| In order to explain the reason of endangerment in Oplopanax elatus of Changbaishan area, the genetic diversity was investigated using RAPD technique.Also,several factors of affecting the rapid propagation in vitro was studied for development and utilization of O.elatus.The optimal PCR system for RAPD analysis was as follows:in a 20.0μL volume,9.8μL ddH2O,250.0μmol·L-1dNTP,2.0 mmol·L-1 MgCl2,1×Buffer,0.4μmol·L-1 primer,10.0 ng template DNA,and 1.5 unit of Taq DNA polymerase.Amplification protocol was initial denaturation of 5 min at 95℃;40 cycles of 45 s at 95℃,1 min at 36℃,and 1.5 min at 72℃and a final cycle of 7 min at 72℃.Among the 120 random primers,16 primers were found to generate distinct DNA fragments,43.62%of 117 RAPD fragments in the samples of O.elatus from 7 different regions were polymorphic,it indicates that O.elatus in Changbaishan area has a lower genetic diversity.The cluster analysis could divide the germplasm into 2 groups:(1) Mt.Laoling of Mt. Changbai,including Baijiyaozi of Tonghua city,Shihu Tonghua country,Manjiang of Fusong country,Bagongli of Linjiang city,and,Dajing of Baishan city;(2) South side of main peak in Mt.Changbai,including Shiwudaogou and Shijiudaogou of Changbai country.The experiments of the rapid propagation in vitro of O.elatus were carried out,the result showed that the favorable proliferation and growth of shoots were found in those inoculums, which were removed apical buds.Moreover,the medium of WPM +1.0 mg·L-1BA+NAA 0.2 mg·L-1+30.0 g·L-1 sugar+7.0 g·L-1 agar and 3/4 WPM+ NAA 0.1 mg·L-1+activated carbon 1.0 g·L-1+30.0 g·L-1 sugar+7.0 g·L-1 agar were suitable for proliferation and rooting of O. elatus in vitro,respectively. |
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