Create Resistance Of New Soybean By Using Bombardment Technology

Soybean is the most important crops in the world,the demanding for soybeans constantlyincrease as the population grows.Disease has been a serious important factors affecting the yieldand quality. root disease include blight,damping-off and wilt blight,according to the survey,due toroot rot caused by soybean production at about10%every year,it has serious impact on thedevelopment of the soybean industry,so people devoted themself to improve the disease resistanceof soybean, traditional breeding methods can solve the soybean disease resistance,it take longer tocultivate new varieties,soybean disease can be reduced to a certain extent through the cultivationand management and spraying pesticides,but the long-term spraying of pesticides is not onlypollute the environment,but will improve the ability of the plant pathogen resistant.With the development of molecular biology, plant disease resistance genes engineering wasstudy deeply,the purpose of transfer of resistance genes to plants to improve plant broad spectrumand long lasting disease resistance.Plant Pathogenic Bacteria hrp (hypersensitive reaction andpathogenicity gene) gene encodes a the harpin protein that can induce plants to produce a series ofresistance responses,it can not only improve broad-spectrum disease resistance but also promoteplant growth.Harpin protein is subject to the hrp gene regulation and secreted by the typeIIIsecretion channel,their common features are:glycine rich,heat stable,protease sensitive.hrpZPstawas cloned from Clove pseudomonas tobacco disease-causing variants (Pseudomonas syringaepv.tabaci) Psta218strains.the gene length is approximately420bp, it can cause an allergic reactionin tobacco-induced plant disease resistance.Somatic embryos is an important target tissue for bombardment transformation method,Somatic embryos was induced mainly through the immature cotyledons.Its induction depend onsoybean genotypes.Different genotype have different induction.There is a difference to the qualityof different soybean genotypes induced somatic embryos.bombardment transformation methodconversion efficiency and simple operation,it is one of important the soybean genetictransformation method.This study construct the PDT-hrpZ plant expression vector, select the41north spring soybeansomatic embryogenesis,The hrpZ gene is into soybean of somatic embryos by using bombardmenttechnology, And the molecular biology detection to transgenic plants.The results showed that:1.Ken Feng23somatic embryos induced the highest rate in the screening of the41northernspring,with great number of somatic embryo on each immature cotyledon,did not producedeformity embryos,furthermore,each somatic embryo exist independently.Kenfeng23is most suitable genotype for soybean transformation.The inductivities of nine cultivars, such asHefeng55,were in the range of70~90%.2.We obtain11regenerated plants,PCR detection results show that the plant were positive.3.Using quantitative real-time PCR detection to transgenic plants that hrpZ were expressed intransgenic plants.