辣椒(CapsicumannuumL.)是世界上重要的经济作物,因其烹饪用途而受到重视。在韩国,炭疽病引起的炭疽病。包括C.truncum,C.gloeosporioides,C.coccodes,C.acutatum,和C.scovillei造成了巨大的经济损失(Kim等人。2008;Oo和Oh2020)。2022年8月,在Yereonggwang的一个田地中观察到一些不同类型的辣椒果实不典型的症状(GPS:35.2579°N,126.4742°E),韩国。疾病症状表现为凹陷,坏死病变,在同心环中形成密集的黑色孢子团。估计的疾病发生率为0.2公顷的田地显示出高达1%的水果受影响。为了分离病原体,收集了六种有症状的辣椒。从病变边缘切下小块(5平方毫米),在70%乙醇中表面灭菌30秒,然后用1%次氯酸钠1分钟,然后在无菌蒸馏水中冲洗三次。将组织块置于马铃薯葡萄糖琼脂(PDA)平板上并在25°C下在黑暗中孵育。3到5天后,对新出现的真菌菌落进行传代培养以获得纯的分离株。总共获得了5个分离株,并初步鉴定为炭疽菌。基于形态特征。七天大的殖民地最初是白色的,在PDA上随着年龄的增长变成浅橙色。油层(在病变上观察)为深棕色,疣糖和隔。分生孢子是圆柱形的,透明玻璃,测量值14.8至19.9×4.2至6.5μm(平均16.7×5.6μm,n=70)大小;压疮为棕色至深棕色,形状不规则。分离株的这些形态特征与Damm等报道的大豆C.sojae形态的特征一致。(2019年)。为了确认分离株的身份,提取DNA,使用以下引物组扩增和测序特定基因区域:ITS(ITS1和ITS4),GAPDH(GDF1和GDR1),ACT(ACT-512F和ACT-783R),TUB(T1和Bt2b),HIS3(CYLH3F和CYLH3R),和CHS-1(CHS-79F和CHS-345R)。将所得序列以登录号(LC830742至LC830766)保藏在NCBIGenBank中。使用ITS的组合序列进行最大似然系统发育分析,GAPDH,ACT,浴缸,MEGAX中的HIS3和CHS-1证实分离株为大豆梭菌,标志着这种病原体在韩国辣椒上的首次报道,以前已知会感染大豆。对伤口和未受伤的健康和成熟的绿色辣椒果实进行了致病性测试(cv。Bicksita)以确认分离的C.sojae的致病性。使用70%乙醇对果实进行表面灭菌,然后用无菌蒸馏水冲洗。使用无菌针头将果实打伤以促进感染。从7天龄的PDA培养物制备分生孢子悬浮液(1×106分生孢子/mL)。通过将10µL分生孢子悬浮液滴在伤口和未缠绕水果的受伤和未受伤部位(4至5)上接种每个水果,分别。对照果实用无菌水接种。每个处理使用总共40个果实,并且重复实验两次。将果实放置在衬有湿纸巾的塑料盒中,以保持高湿度,并在25°C下孵育。接种水果在7天内出现炭疽病症状,而控制和未受伤的水果仍然没有症状。大豆炭疽病成功地从有症状的果实中重新分离,履行科赫的假设,并确认其作为疾病病因的作用。大豆炭疽病已知感染世界各地的豆科物种,如大豆,紫花苜蓿,菜豆,白术和Vignaunguiculata(Damm等人。2019年;Talhinhas和Baroncelli2021),韩国的苍术(Hassan等人。2021年)和中国的辣椒(Zhangetal。2023年)。C.sojae在韩国引起辣椒炭疽病的第一份报告对辣椒种植者来说是一个新的挑战。需要制定和实施综合疾病管理策略,以减轻其影响。
Chili (Capsicum annuum L.) is an economically important crop worldwide, valued for its culinary uses. In South Korea, anthracnose caused by Colletotrichum spp. including C. truncatum, C. gloeosporioides, C. coccodes, C. acutatum, and C. scovillei incurs on substantial economic loss (Kim et al. 2008; Oo and Oh 2020). In August 2022, somewhat different types of symptoms that was not typical on chilli fruits were observed in a field in Yereonggwang (GPS: 35.2579° N, 126.4742° E), South Korea. The disease symptoms appeared as sunken, necrotic lesions with dense black spore masses forming in concentric rings. The estimated disease incidence the 0.2 ha field showing up to 1% of fruits affected. To isolate the pathogen, six symptomatic chilli fruits were collected. Small pieces (5 mm²) were cut from the margins of the lesions, surface-sterilized in 70% ethanol for 30 sec, followed by 1% sodium hypochlorite for 1 minute, and then rinsed three times in sterile distilled water. The tissue pieces were placed on potato dextrose agar (PDA) plates and incubated at 25°C in the dark. After 3 to 5 days, emerging fungal colonies were sub-cultured to obtain pure isolates. A total of five isolates were obtained and initially identified as Colletotrichum spp. based on morphological characteristics. Seven-day old colonies