| In this experiment,colchicine was used as an inducer to induce polyploidy in tetraploid Lyciun ruthenicum tissue culture seedlings.The colchicine concentration was 500 mg·L-1 and 1000 mg-L-1,and the treatment time was 6,12 and 24 hours,the top leaves of the tetraploid pods were cultured by soaking method to obtain the octoploid plant,and the polyploid identification method was perfected to promote the development of the Lycium ruthenicum industry.The results of the study are as follows:1.The survival rate of the colchicine concentration of 1000 mg·L-1 for 12h was a semi-lethal dose.In this experiment,12 octoploid plants were initially screened in the treatment of 12 groups of materials.The colchicine concentration was 1000 mg·L-1,the treatment time was 12h(2-2-4),and the colchicine concentration was 1000 mg·L-1,the treatment time was 24h(2-3-2),identified as octaploid by chromosome.2.Using flow cytometry,the WPB dissociation solution was used to determine the chromosome ploidy of the materials treated,and the known diploid Lycium ruthenicum as the internal reference and the external reference.The results were consistent with the results of chromosome counting.The internal reference was superior to the external reference in analysis and flow cytometer parameter setting.3.When the ploidy of tissue cultured seedlings was increased,the stomatal density,guard cell length and width,and guard cell chloroplast number were significantly different in the leaves.With the increase of ploidy of tissue culture seedlings,the stomatal density of the leaves decreased significantly,the length and width of the guard cells increased significantly,and the number of guard cells increased significantly.In the induction of polyploidy in Lycium ruthenicum,the leaf anatomy can be used as a basis for preliminary identification.4.Different ploidy Lycium ruthenicum have the same suitable growth medium.,formula of medium is MS + ZT 0.15 mg·L-1+ IBA 0.01 mg·L-1,different ploidy Lycium ruthenicum have significant difference in the value of the increment in this medium,diploid 9.15,tetraploid 6.25,octaploid 5.35,5.The difference in rooting rate of different ploidy Lycium ruthenicum is extremely significant,The rooting rate of diploid was 100%at iba concentration of 0-1.0 mg·L-1;The rooting rate of tetraploid was 45%when iba was 1.0 mg·L-1;The highest rooting rate of octoploid was 35%at 1.0 mg·L-1.There are great differences in the rates of different batches of tetraploid and octoploid Lycium ruthenicum On the same rooting medium,and the rooting effect is not stable,Specific reasons need to be further explored. |
