Research On DiaMYB Gene Expression Characteristic And Stress Tolerance In Dendranthema Indicum Var.Aromaticum

Dendranthema indicum var. aromaticum is a kind of new resources of compositae Chrysanthemum, this paper takes Dendranthema indicum var. aromaticum DiaMYB gene and Arabidopsis thaliana as the research object, use has been building a successful PCAMBIA1301-MYB expression vector, dip by agrobacterium-mediated transformation of an Arabidopsis thaliana, the homozygous obtained from the screening transformation of T3 generation of Arabidopsis thaliana three positive strain, the research of transgenic Arabidopsis under drought and salt stress enzyme activity, Chlorophyll content and Photosynthetic Physiological indexes.Building containing fluorescent protein pBI121-MYB-GFP expression vector, and observe the subcellular localization. Real-time fluorescent quantitative analysis turns DiaMYB gene plant in adversity stress under stress related gene expression. In this paper, the main research results are as follows:1. Using Agrobacterium-mediated dip in an Dendranthema indicum var. aromaticum DiaMYB gene into Arabidopsis thaliana, get three positive strain, named, MYB-1, MYB-2 MYB-3, GUS staining and PCR identification, confirm DiaMYB genes have been successfully transferred to Arabidopsis thaliana genome. Real-time fluorescent quantitative PCR methods to analyze a express DiaMYB gene expression quantity of different organs of Arabidopsis thaliana, the results showed that the expression in stem, leaf and flower quantity slightly higher, followed by fruit, expressed in root in the lowest amount. Using DiaMYB building containing green fluorescent protein gene fusion expression vector pBI121-MYB-GFP, use gene marksmanship into onion epidermal cells, showed DiaMYB protein can locate on the nucleus.2. Take T3 generation of transgenic Arabidopsis three strain and wild-type Arabidopsis in high salt stress, the results showed that with the increase of concentration of salt solution, each strain of transgenic Arabidopsis thaliana seed germination rate and seedling survival rate was significantly stronger than the wild type,combination of Superoxide Dismutase (SOD) and Peroxidase (POD) enzyme activity, Chlorophyll a/Chlorophyll b, Carotenoids/chloroPhyll and net Photosynthetic rate(Pn), stomatal conductance (Gs) and intercellular CO2 concentration (Ci) of the numerical results obtained transgenic Arabidopsis each strain showed more stronger than the wild type of salt resistance. High real-time fluorescent quantitative PCR results showed that under salt stress, turn DiaMYB gene in Arabidopsis thaliana RD22, RD29A, RAB18, COR47, the expression of ABA1 volume increased significantly, at the same time, ABI1, HAB1 gene expression has a slight decline amount and everyone is lower than the wild type plants, DiaMYB gene expression quantity increased under salt stress.3. Take T3 generation of transgenic Arabidopsis three strain and wild-type Arabidopsis in drought stress, the results showed that with the increasing of drought stress, each strain of transgenic Arabidopsis thaliana seed germination rate and seedling survival rate was significantly stronger than the wild type, combination of Superoxide Dismutase (SOD) and Peroxidase (POD) enzyme activity, Chlorophyll a/Chlorophyll b, Carotenoids/chloroPhyll and net Photosynthetic rate(Pn), stomatal conductance (Gs) and intercellular CO2 concentration (Ci) of the numerical results obtained transgenic Arabidopsis each strain showed more stronger than the wild type of drought resistance. Real-time fluorescent quantitative PCR results showed that under drought stress, turn DiaMYB gene in Arabidopsis thaliana RD22, RD29A, RAB18, COR47, the expression of ABA1 volume increased significantly, at the same time, ABI1, HAB1 gene expression has a slight decline amount and everyone is lower than the wild type plants, DiaMYB gene expression under drought stress first increases and then decreases.