Determination of Genetic Diversity of Tamarind (Tamarindus indica) Accessions in Eastern Region of Kenya Using Inter Simple Sequence Repeat (ISSR) Markers

Mercy Liharaka Kidaha, Cornelius Wainana Mbathi, Fredah K. Rimberia Wanzala
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Abstract

There is scanty information on tamarind’s genetic diversity in Easter Kenya. The objective of this study was to determine the genetic diversity of 64 tamarind accessions from Eastern Kenya using 12 Inter Simple Sequence Repeat (ISSR) markers. DNA was extracted from the young apical leaves using modified CTAB method and amplified using standard PCR.  The data collected were scored as presence (1) or absence (0) of bands then compared to the 100bp ladder and analyzed using GeneAlex and R softwares. Only seven primers produced reproducible bands. A total of 46 alleles were produced for the 7 loci with an average of 6.6 per loci. Polymorphic information content (PIC) varied from 0.72 to 0.89 and genetic diversity varied from 0.74 to 0.9. The ISSR markers revealed effective polymorphism of 40.87 to 99.46% and the band  sizes varied from 100 to 1000 bp. Analysis of Molecular Variance (AMOVA) depicted high variation within the tamarind populations at 90% and the least variation of 10% among the population. The first 3 components of Principal coordinate analysis (PCoA) contributed 40.83% of the total variation. Hierarchical cluster analysis grouped the tamarind accessions into seven major distinct groups. Tamarind accessions were different within counties with minimal variations among counties, proving that genetic diversity exists among the tamarind accessions in the Eastern region of Kenya. High genetic diversity was evident among Embu accessions and least among Masinga accessions. Diversity in tamarind can be utilized in marker-assisted breeding and primer ISSR17899A and ISSRHB11 can be explored in studying genes that code for various traits in tamarind accessions.
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利用ISSR标记测定肯尼亚东部地区罗望子(Tamarindus indica)遗传多样性
关于肯尼亚东部罗望子遗传多样性的信息很少。本研究的目的是利用12个ISSR标记确定来自肯尼亚东部的64份罗望子材料的遗传多样性。采用改良的CTAB法从幼嫩的顶叶中提取DNA,用标准PCR扩增。收集到的数据被评分为条带存在(1)或缺失(0),然后与100bp阶梯进行比较,使用GeneAlex和R软件进行分析。只有7个引物产生了可重复的条带。7个基因座共产生46个等位基因,平均每个基因座6.6个。多态性信息含量(PIC)在0.72 ~ 0.89之间变化,遗传多样性在0.74 ~ 0.9之间变化。ISSR标记的有效多态性为40.87 ~ 99.46%,条带大小在100 ~ 1000 bp之间。分子方差分析(AMOVA)显示罗望子群体内变异率高达90%,群体间变异率最低为10%。主坐标分析(PCoA)的前3个分量占总变异量的40.83%。分层聚类分析将罗望子材料分为7个不同的主要类群。县内罗望子的遗传多样性存在差异,县间罗望子的遗传多样性最小,说明肯尼亚东部地区罗望子遗传多样性存在。Embu系遗传多样性高,Masinga系遗传多样性低。罗望子的多样性可用于标记辅助育种,引物ISSR17899A和ISSRHB11可用于研究罗望子各种性状的编码基因。
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