ISSR Analysis,Excellent Strain Breeding And New Germplasm Creation Of Lycium Ruthenium

Lycium ruthenium is a perennial shrub of the Solanaceae family.It is widely spread over northwestern China.It is known as the plant "soft gold"and the wild "blue demon girl" because its fruits are rich in various nutrients.In this study,ISSR-PCR was used to analyze the genetic diversity of 24 different types of Lycium ruthenium materials.Taking the current year branches and seeds of Lycium ruthenium in Alashan region as explants,rapid tissue propagation system was established through primary generation,subculture and rooting culture.And the Alashan diploid seeds were treated with colchicine at different concentrations and different treatment time and karyotype were analyzed.The research results can provide the genetic basic theory and technical basis for genetic classification,germplasm resource innovation and chromosome ploidy breeding of Lycium ruthenium in the ruture.The main rindings are as follows:(?) seiected 30 primers from the published ISSR universal primers and selected 11 suitable primers.The genetic diversity of the test materials was analyzed,and a total of 138 clear and bright bands were amplified.including polymorphic bands 131 strips(95.20%).The average number of amplified bands and polymorphic bands of each primer is 12.5 and 11.9,respectively.and the genetic similarity coefficient is between 0.154 and 0.971,the average genetic similarity coefficient is 0.445,it can be divided into 3 categories at the threshold of 0.52.(2)Taking the current year branches of Lycium ruthenium in the Alashan as explants,suitable medium for inducing callus:1/2MS+6-BA1.0 mg/L+NAA 0.2mg/L+5.5g/Lagar+30g/Lsucrose;Adventitious bud propagation medium:WPM+6-BA 1.0 mg/L+NAA 0.1 mg/L+5.5g/L agar+30g/L sucrose;Rooting medium:WPM+IBA 0.5 m/L+5.5g/L agar+30g/L sucrose.(3)Taking the seeds as explants,suitable medium for inducing sterile seedlings:WPM basic medium;suitable medium for sterile seedlings micro-cutting propagation:WPM+IBA 0.5mg/L+5.5g/L agar+30g/L sucrose.(4)Treatment Alashan diploid Lycium ruthenium seeds with 0.2%colchicine for 48h,the autotetraploid material Lycium ruthenium could be obtained,and the highest induction rate is 22.2%.(5)Compared the diploid test material(2n=2x=24=6sm+18m)with the tetraploid plant,the tetraploid have longer leaves,wider leaves,shorter plant heights,slower growth,and significantly larger stomata,lower stomata density,and increased chloroplast numbers.The karyotype formula is 2n=4x=48=12sm+36m,which belongs to "2A" type,its relative DNA content(100)is twice that of the diploid material.