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Construction of Expression Vectors for Key Enzyme Genes in Trehalose Synthesis Pathway, and the Study on Their Transformation in Lisianthus

Author: HeYiMin
Tutor: ChenLiMei
School: Kunming University of Science and Technology
Course: Biochemistry and Molecular Biology
Keywords: Transgenic plants Salt tolerance Genetic engineering Trehalose synthesis Lisianthus
CLC: S681.9
Type: Master's thesis
Year: 2009
Downloads: 37
Quote: 0
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Abstract


Salinity is a major factor in plant growth and crop yield . Trehalose can stabilize biological structures under abiotic stress . However , in most plant trehalose accumulation are low. , Trehalose by trehalose-6 - phosphate synthase (TPS) and trehalose-6 - phosphate phosphatase (TPP) , synthesized in bacteria and yeast . The otsA and otsB is the E. coli trehalose biosynthesis of the two genes . Lisianthus (Eustoma grandiflorum) is a widely used flowers , has a high ornamental value and economic value . The Lisianthus Yunnan cultivated more greenhouses and greenhouse cultivation , are vulnerable to the role of soil salinization , and not conducive to the growth of Lisianthus . This study expects that overexpression of trehalose-6 - phosphate synthase / trehalose-6 - phosphate phosphatase fusion gene , increased salt tolerance in plant breeding of new materials in plants through genetic manipulation . This study through plant genetic engineering techniques to construct the three E. coli trehalose synthesis light - inducible plant gene expression vector ( PRBCs promoter control) : Trehalose - 6 - phosphate synthase ( TPS gene) of the plant expression vector pK2-PrbcS -tps, trehalose-6 - phosphate phosphatase the (tpp gene ) the pH value was brought to 2 - PrbcS - tpp and a the trehalose -6 - phosphate synthase / the phosphatase (tpsp gene ) the PH2- PrbcS - tpspo this study also for Lisianthus antibiotics concentration gradient test and plant hormones concentration gradient test , to determine the best growing conditions for Lisianthus ( differentiation medium was MS 6-BA0.5mg / L IBA0.2mg / L, the rooting medium was MS IBAlmg / L GA 0.2mg / L, cefotaxime sodium concentration of 300mg / L, hygromycin concentration of 10 mg / L ) . Agrobacterium-mediated transformation , we transfected with pH2-PrbcS-tpp the pH2-PrbcS-tpsp two plasmids Lisianthus leaf discs . In this study, the Communists a the turn tpsp gene Lisianthus seedlings 42 , 10 seedlings hygromycin resistance , discovered by genomic PCR detection , integrated in the genome of a plant the tpsp gene . But tpsp gene expression of other molecular level has yet to be detected , it Lisianthus physiological and biochemical indexes also require further analysis .

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CLC: > Agricultural Sciences > Gardening > Ornamental Horticulture ( flowers and ornamental trees) > Annual and biennial flower class > Other
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