Establishment Of Solanum Chinensis Regeneration System And Analysis Of Chromosome Karyotype

Solanum diphyllum L.is a variant of Solanum nigrum L.Since Solanum diphyllum L.has extremely high medicinal and economic value,Solanum diphyllum L.is also regarded as a wild plant with great development value.However,Solanum diphyllum L.is susceptible to seasonal,environmental and other factors during conventional breeding,which limits the development and utilization of Solanum diphyllum L.Therefore,the establishment of the regeneration system of Solanum diphyllum L.and the karyotype analysis of its chromosomes are of great significance for the preservation of the germplasm resources of Solanum diphyllum L.and its medicinal value,as well as for the systematic evolution and gene positioning of Solanum diphyllum L..In this experiment,I used Solanum diphyllum L.as the experimental material to study the effects of induction,differentiation and rooting of Solanum diphyllum L.callus under different media,different hormone types,different concentration ratios and different explants.I also used this method to analyze the effect of seedling time on the transplantation and domestication of test-tube seedlings,screened out the most suitable medium for each stage,and finally established the regeneration system of Solanum diphyllum L..Then,in this experiment,I used Solanum diphyllum L.root tip as experimental material to compare the effects of different pretreatment time,different pretreatment solution concentration and different dissociation time on chromosome preparation,and finally obtain a well-dispersed and clear mid-mitosis cells to further perform karyotype analysis on Solanum diphyllum L..The main findings are as follows:1.Establishment of the regeneration system of Solanum diphyllum L.:The optimal disinfection combination of seeds of Solanum diphyllum L.is 75% ethanol for 30 s plus mercury for 7 min.The pollution rate and germination rate are 26.6% and 88.6%respectively.The optimal disinfection combination of Solanum diphyllum L.leaves is75% ethanol for 30 s + 6 liters of mercury,and the pollution rate and germination rate are 10% and 83.3% respectively;In terms of callus induction,the best explants are theleaves of Solanum diphyllum L.,and the induction effect is superior to cotyledons and hypocotyls.The best medium is MS + KT 2 mg/L+NAA 0.1 mg/L,the recovery rate is96.7%.In terms of callus differentiation,the best explant is the leaves of Solanum diphyllum L.,the differentiation effect is better than cotyledons and hypocotyls.The optimal medium is MS+KT 2 mg/L+NAA 0.2 mg/L,the differentiation rate is 56.7%.In rooting culture,Solanum diphyllum L..The optimal rooting medium is MS+NAA0.1 mg/L and MS+NAA 0.2 mg/L,rooting rate is 100%;in terms of seedling transplanting,the best days for seedling refining is 4 days,and the survival rate reaches 90%.2.Chromosome preparation and karyotype analysis of Solanum diphyllum L.: For preparation,the best combination of root tip pretreatment of Solanum diphyllum L.is0.05% colchicine solution pretreatment for 2 h.At this time,the dyeing dispersion is good and the shape is clear.The optimal dissociation time is 7 min,at which time chromosome dispersion is good and easy to observe.In terms of karyotype analysis,Solanum diphyllum L.is an autotetraploid plant with a chromosome number of 4n=48,a karyotype formula of K(4n)=48=48m,and all chromosomes are central centromere chromosomes.The arm ratio range is 1.05-1.54,and the average arm ratio is 1.27.The relative length range is 4.7%-11.97%,the chromosome length ratio is 2.54,and the arm ratio greater than 2:1 is 0%.The karyotype asymmetry coefficient is55.62%.