Studies On Genetic Diversity Of Several Chemistries Types Of The Cinnamomum Caphora And Related Species

Cinnamomum Caphora(L.)Prest is of high economic value and is used widely.Its main and sideline product is also used widely for architecture, craft, chemical industry, war industry and medical, and so on. In this research, we selected the several Chemical types of the camphor tree and its related species as an object, and analyzed their blood relationship and genetic diversity by the RAPD and ISSR molecular markers. Through the research,several conclusions have be find as follows:1. According to the common procedures of molecular markers of RAPD and ISSR, some methods have been adjusted in analyzing several chemical type and allied species of Cinnamomum Caphora (L.) Prest. And we have exploded a fit of reliable method for extracting DNA from Cinnamomum Caphora (L.) Prest and established a suit of analysis system of RAPD and ISSR which is proper for Cinnamomum Caphora (L.) Prest. The RAPD reaction system in our research followed as: the whole volume of the reaction is 20μL, and the mixture in this reaction included 50ng DNA, 2.0mmol/L MgCl₂, 0.2μmol/L primer, 150μmol/LdNTPs, 1U TaqDNA polymerise and 1×PCR buffer; and the RAPD PCR program was 94℃7min, 1 cycle; 94 ℃ 30s, 37℃ 30s,, 72℃ 90s, 38 cycles; 72℃ 10min, 1cycle. The best reaction system for ISSR in our research followed as: the whole volume of the reaction is 20μL, and the mixture in this reaction included 50ng DNA, 2.0mmol/L MgCl₂, 0.2μmol/L primer, 200μmol/LdNTPs, 0.5U TaqDNA polymerise and 1×PCR buffer; and the PCR program was 94℃7min, 1 cycle; 94 ℃ 30s, specific temperature 45s, 72℃ 90s, 38 cycles; 72℃ 10min, 1cycle.2. Twenty-three primers of 10 base pairs were selected from 112 random primers. They were used to analyze the 35 accessions with RAPD, and 173 bands were generated, among them the number of polymorphic bands was 81, the percentage of polymorphism was equaled to 46.8%. The average number of bands directed by each primer is 7.5; 16 primers, which could amplified bands clearly, were selected from 46 ISSR primers. As a result, 141 bands were generated, among which polymorphic bands number was 78, and the percentage of polymorphism was 55.3%. The average number of bands directed by each primer is 8.8. Through relevant analysis, the results gained through RAPD and ISSR molecular markers were significantly correlated. The conclusion indicated that the two methods can be used to study the blood relationship and genetic diversity in the camphor tree and the ISSR is more effective than the RAPD.