性研究。方法:试剂盒法提取当归试管苗DNA,采用正交试验筛选最佳ISSR-PCR反应条件,利用已建立的ISSR-PCR反应体系分析当归试管苗的遗传稳定性。结果:建立了当归试管苗ISSR-PCR反应的最佳体系(25μL):dNTPs 1.20μL、Mg2+0.9μL、引物1.0μL、模板DNA 1.70μL、Taq酶0.60μL、10×PCR Buffer 2.5μL、ddH2O 17.1μL。结果表明,当归试管苗的DNA条带变异率为0.662%。结论:建立的ISSR-PCR技术体系可用于当归种质资源鉴定,当归试管苗具有很高的遗传稳定性,可作为种苗在生产上使用。
OBJECTIVE: To establish an ISSR-PCR reaction system for test-tube seedlings of Angelica sinensis, and to study the genetic stability of the test-tube plantlets. Methods: The DNA of Angelica Sinensis plantlets was extracted by kit method, and the optimal ISSR-PCR reaction conditions were screened by orthogonal test. The genetic stability of Angelica Sinensis plantlets was analyzed by the established ISSR-PCR reaction system. Results: The optimal system (25μL) for the ISSR-PCR reaction was established: dNTPs 1.20μL, Mg2 + 0.9μL, primer 1.0μL, template DNA 1.70μL, Taq enzyme 0.60μL, 10 × PCR Buffer 2.5μL, ddH2O 17.1 μL. The results showed that the variation rate of DNA bands of Angelica sinensis in vitro was 0.662%. Conclusion: The established ISSR-PCR system can be used for identification of Angelica Germplasm resources. Angelica plantarum has high genetic stability and can be used as seedling in production.