Development and application of oligonucleotide-based chromosome painting for chromosome 4D of Triticum aestivum L.
Development and application of oligonucleotide-based chromosome painting for chromosome 4D of Triticum aestivum L.
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基于寡核苷酸的小麦4D染色体染色技术的开发与应用。
DOI：
10.1007/s10577-020-09627-0
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发表时间：
2020-01-30
影响因子：
2.6
通讯作者：
Wang, Xiue
中科院分区：
文献类型：
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作者：
Song, Xinying;Song, Rongrong;Wang, Xiue
染色体绘画是区分特定染色体（片段）、阐明不同基因组或染色体的遗传关系以及鉴定染色体重排的一种有用技术。染色体或基因组特异性探针的开发是染色体绘制的基础。由于属于同一同源类群的染色体具有高度保守的序列，因此在异源多倍体物种中开发特异性标记同源染色体的可能性一直受到质疑。在本研究中，我们试图通过选择普通小麦变异基因参考基因组中的4d特异性序列，构建小麦染色体4d特异性寡核苷酸探针文库。中国春天（CS, 2n = 6x = 42， AABBDD）。合成库包含27,392个寡核苷酸。利用探针文库绘制Oligo证实了它的特异性，在3个小麦基因型和CS无四染色体系N4AT4D中，只有4D染色体可以绘制。利用Oligo绘制成功地定义了CS缺失系的4D断点，该缺失系涉及4D和两个小麦-海纳迪亚-绒毛4D- 4v易位系。13种小麦亲缘种和一种小麦。用两倍体villlosa进行oligo染色。除两份黄斑天牛资料中的4D外，黄斑天牛资料中的4M。comosa和4U在Ae。伞形植物可以被画上。在四倍体Ae。脑室4D、4M均可染色；但与4D相比，4M的信号强度较小。其他外来种未见染色染色体。这表明D/M/U与基因组H/R/Ss/Sc/Y/P/N/J之间的关系比D/A/B和D之间的关系更密切。我们成功开发的4d特异性寡核苷酸探针文库可作为开发其他同源染色体特异性寡核苷酸探针的模型。
Chromosome painting is a useful technique for distinguishing specific chromosomes (fragments), elucidating the genetic relationships of different genomes or chromosomes, and identifying chromosomal rearrangements. The development of chromosome- or genome-specific probes is fundamental for chromosome painting. The possibility for developing such probes specifically painting homoeologous chromosomes in allopolyploid species has been questioned since that chromosomes belonging to the same homoeologous group share highly conserved sequences. In the present study, we attempted to construct a wheat chromosome 4D-specific oligo probe library by selecting 4D-specific sequences in reference genome of common wheat cv. Chinese Spring (CS, 2n = 6x = 42, AABBDD). The synthesized library contains 27,392 oligos. Oligo painting using the probe library confirmed its specificity, shown by that only chromosome 4D could be painted in three wheat genotypes and CS nulli-tetrasomic line N4AT4D. Oligo painting was successfully used to define the 4D breakpoints in CS deletion lines involving 4D and two wheat-Haynaldia villosa 4D-4V translocation lines. Thirteen wheat relatives and a Triticum durum-H. villosa amphiploid were used for oligo painting. Except the 4D in two Aegilops tauschii accessions, the 4M in Ae. comosa and 4U in Ae. umbellulata could be painted. In tetraploid Ae. ventricosa, both 4D and 4M could be painted; however, the signal intensity of 4M was less compared with 4D. No painted chromosome was observed for the other alien species. This indicated that the relationship among D/M/U was closer than that among D/A/B as well as D with genomes H/R/Ss/Sc/Y/P/N/J. Our successful development of 4D-specific oligo probe library may serve as a model for developing oligo probes specific for other homoeologous chromosomes.