Single spore isolation and purification method of verticillium dahliae

Abstract

The invention relates to a single spore isolation and purification method of verticillium dahliae. The method mainly comprises the following steps: an agar medium cylinder is taken from a water agar plate by the use of a glass capillary with diameter being 1mm and is picked onto a sterilized glass slide with a sterilization needle; a verticillium dahliae strain is activated in a PDA solid medium and the activated strain is cultured in a Czapek-Dox liquid medium; spore concentration is counted under a microscope; and the spore concentration is adjusted to 102-106 cfu to prepare a spore suspension; 1 microlitre of the spore suspension is sucked up and dripped onto the surface of the water agar cylinder on the glass slide; the glass slide is moved into a culture dish for constant temperature incubation at 25 DEG C; the glass slide is detected under the microscope, and the cylinder containing a single conidiospore which has germinated germ tubes is transferred onto a PDA plate to be cultured at 25 DEG C so as to finish single spore isolation. The method is simple to operate, can be implemented by general operation staff and is suitable for mass separation. By the method, separation success rate is high. The method is also suitable for production of other fungi of microconidia.

Description

technical field [0001] The invention belongs to the field of separation and purification of fungi, and relates to a single-spore separation and purification method of Verticillium dahliae. Background technique [0002] Verticillium dahliae (Verticillium dahliae) belongs to the fungus of the subphylum Verticillium genus Verticillium, which has a wide host range and can harm more than 600 kinds of plants in 38 families, such as cotton, soybean, mung bean, sugar beet, sesame and other field crops. And eggplant, pepper, tomato, watermelon, cucumber, melon and other vegetables and fruits. Cotton Verticillium dahliae caused by Verticillium dahliae is particularly harmful. There were 5 major outbreaks in my country in 1993, 1995, 1996, 2002 and 2003, and the annual loss was about 1.2 billion yuan. Therefore, it is particularly important to carry out research on the pathogenic mechanism of Verticillium dahliae and to prevent and control plant diseases caused by Verticillium dahliae...

AI Technical Summary

Problems solved by technology

Although this invention can realize the isolation of single spores of fungi, its isolation operation must be carried out under a compound microscope or dissecting microscope, and it has high requirements for the skills and experience of the operator, and the success rate of its isolation will vary depending on the operating factors of the operator. Great changes have taken place due to the influence of the bacterium, and it is not suitable for the separation of large quantities of fungal unit cells; at the same time, the invention is not ideal for the separation of small conidia, and is not suitable for producing small conidia like Verticillium dahliae Single spore isolation of fungi

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