were initially white, turning light orange with age on PDA. Setae (observed on lesion) were dark brown, verruculose and septate. Conidia were cylindrical, hyaline, and measured 14.8 to 19.9 × 4.2 to 6.5 µm (mean 16.7 × 5.6 μm, n = 70) in size; appressoria were brown to dark brown and irregularly shaped. These morphological characteristics of the isolates agree with those reported for the morphology of C. sojae by Damm et al. (2019). To confirm the identity of the isolates, DNA was extracted and specific gene regions were amplified and sequenced using the following primer sets: ITS (ITS1 and ITS4), GAPDH (GDF1 and GDR1), ACT (ACT-512F and ACT-783R), TUB (T1 and Bt2b), HIS3 (CYLH3F and CYLH3R), and CHS-1 (CHS-79F and CHS-345R). The resulting sequences were deposited in the NCBI GenBank with accession numbers (LC830742 to LC830766). Maximum likelihood phylogenetic analysis using combine sequences of ITS, GAPDH, ACT, TUB, HIS3 and CHS-1 in MEGA X confirmed the isolates as C. sojae, marking the first report of this pathogen on chilli in South Korea, previously known to infect soybean. Pathogenicity tests were conducted on wound and nonwounded healthy and mature-green chili fruits (cv. Bicksita) to confirm the pathogenicity of the isolated C. sojae. The fruits were surface-sterilized using 70% ethanol and then rinsed with sterile distilled water. The fruits were wounded using a sterile needle to facilitate infection. A conidial suspension (1x106 conidia/mL) was prepared from 7-day-old PDA cultures. Each fruit was inoculated by placing a 10 µL drop of the conidial suspension onto the wounded and nonwounded sites (4 to 5) of the wound and unwound fruits, respectively. Control fruits were inoculated with sterile water. A total of 40 fruits per treatment were used and the experiment repeated twice. The fruits were placed in plastic box lined with moist paper towels to maintain high humidity and incubated at 25°C. Anthracnose symptoms developed on the inoculated fruits within 7 days, while control and unwounded fruits remained symptom-free. Colletotrichum sojae was successfully reisolated from the symptomatic fruits, fulfilling Koch\'s postulates and confirming its role as the causal agent of the disease. Colletotrichum sojae is known to infect Fabaceae species worldwide such as Glycine max, Medicago sativa, Phaseolus vulgaris, Atractylodes ovata and Vigna unguiculata (Damm et al. 2019; Talhinhas and Baroncelli 2021), Atractylodes ovata in South Korea (Hassan et al. 2021) and chili pepper in China (Zhanget al. 2023). The first report of C. sojae causing chili anthracnose in South Korea represents a new challenge for chili growers. Integrated disease management strategies need to be developed and implemented to mitigate its impact.