小麦(Triticum aestivum) a和B基因组重复DNA序列的分离、定位和应用。

Zeng Zi-xian, Yang Zu-jun, Liu Cheng, Hu Li-jun, Ren Zheng-long
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引用次数: 0

摘要

利用102对微卫星引物对5个黑麦种、4个小麦种和一个小黑麦品系汾直-1号进行了SSR分析。一个387 bp特异性DNA片段FZ387 (GenBank登录号:引物Xgwm614从小黑麦Fenzhi-1中分离得到EF179137,未在黑麦中扩增。NCBI BLAST结果显示,该FZ387序列与Triticum monococum (AY485644)和Triticum turgidum (AY494981)中Gypsy Ty3-LTR反转录转座子Fatima的部分序列相似性分别为94%和95%。基于FZ387序列的保守区,设计了一对特异性PCR引物FaF和FaR。引物对Xgwm614F和FaR的扩增结果显示,从含有a染色体的种属中获得了一条350 bp的特异条带(命名为A350)。进一步对Langdon Chinese Spring代换系进行PCR,结果发现该片段位于所有A染色体的长臂和短臂上。然而,引物对FaF和Xgwm614R的扩增在含有a和/或B染色体的材料中产生了约350bp的特定DNA带(命名为AB350)。利用这两对引物对小麦野生种及其近缘种进行扩增,结果表明,在中国春(CS)中只发现了A350和AB350。FZ387的序列比较和SSR引物结合区的变异表明,该法蒂玛样元件在小麦基因组中可能存在显著的内部序列多样性。同时,A350和AB350都可以作为A和AB基因组检测的分子标记。
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Isolation, mapping and application of a repetitive DNA sequence in wheat (Triticum aestivum) A and B genomes.
Simple sequence repeat (SSR) analysis was performed on five Secale species, four Triticum species and a Triticale line Fenzhi-1 using 102 pairs of microsatellite primers. A 387-bp specific DNA fragment FZ387 (GenBank accession no. EF179137) was obtained from the Triticale Fenzhi-1 with primer Xgwm614, without amplification in Secale. NCBI BLAST revealed that this FZ387 sequence had 94% and 95% similarity to part of the Gypsy Ty3-LTR retrotransposon Fatima in Triticum monoccocum (AY485644) and Triticum turgidum (AY494981), respectively. A pair of specific polymerase chain reaction (PCR) primers, FaF and FaR, was designed based on the conserved region of this FZ387 sequence. The amplification of primer pair Xgwm614F and FaR revealed that a specific 350-bp band (designation as A350) was obtained from the species containing A chromosomes. Furthermore, PCR on Langdon Chinese Spring substitution lines was performed, and the results found that this segment was located on both long and short arms of all A chromosomes. However, the amplification of primer pair FaF and Xgwm614R gave rise to a specific DNA band of about 350bp (designated AB350) from materials containing A and/or B chromosomes. The wild species of wheat and the relatives were amplified using the two pairs of primers, and revealed that only A350 and AB350 were found in Chinese Spring (CS). Sequence comparison and variation of SSR primers binding regions of FZ387 indicated that significant diversity might exist in the internal sequence of this Fatima-like element among triticeae genomes. Meanwhile, both A350 and AB350 can be used as molecular markers for the detection of A and AB genomes.
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