Development and characteristation of EST-SSR markers for genetic analysis of Casuarina species
Development and characteristation of EST-SSR markers for genetic analysis of Casuarina species
复制标题
用于木麻黄物种遗传分析的 EST-SSR 标记的开发和表征
DOI：
10.26525/jtfs2021.33.4.425
复制
发表时间：
2021
影响因子：
0.9
通讯作者：
Pan Hu
中科院分区：
文献类型：
--
作者：
Zhen Li;Yong Zhang;Pan Hu
简单重复序列（SSR）标记具有孟德尔遗传和共显性表达的特点，在遗传分析中相对于其他分子标记具有更大的优势。大量的木麻黄属植物表达序列标签（expressed sequence tags，EST）已从公共数据库中检索到，possible.to为开发低成本、高效率的EST-SSR标记提供了条件。通过对木麻黄属12，063个EST的分析，从352个单基因中鉴定出367个SSR位点，SSR位点的分布频率为3.0%。二核苷酸和三核苷酸分别占57.77%和34.06%。它们在5种SSR重复基序类型中占绝大多数，其中AG/CT是最主要的SSR重复基序类型。根据非冗余EST-SSR序列设计了79对引物，但只有21对引物在4个不同木麻黄种和8个木麻黄无性系中扩增出多态性条带。木贼从21对引物中筛选出8对引物，对一个雌株的自由授粉后代进行了亲子鉴定。木贼在95%的置信区间内，461个后代中有262个后代的花粉亲本被鉴定为76个雄性个体，而在80%的置信区间内，461个后代中有328个后代的父树被鉴定为84个雄性个体。
Simple sequence repeats (SSR) markers have greater advantages relative to other molecular markers in genetic analysis due to their Mendelian inheritance and co-dominance expression features. Numerous expressed sequence tags (EST) of genus Casuarina, now available from public databases, make it possible.to develop EST-SSR markers with high efficiency at low cost. From the analysis of 12,063 EST derived from casuarinas, 367 SSR loci were identified from 352 unigenes, and the distribution frequency of SSR loci was 3.0%. The di- and trinucleotide accounted for 57.77 and 34.06% respectively. They comprised.overwhelming majority in five types of repeat motifs of SSR, and AG/CT was the most predominant motif type of SSR. A total of 79 primer pairs were designed based on the non-redundant EST-SSR sequences, but only 21 were successfully amplified and generated polymorphic SSR products in four.different Casuarina species and eight clones of C. equisetifolia. Eight primer pairs out of the 21 were applied for paternity analyses of open-pollinated progenies of one female individual of C. equisetifolia. At 95% confidence interval, 76 male individuals were identified as pollen parents of 262 out of 461 progenies, but at 80% confidence interval, 84 male individuals were identified as father trees of 328 out of the 461 progenies.