Study On Tissue Culture Of Pleione Formosana (Orchidaceae)

To build the rapid propagation system of Pleione formosana,the wild plants were introduced into the greenhouse for cultivation,then the capsules from artifical pollnation and wild plants were used as materals were preserved at low temperature of 4? and the seed activity was measured during storage.6-BA and NAA were used as the two factors,and the samples were tested for sterile germination at three levels.The orthogonal design was designed with 6-BA,NAA and TDZ as the three factors;for propagation,the orthogonal experiment of protocorm was designed with 6-BA,NAA and TDZ as the three factors.The orthogonal experiment of protocorm,differentitation was desigued with 6-BA,NAA and TDZ as the three factors.MS and 1/2MS were used as the basic medium,and different concentrations of NAA were used to design experiment of rooting.The method of single and secondary disinfection were used to disinfect the root with 75% alcohol,0.1% mercuric chloride and saturated hypochlorite as disinfectant,the root segments and root grinding fluid were used as inculum respectively.The isolated fungi were co-existing with saved seeds in the mixture of wood chips and bark as equal volum.1,The seed activity was detected at 0 d,1 d,2 d,3 d,4 d,10 d,13 d,20 d,25 d and 30 d,and the seed activity at 0 d was 76.25%,80.84% for 1 d,2 d seed activity was the highest in 2 d,reaching 93.87%,92.24%for 3 d and began to decline,the seed activity was 83.28% at 20 d,and the seed activity decreased to 65.80% at 30 d.The best sowing time was for 2 d,and the sowing was carried out in 20 days.The germination rate of sterile germination was more than 90%.The earliest germination medium was 1/2MS+0.5 mg/L 6-BA+1.0 mg/L NAA,but the young buds were weak;germination after the protocorm to maintain a longer time for 1/2MS+0..5~1.0 mg/L NAA.2,The best protocorm proliferation medium was MS+0.2 mg/L NAA+ 1.5 mg/L 6-BA+50 g/L sucrose,and the multiplication coefficient was 3.150,and the 6-BA,NAA had significant effect on proliferation of protocorms,but not the sucrose.3,.The optimum inflation medium of sucrose was MS+0.5 mg/L 6-BA+ 1.5 mg/L NAA,and the expansion coefficient was 3.205.The optimum inflation medium of hormone was MS+0.5 mg/L 6-BA+1.5 mg/L NAA,the expansion coefficient was 3.205,6-BA and NAA had significant effect on the protocorm enlargement,and TDZ had no significant effect on protoplast enlargement.4,The best differentiation medium was MS+0.6 mg/L TDZ+1.5 mg/L 6-BA+1.0 mg/L NAA,the differentiation rate was 84.1%,6-BA and NAA had significant effect on the differentiation of protocorms.The results showed that the best rooting medium was 1/2MS+0.5 mg/L NAA,the rooting rate was 85.4%.The total number of plants was 120,the total number of rooting was 213,the average number of rooting was 1.8.5,The best disinfection step was this way,which was 75% alcohol 30s?0.1% mercuric chloride 60 s,90s,with grinding inoculation,73 kinds of fungi were found;of which 7 endophytic fungi can not promote seed germination,66 kinds of endophytic fungi can promote seed germination to form protocorms,and 29 can promote seed germination to form yellow-green protocorms,and 37 species can promote seed germination to form green protocorms.All germinated protocorms did not undergo differentiation.