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目的:利用RAPD分子标记技术,对11份荆芥种质资源进行遗传多样性分析,探讨我国荆芥种质资源的遗传多样性。方法:以取自不同主要生产地区的11份荆芥幼苗为材料,CTAB法提取基因组DNA,进行RAPD分析,统计软件DPS(V7.5)分析遗传距离,利用UPSMA方法进行聚类分析;RAPD反应体系为20μL,包括2μL模板DNA,1.6μL d NTPs(2.5 mmol/L),0.4μL Taq DNA聚合酶1 U,2μL 10×PCR buffer,随机引物组合(100μmol/L)2μL,12μL无菌水;RAPD扩增程序:95℃预变性2 min,95℃变性30 s,35℃退火30 s,72℃延伸1 min,循环35次;最后72℃延伸7 min。结果:筛选的14个引物组合共获得36条可重现谱带,其中23条是多态的,多态性百分率为63.89%;聚类分析后,11份种质资源的遗传距离在0.1333~0.7778之间;以λ=0.3作为阈值,可将供试11份荆芥种质资源分成4类,其基于RAPD的聚类结果与材料地域远近的分类结果并不完全一致,一定程度上有相关性,并与引种频繁与否有关。结论:供试荆芥种质资源遗传差异不大,遗传基础较窄,遗传多样性不高。
OBJECTIVE: To analyze the genetic diversity of 11 Nepeta germplasm resources using RAPD molecular markers and to explore the genetic diversity of Nepeta germplasm resources in China. Methods: Eleven Nepeta seedlings from different regions were used as materials. Genomic DNA was extracted by CTAB method and analyzed by RAPD. The genetic distance was analyzed by statistical software DPS (V7.5), and cluster analysis was performed by UPSMA method. RAPD reaction The system consisted of 2 μL of template DNA, 1.6 μL dNTPs (2.5 mmol / L), 0.4 μL Taq DNA polymerase 1 U, 2 μL 10 × PCR buffer, 2 μL random primer combination (100 μM / L) RAPD amplification program: pre-denaturation at 95 ° C for 2 min, denaturation at 95 ° C for 30 s, annealing at 35 ° C for 30 s, extension at 72 ° C for 1 min, cycling 35 cycles and final extension at 72 ° C for 7 min. Results: A total of 36 reproducible bands were obtained, of which 23 were polymorphic and the percentage of polymorphism was 63.89%. After the cluster analysis, the genetic distance of 11 germplasm resources was between 0.1333 ~ 0.7778. With λ = 0.3 as the threshold, 11 samples of Schizonepeta tenuipes could be divided into 4 categories. The clustering results based on RAPD were not completely consistent with the classification results of material areas, and to a certain extent, they were related Sexuality, and with the introduction of frequent or not. Conclusion: The genetic diversity of Nepeta germplasm resources is not large, the genetic basis is narrow, and the genetic diversity is not high.