| 小麦AB基因组中一个重复序列的分离、定位与应用 |
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| 引用本文： | 曾子贤,杨足君,刘成,胡利君,任正隆.小麦AB基因组中一个重复序列的分离、定位与应用[J].农业生物技术学报,2008,16(6). |
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| 作者姓名： | 曾子贤 杨足君 刘成 胡利君 任正隆 |
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| 作者单位： | 电子科技大学生命科学与技术学院,成都,610054 |
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| 基金项目： | 国家自然科学基金重点项目，国家自然科学基金，教育部新世纪优秀人才支持计划 |
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| 摘 要： | 利用102对微卫星引物对5份黑麦（Secale）、4份普通小麦（Triticum aestivum）和1份分枝小黑麦（Triticale）进行SSR分析，引物Xgwm614能在分枝小黑麦中扩增出一个387bp的特异DNA片段（记为FZ387，GenBank登录号为EF179137），而黑麦未能扩增出。序列比对结果显示该片段与一粒小麦（T. monococcum）（AY485644）和栽培二粒小麦（T. turgidum）（AY494981）A基因组中Gypsy Ty3-LTR反转座子fatima的一部分分别有94%和95%同源性。根据序列同源性比对结果，在FZ387内部设计1对特异引物FaF和FaR。引物Xgwm614F和FaR能在含有A基因组的物种中扩增出约350bp的条带（记为A350），而其不含A基因组的物种都未扩增出该条带。利用小麦二体和端体代换系材料对其进行定位，结果显示该片段分布在所有A染色体的长臂和断臂上。此外，引物FaF和Xgwm614R能在含有A、B或AB基因组的物种中扩增出约350bp的条带（记为AB350），而不含AB基因组的材料未扩增出目标条带。利用这两对特异引物对小麦属近缘物种进行PCR扩增，发现只有中国春能够扩增出A350和AB350。序列比对结果和FZ387两侧SSR引物结合区的规律性变化表明该反转座子在进化上可能存在属间多样性和属内相似性。A350和AB350也可以分别作为分子标记检测A染色体和AB染色体。
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| 关 键 词： | 小麦 分子标记 SSR 反转座子 |
| 收稿时间： | 2008-3-27 |
| 修稿时间： | 2008-5-22 |
Isolation, Mapping and Application of a Repetitive DNA Sequence in AB genomes of Wheat (Triticum aestivum) |
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| ZENG Zi-xian,YANG Zu-jun,LIU Cheng,HU Li-jun,REN Zheng-long.Isolation, Mapping and Application of a Repetitive DNA Sequence in AB genomes of Wheat (Triticum aestivum)[J].Journal of Agricultural Biotechnology,2008,16(6). |
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| Authors: | ZENG Zi-xian YANG Zu-jun LIU Cheng HU Li-jun REN Zheng-long |
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| Abstract: | SSR analysis was performed on five Secale species, four Triticum species and one Triticale line with branch-spike by 102 microsatellite primer pairs. A 387 bp specific DNA fragment FZ387 (GenBank Accession No.EF179137) was obtained from the Triticale by primer Xgwm614, while no amplification in Secale. NCBI BLAST revealed that this FZ387 sequence had 94% and 95% similarity to a part of Gypsy Ty3-LTR-retrotransposon fatima in Triticum monoccocum (AY485644) and T. turgidum(AY494981), respectively. A pair of specific PCR primers, FaF and FaR, was designed based on the conserved region of this FZ387 sequence. The amplification by primer pair Xgwm614F and FaR revealed that a specific 350 bp band (designation as A350) was obtained from species containing A chromosome. Furthermore, the PCR on Langdon Chinese Spring substitution lines was performed, and found that this segment was located on both long and short arms of all A chromosomes. However, using a primer pair FaF and Xgwm614R, a specific DNA band with about 350 bp (designation as AB350) from materials containing A or/and B chromosomes was amplified. The wild relatives species of wheat were amplified by the two pairs of primers, and revealed that only A350 and AB350 were obtained in CS. The 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 Triticum genomes. Meanwhile, the A350 and AB350 can be used as molecular markers for the detection of A and AB genomes. |
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| Keywords: | wheat molecular marker SSR retrotransposon |
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