Optimization Of Tissue Culture And Rapid Propagation System Of Hemerocallis Spp.and Haploid Culture Study

In this study,different Genotypes of Hemerocallis spp.were used as materials to optimize the rapid propagation system of Hemerocallis spp.and the selection of explants was broadened,and the rooting culture effect was optimized.In addition,the in vitro induction of haploid induction in the genus Hemerocallis spp.was studied,and the relevant research foundation was established for the creation of DH line of Hemerocallis spp..The test results are as follows:(1)Taking the 'Datong Huanghua' dwarf stem as the main experimental material,the tissue culture effect at different developmental stages was compared by annual sampling.The germination period in late2-3 months was the best sampling period,the induction rate was the highest,and the pollution rate was the lowest.The optimal medium for adventitious bud induction: MS+1.0 mg·L-16-BA+0.1 mg·L-1NAA,the highest induction rate was 88.89%,the induction coefficient was 6.67,and the adventitious bud proliferation medium: MS+2.0 mg·L-16-BA+0.5 mg·L-1NAA;the culture system was verified by the above adventitious bud induction medium,and 14 short stems of different genus Hemerocallis spp.were inoculated,and 12 parts of the material could induce adventitious buds.(2)Inoculate the adventitious buds of 'Datong Huanghua' at different growth stages in different rooting induction medium,inoculate in medium A(MS+0.25 mg·L-1NAA+30 g·L-1 sucrose),adventitious buds(3~5)cm)Plants have the best growth and the highest number of roots(4.33 per plant).(3)The unfertilized ovules were used as explants to study the in vitro gynogenesis of Hemerocallis spp.and the results showed that the unfertilized ovule induction effects were significantly different under different genotypes and different treatments.Of the 29 different genotypes,22 and 122 had better culture effects.The optimal culture conditions were:4? low temperature pretreatment for 24 h,medium MS + 3.0mg·L-12,4-D + 1.0 mg·L-16-BA+1.0 mg·L-1NAA,sucrose concentration 30 g· L-1,dark culture,22 produced dense green callus,induction rate of 66.67%;No.122 induced adventitious buds.(4)An anthocyanin was used as an explant to study the in vitro culture of the genus Hemerocallis spp.The cytological observation of the microspore development period was carried out on 29 materials in the resource sputum.The length of the corresponding genotypes of microspores in different genotypes was different,and the interval was 5.57-8.48 mm.Different genotypes,different culture effects,screened out No.3 in Y1 medium MS + 2mg · L-12,4-D + 2mg · L-1KT + 4mg · L-1VB1 + 60 g · L-1sucrose,callusThe dense green color,the call injury rate is 0.90.In this study,the tissue culture and plant regeneration system of the dwarf shrubs of Hemerocallis spp were constructed,the reproductive coefficient of the genus Hemerocallis spp was improved,and the technical guarantee for the industrialization of the tissue culture of Hemerocallis spp was provided.The in vitro culture study of the gynogen and male nucleus of Hemerocallis spp has obtained phase results,which provides a technical basis for the creation of new haplotypes and double haploid germplasms of Hemerocallis spp,and reduces the heterozygous degree of genus Hemerocallis spp.The study of omics and the acceleration of the breeding process of the genus Hemerocallis spp are of great significance.