Tian Tian, Lin Xiangsheng, Tang Fajiang, Ding Xiawei, Wang Zhang-Ying, Liang Xuelian, Huang LiFei
Guangdong Academy of Agricultural Sciences, Crop Research Institute, Guangzhou, Guangdong, China;
Zhongkai University of Agriculture and Engineering, Guangzhou, Guangdong, China;
Plant Dis. 2025 Jan 1. doi: 10.1094/PDIS-11-24-2340-PDN.
Sweet potato ( (L.) Lam) is a major food crop that is cultivated in southern China (Huang et al. 2020). In February 2024, a survey was conducted in three warehouses of Suixi County (21°3' N, 109°50' E), Dianbai County (21°44' N, 111°6' E) and Chengmai County (19°57' N,109°56' E). Symptoms similar to charcoal rot were found on the storage roots of sweet potato. An inspection of 1,000 kg of sweet potatoes in each of the three warehouses showed that 5.5% (Suixi), 6.8% (Dianbai), and 8% (Chengmai) of the sweet potatoes were affected respectively. The disease began as a firm, moist rot that was reddish brown to brown and initially restricted to the cortex of the storage root. As the decay progressed, the pathogen crossed the vascular cambium, and the pith progressively rotted. Two distinct zones become apparent within the infected tissue. The leading edge continued as an area of reddish-brown decay, and an area of black tissue developed behind the area of active decay. The infected areas often encompassed the entire root, which eventually dried and became hard and mummified (Clark et al. 2013). Three sick sweet potatoes were selected from each warehouse for separation experiment,the early lesions of sweet potato storage root (3 mm × 3 mm) were isolated. Diseased segments were disinfected with 75% ethanol for 30 s, rinsed with 2.5% sodium hypochlorite for 3 min and washed three times with sterile water. The segments were placed on Potato Dextrose Agar (PDA) adjusted to a pH of 6.2. Isolates were purified by hyphal tipping. Ten isolates exhibited moderate growth rates and produced colonies that were white to fluffy black after 4 to 5 days of growth on PDA at 28℃. Microscopic observation showed that there were a large number of ellipsoidal microsclerotia between the hyphae. The microsclerotia were subfusiform, black and approximately 50 to 300 µm long. The morphological features matched the description of (Abd et al. 2019), the causal agent of charcoal rot. Three representative strains of the ten that were isolated from the three warehouses were selected for identification. PCR was performed using the primer sets ITS1/ITS4 to amplify the internal transcribed spacer (ITS) region of the rDNA (White et al. 1990). The PCR products with positive amplicons were sent to Sangon Biotech (Guangzhou, China) for sequencing. The analysis showed that the sequences of the three strains were consistent. Sequences were deposited in GenBank with the accession number PP887785.1, PQ727081.1 and PQ727082.1. Select one of the sequences for analysis (CRI-MP2: PP887785.1), It had 596 bp and was 100% homologous with the reference sequence of the ITS (GenBank accession numbers OR240859.1, OR240847.1, OR240855.1, OR240846.1, and OR240843.1). Further verify whether it is the . The genomic DNA of the three strains was amplified by the specific primer of (Babu et al. 2003), and the specific fragments that were 350 bp long were amplified in all three strains. The pathogenicity was determined using one isolate (Chengmai CRI-MP2). The pathogenicity of the isolate was confirmed by inoculating three healthy storage roots of the sweet potato cultivar Pushu 32 with a mycelial plug (0.5 cm × 0.5 cm) of . An additional three Pushu 32 storage roots were each inoculated with a sterile plug of PDA agar (0.5 cm × 0.5 cm) to serve as controls. All the storage roots were incubated at 25°C. All the inoculated storage roots exhibited the same symptoms observed in the field at 5 days after inoculation, whereas the controls remained asymptomatic. The fungi that were reisolated from the symptomatic roots were morphologically and molecularly identical to , while no pathogens were isolated from the mock inoculated controls. This experiment was repeated three times with similar results and thus, fulfilled Koch's postulates. is a generalist soilborne fungus present all over the world. Under high temperatures and low soil moisture, the fungus has a broad host range and can survive for a long time (Nathalie et al. 2021). The high temperature in southern China is suitable for the cultivation of sweet potato, but it interferes with the control of diseases, which can be challenging. To our knowledge, this is the first report of charcoal rot of sweet potato caused by in southern China. It provides new information and clues for the next step of disease prevention, which is to control the spread of this pathogen.
