| Chinese fir (Cunninghamia lanceolata) is an endemic species in China, which has a lot of good characteristic such as rapid growth, high yield, good reproductive performance, et cetera. With the continuous progress of the forest rights reform in Fujian province, a large amount of natural forest including broad-leaved forest has been destroyed in order to plant the fir forest. Therefore, there is a sharp decline in the germplasm resources of natural fir forest. As the planting history of Chinese fir king is very long, the germplasm resources of natural Chinese fir had scattered in many places, which had increased the difficulties of the management of the germplasm resources of natural fir forest. Fujian is the traditional center production areas of Chinese fir and it is rich in resources of Chinese fir king. Chinese fir king has experienced the natural ordeal for a long time, and during the process,it has record the changes of climate history, which made it be full of great scientific value.Therefore, the study on genetic variation of Fujian Chinese fir trees in Fujian Chinese fir center production area can provide an important evidence to reveal the distribution patterns of Chinese fir trees in Fujian province.Yu Xin-tuo, etc. had investigated the growth condition of the 20 Chinese fir king in Fujian Province in 1979, but 30 years later, there is no relevant reports about the resources distribution, particularly in relation to the number of Chinese fir king, growth conditions and their genetic diversity is also scarce. For this reason, this study investigates the number of Chinese fir king, growth conditions and distributing, and compares the genetic diversity of the existing 44 different geographical provenances of Chinese fir king by ISSR molecular marker technique, and analyzes Eco-fir and Genetic variation of genetic about Chinese fir king, At the same timeanalyzes samples for genetic relationship,using genetic dissimilarity coefficient for different provenances, which revealed a different provenances Genetic variation of Chinese fir king and population distribution patterns, providing the molecular level evidences on, the range of Fujian fir Center production and population distribution patterns for a reasonable division of Chinese fir provenances, breeding programs to develop and germplasm resource management but also providing a scientific basis for Chinese fir seed source germplasm gene pool for the establishment and protection of ancient and famous trees The major findings are as follows:1,The comprehensive survey about Chinese fir king germplasm resources indicates that: accirding to the standard about Chinese fir king, its diameter at breast height, tree height and plant volume respectively over 1m, 30m, and 10m3, the old and big fir can be called Chinese fir king.There are 44 Chinese fir king germplasm resources currently. Mainly distributing in western Fujian, northern Fujian and Taiwanese, eastern regional distribution at least, of which the number of western Fujian is largest,17;Taiwanese,11; northern Fujian,8; central Fujian,6; eastern Fujian,2.The surveyed 44 Chinese fir king are belong to the scope of National fir trees protection, the first level 16,the second level 9,the third level 19.2,Three is a relatively large differences in growth conditions of different Chinese fir king germplasm.In the central production region of Northern Fujian, the growth condition of different Chinese fir king on volume, has the greatest difference, coefficient of variation reached 0.68; In the Fujian fir ordinary production region, the growth condition of different Chinese fir king on high living branch, has the greatest difference, the coefficient of variation reached 0.71;In the Fujian fir boundary production region, the growth condition of different Chinese fir king on volume, has the greatest difference, the coefficient of variation reached 0.37;In the central production region of Northern Fujian, the Fujian fir ordinary production region, and the Fujian fir boundary production region, the average growth condition f different Chinese fir king on crown, has the greatest difference, the coefficient of variation reached 0.51;the two surveys separated by 30 years show that a part of Chinese fir king grows slowly, their growth condition are at the saturation.3,The DNA extraction of Chinese fir king has stabled. This is very difficulty because of the high level of secondary compounds and their different distribution, for example, polysaccharides, pigments and phenols. The improved SDS method which is suited for Chinese fir king,the extracted DNA is suitable for ISSR-PCR.4,A valid reaction system suitable for the ISSR analysis about Chinese fir king was established (20μL,total reaction buffer), composed by 75ng template DNA,150μmol/L dNTP, 0.4umol/L premier, Mg2+1.5mM, 1.5 U Taq polymerase;The PCR parameter is:94℃pre-denature for 7 min, then 45 cycle of 30 second at 94℃for denaturation of 40 second at 55℃for annealing,of 2 min at 72℃for extension ,finally extension at 72℃for 7 min ,the product of PCR was kept at 4℃.5,Twenty-seven primers which are selected from one hundred primers tested on the base of the number and frequency of polymorphisms are utilized in analyzing genetic diversity and relationship of Chinese fir king.They had 226 excellent amplification DNA, One hundred and ninety-eight Polymorphic loci are amplified and the average percentage of polymorphic sites are 87.61﹪, it showed rich polymorphism.6,According to Nei-Li method, it calculated that the genetic distance range of 44 different Chinese fir king germplasm was between 0.0891 and 0.6435, the average genetic distance 0.3829,22 and 27 have the largest genetic distance 0.6435, 27 and 29 have the least genetic distance,only 0.0891.Description ISSR markers can effectively reveal that the genetic differentiation of different Chinese fir king provenances is not at a low level.7,The results of the ISSR-PCR and UPGMA clustering analysis showed that the genetic diversity of Chinese fir king was not at a low level, the result of germplasm genetic distance clustering is basically consistent with the geographical distribution, which shows that geographical distance and differences in their environment different conditions may be the important factor of germplasm genetic variation. |
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