芸薹根肿菌(Plasmodiophora brassicae)单孢分离接种及生理小种的鉴定

Separate inoculation of single resting spores and identification of Plasmodiophora brassicae races

芸薹根肿菌(Plasmodiophora brassicae)单孢分离接种方法曾有报道,本试验改进了国外单孢分离技术。将休眠孢子悬浮液稀释到1个/0.5μL,滴在一块消过毒的载玻片上,放置在显微镜下反复检查,以确保它精确只含有一个孢子。用微管将孢子吸入接种在2日龄白菜幼苗根毛上,营养液培养3周。为使其更接近田间发病环境条件,后期将疑似发病根在无菌土中继续进行病菌增繁。该方法能够有效地加快病菌繁殖感染,提高单孢分离接种成功率。对不同地区病菌进行大量单孢分离接种。本试验对3个地区的病菌单孢接种共950株,接种成功27株,使用Williams根肿病菌鉴别寄主鉴定,共9个生理小种。

Plasmodiophora brassicae single-spore separate inoculation method has ever been reported. This trial improved foreign method of single-spore isolation. The dormant spore suspensions were diluted to 1/0.5 μL, and dropped on a sterile glass slide. Then it was placed under a microscope to check it repeatedly and make sure it contains only one spore. The spores were then inhaled into the microtube and inoculated on the two-day-old cabbage seedling＇s root hairs, with greenhouse culture in the nutrient solution for 21 days. To bring it closer to the conditions of field incidence, the suspected infected root was later developed in the sterile soil to improve the proliferation of P. brassicae. The method could effectively speed up the infection and breeding of P. brassicae, and promote the success rate of separate inoculation. This separate inoculation experiment was carried out using a large number of single spores of different physiological pathogens. The number of pathogen inoculated in the three regions added up to 950, and 27 of them worked. The Williams root swelling P. brassicae helped to isolate 9 physiological races.