In 1989, Leymus racemoses DNA was introduced into a spring wheat cultivar “761” through pollen tube Pathway. In the resulted F3 progenies, apparent trait changes were observed. In the F4 progenies, many lines, posse...In 1989, Leymus racemoses DNA was introduced into a spring wheat cultivar “761” through pollen tube Pathway. In the resulted F3 progenies, apparent trait changes were observed. In the F4 progenies, many lines, possessing longer ears, more grains per ear, higher 1000 grain weight and higher protein content, were obtained. We reasoned that some genetic components of Leymus racemoses might have been transferred and integrated into the wheat genome. In order to get molecular proof for above reasoning, RFLP, RAPD and storage protein analysis were carried out. The following results were obtained. 1. RFLP analysis. Four repetitive DNA sequence (pHv7161,pHv7179, pHv7191 and pHv7293) clones from barley ( Hordeum vulgare) genome were used for molecular hybridization using the genome DNA of Leymus racemoses (donor), spring wheat 761 (receptor) and the putative transformed wheat lines. Our results revealed some bands, common to Leymus racemoses and the putative transformed wheats, were absent in spring wheat 761. 2. RAPD analysis. 160 operon primers were used to amplify polymorphic bands using genomic DNA of Leymus racemoses, spring wheat 761 and the putative transformed wheats. 20% of the primers was polymorphic. The calculated genetic relationship based on the RAPD analysis were:35% between Leymus racemoses and spring wheat 761, 90% between the putative transformed wheats and spring wheat 761. 3. Storage protein analysis. After comparison of the patterns of gliadin and glutenin by SDS PAGE among Leymus racemoses, spring wheat 761 and the putative transformed wheats, we found that, in gliadin analysis, a new band appeared in the putative transformed wheats with an electrophoretic mobility similar to that of a gliadin polypeptide of Leymus racemoses .In glutenin analysis, a few high molecular weight glutenin subunits appeared in the putative transformed wheats, which might be derived from Leymus racemoses. These subunits may have direct impact on flour processing properties, and are worthy of further investigations.展开更多
文摘In 1989, Leymus racemoses DNA was introduced into a spring wheat cultivar “761” through pollen tube Pathway. In the resulted F3 progenies, apparent trait changes were observed. In the F4 progenies, many lines, possessing longer ears, more grains per ear, higher 1000 grain weight and higher protein content, were obtained. We reasoned that some genetic components of Leymus racemoses might have been transferred and integrated into the wheat genome. In order to get molecular proof for above reasoning, RFLP, RAPD and storage protein analysis were carried out. The following results were obtained. 1. RFLP analysis. Four repetitive DNA sequence (pHv7161,pHv7179, pHv7191 and pHv7293) clones from barley ( Hordeum vulgare) genome were used for molecular hybridization using the genome DNA of Leymus racemoses (donor), spring wheat 761 (receptor) and the putative transformed wheat lines. Our results revealed some bands, common to Leymus racemoses and the putative transformed wheats, were absent in spring wheat 761. 2. RAPD analysis. 160 operon primers were used to amplify polymorphic bands using genomic DNA of Leymus racemoses, spring wheat 761 and the putative transformed wheats. 20% of the primers was polymorphic. The calculated genetic relationship based on the RAPD analysis were:35% between Leymus racemoses and spring wheat 761, 90% between the putative transformed wheats and spring wheat 761. 3. Storage protein analysis. After comparison of the patterns of gliadin and glutenin by SDS PAGE among Leymus racemoses, spring wheat 761 and the putative transformed wheats, we found that, in gliadin analysis, a new band appeared in the putative transformed wheats with an electrophoretic mobility similar to that of a gliadin polypeptide of Leymus racemoses .In glutenin analysis, a few high molecular weight glutenin subunits appeared in the putative transformed wheats, which might be derived from Leymus racemoses. These subunits may have direct impact on flour processing properties, and are worthy of further investigations.