甘薯((L.) Lam)是中国南方种植的主要粮食作物(Huang等人,2020年)。2024年2月,在遂溪县(北纬21°3′,东经109°50′)、电白县(北纬21°44′,东经111°6′)和澄迈县(北纬19°57′,东经109°56′)的三个仓库进行了一项调查。在甘薯的贮藏根上发现了类似炭腐病的症状。对三个仓库中各1000千克甘薯的检查表明,受影响的甘薯比例分别为5.5%(遂溪)、6.8%(电白)和8%(澄迈)。该病最初表现为坚实、湿润的腐烂,颜色从红棕色到棕色,最初局限于贮藏根的皮层。随着腐烂的进展,病原菌穿过维管形成层,髓部逐渐腐烂。在受感染的组织内出现了两个明显的区域。前沿继续作为红棕色腐烂区域,在活跃腐烂区域后面形成黑色组织区域。受感染区域常常覆盖整个根,最终根变干、变硬并木乃伊化(Clark等人,2013年)。从每个仓库中挑选三个患病甘薯进行分离实验,分离甘薯贮藏根的早期病斑(3毫米×3毫米)。将患病组织块用75%乙醇消毒30秒,用2.5%次氯酸钠冲洗3分钟,并用无菌水冲洗三次。将组织块放置在pH值调至6.2的马铃薯葡萄糖琼脂(PDA)上。通过菌丝尖端法纯化分离物。十个分离物表现出中等生长速率,在28℃下于PDA上生长4至5天后产生白色至蓬松黑色的菌落。显微镜观察表明,菌丝之间有大量椭圆形微菌核。微菌核呈亚梭形,黑色,长约50至300微米。形态特征与炭腐病病原菌(Abd等人,2019年)的描述相符。从十个分离物中挑选出从三个仓库分离得到的三个代表性菌株进行鉴定。使用引物对ITS1/ITS4进行PCR扩增rDNA的内部转录间隔区(ITS)(White等人,1990年)。扩增出阳性条带的PCR产物送往上海生工生物工程股份有限公司(中国广州)进行测序。分析表明,这三个菌株的序列一致。序列已存入GenBank,登录号为PP887785.1、PQ727081.1和PQ727082.1。选择其中一个序列进行分析(CRI-MP2:PP887785.1),其长度为596 bp,与炭腐病菌ITS的参考序列(GenBank登录号OR240859.1、OR240847.1、OR240855.1、OR240846.1和OR240843.1)100%同源。进一步验证其是否为炭腐病菌。使用炭腐病菌的特异性引物对这三个菌株的基因组DNA进行扩增,在所有三个菌株中均扩增出长度为350 bp的特异性片段。使用一个分离物(澄迈CRI-MP2)测定致病性。通过用该菌的菌丝块(0.5厘米×0.5厘米)接种三个健康的甘薯品种‘普薯32’的贮藏根来确认该分离物的致病性。另外三个‘普薯32’贮藏根分别接种PDA琼脂无菌块(0.5厘米×0.5厘米)作为对照。所有贮藏根在25℃下培养。接种后5天,所有接种的贮藏根均表现出在田间观察到的相同症状,而对照仍无症状。从有症状的根中重新分离得到的真菌在形态和分子水平上与炭腐病菌相同,而从模拟接种对照中未分离到病原菌。该实验重复三次,结果相似,从而满足了柯赫法则。炭腐病菌是一种遍布全球的土传真菌。在高温和低土壤湿度条件下,该真菌寄主范围广泛,且能长期存活(Nathalie等人,2021年)。中国南方的高温适合甘薯种植,但这干扰了病害防治,可能具有挑战性。据我们所知,这是中国南方由炭腐病菌引起的甘薯炭腐病的首次报道。它为下一步病害防治,即控制该病原菌的传播提供了新的信息和线索。