红芽芋低温疗法脱毒苗遗传变异的AFLP检测

AFLP Detection of Genetic Variation of Virus-Free Plantlets by Cryotherapy in Jiangxi Yanshan Red Bud Taro

为探明红芽芋低温疗法脱毒苗的遗传变异,本研究利用AFLP荧光标记和毛细管电泳分离技术对其进行分析。研究表明,利用10对引物组合对江西铅山红芽芋4种低温疗法脱毒苗和大田苗30个样本进行扩增,扩增后的多态性位点数在49~70范围之内。多态性百分率最低是E36M72,为89.23%,多态性百分率最高是E59M66,为100%;观测等位基因数Na在1.892 3~2.000 0的范围之内,有效等位基因数Ne在1.616 6~1.908 2的范围之内;Nie's基因多样度H的范围为0.357 4~0.473 8,Shannon信息指数I的范围为0.526 8~0.666 2;依据其荧光AFLP扩增结果进行编码组合,构建了江西铅山红芽芋AFLP组合编码指纹图谱。4种低温疗法脱毒苗和大田苗30个样本均出现了AFLP变异位点,每个样本变异位点平均出现的数量为8.73~23.30;4种低温疗法脱毒苗和大田苗30个样本的遗传相似系数为0.471 8~0.796 2,样本间的相似度不是很高,低温疗法脱毒存在一定程度上的变异。在遗传相似系数0.605 1处,4种低温疗法脱毒苗和大田苗30个样本被分为4大类。第Ⅰ类包括样本1和样本30,第Ⅱ类包括样本27和样本29,第Ⅲ类包括样本26和样本28,剩下的样本均归于第Ⅳ类。表明低温疗法脱毒造成了江西铅山红芽芋一定程度的遗传变异。本试验结果为江西铅山红芽芋茎尖低温疗法脱毒提供一定的理论依据。

In ord er to verify genetic variation in virus-free plantlets by cryotherapy of Jiangxi Yanshan red bud taro,the study applied AFLP fluorescence labeling and capillary electrophoresis to analyze the material.The results showed that 30 samples of 4 kinds of virus-free plantlets by cryotherapy and field seedling of Jiangxi Yanshan red bud taro were amplified based on 10 pairs of primer combinations,and the polymorphic loci after amplification were within the range of 49～70.The polymorphism percentage of E36 M72 was the lowest（89.23%）,and the that of E59 M66 was the highest（100%）.The number of observed alleles（Na） was within the range of 1.892 3～2.000 0,and the number of effective alleles（Ne） was within the range of 1.616 6～1.908 2.Nie ＇s gene diversity H was within the range of 0.357 4～0.473 8,and the Shannon information index I was at the scopes of 0.526 8～0.666 2.According to the amplification results of fluorescence AFLP,the research encoded and constructed the AFLP encoding DNA fingerprinting of Jiangxi Yanshan red bud taro.AFLP mutation sites were found in 30 samples of 4 kinds of virus-free plantlets by cryotherapy and field seedlings.The average number of variation sites in each sample was 8.73～23.30.The genetic similarity coefficient of 30 samples of 4 kinds of virus-free plantlets by cryotherapy and field seedlings was 0.471 8～0.796 2.The similarity between samples was not very high,and there was a certain degree of variation in virus-free plantlets by cryotherapy to some extent.At the genetic similarity coefficient of 0.605 1,30 samples of 4 kinds of virus-free plantlets by cryotherapy and field seedlings were divided into four categories.Group Ⅰ included sample 1 and sample 30,and group Ⅱ included sample 27 and sample 29.Group Ⅲ included sample 26 and sample 28,and the remaining samples were all ranked into group Ⅳ.These results suggested that virus-free by cryotherapy could lead to a certain degree of genetic variation in Jiangxi Yanshan red bud taro,which might provide a theoretical basis for virus-free by cryotherapy on shoot tip of Jiangxi Yanshan red bud taro.