Agrobacterium Tumefaciens-mediated Transformation Of Antisense-LEAFY Gene Into Platanus Orientalis L.

Platanus orientalis L. is a tree of strong stem, good effects of green and its leaves have great capability to resist poisonous gas and insects and become a famous street tree of the world, so that it be planted in many cities of our country. But its seed's fur not only pollute surroundings but also provoke disease of breathing system and skins and bring very inconvenience to citizen's living. So scholar of home and abroad proceed a great research to solve the problem of pollution of seed's fur. With the boom of modern science and technology, gene engineering provide new methods to solve these problems.Factors affecting the transformation of P. orientalis by Agrobacterium tumefaciens are studied and the optimum transformation conditions are defined. The genetic transformation system of P. orientalis is originally established. Method of extraction DNA from leaves of P. orientalis is selected. The transformation of P. orientalis is verified by PCR and GUS histochemistry test. The following is the results of experiments:1. The Agrobacterium tumefaciens is made of the engineering Agrobacterium for transformation of P. orientalis. The expression vector pBI121 contains anti-LEAFY gene, and there are a position, which can be defied by HindIII. We obtain the anticipative DNA segement from plasmid is digestioned by HindIII and EcoRI. The anti-LEAFY gene can be examined by PCR as it is transferred into Agrobacterium tumefaciens.2. In this paper dealt with effects of different antibiotics on in vitro plantlet regeneration of P. orientalis. Induction frequencies of the calli, shoots and rootings with the leaves were reduced with Ka and Cef in the media. The rootings of their explants and calli, shoots were inhibited with 15mg·L^-1 and 40 mg·L^-1 of Ka concentrations in proliferation and rooting media respectively; So were that with Cef concentrations of 500 mg·L^-1 and 800 mg·L^-1 in those media. There existed no leaf callus and shoot formations on the proliferation media with 30 mg·L^-1 and 400 mg·L^-1 of Ka and Cef. This would be referenced to the manipulation of gene engineering of P. orientalis.