Study On Germplasm Diversity Of Olive From QingChuan Of GuangYuan By RAPD And Morphologic

This study relied on school-county cooperated item. In 2005, all around investgation on the olive resource of Qingchuan of SiChuan, and cull excellent trees. This paper aimed at the analysis on Olive resources which belong to the county of QingChuan by using Morphological and RAPD technique. Investigation and selectness excellent trees relate to seven villages and towns of QingChuan: ShaZhou Town, YingPan Town, MuYu Town, YaoDu Town, DongShui Town and BanQiao Town. For the selected 188 trees, construe the intrinsic relation among morphological characters by Principal Component Analysis, and seeking after fitting the reaction system and amplified program of RAPD. Two hundreds primers were the first selected, obtained 14 primers which had better polymorphism. These primers could be used for UPGMA(unweighted pair group method arithmetic average), established tree plot. The results were as follows:1. Description and Statistic 13 morphological characters showed that: coefficients of differentiation of the height and thickness of trunks were largest among all, of the height of trees and length of leafstalks were second largest, for the rest, all were only less than 20 percent. In addition, the order of coefficients of differentiation from big to small was: the leafage shape, fruits shape, stone shape. These showed that stone shape index was a steady guide line.2. Factor analysis of morphological characters indicated that: could distill four principal component. These four components accounted for 75.636% of the total variance. Among Fac-1, variables such as the height and thickness of minks, trees shape explained the largest portion of the variance. So, Fac-1can be said tree shape index. Among Fac-2, variables such as indexes of fruit shape and pit shape explained the largest portion of the variance. So, Fac-2 was fruit shape guide line. On Fac-3 the largest scores were due to characters associated with the shape of leafage. So, Fac-3 was the index of the shape of leafage. Also, on Fac-4 the largest scores were due to characters associated with the force and the height of trees. Called fac-4 as the index of potential of tree.3. Correlation analysis of indexs indicated that: coefficient of fruit shape, coefficient of pit shape, coefficient of leafage shape and length of leafstalk are important indexs for Olive of variety distinguish and choice of better Germplasm Resources. However, the height of trees, the force of trees, the shape of trees, the shape of crown of a tree, the thickness and the height of trunks are only reference. These indexs don't can be indexs of variety distinguish and choice of better Germplasm Resources for Olive. 4. CTAB method was adopted to isolate genomic DNA from Olive leaf. The result indicate that obtained DNA which has better integrality and equal concentration, was much idealer to the further research of RAPD marker in Olive.5. By group for the PCR amplification program and the reaction system in Olive, approve that an optimal reaction system suitable for the assay and usage of RAPD in Olive, and establish base on biological investigation for the future.PCR amplification program was denature in advance at 94℃for 3min, denature 94℃for 1min, anneal at 36℃for 1min, extend at 72℃for 1min 30s, followed 38cycles. Then extend at 72℃for 10min and store at 4℃.The reaction system(25μl) was consisted of dNTP 0.54μl (10mM), Mg²⁺ 2.4μl(25mM), Taq Enzyme 0.4μl ( 2.5U/ul ), Tap Reaction Buffer 2μl (10×), Primer 0.3μl (51 pm/μl), DNA 2μl (20ng/μl) and ddH₂O 12.36μl。6. 200 primers were first selected. 14 primers were decided to apply to formal amplification. These primers had better polymorphism and could get clear repeat bands. 149 bands were amplification by fourteen primers in Olive, 147 polymorphic bands were detected, representing of 98.66% of polymorphic bands(PPB). These bands length ranged from 150bp to 2000bp. Per primer produced 11 bands. Showed Olive came from QingChuan has great rich genetic diversity.7. Among 188 sample, the RAPD-based genetic distance ranged from 0.02 to 0.68, with the average of 0.29. Cluster analysis by unweighted pair group method of arithmetic means (UPGMA) suggested that 188 samples could be divided into four groups with genetic similarity from 0.70. The clustering result showed the variety singleness in some planting area, Lacked considerer in logical arrange in pairs or groups about variety.