Tissue Culture Of Pinus Massoniana's Seedling Stems And Spring Shoots

Pinus massoniana Lamb is a important fast-growing local conifer which has a high comprehensive utilization value in the south of China.but there has some problems,the seed orchard'yield was low and the rooting of cutting had age benefit,establish rapid propagation frmaework can lay a nicer basis for Pinus' cultivation,technology of somatic embryogenesis and the geneic transformation.This article maked seedling stems which came from Masson pines seed orchard of Guiyang Hunan province, and spring shoots of superior trees from Hunan Botanical Garden as the starting explants.the results showed:(1)the suitable culture medium of seedling stem's callus was modified GD+4 mg/L 2,4-D+2 mg/L 6-BA+500 mg/L CH+450mg/L Gln+30 g/L Sucrose,the callus'maximum diameter was 8.5mm; the suitable culture medium of spring shoots' callus was DCR+0.5 mg/L NAA+2 mg/L 2,4-D+1.5 mg/L 6-BA+0.2 g/L Vc+500 mg/L CH+450mg/L Gin+30 g/L Sucrose,Callus looks crystal clear, surface was a little yellow, and had a little bit sticky.(2).Added concentration of 3 g/L inositol in the induction medium can solution the problem of browing in the callus subculture., you can maintain the original morphology of callus more than five generation. the ratio of auxin and cytokinin in 1-3 can maintain and proliferate calluses, In this hormone concentration falling within the scope of no significant differences between treatments,2,4-D concentration in the 4 mg/L,6-BA concentration of 2 mg/L were the best for the Masson pine seedling stem's callus between subculture; the best one of the subculture about spring shoots' callus were add add 0.5 mg/L NAA,2 mg/L 2,4-D,1.5 mg/L 6-BA. How to prevent the callus turned brown is the basis of solution of the key problems of embryogenesis that preservation the embryogenic callus and promoting germination of the somatic embryo.(3)This is the first time using the spring shoots of superior trees as the explants to induce adventitious buds,and had two adventitious buds induced with the medium was DCR+1 mg/L KT+0.5 mg/L TDZ+500 mg/L CH+3 g/L Inositol+30 g/L Sucrose,it also validated Masson pine adventitious buds could be reduced by indirect organogenesis.(4)It's relatively easy to establish Masson pine regeneration system by using seedling stem with cotyledon as the explants. After four subculture,root induction rate was 21.4% by using the medium GD+0.5 mg/L IB A+0.05 mg/L 6-BA+30 g/L Sucrose;the root induction rate was 57.1% by using such combination medium modified GD+0.5 mg/L NAA+20 g/L Sucrose.