一种基于菌丝悬浮液的核盘菌(Sclerotinia sclerotiorum)接种方法的建立

Establishment of an inoculation technique system for Sclerotinia sclerotiorum based on mycelial suspensions.

通过分析核盘菌(Sclerotiniasclerotiorum)菌丝培养时间、菌丝体浓度以及接种后的保湿时间等因子对接种后油菜植株发病的影响,建立了一套有效的基于菌丝悬浮液的核盘菌喷雾接种方法体系.主要步骤包括:从在平板活化3 d后核盘菌菌落边缘打取菌碟(Φ6 mm),按10块?100 mL-1加入液体培养基中,25℃、200 r?min-1下振荡培养4 d,用匀浆机匀浆30 s制成菌丝悬浮液,终浓度调至600 nm波长下吸光度值为1.0～2.0,用空压机喷雾接种油菜植株,保湿48 h.该方法适用于油菜苗期对菌核病的抗病性评价,还可用于立枯丝核菌(Rhizoctoniasolani)的接种研究.

We investigated the effect of Sclerotinia sclerotiorum mycelial culture time,concentration of mycelial suspension for inoculation,and time for keeping high relative humidity after inoculation on disease development in the inoculated rape plants,thereby established an effective spray inoculation technique system based on mycelial suspension for S.sclerotiorum.The main steps included： chose the newly grown mycelium that located on the most outside of the margins of 3-day-old colonies,which was added to the liquid culture medium by 10 cakes（6 mm at diameter） per 100 mL,shaking-cultured at 25 ℃ and 200 r·min-1 for four days,homogenized using a homogenizer for 30 seconds to form mycelial suspension,diluted it to a final concentration with an optical density at 600 nm of 1.0-2.0,which were then sprayed using a small air compressor on the rape leaves.The inoculated plants were grown under a high relative humidity for 48 hours.This inoculation system is valuable for evaluation of rape seedling resistance to S.sclerotiorum,and is suitable for Rhizoctonia solani inoculation study as well.