A method for improving the quality of DNA extraction of the coin spot bacteria genome

Abstract

The invention discloses a method for improving the extraction quality of sclerotinia homoeocarpa genome DNA. The method comprises the following steps that an MM culture medium containing antibiotics is added into a culture vessel, sterilized glass paper is laid on the surface of the culture medium, and hyphae are picked to be transferred to the middle of the culture medium and cultured for 7-14 d under the dark condition at the temperature ranging from 25 DEG C to 30 DEG C; 20-50 mg of the hyphae on the surface of the glass paper are scraped to be put into a centrifuge tube, ventilating drying is conducted, steel balls are added into the centrifuge tube, and the hyphae are frozen with liquid nitrogen and then ground; genome DNA is extracted through a classical CTAB method and dissolved into a TE buffer solution, standing is conducted for 24-48 h, centrifuging is conducted, and impurities which are not dissolved into the TE buffer solution are removed. According to the method, the content of polysaccharide in sclerotinia homoeocarpa hypha tissue is decreased by adopting the MM culture medium, and therefore the extraction quality of pathogenic bacterium genome DNA is improved; expensive instruments and equipment or DNA extracting kits are not needed, the DNA extraction quality is significantly improved, the stability is good, and the method is economical and efficient.

Description

Technical field [0001] The invention belongs to the field of plant pathogenic microorganisms, and relates to a method for improving the extraction quality of genomic DNA of dollar spot pathogen. Background technique [0002] Sclerotinia homoeocarpa F.T. Bennett is an important plant pathogenic fungus on turfgrass. It can infect almost all cold-season and warm-season turfgrasses and other grasses. In 1932, the currency spot disease caused by the pathogen infection was first reported in the United States. Currently, currency spot disease has been widely distributed worldwide. In our country, the pathogen has appeared on a variety of lawn grasses in more than 20 provinces. Coin spot disease mainly has two peaks of incidence in early summer and early autumn. When the soil temperature is higher than 18℃ and the humidity at night is high, coin spot disease begins to develop. At the beginning of the disease, a coin-sized nearly round spot is formed on the lawn, which is serious. At ti...

AI Technical Summary

Problems solved by technology

In the prior art, PDA is generally used to cultivate coin spot bacteria. When the pathogenic bacteria are cultivated on the medium, because the medium provides the pathogenic bacteria with a large amount of sugar, the polysaccharide content in the bacteria is relatively high, and the level of polysaccharide produced between different strains exists. difference, and the polysaccharide content directly affects the extraction efficiency of genomic DNA

When using conventional DNA extraction methods or kits commonly used in the market to extract the genomic DNA of coin spot bacteria, it is often due to the presence of a large number of polysaccharides in the mycelium tissue, which seriously affects the yield of DNA, and some methods for removing polysaccharides are also difficult to extract. Effective separation of polysaccharides and target DNA in the DNA extraction process of leaf spot fungus

As a result, when carrying out subsequent experiments related to molecular biology, especially when conducting research related to population biology or genomics, it is often difficult to obtain stable and high-quality genomic DNA, which seriously affects the progress of the experiment

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