Xie Jiangtao, Liu Xue, Lu Junqian, Qin Zaili, Yu Naqin, Zeng Xiang-Yu, Tian Fenghua
Guizhou University, Huaxi District, Guiyang City, Guizhou ProvinceGuiyang, China, 550025;
Guiyang, China;
Plant Dis. 2023 Sep 12. doi: 10.1094/PDIS-02-23-0322-PDN.
Auricularia cornea is a widely cultivated mushroom in China, which has high medicinal values such as hemostaticity, analgesia, antioxidation and anti-tumor (Wu et al., 2019). In 2022, an investigation on edible mushroom diseases in Guizhou Province observed a suspected cobweb disease in an A. cornea growing factory, with up to 30% incidence. The pathogen first produced flocculent hyphae on the surface of the fruiting body of A. cornea, and then developed spider web-like aerial hyphae, covering the entire fruiting bodies. It hinders the normal growth of A. cornea, resulting in deformity and rot of the fruiting bodies. These symptoms seriously affect the quantity and quality of mushroom yields and cause huge economic losses. Three fungal isolates (GUCCX001, GUCCX002 and GUCCX003) were recovered from the diseased mushroom fruiting bodies and purified through single spore isolation. The colonies of three isolates spread rapidly on PDA, reaching 79-82 mm in seven days. The flocculent mycelium was whitish, and its reverse turned from yellowish to amber after 14 days. The branched conidiophores arising from aerial mycelia were septate and each cell contained several denticulate conidiogenous loci. Each denticle contained a single conidium. Conidia were observed at the tip of conidiophore branches and were 0-1-septate, oval or spherical, transparent, 5.2-11.3 × 11.7-18.7 μm (n = 35). Chlamydospores were visible as 3-4 thick-walled cells at the tip of lateral hyphal branches. Three isolates were tentatively identified as H. mycophilus based on their morphological characteristics similar to those described by Rogerson and Samuels (1993). The sequence of internal transcribed spacer (ITS) region (primers ITS5/ITS4) (Rehner and Samuels, 1994) and nuclear ribosomal large subunit (LSU) region (primers LR0R/LR5) (Vilgalys and Heste, 1990) of GUCCX001 (ITS: OP777905; LSU: OQ152071), GUCCX002 (ITS: OP862872; LSU: OQ152072) and GUCCX003 (ITS: OP862873; LSU: OP862873) were 99%-100% similar to H. mycophilus CBS 175.56 (ITS: MH857567; LSU: MH869110). Fifteen healthy fruiting bodies of A. cornea were inoculated by spraying spore suspension (106 conidia/mL) of the three isolates and five healthy fruiting bodies were sprayed with sterile water as control. All inoculated fruiting bodies were kept at 25 ℃. After three days, fruiting bodies of A. cornea treated with the spore suspension exhibited the same symptoms of cobweb as in the factory, while no symptom appeared in the control. Pathogens re-isolated from diseased fruiting bodies were confirmed to be H. mycophilus based on morphological characteristics, which fulfills the Koch's postulate. Zeng et al. (2017) reported H. mycophilus on the fruiting bodies of Auricularia sp. as a new record in Guangdong, China. H. mycophilus caused cobweb disease on A. auricula (Liu et al., 2020), A. cornea var. Li. (Cao et al., 2023) and A. heimuer (Zhang et al., 2023). To our knowledge, this is the first report of cobweb disease in A. cornea caused by H. mycophilus in Guizhou, China. Our findings will provide a basis for correct diagnosis and management of cobweb diseases on A. cornea.
皱木耳是中国广泛栽培的一种蘑菇,具有止血、镇痛、抗氧化和抗肿瘤等较高的药用价值(Wu等人,2019)。2022年,一项对贵州省食用菌病害的调查在一个皱木耳种植厂观察到一种疑似蛛网病,发病率高达30%。病原菌首先在皱木耳子实体表面产生絮状菌丝,然后发育成蛛网状气生菌丝,覆盖整个子实体。它阻碍了皱木耳的正常生长,导致子实体畸形和腐烂。这些症状严重影响蘑菇产量的数量和质量,并造成巨大的经济损失。从患病蘑菇子实体中分离出三株真菌菌株(GUCCX001、GUCCX002和GUCCX003),并通过单孢分离进行纯化。三株分离物的菌落在PDA上迅速蔓延,7天内达到79 - 82毫米。絮状菌丝体为白色,14天后其背面从淡黄色变为琥珀色。从气生菌丝产生的分枝分生孢子梗有隔膜,每个细胞含有几个具细齿的产孢位点。每个细齿含有一个分生孢子。分生孢子在分生孢子梗分支的顶端观察到,0 - 1分隔,椭圆形或球形,透明,5.2 - 11.3×11.7 - 18.7μm(n = 35)。厚垣孢子在侧生菌丝分支的顶端可见为3 - 4个厚壁细胞。根据其形态特征与Rogerson和Samuels(1993)描述的相似,三株分离物初步鉴定为嗜菇毁丝霉。GUCCX001(ITS:OP777905;LSU:OQ152071)、GUCCX002(ITS:OP862872;LSU:OQ152072)和GUCCX003(ITS:OP862873;LSU:OP862873)的内部转录间隔区(ITS)序列(引物ITS5/ITS4)(Rehner和Samuels,1994)和核糖体大亚基(LSU)区域(引物LR0R/LR5)(Vilgalys和Heste,1990)与嗜菇毁丝霉CBS 175.56(ITS:MH857567;LSU:MH869110)的相似度为99% - 100%。用三株分离物的孢子悬浮液(106个分生孢子/毫升)喷雾接种15个健康的皱木耳子实体,并用无菌水喷雾5个健康子实体作为对照。所有接种的子实体保持在25℃。三天后,用孢子悬浮液处理的皱木耳子实体表现出与工厂中相同的蛛网病症状,而对照中未出现症状。从患病子实体中重新分离出的病原菌根据形态特征确认为嗜菇毁丝霉,这满足了科赫法则。Zeng等人(2017)报道嗜菇毁丝霉在木耳属子实体上是中国广东的一个新记录。嗜菇毁丝霉在黑木耳(Liu等人,2020)、皱木耳变种Li.(Cao等人,2023)和毛木耳(Zhang等人,2023)上引起蛛网病。据我们所知,这是中国贵州首次报道嗜菇毁丝霉引起的皱木耳蛛网病。我们的研究结果将为正确诊断和管理皱木耳的蛛网病提供依据。