Study On Tissue Culture And Rapid Propagation Of Ligularia Jamesii From Changbai Mountain

The experiment used the seeds of Ligularia jamesii as material to getting aseptic seedling first, then take the stem tip and leafstalk from aseptic seedling as exophyte, 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 consistency of growth regulator, consistency of sucrose, appending of active carbon, culture temperature to isolated culture, and using spssl3.0 to doing significance analysis of test result, the results were as follows:(1)Aseptic seedling culture:Cold stratification could cut down the time of Ligularia jamesii seeds bourgeon, the seeds got the best bourgeon situation after 4℃fridge stratification for 13days. The best sterilizing method was that:after divesting the epicarp of the seeds, dip in 75% alcoholic solution for 10s, then dip in 0.1% HgCl2 (with tween-80)solution for 2min,wash down for 6-7 times with sterile water. The best culture medium for seed germination was MS+ 6-BA(0.02mg/L)+ sucrose (30g/L), pH5.80,the best pregermination temperature was 15℃, illumination time of one day was 14h, germination rate was 83%, the plant height was about 3 cm, with wide and round cotyledon.(2)Stem tip inducing culture:Cut the 0.2cm stem tip of Ligularia jamesii as exophyte, the best induced medium was MS+6-BA(3.0mg/L)+NAA(0.3mg/L), appending 35g/L sucrose, inductivity of plumelet was 90.1%, the average plumelet number of per stem tip was 4.1, the plumelet growth well.(3) propagation culture of plumelet induced from stem tip:the plumelet induced from stem tip had the high multiplication capacity, separated them and then put them into propagation culture medium. The best propagation culture medium was MS+6-BA(2.5mg/L)+NAA(0.1mg/L), the average multiplication coefficient was4.5,the plant growed well. The best subculture cycle was 30days, we founding that, when propagation culture for the seventh time, the plumelets growing well, rapid propagation coefficient of stem tip tissue culture was more than 600.(4)Callus inducing culture:the best exophyte for inducing callus was leafstalk, callus induction was 99.5% with the culture medium of MS+6-BA(3.0mg/L)+NAA(0.2mg/L)+sucrose (35g/L), the callus was emerald green, compact, and could produce some adventitious buds without malformation.(5)Adventitious buds producing culture:Cut off the bad part of the callus, and cut into 1cm2, then put them into multiplication medium. The best multiplication medium was MS+6-BA(1.5mg/L)+NAA(0.1mg/L), which could produce a amount of adventitious buds, the inductivity was 98.7%, the adventitious buds number of the per piece callus was 7.8, and the adventitious buds growing well.(6)The best rhizogenesis medium was 1/2MS+NAA(0.5mg/L)+IBA(0.2mg/L), raised the sucrose concentration to 40g/L, could got sound seedling; Adding 2.0g/L active carbon, could cut down the time of rhizogenesis, the rooting rate could got 99.7% under the best culture condition, the descending axis was white, thick with fibril, viridity and growing well.(7) Domestication and transplant:The best transplant medium was mixed of garden mould:perlite:grass peat=1:1:2(volume ratio), irrigated 0.1% sanmate to medium after transplant, protected the overground part with transparent plastic cup to preserve moisture. Put them into an environment of 20℃and with plenty sunshine. Protect the plants in the noon out of direct sunlight at the beginning of transparenting. Transparent survival percent could get to 100%, the plants were emerald green, robust and growing well.