Study On Tissue Culture And Rapid Propagation Of Mukdenia Rossii (Oliv.) Koidz

The study used the seeds of Mukdenia rossii (oliv.) Koidz as material to getting aseptic seeding first, then took the petioles、leaf and tender bud from aseptic seeding as exophyte to establish the system of primary culture, subculture proliferation, rooting culture, domestication and transplant culture, studied the effect of kinds of basic culture medium, the kinds and concentration of growth regulator, consistency of sucrose, appending of active carbon, culture temperature to isolated culture. A method which was fit for rapid propagation of Mukdenia rossii (oliv.) Koidz was summarized to improve the technology of plants regeneration in vitro and lay a theoretical foundation for the using of Mukdenia rossii (oliv.) Koidz.1. Selection of exophytesThe leaf、petiole、pedicels、tender bud-root and bud in the first fifteen days of May and the mature seed were chosen as exophyte。The filter paper with the seeds of Mukdenia rossii (oliv.) Koidz was folded into2cm2. Other explants were cut into1～2cm. The exophytes were sterilized in0.1%HgCl2for5minutes and then washed with sterile distilled water for6～8times under aseptic operation. The results showed that the seed culture had the highest rate of success and got the most exophytes. The seedlings had no variation phenomenon in that condition.2. Getting aseptic exophytesThe filter paper with the proper and cleaned seeds of Mukdenia rossii (oliv.) Koidz was folded into2cm2. The seeds were sterilized in0.1%HgCl2for6minutes and then washed with sterile distilled water for6～8times under aseptic operation. It had the highest rate of germination of97.55%in that condition. So this sterilization method was the best one. It had the shortest germination time and best effect when putting the seeds after sterilization on Ms+6-BA0.5mg/l. The results show that lower concentration of6-BA was good to build sterility system of Mukdenia rossii Koidz. So the best combination of culture medium was Ms+6-BA0.5mg/l.3. Inducing callusIt can be found that tender bud had the highest callus induction rate and best quality by testing the petioles、leaf and tender bud from aseptic seeding. So the tender bud was the best exophyte for callus induction. The tender bud had the highest callus induction rate by using different kinds and concentration of6-BA、NAA and2,4-D. The best combination of culture medium was Ms+6-BA1mg/l+2,4-D1mg/l under the sucrose concentration of3%. The callus was emerald green, compact. The callus induction rate was the highest. 4. Adventitious bud regeneration of the callusPut the callus which was cut into0.5cm2on Ms+6-BA2mg/l+NAA0.2mg/l. There were most adventitious buds and the plants grown well after20days.5. Formation of adventitious budsMs+6-BA1mg/l+NAA0.1mg/l was found good for proliferation of adventitious buds, the multiplication rate being the highest, the number of adventitious buds being the most and the plants grown well. It can meet the purpose of the rapid propagation.The results showed that the very subculture cycle was20-25days by testing different subculture cycles. It had the best cultivation effect in that condition.6. Root inductionPut the adventitious buds which were3-4cm on the rooting medium with different concentration of auxins and Activated Carbon. The results showed that the optimal medium for root induction was1/2MS and its root-inducing rate reached as high as100%. The average root length was2.5cm and the plants grown well.7. Domestication and transplantsPut the tissue culture seedlings in different kinds of growing media and temperature environment. The results showed that the best growing media was perlite is v:v=1:1and the best temperature was20±2℃with enough sunlight.