| 杨 骄，刘志涛，陈健泳，王艳芝，庄丽芳，亓增军.寡核苷酸探针套涂染结合基因组原位杂交和分子标记分析准确鉴定小麦-百萨偃麦草异源易位系的研究[J].麦类作物学报,2018,(3):253 |
| 寡核苷酸探针套涂染结合基因组原位杂交和分子标记分析准确鉴定小麦-百萨偃麦草异源易位系的研究 |
| Accurate Characterization of Wheat-Thinopyrum bessarabicum Alien Chromosome Translocations Using Oligonucleotide Multiplex Painting Combined with Genomic in situ Hybridization and Molecular Marker Analysis |
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| DOI：10.7606/j.issn.1009-1041.2018.03.01 |
| 中文关键词: 小麦-百萨偃麦草易位系 电离辐射 寡核苷酸探针套涂染 分子标记 物理作图 |
| 英文关键词:Wheat-Thinopyrum bessarabicum translocations Ionizing radiation Oligonucleotide multiplex Molecular marker Physical mapping |
| 基金项目:四川省科技计划项目（应用基础计划类型）（2016JY0070）；国家重点研发计划项目（2017YFD0100900） |
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| 中文摘要: |
| 培育小麦异源易位系是转移和利用外源基因的有效途径，快速准确的染色体鉴定方法有利于提高染色体工程效率。本研究以电离辐射诱致的2个普通小麦-百萨偃麦草染色体5J易位系（NAU16YJ127和NAU16YJ124）为例，分析了综合利用本课题组前期开发的寡核苷酸探针套［包括(GAA)10、pSc119.2-1、pAs1-1、pAs1-3、AfA-3和AfA-4六个寡核苷酸探针］涂染，结合基因组原位杂交和分子标记等方法鉴定小麦异源易位系的效果。结果表明，NAU16YJ127（2n=6x=42）包含一对小麦-百萨偃麦草易位染色体T5JS·5JL-5AL#1，同时在染色体6A和2D长臂顶端也发生了明显的相互易位；NAU16YJ124（2n=6x=44）附加了一对小麦-百萨偃麦草易位染色体，同时包含一对6A长臂缺失染色体del6AL，其缺失区段易位到5JL近端部，形成T5JS·5JL-6AL，其他染色体未见明显变化。利用21个5J特异分子标记（19个位于长臂和2个位于短臂）分析发现，两个易位系均含有全部5J短臂标记，同时T5JS·5JL-5AL#1含有12个长臂标记，而T5JS·5JL-6AL仅含有2个长臂标记，确证两个易位系断点不同，并将21个标记定位于4个不同区段，其中位于5J短臂的2个标记位于1个区段，位于5J长臂的19个标记位于3个区段（5JL-1~3），分别包含2、10和7个标记；发现3个为可以同时识别5J和5A的共显性标记，其中NAU16YJ124含有全部5A标记，而NAU16YJ127仅包含其中2个标记，进一步验证了细胞学鉴定结果。本研究结果表明，寡核苷酸探针套涂染、基因组原位杂交和分子标记分析相结合为准确鉴定小麦异源易位系提供了新方法。 |
| 英文摘要: |
| Development of wheat alien translocations is an important approach to transfer and utilize the alien genes.Fast and accurate chromosome identification method will improve the efficiency of chromosome engineering.In this paper, by using two irradiation-induced wheat-Thinopyrum bessarabicum alien translocations as materials,oligonucleotide multiplex［including(GAA)10, pSc119.2-1, pAs1-1, pAs1-3, AfA-3, and AfA-4 previously developed in our lab］painting, genomic in situ hybridization(GISH) and molecular marker analysis were combined to accurately characterize wheat alien translocations.The results revealed that NAU16YJ127(2n=6x=42) contained not only a pair of wheat-Th.bessarabicum translocation T5JS·5JL-5AL#1, but also two pairs of wheat-wheat reciprocal translocations between 2D and 6A.NAU16YJ124(2n=6x=44) not only contained a pair of wheat-Th.bessarabicum translocation T5JS·5JL-6AL, but also contained a pair of chromosome 6A deletion(designated del6AL).Amplification of 21 chromosome 5J-specific markers of Th.bessarabicum including 19 on 5JL and 2 on 5JS showed that both two lines contained all 2 short arm markers but had different number of long arm markers of 5J, for example, NAU16YJ127(T5JS·5JL-5AL#1) contained only 12 long arm markers, while NAU16YJ124(T5JS·5JL-6AL) only had 2 long arm markers, confirming the size difference of the alien segments involved in the two lines.The 21 markers were then allocated to 4 different blocks of 5J, which included one block in 5JS with 2 markers and three blocks in 5JL designating 5JL-1-3 with 19 markers, each block contained 2, 10 and 7 specific markers.Among 21 markers, 3 were co-dominant which can be used to discriminate 5A and 5J simultaneously.NAU16YJ124 contained all but NAU16YJ127 contained only 2 of these co-dominant markers,which further confirmed the chromosome constitutions of the two lines based on cytological analysis.This result indicated that combination of using oligonucleotide multiplex painting, GISH and molecular marker analysis provides an accurate method for chromosome characterization of wheat alien translocations. |
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