The Use Of Phosphomannose-isomerase As A Selective Marker To Recover Transgenic Popular Via Agrobacterium Transformation

A new method for the selection of transgenic popular plants without the use of antibiotics or herbicides has been developed. Although mannose itself has no adverse effects on plant cells, it leads to an accumulation of mannose-6-phosphate, which depletes intracellular stores of inorganic phosphate. This results in an inhibition of plant cell growth. The selection system uses the Escherichia coli pmi gene that encodes phosphomannose isomerase (PMI). Transgenic plants carrying the pmi gene can detoxify mannose-6-phosphate by conversion to fructose-6-phosphate, an intermediate of glycolysis, via the PMI activity. They differ from conventional selection techniques as they are based on supplementing the transgenic cells with a metabolic advantage rather than killing the non-transgenic cells. In many cases, these new selection systems have been found to be superior to conventional methods. In this paper, we introduce our study on the cloning of The phosphomannose isomerase (PMI) gene and the construction of expression vectors containing of pNOV2819-GFP and pNOV2819-NP1. The main contents are as follows : 1. The phosphomannose isomerase (PMI) gene from Escherichia coli has been cloned and consitutively expressed in vector pMBIA1301--hyg. 2.The PMI gene was transferred to NL895 by Agrobactrium-mediated transformation, which allowed the selection of transgenic plants with mannose as selective agent. the integration and expression of the transgene was confirmed by the GFP. 3. Poplar is a very important forest tree speice. In order to improve the anti-disease ability of poplar, pNOV2819-GFP, pNOV2819-NP1were transferred into the poplar hybrid clone (Populus deltoides X P. simonii) NL-80106 by infected with A. tumefaciens. 4. Black Yang X Simon poplar of America of Israel (Populus deltoides X P. Simonii), fine asexual department NL-895 Yang receptor material, is it is it is it transform to go on to lead to lie between through root cancer agricultural bacillus EHA105 (pNOV2819-GFP , pNOV2819-NP1 ). Have got the transgenic bud after a batch of mannoses are screened . Carry on fluorescence measure to it , prove pNOV2819-GFP change poplar over to already , and transgenosis split up bud can a grows at the culture in mannose.