Development Of SSR Molecular Marker Of Transcrip Tome And Construction Of Vitro Regeneration System In Bougainvillea

In order to clarify the overall characteristics of the SSR,develop the S SR primers for molecular breeding,and excavate the genes related to the development of bract of Bougainvillea,the experiment analyzed the transcriptome sequencing results then analyzed the SSR locus information of the B.specta-bilis cv 'Speciosa' transcriptome by MIS A,and designed primers by Primer 3.50 pairs of primers were selected to analyze the transferability and diversity of 6 different species and hybrids.The result showed that a total of 79,210 SSRs located in 60,318 unigenes with the frequency of 44.91%and a mean distance of 11.99 kb per one in 'Speciosa' Mononucleotide and dinucleotide were major types with the percent-ages of 76.37%and 6.77%,respectively.A/T and AG/CT were most frequent motifs in monnucleotide and dinucleotide repeats,accounting for 71.66%and 6.77%,respectively.In 50 pairs of primers selected,42 of which could amplify clear and stable target bands with a valid rate of 84.00%.The average of transferable rate between different varieties are 85.86%and 33 primers showed polymorphism with He and PIC mean values of 0.5548 and 0.4603,respectively.In order to achieve the purpose of germplasm innovation of Bougainvillea,induce sufficient callus and multiple species at the level of tissue culture,we took young stems and leaves grown from B.glabra cv 'Singapore Pink' and B.specto-peruviana cv 'Mrs.Butt';hypocotyls and cotyledons from B.glabra cv 'Formosa' and B.spectabilis cv 'Speciosa' after two weeks of sowing.MS was the basic medium added by different hormone-level,which compared to find out the best callus of growth-promoting auxin and cytokinin ratios,and then proliferated with the same medium to obtain sufficient callus.The results showed that the stem segments and hypocotyls ofBougainvillea were more suitable for callus induction and proliferation.Above all,the medium which were suitable in concentration for the callus of'Singapore Pink' was MS+NAA 0.2mg/L+6-BA 1.0mg/L,and the medium suitable for the callus of 'Mrs.Butt' was MS+NAA 0.2mg/L+6-BA 0.5-1.0mg/L.The medium was the best when the concentration for callus culture of 'Formosa' hypocotyl was MS+IBA 0.1mg/L+6-BA 2.0mg/L,and the medium of hypocotyl callus suitable for 'Speciosa' has not been found.