Development of ILP-markers for Aegilops tauschii and their application in molecular genetic analysis

Faktori eksperimental'noi evolucii organizmiv Pub Date : 2022-09-01 DOI:10.7124/feeo.v30.1455

S. Hordynskyi, A. Postovoitova, A. Rabokon, Y. Pirko, Y. Blume

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Abstract

To develop marker systems based on the study of intron length polymorphism of various genes (ILP) in Ae. tauschii, to test the possibility of their use for genetic differentiation of Ae. tauschii and Ae. biuncialis. Methods. The NCBI database was used to obtain EST sequences, online tools CD-HIT, BLAST and Primer3Plus (for the development of primers). The polymerase chain reaction (PCR) method with the developed primers was done. Amplified fragments were separated by non-denaturing polyacrylamide gel electrophoresis and stained with silver. Results. Molecular markers Aet_ILP1, Aet_ILP6 were developed and tested for different genotypes of Ae. tauschii and Ae. biuncialis. No intraspecific differentiation was observed in all studied samples, but amplicons of introns of different species differed significantly. It was also found that the studied species had one copy of the HO222074.1 gene and 1 to 3 copies of the CX244643.1 gene. Conclusions. The obtained results indicate a low level of intraspecific variability of the developed ILP markers and the possibility of their use for interspecific differentiation of Ae. tauschii and Ae. biuncialis.

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黄斑鱼ilp标记的开发及其在分子遗传分析中的应用

目的:建立基于伊蚊各基因内含子长度多态性(ILP)研究的标记系统。tauschii，以测试它们用于伊蚊遗传分化的可能性。牛头和Ae。biuncialis。方法。利用NCBI数据库获取EST序列，在线工具CD-HIT、BLAST和Primer3Plus(用于开发引物)。用所建立的引物进行了聚合酶链反应(PCR)。扩增片段用非变性聚丙烯酰胺凝胶电泳分离，并用银染色。结果。建立Aet_ILP1、Aet_ILP6分子标记，对不同基因型的伊蚊进行检测。牛头和Ae。biuncialis。所有研究样本均未观察到种内分化，但不同物种的内含子扩增量差异显著。还发现所研究的物种具有1个拷贝的HO222074.1基因和1 ~ 3个拷贝的CX244643.1基因。结论。所获得的结果表明，所开发的ILP标记具有低水平的种内变异性，并且可能用于Ae的种间分化。牛头和Ae。biuncialis。

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Faktori eksperimental'noi evolucii organizmiv

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