Method for inducing phytophthora parasitica var. nicotianae to generate zoosporangium and release spore

Abstract

The invention discloses a method for inducing phytophthora parasitica var. nicotianae to generate zoosporangia and release spores. The method comprises steps of: culturing phytophthora parasitica var. nicotianae mycelia on an oat medium prepared in a narrow mouth conical flask; preparing an induction liquid for generation of zoosporangia by selecting roots of tobacco seedlings, with 6-7 pieces of true leaves completely expanding, of a black shank infected variety; inducing the mycelia with the induction liquid to generate zoosporangia and to release spores. According to the invention, a characteristic that a black shank infected tobacco variety is more easily to generate affinity reaction with phytophthora parasitica var. nicotianae is employed, and a filtrate of seedling roots of the black shank infected tobacco variety is used as the induction liquid, so as to induce the cultured mycelia to generate zoosporangia. Proved by tests, the invention has a short induction time, high zoospore release rate, high obtained zoospore content; and scraping mycelia for culture is not needed. Therefore, the invention is of less pollution and easy to operate.

Description

technical field [0001] The invention belongs to the technical field of microorganism cultivation, and in particular relates to a method for inducing Phytophthora nicotianae to produce zoosporangia and release spores with good effect, high efficiency and low pollution rate. Background technique [0002] Phytophthora tobacco ( Phytophthora parasitica var . nicotianae ) is the pathogen of tobacco black shank. Under natural conditions, zoospores are mainly used to infect tobacco plants from the roots or stem bases. In artificial culture, it can be cultured by culturing hyphae, inducing zoosporangia, and low-temperature treatment. The zoosporangium releases zoospores, thereby obtaining a greater number of zoospores. Commonly used induction methods are: using oat plate to culture mycelium, 0.1% KNO 3 Solution infiltration can induce Phytophthora nicotianae to produce zoosporangia[Gooding G V, Lucas G B. Factors influencing sporangail formation and zoospore activity in Phytoph...

AI Technical Summary

Problems solved by technology

It can also be taken: after Phytophthora nicotianae grows mycelium on oat medium, drop the Peeter's culture solution of soil leaching liquid to induce zoosporangia, after low temperature treatment, then cultivate at an appropriate temperature to produce a large number of zoospores [Yang Jianqing , Jiang Tong, Chen Xueping. Research on the cultivation of Phytophthora nicotiana and the method of mass production of zoosporangia and zoospores. Plant Protection, 2001, 27(4):12-14.], but because the soil leachate was not strictly sterilized Bacteria and Petri culture fluid are sterilized by moist heat, the effect of inducing zoosporangia is unstable and easy to contaminate

Chinese invention patent ZL 200410040934.4 also discloses a sporulation culture method of Phytophthora nicotiana, which adopts the selection of sesame medium or rye medium as the sporangia medium of Phytophthora nicotiana when separating Phytophthora nicotiana, Inoculate the activated Phytophthora nicotianae on the sesame medium and rye medium plate, culture in the incubator at 28°C for 14 days, scrape the mycelium, and use 0.1% KNO 3 After infiltration, the number of sporangia and the size of the sporangia were measured after 5-8 days, and the sporangia were treated at 6-11°C for 15-25 minutes to release zoospores, but the induction time of this method is relatively long, and the release rate of zoospores needs to be improved