用4 μg/mL 4 ′,6-二脒基-2-苯基吲哚(DAPI)对细胞核染色,采用激光共聚焦显微成像技术观察粗腿羊肚菌(Morchella crassipes)子囊孢子的萌发过程.新鲜的粗腿羊肚菌子囊孢子呈椭圆状,大小13.0~15.3 μm×21.5~24.0 μm,成熟子囊孢子的细胞核数量为18~32个;CYM液体培养基,23 ℃培养6 h后子囊孢子体积逐渐膨大至初始体积的1.7~2.9倍,泡囊化现象明显;6~12 h开始形成芽管,泡囊逐渐消失,出芽点在椭圆形细胞的一端形成或两端先后形成,数量不等的细胞核由子囊孢子内迁至芽管中,形成多核丝状;芽管生长至20~30 μm后,在出芽点与孢子连接处形成隔膜,12~24 h后,随着芽管的持续生长,形成新的隔膜和菌丝细胞,菌丝细胞多核;芽管及初生菌丝直径6~8 μm,约为子囊孢子直径的1/3;24 h内没有观察到初生菌丝之间发生细胞融合现象.%Ascospores germination and nuclear behavior (stained by 4 μg/mL DAPI) was observed under confocal laser scanning microscope.Fresh ascospores(ellipsoid,13.0~15.3 μm×21.5~24.0 μm,18~32 nuclei in each spore) could expanded to 1.7~2.9 fold in volume when incubated in CYM broth(23 ℃) for six hours with distinctive vesiculation, and formed germ tubes from one or both sides at oval ends during six to twelve hours incubation accompanied by visicles disappearing and nuclei migrating.Germ tubes extended further to form primary hypha(6~8 μm in width), septum was observed at joint of tube and spore when germ tube extending to 20 to 30 μm length, but no cell fusion between primary hypha was detected within 24 h of incubation.
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