The Tissue Culture Of Two Kinds Of Lilies And The Chromosomal Deed During Tissue Culture

This article research the tissue culture and cloning propagation and the chromosomal variation of two kinds of lilies —Lilium davidii Var.Unicolor(Hoog)Cotton and Lilium brownii F.E.Brown ex miellez.At the same time, we also observed the nuclear and the B-chromosome. The tissue culture and cloning propagation include the establishment of aseptic culture, bud induction and proliferation, selection of suitable hormone , rooting and acclimatization. The chromosomal variation include the change of chromosome number mainly.When the explants were collected in the spring, high induction rate can be achieved. Brushing the dust and washing the explants under ranning tap water to remove some of the dirt and contaminations, in 75%CH3CH2OH for 30 second and in 0.1%Hgcl2 for 13 minutes, rinsed 5-6 times with sterile water. The two kinds of explants were cultured on Murashige&Skoog medium(MS) added with 0.6-1.0mg/lN-benzyladenine(BA)+0.2mg/l naphthalane acetic(NAA)and added with 0.1-0.5mg/lBA+0.1-0.2mg/lNAA or indole-3-acetic(IAA).The induction rate can reached 70-100%.The best explants were the parts that were below and linked the scales.During the buds poliferation,the best medium of L.davidii Var.Unicolor(Hoog)Cottonis MS+0.5mg/lBA+0.1mg/lNAA, the best medium of L.brownii F.E.Brown ex miellez is MS+1.0mg/lBA+0.1mg/lIAA. It good to keep the culture at 23-25℃,photoperiod with a light intensity of 2,000Lx,keep 16 hours lighting and 8 hours darking or 12 hours lighting and 12hours darking on culture, to subculture every 28-32days.The poliferation can be reached 4-5 times.One-half MS basal medium added 0.1-0.2mg/lNAA or IAA can make the root long and strong. The activated charcoal is good to root formation and growth. When the length of root is 1-1.5cm,lilies can be acclimatized. If the living condition is good, the survival rate were over 80%During the tissue culture, we found the chromosomal variation in cells of two kinds of lilies. The variation rate of callus of L.davidii Var.Unicolor(Hoog)Cotton is 54.4%and the L.brownii F.E.Brown ex miellez is49.4%, the chromosome number varied from 9-50and 12-50;the variation rate of buds of L.davidii Var.Unicolor(Hoog)Cotton and L.brownii F.E.Brown ex miellez are 17.5%and 19.5%,these show that the variation rate of callus is higher than buds. The hormone can affect the deeds of the chromosome. The variation rate caused by single hormone is lower than two kinds of hormone, the kind and intensity of hormone is different, the variation rate is different. We think that No.5and No.11 are best medium to two kinds of lilies when we thought over the induction rate and variation rate. The times of subculture should be limited because we found when the subculture times is increasing, the variation rate is higher and the differentiation capacity is lower. The best times of L.davidii Var.Unicolor(Hoog)Cotton is 4-5,the other is 6 times.We observed the cells of root-tip, callus and bud of two kinds of lilies, we could not find the B-chromosome in cells of L. brownii F.E. Brown ex miellez, but find it in L.davidii Var.Unicolor(Hoog)Cotton,the number is 1-2.