偃麦草基因组特异重复序列的分离与应用

doi:10.3864/j.issn.0578-1752.2016.19.002

Abstract

Abstract: 【Objective】As a tertiary gene pool of wheat, Thinopyrum genus has many useful traits, such as tolerance to both abiotic and biotic stresses. DNA repetitive sequence is one major component for genomes of most eukaryotes. Isolation of genome specific repeats is very helpful for understanding of sub-genome component of polyploid species, their evolution and utilization in crop improvement. To identify Th. ponticum chromatin in wheat genetic background more accurately and efficiently, genome specific repetitive sequences were cloned from Th. ponticum.【Method】Screening the plasmid library of Th. ponticum via dot-blot hybridization, combining with fluorescence in situ hybridization (FISH) analysis of the wheat-Th. ponticum partial amphiploids, Th. ponticum genome specific repetitive sequences were isolated and their distribution patterns on chromosomes were characterized by FISH. The Th. ponticum genome specific repeats were analyzed by Repeat Masker, Triticeae Repeat Sequence Database and NCBI. A set of PCR primers was designed for detecting Th. ponticum chromosomal fragments. Wheat-Th. ponticum hybrid derivatives were employed to testify the genome specificity and effectiveness of these repeats in identifying Th. alien fragments in wheat genetic background. 【Result】Seven Th. ponticum genome specific repetitive sequences were obtained. Their FISH signals dispersively covered all chromosomes in Th. ponticum and Th. intermedium. Furthermore, they could discriminate chromosomes of Th. ponticum and Th. intermedium from wheat chromosomes without wheat DNA block by FISH. This was further proved in wheat-Th. ponticum substitution lines and translocation lines. Ten PCR primers were developed for efficient amplification of Th. ponticum segments in wheat. Their detection efficiency ranged from 73.3% to 95% in 109 wheat-Th. ponticum derivatives. 【Conclusion】Th. ponticum genome specific repetitive sequences were isolated, which can be used for detection of wheatgrass fragments in wheat by FISH or PCR amplification.

Key words: T. aestivum, Th. ponticum, genome specific repetitive sequences, FISH, PCR

YAO Han, TANG Cai-guo, ZHAO Jing, ZHENG Qi, LI Bin, HAO Chen-yang, LI Zhen-sheng, ZHANG Xue-yong. Isolation of Thinopyrum ponticum Genome Specific Repetitive Sequences and Their Application for Effective Detection of Alien Segments in Wheat[J].Scientia Agricultura Sinica, 2016, 49(19): 3683-3693.

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Isolation of Thinopyrum ponticum Genome Specific Repetitive Sequences and Their Application for Effective Detection of Alien Segments in Wheat

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