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小麦品系CH5383是源于中间偃麦草(Thinopyrum intermedium)的兼抗多种小麦病害的新种质。为明确其抗性来源和外源DNA片段的渗入位置,采用基因组原位杂交(GISH)和荧光原位杂交(FISH)对CH5383进行分析,GISH分析未发现外源信号,FISH结果观察到CH5383的3BL染色体端部与对照小麦(Triticum aestivum)品种中国春相比有明显的重组信号,初步推断CH5383染色体3BL端部可能有中间偃麦草DNA片段插入。用135对PLUG(PCR-based landmark unique gene)标记对CH5383、中国春和多个近缘物种进行扩增分析,发现位于3B染色体长臂端部的引物TNAC1383,能在CH5383和中间偃麦草中扩增出大约1 300 bp的特异DNA产物。从而进一步证实,CH5383是一个小麦-中间偃麦草小片段渗入系,携带抗条锈病基因的外源中间偃麦草DNA渗入片段位于3B染色体端部0.81-1.00区段。CH5383可以作为优异的小麦抗病育种新种质加以利用。
The wheat line CH5383 is a new germplasm derived from Thinopyrum intermedium against multiple wheat diseases. In order to clarify the source of resistance and the infiltration position of foreign DNA fragments, CH5383 was analyzed by genomic in situ hybridization (GISH) and fluorescence in situ hybridization (FISH). No external signal was detected by GISH. FISH results showed that CH5383 There was a significant recombination signal at the end of chromosome 3BL compared with that of Chinese wheat (Triticum aestivum). It was preliminarily concluded that there may be some insertion of DNA fragment from chromosome 3BL of CH5383 on the end of chromosome 3BL. Using PCR-based landmark unique gene (PLUG) marker to amplify CH5383, Chinese spring and several related species, we found that the primer TNAC1383 located at the end of the long arm of chromosome 3B could amplify in CH5383 and middle- Add about 1 300 bp of specific DNA product. It was further confirmed that CH5383 was a small wheat-Thinopyrum intermedium infiltration system, and the DNA fragment of exogenous Tricholoma matsutake carrying the stripe rust resistance gene was located in the segment of chromosome 0.81-1.00 at the end of chromosome 3B. CH5383 can be used as excellent germplasm of new wheat breeding.