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椰子(Cocos nucifera L.)多态微卫星的分离与表征

Isolation and characterization of polymorphic microsatellites in Cocos nucifera L.

作者信息

Rivera R, Edwards K J, Barker J H, Arnold G M, Ayad G, Hodgkin T, Karp A

机构信息

Philippine Coconut Authority, Zamboanga Research Centre, Philippines.

出版信息

Genome. 1999 Aug;42(4):668-75. doi: 10.1139/g98-170.

Abstract

Microsatellites or simple sequence repeats (SSRs) were isolated from coconut (Cocos nucifera) and tested for polymorphism on restricted germplasm. Sequencing of 197 clones from a cv. Tagnanan Tall-enriched genomic library showed that 75% contained a microsatellite, of which 64% were dinucleotide (GA/CT, CA/GT and GC/CG), 6% were trinucleotide, and 30% were compound repeats. Of 41 primer pairs tested on Tagnanan Tall genomic DNA, 38 gave the expected size product, two amplified two loci, and another gave a multilocus pattern. On 20 coconut samples, the 38 SSRs detected 198 alleles (average: 5.2 alleles per microsatellite). Genetic diversity (D = 1 - sigma pi2) values ranged from 0.141 to 0.809. Heterozygotes were present at high frequencies among some dwarf samples. Analysis of similarity matrices based either on shared alleles at each locus (simple matching coefficient) or on allele bands across all loci (Jaccard coefficient) showed similar results. Dwarfs grouped separately from talls and showed less genetic diversity. In a wider test on 40 samples, 8 SSRs detected 64 alleles (average: eight alleles per microsatellite). These results indicate the high potential of microsatellites to detect genetic diversity in coconut germplasm.

摘要

从椰子(Cocos nucifera)中分离出微卫星或简单序列重复(SSR),并在有限的种质资源上测试其多态性。对来自一个塔格纳南高大种富集基因组文库的197个克隆进行测序,结果表明75%的克隆含有微卫星,其中64%为二核苷酸(GA/CT、CA/GT和GC/CG),6%为三核苷酸,30%为复合重复。在塔格纳南高大种基因组DNA上测试的41对引物中,38对产生了预期大小的产物,2对扩增出两个位点,另一对产生了多位点模式。在20个椰子样本上,38个SSR检测到198个等位基因(平均每个微卫星5.2个等位基因)。遗传多样性(D = 1 - sigma pi2)值范围为0.141至0.809。在一些矮种样本中杂合子频率较高。基于每个位点共享等位基因(简单匹配系数)或所有位点的等位基因条带(杰卡德系数)的相似性矩阵分析显示了相似的结果。矮种与高大种分开聚类,且遗传多样性较低。在对40个样本进行的更广泛测试中,8个SSR检测到64个等位基因(平均每个微卫星8个等位基因)。这些结果表明微卫星在检测椰子种质资源遗传多样性方面具有很高的潜力。

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