The Establishment And Optimization Of Hybrid Phalaenopsis Tianxiang L04 Tissue Culture Technical System

Phalaenopsis possesses some excellent characteristics ,such as the great flower, radiantly beautiful color , the very unique shape and elegant looks of its flower, a long blooming date and so on.It's one of the most widespread and welcome kind in Orchidaceae plant. In the past , Since the selected parent plants for cross breeding lack fragrance gene,descendant nurture often do not has fragrance.But the Chinese feel minor defect in something otherwise perfect unavoidably because they are accustomed to appreciate color ,fragrance and flavor'of Chinese Cymbidium. In this experiment ,the selected Phalaenopsis hybrid Tianxiang L04 was nurtured by Zhengzhou City Flower Engineering Research Center,belongs to intergeneric hybrids from Phalaenopsis ,Hygrochilus and Cymbidium.besides the characteristics above -mentioned, it has Its own unique fragrance ,longer flowering,The whole inflorescence can be open 4-5 months. It has very good ornalmental value and market prospects. To speed up its Market applications , We study tissue culture techniques system of Phalaenopsis hybrid.In this experiment we used a length of pedicel with axillary bud as explant to induce clustered shoots and pedicel as explant to induce PLB, then undertaking proliferation cultivation,at last inducing rooting. This experiment based on orthogonal experiment and single-factor experiment, and test results of the analysis of variance and multiple comparisons, at last we definite a series of phalaenopsis'tissue culture techniques . The results were as fellows:(1) The optimization temperature for culture :Day 25±1℃.12 hours illumination everyday,and about 2200 Lux illumination intensity ,70% relative humidity.(2) The inducement medium of the axillary bud of pedicle: 1/2MS+6-BA0mg/L 1 + KT1.0mg/L+NAA0.05mg/L+ AC0.5g/L was the most suitable medium. After 30 days culturing, the inducing rate could reach 99%.(3) The proliferation of clustered shoots: 1/2MS improvement 1+6-BA7mg/L +KT1.5mg/L+ peptone 2g/L was the most suitble medium, After 60 days culture, each explant can obtain 5.12 shoots on average. The optimized condition of medium for clustered shoots:the best AC concertration is 1g/L,the best CM concertration is 30ml/L ,the best sugar concetration is 25g/L.(4) Radicate: The most suitble medium is 1/2MS improvement2+NAA0.3mg/L +IBA0.7mg/L+AC1.5g/L,after 60 days culture,every shoot can obtain 3.87 roots. The improvement condition of the medium is: the peptone concertration is 1.5g/L, the murphy concertration is 30g/L,the sugar concetration is 25g/L. (5) Pedicel induced PLB: the most suitble medium is 1/2MS + 6 - BA5mg/ LNAA0.05mg/L + KT0.3mg/L+AC1.0g/L ,after 45 days the inducing rate can reach 69.3%.(6) The proliferation of the PLB: the most suitble medium is 1/2MS improvement 1+6-BA3mg/L+KT0.5mg/L+peptone1.5g/L.After 60 days culture, the multiplication rate of each PLB could reach 7.99. The optimized condition of proliferation medium is:the AC dentisy 1.0g/L, the CM concertration is40 ml/L,the sugar concetration is 20g/L.(7) The differention and rooting of PLB :the most suitble medium is 1/2MS improvement2+NAA0.5mg/L+IBA0.7mg/L +AC1.5g/L,each PLB can obtain 4.12 roots. The optimized condition is :the peptone concertration is 1.5g/L, the murphy concertration is 30g/L, the sugar concetration is 25g/L.(80 AC was effect for preventing the explants from browning to a certain extent. the suitble concertration for inducing axillary buds is 0.5g/L , proliferation of culster -ed shoots is 1g/L , and radicating is 1.5g/L. 1.0g/L was the suitable concentration for the PLB's inducement and prolifeation and 1.5g/L was the suitable concentration for thePLB's differentiation and rooting.9. The shoot grew very well in lichenes, the survival rate could reach 100%.