Leonardo de Jesus Machado Gois de Oliveira, Cleude Mayara França Dos Santos, Gustavo Henrique Silva Peixoto, Larisse Raquel Carvalho Dias, Edson Pimenta Moreira, Erlen Keila Candido E Silva, Thaís Ribeiro Santiago, Antonia Alice Costa Rodrigues
{"title":"巴西香菜枯萎病病原菌尖孢镰刀菌首次报道。","authors":"Leonardo de Jesus Machado Gois de Oliveira, Cleude Mayara França Dos Santos, Gustavo Henrique Silva Peixoto, Larisse Raquel Carvalho Dias, Edson Pimenta Moreira, Erlen Keila Candido E Silva, Thaís Ribeiro Santiago, Antonia Alice Costa Rodrigues","doi":"10.1094/PDIS-10-24-2215-PDN","DOIUrl":null,"url":null,"abstract":"<p><p>Coriander (<i>Coriandrum sativum</i> L.), an aromatic leafy vegetable from the Apiaceae family, is globally renowned for its flavorful leaves and medicinal properties (Mahleyuddin et al. 2022). In Brazil, coriander cultivation faces challenges like seedling damping-off and leaf burn, symptoms frequently linked to Fusarium wilt, caused by <i>F. oxysporum</i>, in other countries (Koike and Gordon, 2005; Li et al., 2021). Since 2018, a rise in leaf chlorosis, wilting, and mortality has been noted in coriander cultivars Verdão and Verdão Super in Maranhão, Brazil, particularly in the municipalities of São Luís (2°34'01\"S, 44°16'00\"W) in August 2018 and Formosa (4°50'60\"S, 46°44'00\"W) in March 2021, affecting up to 70% of 20-25-day-old field plants. To investigate it further, ten symptomatic plants were randomly collected from Formosa and ten from São Luís, then sent to the Plant Pathology Laboratory at the State University of Maranhão. From symptomatic tissues of each sample, vascular fragments (1 x 1 cm) were disinfected (70% alcohol, 1% sodium hypochlorite, and sterile water) and cultivated on Potato Dextrose Agar, then incubated (25°C - 12 h light/dark cycle). After five days, rapidly growing pink colonies appeared in all isolated fragments, and then microscope slides were prepared to observe the fungal structures, where short monophialides and unicellular, elliptical microconidia were observed, measuring 14-25 µm in length (mean = 20.55 µm) and 36.4-55.4 µm in width (mean = 48.6 µm) (n = 20). Macroconidia, slightly curved with three septa, measured 116-467 µm in length (mean = 370 µm) and 28.8-51.3 µm in width (mean = 42.78 µm) (n = 20). Chlamydospores were also observed terminal or intercalary in hyphae. Single-spored isolates were molecularly identified and tested for pathogenicity. DNA was extracted using Zymo Research® kit, and TEF-1α, ITS, and IGS regions were amplified using PCR. The primers used were EF1-728R/EF2 for TEF-1α (Carbone & Kohn, 1999), ITS1/ITS4 for ITS (White et al., 1990), and Inl11/FoIGS-R for IGS (O'Donnell et al., 2009). Amplicons were sent to ACTGene company and sequenced by Sanger, submitted to GenBank, and analyzed phylogenetically. The accession numbers are MGSS 305 (OR102500, OR859840, PQ468316), MGSS 415 (OR102501, OR859842, PQ468317), and MGSS 416 (OR102502, OR859841, PQ468318). Bayesian analysis confirmed that the isolates belong to the <i>F. oxysporum</i> species complex. Pathogenicity tests involved two coriander varieties (Verdão and Verdão Super), totaling 200 plants, with 25 per isolate (MGSS 305, MGSS 415, MGSS 416 and control)/ variety combination. Thirty-day-old seedling roots were dipped in a conidial suspension of 1 x 106 conidia/mL for 20 min; while the control group was treated with sterile