| Lilium regale Wilson.has a large corolla and beautiful flower type.In lily breeding research,it is often used as a parent for cross-breeding to breed new varieties.However,under natural conditions,Lilium regale Wilson.is a shady plant,and the stems are often too high and easy to lodging.To solve this problem,at the same time expand the Lilium regale Wilson.germplasm resource bank.In this study,different materials of Lilium regale Wilson.were used as explants for polyploid induction research,Lilium regale Wilson.tissue cultured seedling scales and seeds were used as materials,colchicine was used as an inducer,different concentration gradients were set,and time was used as an investigative factor to induce polyploidy of Lilium regale Wilson.after differentiation culture.Then through morphology,flow cytometry analysis,root tip chromosome compression,stomata density and size identification,chlorophyll content identification and shape index analysis,etc.,the mutant plants were identified and analyzed,and finally the tetraploid Lilium regale Wilson.plant was obtained.Results of mutagenesis using scales as materials:Lilium regale Wilson.scales were cultured in the differentiation medium for 40 days,and 35% of the leaves showed hypertrophy,widening,and darkening of leaf color.After successive generations,subculture and flow cytometry analysis,the material with changes in the leaves showed peaks at positions 80,80 and 160,and 160,respectively.In the control group,diploids only had absorption peaks at position 80.This means that the80 position is a diploid plant,the 80 and 160 positions are a chimera plant,and the 160 position is a tetraploid plant.Subculture was carried out at positions 80 and 160,and positions 160 respectively.Chromosome pressing was performed at 30 days.Each root tip chromosome pressing was performed to observe several cell phases.The results showed that the number of chromosomes of the chimeric plant was present.2n=2x=24,there is also 2n=4x=48,and the number of chromosomes in tetraploid plants is 2n=4x=48.The control group was subjected to root apical compression,and the number of chromosomes was 2n=2x=24.The stomata density and size of the diploid and tetraploid plants obtained from the scale induction showed that the average number of tetraploid stomata was 33 fewer than that of the diploid under the 10×10 fields of view.Under the 10×40 field of views,the tetraploid the average plant length is 19.67μm longer than diploid,but there is no significant difference in stomata wide.The best induction method: the best treatment concentration of scale is 0.025%,the best treatment time is 2h,and the induction rate of scale tetraploid is 16.67%.Results of mutagenesis using seeds as materials:Seeds of Lilium regale Wilson.were used as materials,and after germination treatment in the culture medium until the seeds were broken,the colchicine solutions of different concentrations were used as mutagens,and different times were used as investigating factors.After 60 days of differentiation and culture,about 40% of the leaves showed hypertrophy,widening,and darkening of leaf color.After successive generations,subculture and flow cytometry analysis,the material with changes in the leaves showed peaks at positions 80,80 and 160,and 160,respectively.In the control group,diploids only had absorption peaks at position 80.This means that the 80 position is a diploid plant,the 80 and 160 positions are a chimera plant,and the 160 position is a tetraploid plant.Subculture was performed at positions 80 and 160,and positions 160,respectively,and chromosome pressing was performed at 30 days.Each root tip chromosome pressing was performed to observe several cell phases.The results showed that the number of chromosomes of the chimeric plant was present.2n=2x=24,there is also 2n=4x=48,and the number of chromosomes in tetraploid plants is 2n=4x=48.The control group was subjected to root apical compression,and the number of chromosomes was 2n=2x=24.The stomata density and size of the diploid and tetraploid plants obtained by seed induction showed that under the 10×10 field of view,the average number of tetraploid stomata was 20 less than that of the diploid,and under the 10×40 field of view,the tetraploid The average plant length is 19.01μm longer than diploid,and there is no significant difference in stomata width.The best induction method: the best seed treatment concentration is 0.1%,the best treatment time is 36 h,and the seed tetraploid induction rate is 12.00%.The chlorophyll content of Lilium regale Wilson.was identified,diploid and tetraploid plants of Lilium regale Wilson.were randomly selected,and leaves with the same growth states were selected.The results showed that the chlorophyll content a of tetraploid plants was 1.53mg/g higher than that of diploid plants.The content of chlorophyll b is 0.64mg/g higher than that of diploid plants,and the total chlorophyll content is 2.17mg/g higher than that of diploid plants.The appearance index of Lilium regale Wilson.was identified,and diploid and tetraploid plants of Lilium regale Wilson.were randomly selected.The results showed that the average diameter of the bulbs of the tetraploid plants was 9.25 mm larger than that of the diploid plants;the average thickness of the scales of the diploid plants was2.79 mm,the tetraploid plant is 1.23 mm thicker;the average leaf width of the diploid plant is 9.01 mm,the average leaf width of the tetraploid plant is 4.53 mm wider than the diploid plant;the average diameter of the petiole of the diploid plant is 1.35 mm,And the average diameter of petioles of tetraploid plants is 1.76 mm,which is 0.41 mm larger than that of diploid plants.In this study,different materials of Lilium regale Wilson.were used as explants to study polyploidy induction,and colchicine was used to induce them.The obtained plants were identified and analyzed by several different methods,and the diploid of Lilium regale Wilson.was compared.The difference between tetraploid and tetraploid,and finally the tetraploid plant of Lilium regale Wilson.was successfully obtained. |
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