Wu Yanpan, Zhang Fuhai, Zhang Shaohui, Zhang Jun, Zhao Sujing, Qiu Zongbo, Zhu 朱 Mo 墨
Henan Normal University, College of Life Sciences, Xinxiang, Henan, China;
Kaifeng Agricultural Technology Extension Station, Kaifeng, China;
Plant Dis. 2025 Jun 3. doi: 10.1094/PDIS-03-25-0477-PDN.
Ipomoea batatas (sweet potato) is an annual herb originating from South America and the large and small Antilles. It is widely cultivated in tropical and subtropical regions around the world and is widely grown in most parts of China. As an edible plant with rich nutrition, I. batatas has high economic and medicinal value (Suhendy et al. 2023). In September 2023, black rot-like disease signs and symptoms were observed on the roots of I. batatas in a farmland (about 6667 m2) located in Kaifeng city, Henan Province, China. The roots showed irregular brown or dark spots on the surface, extending to the internal center, and brown to black necrosis. Additionally, above-ground parts of infected plants showed symptoms such as yellowing and wilting of leaves, brown spots on stems, and stunted growth. Around 80 % of monitored plant roots (n = 200) exhibited the symptoms. Infected roots were cut into pieces and then placed on potato dextrose agar (PDA) (light/dark, 16 h/8 h; temperature, 18°C). After a period of 3-10 days, single hyphal tips of each fungal colony were placed on PDA and incubated for another 5-10 days (Paul et al. 2021). Colonies of the fungal pathogen on PDA reached 50 mm in diameter within 7 days, dark gray on the inner side, dark brown extending to the edge, irregular round edge, with abundant aerial mycelium, cotton-like, irregularly convex upward, undulating. The conidia were brown, ellipsoid to oval, 8 to 19 × 3 to 6 μm (n = 50). Morphologically similar isolates with characteristics consistent with those of Curvularia spicifera (Cui et al. 2020) were recovered from 87% of symptomatic root tissues (n=100). The internal transcribed spacer (ITS) region and glyceraldehyde-3-phosphate dehydrogenase (GAPDH) gene from three independent isolates (CSZM202101, CSZM202102, and CSZM202103) of the fungus were amplified and sequenced with primers ITS1/ITS4 (Seliger et al. 1990) and GAPDH1/GAPDH3R (Bradshaw et al. 2022) according to a previously reported method (Zhu et al. 2022). The resulting sequences were deposited in GenBank (Accession No. OR885691, PV056889, PV056568, PQ839726, PV072835 and PV02836). BLASTn analysis showed that the ITS and GAPDH sequences had 100% (ITS, 516/516; GAPDH, 508/508) identity with C. spicifera (OQ845826 and CBS 274.52 JN192387) from maize (Ram et al. 2024) and pearl millet (S. et al. 2024), respectively. The phylogenetic analysis clearly illustrated that these isolates clustered with the sequences of a representative reference strain of C. spicifera(CBS 274.52, GenBank accession number JN192387). Therefore, the morphological, molecular and phylogenetic analysis indicated that the pathogen was C. spicifera. To complete Koch's postulates, pathogenicity experiments were carried out by inoculating spore suspension (106 spores mL-1) into the wounded roots (n=10) of I. batatas plants. Sterile distilled water treated wounded roots (n=10) served as control. One to two days after inoculation, visible mycelia were produced at the inoculation sites of I. batatas. Inoculated roots showed black rot signs 21 days post-inoculation; controls were unaffected. This was confirmed in three repeated pathogenicity tests. To our knowledge, this is the first report of black rot caused by C. spicifera on I. batatas in China. The emergence of the black rot pathogen could harm valuable food crops and reduce agricultural productivity in China. Accurate identification of the black rot fungus is essential for devising effective disease management strategies and supporting future control of C. spicifera in China.
甘薯是一种一年生草本植物,原产于南美洲以及大小安的列斯群岛。它在世界各地的热带和亚热带地区广泛种植,在中国大部分地区也广泛栽培。作为一种营养丰富的可食用植物,甘薯具有很高的经济和药用价值(苏亨迪等人,2023年)。2023年9月,在中国河南省开封市一块面积约6667平方米的农田里,观察到甘薯根部出现类似黑腐病的症状。根部表面出现不规则的褐色或深色斑点,并延伸至内部中心,呈现褐色至黑色坏死。此外,受感染植株的地上部分出现叶片发黄和枯萎、茎部出现褐色斑点以及生长受阻等症状。大约80%受监测的植株根部(n = 200)表现出这些症状。将受感染的根切成小块,然后置于马铃薯葡萄糖琼脂(PDA)培养基上(光照/黑暗,16小时/8小时;温度,18°C)。3至10天后,将每个真菌菌落的单个菌丝尖端置于PDA上,再培养5至10天(保罗等人,2021年)。该真菌病原体在PDA上的菌落在7天内直径达到50毫米,内侧为深灰色,深褐色延伸至边缘,边缘不规则圆形,有丰富的气生菌丝,呈棉絮状,向上不规则凸起,呈波浪状。分生孢子呈褐色,椭圆形至卵形,8至19×3至6微米(n = 50)。从87%有症状的根组织(n = 100)中分离出形态上相似且特征与尖孢弯孢霉(崔等人,2020年)一致的菌株。根据先前报道的方法(朱等人,2022年),使用引物ITS1/ITS4(塞利格等人,1990年)和GAPDH1/GAPDH3R(布拉德肖等人,2022年)对该真菌的三个独立分离株(CSZM202101、CSZM202102和CSZM202103)的内部转录间隔区(ITS)和甘油醛-3-磷酸脱氢酶(GAPDH)基因进行扩增和测序。所得序列保存在GenBank中(登录号:OR885691、PV056889、PV056568、PQ839726、PV072835和PV02836)。BLASTn分析表明,ITS和GAPDH序列分别与来自玉米(拉姆等人,2024年)和珍珠粟(S.等人,2024年)的尖孢弯孢霉(OQ845826和CBS 274.52 JN192387)具有100%(ITS,516/516;GAPDH,508/508)的同一性。系统发育分析清楚地表明,这些分离株与尖孢弯孢霉代表性参考菌株(CBS 274.52,GenBank登录号JN192387)的序列聚类在一起。因此,形态学、分子和系统发育分析表明,病原体为尖孢弯孢霉。为了完成科赫法则,通过将孢子悬浮液(106个孢子/毫升)接种到甘薯植株的受伤根部(n = 10)进行致病性试验。用无菌蒸馏水处理的受伤根部(n = 10)作为对照。接种后1至2天,甘薯接种部位产生可见菌丝体。接种后21天,接种的根部出现黑腐症状;对照未受影响。这在三次重复的致病性试验中得到证实。据我们所知,这是中国首次报道尖孢弯孢霉引起甘薯黑腐病。黑腐病原体的出现可能会损害有价值的粮食作物,并降低中国的农业生产力。准确鉴定黑腐真菌对于制定有效的病害管理策略以及支持中国未来对尖孢弯孢霉的防治至关重要。