Establishment Of A Multipurpose T-DNA Tagging System Andcreation Of Populus Trichocarpa Mutant Library

Mutant is the live individual with hereditable change in one or some genes ,it has direct reason relation to the mutant gene .it can play an important role to explain function of gene and has important science value. With the rapid development of the gene sequencing technology,people have accomplish three important model plant genome sequencing, such as Arabidopsis thaliana,rice and populus.Following the completion of the genome sequencing of poplar, the international focus is now to identify the function of the predicted genes in the poplar genome. The obvious method of determining gene function is to explore mutants of each gene and study the effects of the mutations on poplar plants. Insertional mutagenesis using T-DNA as tags has been performed with Arabidopsis and rice, and many novel genes have been isolated.Populus has been considered as a model for genomic study of forest trees because of its rapid growth,prolific sexual reproduction,ease of cloning,small genome,facile transgenesis, and tight coupling between physiological traits and biomass productivity.Following the completion of the genome sequencing of poplar(populus trichocarpa),the main task of poplar genome research has been shifted to functional genome study. The most direct and efficient way of functional genome research is to analyze phenotypes through a large number of mutants population generated by T-DNA tagging. In last decade, a number of T-DNA tagging system have been developed, such as gene trapping, activation tagging, multi-purpose T-DNA tagging and inducible tagging etc. Among them, multipurpose tagging is the most efficient strategy, which combines several purposes in one system。The aim of this thesis is to generate a population of T-DNA insertional mutants via Agrobacterium-mediated transformation and make some analysis of important mutants to supply technique and material to the research of forest tree functional genomics. The major results obtained are as follows:(1) We have constructed the pCAS05 vector which is sutiable for functional gneomic of populus study(2) Construct the leaf regeneration system of Populus cathayana: medium of inducing indefinite bud: MS+6-BA1.0+IBA0.05+NAA0.05 + sucrose 30g/L medium of inducing bud rooting: 1/2MS+IBA0.3mg/L+ sucrose 30g/L(3) Explore the Populus cathayana leaf transforming system by Agrobacterium- mediated: a. Pressure of select: Kan 15g/L b. The best concentration of Agrobacterium: OD6000.4~0.5 c. The best time of dipping: 10min(4) We get many transformed plants with strange phenotypes...