| DNA molecular markers is a new advanced technology applied in biological and biotechnological research. It has a number of advantages of over other markering technologies of morphology, cell cytology and biochemistry. Because this technology works directly on DNA, the genetic basis of plant, it has no restriction of plant develop stage, covering of all genome, allelic identification with numerous polymorphisms, simple and quick detection, less environment interaction, and well experimental repeatability. At present, there are four major methods, RFLP, RAPD, AFLP and SSR to detect the DNA variance. Currently, the technology of DNA molecular markers are being widely applied in the areas of plant research in germplasm classification and reservation, genetics study, gene mapping and marker assistant breeding. It has been shown that the method called molecular identification of seed purity and authenticity has been put forward and is considered to have high accuracy, reliability and low cost. Therefore, the method is becoming a developing trend to identify seed purity and authenticity. And also it will provide the managers and breeders with legal evidence to get rid of the assumed and imitation seeds to be brought in the market and to protect plant proprietary rights.In this study, five super hybrid rice combinations including the new promising super hybrid rice combination which will be released in the near future in China and their parental lines were tested by means of SSR analysis. The purpose of the research is: 1) to make the distinguishment of these hybrid rice combinations and their parents; 2) to establish a set of screening pictures and DNA fingerprint map concerning these super hybrid rice combinations and their parents; 3) to find a convenient method for identification of rice seed purity.In this study, 144 SSR primer pairs distributing in 12 rice chromosomes were used to detect the experimental materials. The result showed that most of primer pairs used amplified visible DNA banes and 47 of these showed evidence of polymorphism. (1)Among all these primers, RM337 and RM 154 produced polymorphic patterns in four or more of the tested experimental materials respectively, and they could distinguish among almost rice genotypes tested. (2)24 of 47 primer pairs which each two are in one of rice chromosome were polymorphic totally with a mean of 3.25 SSLP markers per primer pair and were ultimately selected as criterion primers to set up the screening pictures. (3)For most of primer pairs, FI hybrids mainly showed complementary pattern of both parents, which could be very useful to distinguish difference of FI and their male and female parents. It could be regarded as one of method to identify seed purity. On the other hand, 5 primer pairs were selected as special primer pairs for five hybrid rice combinations respectively. Primer pair of RM250 was used for identification the purity with single seed of HYS-1/R105. (4)By combining the rapid, simple method on DNA extraction, it is suggested that SSR technique has wide prospective in variety authentication and purity identification.In this study, several problems were discussed:(1) SSR analysis is suitable for variety authentication and purity identification.(2) For three methods used to extract rice total DNA in ths study, the simple method with acid and alkali for DNA extraction has wide prospective in variety identification.(3) Because of the limitation of experimental condition, it was hard to find the primer for distinguishment between 9311 and 0293.(4) It had a good result to get perfect banes under the condition of electrephoresis with 100-120 volt/cm and 2.5EB staining.It is suggested that the result obtained in this study could provide the legal evidence to protect plant proprietary rights of the new super hybrid rice combinations breeded recently. |
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