Studyof Early Flowering Genetic Transformation System Of Grape Hyacinth

Grape hyacinth(Muscari Miller)has a wide variety of blue flowers which makes it become an ideal material to study the formation mechanism of blue flowers in monocotyledonous.However,the dispute of ploidy of its common cultivars and long juvenilityhave serious restricted the progress of its research and genetic breeding.Six typical grape hyacinth varieties were used in this study to find the suitable materials for the study,and in two ways :in vitro flowering and early flowering gene to shorten the juvenility,and providing a reference for the breeding of grape hyacinth.The results are as follows: 1.Ploidy identification of six grape hyacinthM.armeniacum(cobalt blue),M.aucheri 'Ocean Magic'(pastel blue),M.armeniacum 'Dark Eyes'(positive blue),M.azureum(sky blue with white stripe),M.aucheri ‘White beauty'(white)and M.'Pink Surprise'(pink)were identified with flow cytometry.The results showed that M.armeniacum 'Dark Eyes' and M.'Pink Surprise' were diploid,M.aucheri ‘White beauty' and M.aucheri 'Ocean Magic' were triploid,M.armeniacum was tetraploid.2.Study on in vitro flowering of grape hyacinthUsing Scapes,pedicels,flower budsat different developmental stages,tepals,and anthersof M.armeniacum,M.azureum and M.aucheri ‘White beauty' as explants,cultured indifferent hormone concentrations.The results showed that only M.armeniacum was regenerated of floral buds,which then differentiated into tepals,whereas the other species only callus and vegetative budscould be observed.The concentration of6-BA was 2 mg·L-1.Only the colouringbud can induce flower buds.3.Genetic transformation of grape hyacinth with early flowering geneEarly flowering gene AtFT and AtSOC1 was cloned from Arabidopsis thaliana,and homologous gene MaSOC1 was cloned from M.armeniacumwhich was 630 bp in length.Real-time quantitative results shows that the expression level of MaSOC1 was increased in uncoloured flower buds(S1-S2),then decreased in coloring period and blossom.In other parts of the grape hyacinth,the expression level of MaSOC1 wasextremely high in bulbus,root comes second and relatively low in leaf.Then constructthe plant expression vector with AtFT,AtSOC1 andMaSOC1,Agrobacterium-mediated transform flower bud of M.armeniacum.Use resistant plants for gus histochemicalandfind4 strains of transgenic plant.