| Stripe rust (Puccinia striiformis Westend f. sp. tritici), caused by Puccinia strriformis Westend f. sp. tritici Eriks, is one of the most important disease of wheat (Triticum aestivum L.), which resulted in massive wheat yield loss in several epidemics in China. Using resistant varieties is the most economic, effective and environment-friend method. It is especially important to use the variety of stripe rust resistance in Gansu province because the new physiological race of stripe rust always originated from Longnan and Tianshui area of Gansu province, and Longdong area is a bridge-area where stripe rust could rapidly spread around to lower reaches of Yellow River and Huaihe river.In order to make the best use of germplasm of wheat stripe rust resistance and to develop new resistant variety for wheat, 69 germplasms of wheat stripe rust resistance were employed to analyze genetic diversity by 9 polymorphic SSRs closely linked to stripe rust resistance genes on chromosome 1B, 4D and 6A. In addition, the comparison was performed between CTAB and SDS to select a powerful method for DNA extraction, afterwards a comparison of the methods of PAGE Electrophoresis and Silver Staining between Sanguinetti-PAGE and Bassam-PAGE silver staining was conducted. The results are as follows:1.For genetic diversity analysis, 87 alleles were detected at 9 loci, and the variation ranged from 6 to 15 alleles with an average of 10.778. The PIC ranged from 0.515 to 0.874 with an average of 0.725. The results of cluster analysis based on SSR data indicated that the gene pool of germplasm of wheat stripe resistance was relatively narrow because the genetic similarity (GS) ranged from 0.600 to 0.985. Sixty-nine cultivars (lines) were clustered into 12 groups. And the results of cluster analysis were basically consistent with the pedigrees of these cultivars (lines).2.In order to select a more useful method of micro-DNA extraction for wheat seedling, the quality of DNA extracted by CTAB and SDS were compared. The results indicated that two DNA extraction methods of CTAB and SDS both could produced quality DNA, but CTAB took as long as twice time than SDS. So SDS method was more effective to some researches that the quality of DNA is low, such as Analysis of Genetic diversity and Gene mapping.3.A comparison of the sensitivity of Sanguinetti and Bassam silver staining for detection of SSR-PCR. Sanguinetti silver staining characterized weak background, sensitivity, time saving. The staining procedure of Bassam silver staining were more complex, meanwhile desire agentia to reach ACS grade; Because of dyeing agentia was made in china which could not satisfy the requirement of Bassam silver staining, this method did not dye successfully in the testing. |
