Primary Study On The Function Of GHLDOX And Plant Regeneration In Natural Colored Cotton (Gossy Pium Hirsutum L.)

Object:Naturally Colored Cotton is special type cotton, which contains pigment in cotton fiber. As the advance of the people's living standard, more and more need for naturally products because it free pollute and health giving. So naturally colored cotton has been the force of attention. But in the fiber's longer, thinner and stronger of the naturally colored cotton can't march for white cotton fibers. So, improvement the quality of cotton fiber and enrich the color of naturally colored cotton is the main emphasis for cotton breeding in the whole world.In this paper:(1) We isolate and analyze function of correlative flavonoid structural genes (GhLDOX), which encoded the leucoanthocyanidin dioxygenase were involved in the pigmentation in cotton frber. (2)Naturally colored cotton Xincaimian No.5 and and Xincaimian No.7 were used as explants, MS salts and B5 vitamins (MSB) medium supplemented with different phytohormone combinations used to callus induction and differentiation, established the regeneration protocol of XinCai7 via somatic embryogenesis.Methods:(1)We get the full length of Ghldox via RACE-PCR,which encoded leucoanthocyanidin dioxygenase;(2)In order to investigate the function of GhLDOX, the gene was cloned into the Pzp 35S vector, then the combined plasmid was transformed into tobacco;(3)The combined plasmid of pET28a-Ghldox was cloned into the E.coilD3,37℃induced to protein by the IPTG;(4)The naturally colored cotton XinCai 5 and 7 as the materials, MS salts and B5 vitamins (MSB)medium supplemented with different phytohormone combinations used for callus induction and differentiation, established the regeneration protocol of XinCai7 via somatic embryogenesis.Results:(1)The full length of GhLDOX is 1323 bp,the ORF is 1062bp and encoding 354 amino acids,the blast analysis implied it belongs to leucoanthoxyanidin dioxygenase members; (2)By transformed Pzp35S-Ghldox vector into tobacco,get transgenic tobacco;(3)By transformed pET28a-Ghldox vector into E.coilD3,37℃induced by the IPTG two hours respectivly,we can found induced albume in the E.coilD3;(4)We established the regeneration protocol of XinCai7 via somatic embryogenesis.Conclusion:(1)The blast analysis vis NCBI implied it belongs to leucoanthoxyanidin dioxygenase members;(2)By transformed pET28a+-GhLDOXvector into E.coilD3,37℃induced by the IPTG two hours respectivly,and its expression profile reached to the peak when six hours. (3)We established the regeneration protocol of XinCai7 via somatic embryogenesis, laying a foundation for using Agrobacterium mediated transformation to further improve the quality of Naturally Colored Cotton.