| Eustoma Gradiflorum is Gentianaceae biennial herbs, be produced in central region in America at first, Japan educated this kind of plant first in 1935, Eustoma Gradiflorum did not develop quickly until be close to 20 years.It has the color abundant, the blossom beauty etc. characteristics, be the cut new talent flowers, be liked by the people more and more, the year of world sells the sum 110,000,000 $, the row cut flowers ranks the seventh.But Eustoma Gradiflorum seed is small, growing up difficulty, so build up the demand that it breeds the system quickly to make it full to satisfy the market to have the important meaning;At the same time because many irregularities that normal regulations breeding made through a new species of the genetic engineering method seed selection became an inevitable trend.Don't bear the dry characteristics according to the pleased wetness of Eustoma Gradiflorum, this experiment choice turns into the anti- drought and the anti- salt alkali genes, making it have the stronger anti- negative, to raise its adverse circumstances of live the rate contain bigger meaning.Because the value of the flower mainly is it appreciates the value, so turn the gene flower commercial were easy, but not the elephant food is so the pressure of the situation society, all countries scientistses will make use of the genetic engineering flower breeding in the important research category.In several years of aftertime, estimate some nations to turn the gene flower industry, in our country, science and technology department had already turn the gene flower to list as 863 research items in 2002, and obtained the outstanding result. The flower anti- genetic engineering in the research result of the report way was not much.The researcher of the Australia duct the coat protein( CP) gene of the yellow spot virus into the carnation, acquiring the transgenic plant.Eustoma was researched less, but convert the Eustoma plant aspect in the agriculture rod bacteria almost blank.This thesis plants the body outside with the different part conduct and actions of Eustoma ,compare different media, the hormone category and density, the environment factor etc. establishment of the system Eustoma quickly regenerate to study meticulously.At the same time this thesis still makes use of the moment of Gus gene of β - glucose to express to harm the built-up influence factor to carry on the research with blue several gricultures of spot rod bacteria conversion Eustoma plant of stable expression more. after analysis builds up its better conversion condition .Agriculture rod bacteria conversion Eustoma that passes the anti-dry gene and the anti- salt alkali genes harms the organization more, and differentiation a plant.PCR examination proved that the gene had already integrated the plant.research result were following few aspectses:0.1% Hgcl2 and 3% Naclo soak the plant 3 mins,5 mins,10 mins,15 mins to put out thegerm result comparing the detection:70% alcohol soak 30 ses and 0.1% Hgck soak 10 minutes was the best way, the free from pollution rate attains 100%, the death rate is 2%. When the basic media is MB,IBA2.0mg/ L,6-0.5 mgs/ L of BA, GA2.0mg/ L, the inducement rate is tallest.Among them, the IBA has t the function of increasing the number of tillering within the scope of small in the 2.0 mg/ L densitythe different hormone ,different density combination of 6-BA, KT, NAA, IBA e influence of organize the occurrence to the Estoma.The different from the hormone of the density combine the big part and can make the outside plant the body to take off to divide, creation quality of different category harms the organization more, but when BA density is 1.0 mgs/ L, half of the callus formation the water immerses, the glass turn more serious, while being its density to reduce to the 0.5 mgs/ L the phenomenon cancellation.In induce process,the density of 6-BA should<1.0 mgs/ L, density of KT<2.0 mgs/ L.When the density of NAA is a 0.2 mgs/ L, more callus occurrence result best.As a result discover that use the MS ,the density of 2,4-Ds are 0.5 mg/ Ls, the density of NAA is zero, more callus was yellow, the quality is more loose and soft, increase heavy most quick.Take MS as the basic media, the PH value adjusts to 6.0, being 6 — BAs and KTs alone with the 0.5 mg/ L density usage, it divides the seedling of normal, leafs slice is thick green and fatty big, but when density over 1.0 mgs/ L, the glass of the young sprout turned severity.When the PH is worth to 6.3, the light shines on the strength as 3000 hz, density of ZT,NAA distinguishes for the 0.5 mg/ L and 0.1 mg/ L, leaf the dish bud divides the rate tallest, long certainly best.In the process of the agriculture rod bacteria invade dye the Eustoma more harm organize inherit convert, the dissimilarity invades to dye time to the conversion the influence of the efficiency is more obvious, dying from the GUS of as a result see, be to invade to dye time as 5 minutes, convert the efficiency tallest, be to invade to dye time over the 7 mins, more harm to organize the pollution severity, even can't become to live.Germ liquid density to be worth to 0.5 for the OD600, the agriculture rod bacteria infection the efficiency tallest, the germ liquid density is 0.1, infect result to descend obviously, but when the density OD value over 0.5, harming to organize the death rate exaltation more.It was proved by PCR analysis that the target gene had been integrated into the genome of regenerated plants.Rate of PCR transformation was 4.9%.
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