Fluorescence in situ hybridization with multiple repeated DNA probes applied to the analysis of wheat-rye chromosome pairing
Fluorescence in situ hybridization with multiple repeated DNA probes applied to the analysis of wheat-rye chromosome pairing
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DOI：
10.1007/s001220050422
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发表时间：
1997-03-01
影响因子：
5.4
通讯作者：
Ceoloni, C
中科院分区：
文献类型：
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作者：
Cuadrado, A;Vitellozzi, F;Ceoloni, C
应用多探针荧光原位杂交技术对普通小麦×黑麦杂种的减数分裂染色体进行了研究。所用探针包括黑麦的pSc 74和pSc 119.2（后者也与小麦杂交，主要是B基因组染色体），Ae. squarosa pAs 1探针，其几乎只在D基因组染色体上杂交，以及小麦rDNA探针pTa 71和pTa 794。用这些探针的二乘二组合进行同时和连续FISH，可以明确鉴定所有黑麦（R）和大多数小麦（W）染色体，无论是未配对的还是参与配对的。因此，不仅可以很容易地区分小麦-小麦和小麦-黑麦的关联，这已经是可行的，通过单独使用黑麦特异性的pSc 74探针，但个别配对伙伴也可以被识别。在所观察到的小麦-黑麦配对中，平均约占同一杂交组合的6个杂种植株中检测到的总配对的7%至11%，大多数涉及B基因组染色体（约70%），而D（约17%）和A（14%）基因组染色体的程度要小得多。黑麦臂1 RL和5 RL表现出最高的配对频率（超过30%），其次是2 RL（11%）和4 RL（约8%），所有其他臂的值低得多。在分析的样品中从未观察到2 RS和5 RS配对。观察到1 RL、1 RS、2 RL、3RS、4 RS和6 RS染色体臂仅与同一同源群的小麦染色体结合。4 RL（与6 BS和7 BS配对）和6 RL（与7 BL配对）的情况正好相反。另一方面，5 RL在超过80%的情况下与4 WL臂或其片段配对，其余情况下与5 WL配对。涉及小麦染色体属于一个以上的同源异型群的配对的其他情况下发生3RL，7 RS和7 RL。这些结果，同时增加了支持以前的证据存在的几个易位在黑麦基因组相对于小麦，表明，FISH与多个探针是一种有效的方法，通过它来研究染色体行为的基本方面在减数分裂，如种间配对。从这种方法获得的知识类型预计将对小麦和相关小麦族的理论和应用研究产生重大影响。
Fluorescence in situ hybridization (FISH) with multiple probes has been applied to meiotic chromosome spreads derived from phlb common wheat x rye hybrid plants. The probes used included pSc74 and pSc 119.2 from rye (the latter also hybridizes on wheat, mainly B genome chromosomes), the Ae. squarosa osa pAs1 probe, which hybridizes almost exclusively on D genome chromosomes, and wheat rDNA probes pTa71 and pTa794. Simultaneous and sequential FISH with a two-by-two combination of these probes allowed unequivocal identification of all of the rye (R) and most of the wheat (W) chromosomes, either unpaired or involved in pairing. Thus not only could wheat-wheat and wheat-rye associations be easily discriminated, which was already feasible by the sole use of the rye-specific pSc74 probe, but the individual pairing partners could also be identified. Of the wheat-rye pairing observed, which averaged from about 7% to 11% of the total pairing detected in six hybrid plants of the same cross combination, most involved B genome chromosomes (about 70%), and to a much lesser degree, those of the D (almost 17%) and A (14%) genomes. Rye arms 1RL and 5RL showed the highest pairing frequency (over 30%), followed by 2RL (11%) and 4RL (about 8%), with much lower values for all the other arms. 2RS and 5RS were never observed to pair in the sample analysed. Chromosome arms 1RL, 1RS, 2RL, 3RS, 4RS and 6RS were observed to be exclusively bound to wheat chromosomes of the same homoeologous group. The opposite was true for 4RL (paired with 6BS and 7BS) and 6RL (paired with 7BL). 5RL, on the other hand, paired with 4WL arms or segments of them in more than 80% of the cases and with 5WL in the remaining ones. Additional cases of pairing involving wheat chromosomes belonging to more than one homoeologous group occurred with 3RL, 7RS and 7RL. These results, while adding support to previous evidence about the existence of several translocations in the rye genome relative to that of wheat, show that FISH with multiple probes is an efficient method by which to study fundamental aspects of chromosome behaviour at meiosis, such as interspecific pairing. The type of knowledge attainable from this approach is expected to have a significant impact on both theoretical and applied research concerning wheat and related Triticeae.