water (Reis et al., 2004). Plants were grown in 3 L pots with sterilized soil, in a greenhouse at 25 ± 2°C, 60% RH, and a 12 h photoperiod. After 21 days, wilt incidence occurred in the inoculated plants: MGSS 415 and MGSS 416 infected both varieties, where we observed an incidence between 80% and 92% in Verdão and 96 and 100% in Verdão Super, and MGSS 305 showed less than 20% incidence in both varieties. Control plants remained healthy. Reisolation confirmed the pathogen via morphology, molecular data (Figures 1 and 2), and pathogenicity, identifying it as <i>F. oxysporum</i> f. sp. <i>coriandrii</i> (Gilardi et al. 2017; Koike and Gordon 2005). The experiment was conducted twice at different times, and similar results were observed. This finding underscores the need for further research and the development of mitigation strategies to protect this crop.</p>","PeriodicalId":20063,"journal":{"name":"Plant disease","volume":" ","pages":""},"PeriodicalIF":4.4000,"publicationDate":"2025-06-10","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":"0","resultStr":"{\"title\":\"First report of <i>Fusarium oxysporum</i> f. sp. <i>coriandrii</i> causing Fusarium wilt in Coriander in Brazil.\",\"authors\":\"Leonardo de Jesus Machado Gois de Oliveira, Cleude Mayara França Dos Santos, Gustavo Henrique Silva Peixoto, Larisse Raquel Carvalho Dias, Edson Pimenta Moreira, Erlen Keila Candido E Silva, Thaís Ribeiro Santiago, Antonia Alice Costa Rodrigues\",\"doi\":\"10.1094/PDIS-10-24-2215-PDN\",\"DOIUrl\":null,\"url\":null,\"abstract\":\"<p><p>Coriander (<i>Coriandrum sativum</i> L.), an aromatic leafy vegetable from the Apiaceae family, is globally renowned for its flavorful leaves and medicinal properties (Mahleyuddin et al. 2022). In Brazil, coriander cultivation faces challenges like seedling damping-off and leaf burn, symptoms frequently linked to Fusarium wilt, caused by <i>F. oxysporum</i>, in other countries (Koike and Gordon, 2005; Li et al., 2021). Since 2018, a rise in leaf chlorosis, wilting, and mortality has been noted in coriander cultivars Verdão and Verdão Super in Maranhão, Brazil, particularly in the municipalities of São Luís (2°34'01\\\"S, 44°16'00\\\"W) in August 2018 and Formosa (4°50'60\\\"S, 46°44'00\\\"W) in March 2021, affecting up to 70% of 20-25-day-old field plants. To investigate it further, ten symptomatic plants were randomly collected from Formosa and ten from São Luís, then sent to the Plant Pathology Laboratory at the State University of Maranhão. From symptomatic tissues of each sample, vascular fragments (1 x 1 cm) were disinfected (70% alcohol, 1% sodium hypochlorite, and sterile water) and cultivated on Potato Dextrose Agar, then incubated (25°C - 12 h light/dark cycle). After five days, rapidly growing pink colonies appeared in all isolated fragments, and then microscope slides were prepared to observe the fungal structures, where short monophialides and unicellular, elliptical microconidia were observed, measuring 14-25 µm in length (mean = 20.55 µm) and 36.4-55.4 µm in width (mean = 48.6 µm) (n = 20). Macroconidia, slightly curved with three septa, measured 116-467 µm in length (mean = 370 µm) and 28.8-51.3 µm in width (mean = 42.78 µm) (n = 20). Chlamydospores were also observed terminal or intercalary in hyphae. Single-spored isolates were molecularly identified and tested for pathogenicity. DNA was extracted