Study On Germplasm Evaluation Of Paphiopedilum And Aseptic Seeding And Tissue Culture Technology Of Paphiopedilum Pacific Shamrock

Paphiopedilum plants have become popular high-end flowers in the flower market because of their unique flowers and colors.The Paphiopedilum Emerald Furture‘Galaxy'become the world's mainstream commercial varieties.At present,there are many varieties of Paphiopedilum,but the evaluation system of these varieties is not perfect.Because Paphiopedilum seeds have no endosperm,the seed setting rate and germination rate under natural condition are very low,so there are a lot of researches on aseptic germination of Paphiopedilum,but there are few studies on the Paphiopedilum pacific.In addition,although there are many studies on the effect of growth regulators on the proliferation of procorm,the results of the study vary greatly due to the variety studied,and the study of adventitious buds induced by explants is not mature in the field of Paphiopedilum.Therefore,12 wild aristolochia varieties were tested and evaluated by analytic hierarchy process(AHP).and the best pollination time suitable for aseptic sowing,the best culture medium,the optimal hormone ratio of procorm proliferation and inducing adventitious bud explants was screened out by using Paphiopedilum pacific as the test material.The results are as follows:?By the AHP,12 Paphiopedilum species could be classified 3 grades.The first grade included ‘P.philippinense'?P.tigrinum'? ‘P.hirsutissimum'?‘P.purpuratum'? ‘P.dianthum'? ‘P.henryanum'? ‘P.malipoense',with the highest ornamental value.The second grade included ‘P.ameniacum' and ‘P.concolor',the ornamental value was general.The third grade included ‘P.emersonii' and ‘P.micranthum',ornamental value was low.Therefore,in the actual production,the first grade varieties could be the first choice.? The pollination time had a great effect on the aseptic germination rate of Paphiopedilum seeds.It took about 300 days for the seeds to mature from pollination to maturity.The hybrid seeds of 150~315 days were collected.On the medium of 1/4MS+ banana mud 25 g/L+ AC 0.5 g/L +agar 5.5 g/L+sucrose 10 g/L,it was found that the germination rate of seeds increased gradually from 150 days and the germination rate of seeds at 150 days was 7.63%.The germination rate of seeds was 90.29% at 255 days.After 255 days,the germination rate decreased gradually,and only 21.21% at 315 days.?The type of basal medium also had a great effect on the germination rate of Paphiopedilum seeds.The seeds of Paphiopedilum were seeded aseptically in MS,1/2MS,1/4MS and 1/8MS media for 255 days after pollination,and found that the highest germination rate was 82.67% in 1/4MS medium,and the germination rate was 67.78% in 1/8 MS medium.In high concentration MS medium,the germination rate was the worst,and the germination rate was only 7%.?Different hormone concentration and ratio had a great effect on the proliferation of procorm of Paphiopedilum pacific.The green protocorms germinated from Paphiopedilum pacific were cultured in 1/4MS medium with different concentrations of 6-BA and NAA.The results showed that the procorm proliferation effect was the best under the hormone ratio of 6-BA5.0 mg/L +NAA0.5 mg/L,and the growth rate was the best,the multiplication multiple was 3.83,but the procorm proliferation effect was the worst when 6-BA3.0 mg/L was used,and the growth rate of the protocorm was worst,browning is serious.?In the study of adventitious bud induction from the explants of Paphiopedilum pacific,the flower buds,stem tips,leaves,root tips and aseptically germinating seedlings of Paphiopedilum pacific were inoculated into 1/4MS+6-BA5.0 mg/L+NAA0.5 mg/L medium as explants.The results showed that the root of aseptically seeded Paphiopedilum seedlings was easily induced to germinate buds,but the adventitious buds were not induced successfully by explants such as leaves,stem tips,flower buds,etc.All explants were browned,and adventitious buds were not induced successfully.In this experiment,12 kinds of wild Paphiopedilum were evaluated,aseptic seeding and tissue culture were studied.The evaluation system and aseptic germination and tissue culture system of Paphiopedilum were established.It provides a theoretical basis and scientific guidance for the breeding and tissue culture of Paphiopedilum.