Molecular Signs And Genetic Analysis Of Cunninghamia Lanceolata(Lamb) Hook And Submarginal Species

Cunninghamia lanceolata (Lamb) Hook, Cunninghamia lanceolata (lamb.) Hook. Var.konishii Hayata, Cryptomeria fortunei Hooibrenk ex Otto et Dietr and Taiwania floudiana Gaussen are important trees species of Taxodiaceae for timber plantation in south China. C. lanceolata (Lamb) Hook distribute widely with long planting history in China. C. lanceolata (lamb.) Hook. Var.konishii Hayata distribute in hilly region of north and middle of Taiwan. For geographic isolation of mainland and Taiwan,C. lanceolata (lamb.) Hook. Var.konishii Hayata and C. lanceolata (Lamb) Hook have diaparity to certent from external morpha, type of nucles in chromosome and so on, then, the classified relationship of them exist disagreement in long term. The ideas about relation of T. floudiana Gaussen and T.cryptomerioides Hayata aren't unitive from 1906 when Taiwania established. Some scholars think that the genus include two species of T. floudiana Gaussen and T.cryptomerioides Hayata, others consider that one species is involeved in the genus because the T. floudiana Gaussen and T.cryptomerioides Hayata are the same species.The relationships of C. lanceolata (lamb.) Hook. Var.konishii Hayata and C. lanceolata (Lamb) Hook was studied in comparison with relationships of C. lanceolata (Lamb) Hook and T. floudiana Gaussen, C. lanceolata (Lamb) Hook and C. fortunei Hooibrenk ex Otto et Dietr and geographic groups of C. lanceolata (Lamb) Hook by RAPD molecular sign from molecular level. The relation of T. floudiana Gaussen and T.cryptomerioides Hayata was analyzed through RAPD from molecular level in comparison with geographical groups of C. lanceolata (Lamb) Hook. At the same time, the hereditary variation about geographical groups of C. lanceolata (Lamb) Hook and about clones of C. lanceolata (Lamb) Hook were researched by RAPD. This research can offer the molecular reference and reliable method for the reasonable partition and management about group of C. lanceolata (lamb.) Hook. Var.konishii Hayata, geographic groups of C. lanceolata (Lamb) Hook and clones of C. lanceolata (Lamb) Hook.The results are as follows:1. The DNA extraction and purification for conifer of Taxodiaceae is very difficulty because of the high level of secondary compounds, for example, pigment, oxybenzene and polysaccharide. The improved SDS method which is suited for conifer of Taxodiaceae isput forward in this study.2. The reliable RAPD analysis system is establishes The reaction volume is 20ul and reactionmixture consist with 5ng/ul DNA, 150uM of dNTPs, 1.8mM Mg2+, 0.5uM random primer and 1U Taq polymerase. RAPD program is 5 min at 94 C for predenaturation, then 38 cycles of 30 seconds at 94 C for denaturation, of 30 seconds at 36 C for annealing, of 90 seconds at 72 C for extension, finally extension at 72 C for 7 min.3. Twenty-one primers which selected from one hundred primers tested on the base of thenumber and frequency of polymorphisms are utilized in analyzing the groups relationships of C. lanceolata (lamb.) Hook. Var.konishii Hayata, C. lanceolata (Lamb) Hook, C. fortunei Hooibrenk ex Otto et Dietr and T. floudiana Gaussen. Nineteen primers are selected to analyze hereditary variation about geographical groups of C. lanceolata (Lamb) Hook. By the way, other nineteen primers are selected to identify clones of C. lanceolata (Lamb) Hook.4. UPGMA cluster analysis was carried out using software BiolD++. Bendrogram wasconstructed based on the RAPD data. (l)The result indicate that this partition that C. lanceolata (lamb.) Hook. Var.konishii Hayata is a variety of C. lanceolata (Lamb) Hook is more reasonable than others. (2) The outcome suggests that T. floudiana Gaussen and T.cryptomerioides Hayata are one species. (3) The result on molecule level of manifest that the division of geographical groups of C. lanceolata (Lamb) Hook by molecular signs and the division by traditional geographical zone and character-growth are accordant. (4)The molecular signs can identify clones of C. lanceolata (Lamb) Hook through a few primers and signs.Concl...