Studies On The Tissue Culture And Polyploidy Induction In Six Species Of Primula

Primula(Primula L.) plant germplasm resources are abundant in China,they have wide variety patterns,bright colors,and high ornamental value.Over the years,our research group have done introduction and cultivation,the results showed that most high altitude Primula species have poor cultivation adaptability,especially they can't surpass the summer,that increased the preserve difficulty of Primula plant resources.Thus,tissue culture and in vitro preservation of germplasm resources strategy are very important for the collection and breeding of Primula.The research studies on the tissue culture of the six species P.bulleyana,P.beesiana,P.cockburniana,P.polyneura,P. sinopurpurea and P.sikkimensis.Meanwhile,in order to improve the Primula ornamental value,In this paper,seeds,callus,cluster buds of the mentioned species were treated by colchicine for polyploid inducing..The results are indicated as follows:1.The suitable disinfection method of seed of.six Primula species which are preserved for 2-5 years.Through improving the germination conditions,the germination rate of P.sikkimensis., P.sinopurpurea,P.bulleyana are:70.49%,45.09%,44%.2.A whole tissue culture of the six species of Primula has been founded..Make callus induction with leaf,the best medium is:1/2MS+2.4-D2.0mg/l+TDZ1.0mg/l,the callus rate is:P. bulleyana25.90%,P.beesiana23.10%,P.cockburniana 19.82%,P.polyneura19.23%,P.sinopurpurea 19.17%and P.sikkimensis16.35%.The best clusterbuds induction medium is:1/2MS+6-BA2.0mg/L+ NAA0.5-1.5mg/L,the induction rate is:52.78%,74.42%,32.62%,40.23%,62.09%和28.28%.The rooting medium:1/2MS+0.2mg/L NAA,root germination is 100%.3.The seeds,callus and clustered buds were treated with colchicines by soaking,or adding into the medium to induce polypoids.The results showed:immersion injury on callus more directly,and high mortality.The aberrance rate of the six Primula species being up to:16.85%,16.69%,17.28%,15.69%, 15.20%,18.59%,when they are treated with 0.8%colchicinesfor 12days.4.Established the optimization system of chromosome sectioning of six Primula species. compared the different methods of pretreatment and dissociations in chromosome sectioning of six species.The results showed that pretreatment with 0.002mol/L8-hydroxyquinoline3h of:P.bulleyana and P.beesiana.P.cockburniana,P.polyneura,P.sinopurpurea and P.sikkimensis.are pretreated for 3.5h.After 24h immobilization in 4℃refrigerator,P.cockburniana and P.sikkimensis were dissociated by 50%glacial acetic acid:1mol/LHCL=1:1 for 5min,other four species were dissociated for 2min,and dyeing,obtained good chromosome sectioning.The optimization system of chromosome sectioning made foundation for Chromosome karyotype analyzation.5.Through the chromosome counting method,morphological variation of the identification of polyploid plants,the results showed:P.bulleyana(2n=4x=44),P.beesiana(2n=4x=44),P.polyneura (2n=4x=44),P.sinopurpurea(2n=4x=44),P.cockburniana(2n=2x=22),and P.sikkimensis(2n=2x=18)To sum up,the tissue culture and induced polyploid research on the six Primula species,it offered special reference to the introduction and germplasm conservation of the wild Primula,and also reference to other Primula plants breeding research.