using Zymo Research® kit, and TEF-1α, ITS, and IGS regions were amplified using PCR. The primers used were EF1-728R/EF2 for TEF-1α (Carbone & Kohn, 1999), ITS1/ITS4 for ITS (White et al., 1990), and Inl11/FoIGS-R for IGS (O'Donnell et al., 2009). Amplicons were sent to ACTGene company and sequenced by Sanger, submitted to GenBank, and analyzed phylogenetically. The accession numbers are MGSS 305 (OR102500, OR859840, PQ468316), MGSS 415 (OR102501, OR859842, PQ468317), and MGSS 416 (OR102502, OR859841, PQ468318). Bayesian analysis confirmed that the isolates belong to the <i>F. oxysporum</i> species complex. Pathogenicity tests involved two coriander varieties (Verdão and Verdão Super), totaling 200 plants, with 25 per isolate (MGSS 305, MGSS 415, MGSS 416 and control)/ variety combination. Thirty-day-old seedling roots were dipped in a conidial suspension of 1 x 106 conidia/mL for 20 min; while the control group was treated with sterile water (Reis et al., 2004). Plants were grown in 3 L pots with sterilized soil, in a greenhouse at 25 ± 2°C, 60% RH, and a 12 h photoperiod. After 21 days, wilt incidence occurred in the inoculated plants: MGSS 415 and MGSS 416 infected both varieties, where we observed an incidence between 80% and 92% in Verdão and 96 and 100% in Verdão Super, and MGSS 305 showed less than 20% incidence in both varieties. Control plants remained healthy. Reisolation confirmed the pathogen via morphology, molecular data (Figures 1 and 2), and pathogenicity, identifying it as <i>F. oxysporum</i> f. sp. <i>coriandrii</i> (Gilardi et al. 2017; Koike and Gordon 2005). The experiment was conducted twice at different times, and similar results were observed. 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引用次数: 0
摘要
芫荽(Coriandrum sativum L.)是一种芳香的叶类蔬菜,来自Apiaceae家族,以其美味的叶子和药用特性而闻名于世(Mahleyuddin et al. 2022)。在巴西,香菜种植面临着幼苗枯萎和叶片烧伤等挑战,这些症状通常与其他国家由尖孢镰刀菌引起的枯萎病有关(Koike和Gordon, 2005;Li等人,2021)。自2018年以来,巴西maranh的香菜品种verd o和verd o Super出现了叶片萎黄、萎蔫和死亡率上升的现象,尤其是2018年8月的s o Luís(2°34'01"S, 44°16'00"W)和2021年3月的Formosa(4°50'60"S, 46°44'00"W),影响到20-25日龄的田间植物的70%。为了进一步调查,我们从福尔摩沙和 o Luís随机收集了10株有症状的植物,然后送到马兰荷州立大学的植物病理学实验室。从每个样品的症状组织中,将血管碎片(1 × 1 cm)消毒(70%酒精,1%次氯酸钠和无菌水),然后在马铃薯葡萄糖琼脂上培养,然后在25°C - 12 h明暗循环中孵育。5天后,所有分离片段均出现快速生长的粉红色菌落,然后制作显微镜载玻片观察真菌结构,其中观察到短的单孢子和单细胞椭圆形微分生孢子,长度为14-25µm(平均为20.55µm),宽度为36.4-55.4µm(平均为48.6µm) (n = 20)。大分生孢子长116 ~ 467µm(平均370µm),宽28.8 ~ 51.3µm(平均42.78µm) (n = 20)。在菌丝中也观察到衣孢子的顶生或隔生。单孢子分离物进行了分子鉴定和致病性测试。使用zimo Research®试剂盒提取DNA,用PCR扩增TEF-1α、ITS和IGS区域。所使用的引物是用于TEF-1α的EF1-728R/EF2 (Carbone & Kohn, 1999),用于ITS的ITS1/ITS4 (White等,1990),用于IGS的in11 /FoIGS-R (O'Donnell等,2009)。扩增子送到ACTGene公司,由Sanger测序,提交GenBank,进行系统发育分析。加入号为MGSS 305 (OR102500, OR859840, PQ468316), MGSS 415 (OR102501, OR859842, PQ468317)和MGSS 416 (OR102502, OR859841, PQ468318)。贝叶斯分析证实分离物属于尖孢菌属复合体。致病性试验涉及两个香菜品种(verd o和verd o超级),共200株,每个分离物(MGSS 305、MGSS 415、MGSS 416和对照)/品种组合25株。将30天大的幼苗根浸泡在1 × 106个分生孢子/mL的分生孢子悬浮液中20 min;对照组用无菌水处理(Reis et al., 2004)。植株生长在3 L盆栽中,土壤消毒,在温室中25±2°C, 60% RH, 12 h光周期。21天后,接种植株发生了枯萎病:MGSS 415和MGSS 416侵染了两个品种,我们观察到verd 0的发病率在80% - 92%之间,verd 0超级的发病率在96% - 100%之间,MGSS 305在两个品种的发病率都低于20%。对照植物保持健康。再分离通过形态学、分子数据(图1和2)和致病性证实了该病原体,鉴定为F. oxysporum F. sp. coriandrii (Gilardi et al. 2017;Koike and Gordon 2005)。实验在不同的时间进行了两次,观察到相似的结果。这一发现强调需要进一步研究和制定缓解战略,以保护这种作物。
First report of Fusarium oxysporum f. sp. coriandrii causing Fusarium wilt in Coriander in Brazil.
