Application Of The Real-Time Q PCR To Analyze The Community Biodiversity Of Two Soil Fungi In Quantity Changing Wheat Field

Because of the application of transgenic technology, the area of biotech crops has been increased constantly, evaluating the safety of genetically modified crops is particularly important. Rhizosphere soil microbes is an important participant in the ecosystem, the growrh and decline of the rhizosphere soil microbes affect the occurrence of plant diseases. In this experiment, absolute quantification to Fusarium and Polymyxa graminis of the transgenic disease-resistance and drought-resistant wheat in rhizosphere soil, and establishing the Real-Time Q PCR detective system. With this reaction system, absolute quanification the Fusarium and Polymyxa graminis in transgenic wheat root soil throughout its growing period. Because of the complication of the microorganisms in soil, and soil fungal DNA extraction is a hard problem on fungal molecular ecology. Using conventional methods is difficult to be a pure culture, and can not reflect the changes in morghology of all microorganisms effectively, the fungal by pure culture can not reflect the real content by the natural state. So we adopt molecular biology to extract the total DNA of soil fungi and quantitative the Fusarium and Polymyxa graminis with Real-Time Q PCR to abtain the copies of the two strains, then we can observe the changes of two strains in soil of transgenic wheat root.Using the different methods to extract the fungal DNA in soil and find the best quality and clear bands DNA extraction with kit â…¡ (UltraCleanTM soil DNA Isolation Kit).Quantitive the Fusarium and Polymyxa graminis in soil of wheat root with Real-Time Q PCR, comparing to normal wheat and we can see that the transgenic wheat has little inhibition to Fusarium in soil. Fusarium in wheat throughout the growing period showed a normal distribution curve. Which turn TaDREB4gene drought-resistant wheat(TB4) strains and turn Tri101(DS) gene wheat strains has significantly inhaibited. High resistance to wheat yellow mosaic disease turned WYMV-Nib8gene wheat(N12-1) strains has no significant inhibition of Fusarium. But the high resistance to wheat yellow mosaic disease turned WYMV-Nib8gene wheat (N12-1) strains has significant inhibition of Polymyxa graminis, and the other two transgenic wheat varieties has no significant inhibition. The result has matched the disease resistance principle of high resistance to wheat yellow mosaic disease turned WYMV-Nib8gene wheat.The test results obtained are wheat year growth data, we need the results that the wheat growth data in3-5years to analysis.