Study On Polyploid Of Lilium Pumilum DC. Test Tube Seedlings Induced By Colchicines

Lilium pumilum DC.is a plant in the Lilium of the Liliaceae.In the wild,Lilium pumilum DC.stems are thin and prone to lodging.In order to solve this problem,this study used nanniwan wild Lilium pumilum DC.scales as material,and used tissue culture technology to establish asexual reproduction line.Polyploid induction was carried out by using small bulbs and scales obtained from tube seedlings with rapid propagation as materials,mixed culture method and soaking method,and colchicine concentration and time as investigation factors.The specific work is as follows:1.Establishment of Lilium pumilum DC.tissue culture fast propagation cloneIn the primary culture,Lilium pumilum DC.scales were used as the material and 6-BA concentration was used as the investigation factor.The ability of scales to differentiate into small bulbs was significantly different in different new media,in which MS+ 6-BA 1.0 mg/L+NAA 0.01 mg/L+AC 2 mg/L,and pH 6.8 medium had the strongest differentiation ability,with the differentiation rate of small bulbs =84%.In the subculture,the differentiated bulbs were used as the material and 6-BA and IBA were used as the investigation factors.In the new medium with different hormone combinations,the ability to differentiate new small bulbs was significantly different,among which MS + 6-BA 1.0 mg/L+IBA 0.2 mg/L,and pH 6.8 medium had the strongest differentiation ability,with the proliferation coefficient =7.3.2.Polyploid inductionThe test tube seedling small bulbs were obtained from the rapid propagation generation as the material,and the colchicine concentration and the treatment time were used as the factors for the study by the mixed cultivation method.The colchicine concentration was designed at five levels and the cultivation time was designed at 4 levels for testing.The results showed that mutant plants appeared in each treatment group.After data analysis,the suitable combination of colchicine 60 mg/L and induction 30 d was selected to induce Lilium pumilum DC.tube seedlings by mixed culture method.The variation rate was 46.7%.Tube seedlings scales as materials,by soaking method,with colchicine concentration and immersion time to examine the factor,concentration of colchicine designed a three level,set up four levels of incubation time,after 30 d statistical results show that the processing of some scales dry dead,not the scales of death part of differentiate into small bulb,including the variation of plant.After data analysis,colchicine 0.1% was selected as the suitable treatment combination for soaking Lilium pumilum DC.test tube seedling scales induced by soaking for 12 h,and the mutagenesis rate was up to 44%.3.Variation analysisThe mutant plant lines and diploid test-tube plantlets produced by mutagenesis were used as materials,and the differences were identified by morphological observation method,stomatal method and chromosome method.Larger,thicker,larger stomata,smaller stomata density.Among them,the average leaf length of the mutant plant line induced by the hybridization method was 1.27 times that of the diploid plant,the average width was 1.66 times,and the average thickness was 1.61 times.The average leaf length of the mutant plant line induced by the soaking method was 1.18 times that of the diploid plant,the average width was 1.68 times,and the average thickness was 2.33 times.The stomata length of the mutant plant line induced by the mixing method was 1.25 times that of the diploid,and the width was 1.56 times.The stomatal density of the diploid plant was 1.93 times that of the mutant plant line.The stomata length of the mutant plant line induced by the soaking method was 1.24 times that of the diploid,and the width was 1.86 times.The stomatal density of the diploid plant was 1.41 times that of the mutant plant line.Using two mutant plant lines and diploid test-tube seedlings as materials,the cytogenetic differences between the mutant plant lines and the diploid plant lines were studied by the method of acid hydrolysis and hypotonicity.The chromosome number of diploid test-tube plantlets was 2n = 2x = 24.No tetraploid plants with 2n = 2x = 48 chromosomes were found in the mutant plant lines produced by mutagenesis.