Coriander (Coriandrum sativum L.), an aromatic leafy vegetable from the Apiaceae family, is globally renowned for its flavorful leaves and medicinal properties (Mahleyuddin et al. 2022). In Brazil, coriander cultivation faces challenges like seedling damping-off and leaf burn, symptoms frequently linked to Fusarium wilt, caused by F. oxysporum, in other countries (Koike and Gordon, 2005; Li et al., 2021). Since 2018, a rise in leaf chlorosis, wilting, and mortality has been noted in coriander cultivars Verdão and Verdão Super in Maranhão, Brazil, particularly in the municipalities of São Luís (2°34'01"S, 44°16'00"W) in August 2018 and Formosa (4°50'60"S, 46°44'00"W) in March 2021, affecting up to 70% of 20-25-day-old field plants. To investigate it further, ten symptomatic plants were randomly collected from Formosa and ten from São Luís, then sent to the Plant Pathology Laboratory at the State University of Maranhão. From symptomatic tissues of each sample, vascular fragments (1 x 1 cm) were disinfected (70% alcohol, 1% sodium hypochlorite, and sterile water) and cultivated on Potato Dextrose Agar, then incubated (25°C - 12 h light/dark cycle). After five days, rapidly growing pink colonies appeared in all isolated fragments, and then microscope slides were prepared to observe the fungal structures, where short monophialides and unicellular, elliptical microconidia were observed, measuring 14-25 µm in length (mean = 20.55 µm) and 36.4-55.4 µm in width (mean = 48.6 µm) (n = 20). Macroconidia, slightly curved with three septa, measured 116-467 µm in length (mean = 370 µm) and 28.8-51.3 µm in width (mean = 42.78 µm) (n = 20). Chlamydospores were also observed terminal or intercalary in hyphae. Single-spored isolates were molecularly identified and tested for pathogenicity. DNA was extracted using Zymo Research® kit, and TEF-1α, ITS, and IGS regions were amplified using PCR. The primers used were EF1-728R/EF2 for TEF-1α (Carbone & Kohn, 1999), ITS1/ITS4 for ITS (White et al., 1990), and Inl11/FoIGS-R for IGS (O'Donnell et al., 2009). Amplicons were sent to ACTGene company and sequenced by Sanger, submitted to GenBank, and analyzed phylogenetically. The accession numbers are MGSS 305 (OR102500, OR859840, PQ468316), MGSS 415 (OR102501, OR859842, PQ468317), and MGSS 416 (OR102502, OR859841, PQ468318). Bayesian analysis confirmed that the isolates belong to the F. oxysporum species complex. Pathogenicity tests involved two coriander varieties (Verdão and Verdão Super), totaling 200 plants, with 25 per isolate (MGSS 305, MGSS 415, MGSS 416 and control)/ variety combination. Thirty-day-old seedling roots were dipped in a conidial suspension of 1 x 106 conidia/mL for 20 min; while the control group was treated with sterile water (Reis et al., 2004). Plants were grown in 3 L pots with sterilized soil, in a greenhouse at 25 ± 2°C, 60% RH, and a 12 h photoperiod. After 21 days, wilt incidence occurred in the inoculated plants: MGSS 415 and MGSS 416 infected both varieties, where we observed an incidence between 80% and 92% in Verdão and 96 and 100% in Verdão Super, and MGSS 305 showed less than 20% incidence in both varieties. Control plants remained healthy. Reisolation confirmed the pathogen via morphology, molecular data (Figures 1 and 2), and pathogenicity, identifying it as F. oxysporum f. sp. coriandrii (Gilardi et al. 2017; Koike and Gordon 2005). The experiment was conducted twice at different times, and similar results were observed. This finding underscores the need for further research and the development of mitigation strategies to protect this crop.
期刊介绍:
Plant Disease is the leading international journal for rapid reporting of research on new, emerging, and established plant diseases. The journal publishes papers that describe basic and applied research focusing on practical aspects of disease diagnosis, development, and management.