Tobacco Variety Improvement Using The Pollen-tube Pathway (K809,MD801,BD804)

In order to optimize the system of pollen-tube pathway technique,Plasmid pIG121Hm containing reported gene gus A and plant-selected gene npt-11 was introduced into tobacco'Basma'via the pollen-tube pathway at different times with different concentration by different ways.Seeds were harvested and screened in 500μg/ml kanamycine solution.Highest ratio(23.5%)of genet ransformation was discovered when 500μg/ml plasmid pIG121Hm DNA was introduced into ovary for 10 hours after pollination by injection.In order to improve aroma quality of tobacco,the genomic DNA of several tobacco germplasms:Ti245,Ti1068,6251 were introduced into K326 together by pollen tube pathway technique.A novel line of high aroma quality,K809 were obtained after 3 year's selection.Compare to K326,there were significant changes of morphology,leaf trichome density and aromatic components discovered in K809.The result of smoking proved K809 is of better aroma quality than K326.RAPD analysis showed that at least some DNA fragments of different donor were integrated into the genome of aroma quality K809.It indicated that different donors contributed to the modification of aroma quality.which its progeny could through gamogenesis in field production.In order to breed quality tobacco variety of curative function,hairy datura (Datura innoxia Mill),a traditional medicinal plant for curing cough and asthma through burning and fumigating was obtained to hybrid with tobacco(Nicotiana tabacum L.cv.Basma)by total DNA transformation way.A new hybrid descendent, designated as TD801 and MD804,were obtained after 3 years' selection.TD801 inherited certain excellent characters from both tobacco and hairy datura pheno typically.Chemical analysis of TD801 and tobacco showed less noticeable changes in other chemical contents,such as of sugar,protein and nicotine.The typical medicinal substances of hairy datura,e.g.atropine and scopolanine,were detected only in hybrid TD801 but in tobacco.75 random DNA markers were used to detect the genomes of tobacco,hairy datura and hybrid TD801 with the objective to assay their genetic diversity.Total 454 bands were produced in TD801 and 21 bands(4.6%)were discovered only in hairy datura but in tobacco,which proved that some DNA fragments of hairy datura were integrated into TD801 genome.This revealed a potential application of TD801 plants as special tobacco variety that can help to protect to respiration system when smoked.MD804 inherited certain excellent characters from both tobacco and corn mint phenotypically.Chemical analysis of MD804 and tobacco showed less noticeable changes in other chemical contents,The typical medicinal substances of corn mint e.g.Menthol,was detected only in hybrid MD804 but in tobacco.78 random DNA markers were used to detect the genomes of tobacco,corn mint and hybrid MD8041 with the objective to assay their genetic diversity.Total 358 bands were produced in MD804 and 19 bands were discovered only in corn mint but in tobacco,which proved that some DNA fragments of corn mint were integrated into MD804 genome.