Wild Rice And Wild Rice Stem Pregnant Black Powder Symbiotic Bacteria Cultured In Vitro And Rapid Detection Technology

As one of the feature vegetables originated from China and Southeast Asia, Zizania caduciflora L. belongs to perennial aquatic vegetables. Under the feasible condition some segments of stems swell quickly after accreting with Ustilago esculenta P. Henn. While the harvest from culture the Zizania caduciflora L. become better, the quantity and the culture area become larger. As a result of this, more scientists are being attracted in this filed. Due to the past researches, it is a real hard work to culture the Ustilago esculenta in vitro in order to get enough mycelia for experiments.With the32main cultured Zizania caduciflora L. in China as the basic experimental materials, we used the Random amplified polymorphic DNA (RAPD) to selecte them. And we used a DNA market technic to rapid detected Ustilago esculenta. Base on the in vitro ways we just founded to culture Ustilago esculenta by utilizing the liquid fermentation, we detected the main endogenous hormones continuously, and found some of their general rules. The results as the follows:12primers were screened from70arbitrary primers to study the DNA polymrphism of32varieties of Zizania caduciflora L. collected in China. Totally75DNA fragments were generated, and average number of DNA band produced by each primer was6.2,21of them varieties, occupied about28%. Similarity coefficients and genetic distances between varieties were calculated, and the clustering analysis of amplifying DNA band showed that32varieties were divided into7clustering groups and21collected had a close similarity, occupied about65.63%from the total. This study showed that, most of our nation’s main Zizania caduciflora L. have a close similarity.A rapid dot-ELISA for detecting Ustilago esculenta’s DNA was established. Using a pair of PCR primers, one was labeled with an extra Biotin on its end and the other one was normal, all the productions of PCR will be labeled with Biotin. A detector labeled FITC hybridized with the PCR products, they got both Biotin and FITC labeled. The hybridizer will be fixed on the NC membrane’s surface by Anti-FITC. Alkaline phosphatase-conjugated streptavidin and its substrate are added to the NC membrane in order to show the results. This method is considered to be a specific, sensitive and rapid for detecting the U. esculenta from different Zizania latifolia Turcz. It can detect the DNA even till0.2pg/ml.The liquid fermentation by Ustilago esculenta was investigated. The effect of culture medium, pH, incubation volume, agitation ratio, growth temperature and growth time on the production of the fungi was studied. The results showed that the optimal culture conditions were: PDA medium, pH6, agitation ratio130r/min, fermentation temperature28℃, inoculation volume5%, and fermentation time6d.Under the condition of liquid fermentation, we utilized the enzyme linked immunosorbent assay (ELISA) to detect Ustilago esculentd’s5main endogenous hormones from the mycelia and its ermentation solution, including GA3(Gibberellin), IAA (Indoleacetic), ABA (Abscisic acid), Z (Zeatin)&ZR (Zeatin riboside). The resluts showed some general rules of the hormones varities attend by the fermented time.