P. Balanagouda, C. T. Ganesh, P. Kotari, R. Rathinavelu
Subabul (Leucaena leucocephala) is a perennial, fast-growing, non-climbing tree originating from tropical America. The species is economically important for the paper and pulp industry supplying 35–40% wood required for these industries in India (Orwa et al., 2009; Global Invasive Species Database, 2023). In March 2022, more than 30% of three-year-old subabul trees exhibited wilting and oozing symptoms in many farmers’ plantations in Khammam, Telangana, India. The affected plants initially showed yellowing of the leaves and wilting symptoms (Figures 1a-b). Symptoms of gummosis were also observed subsequently (Figures 1c-e). Cross-sections of affected roots in nursery-grown trees revealed brownish discolouration and rotting. Diseased roots and bark sections were surface-sterilised in 75% ethanol for 15 seconds, followed by sodium hypochlorite (1%) for 45 seconds and rinsed three times in distilled water. After drying, samples were placed onto potato dextrose agar (PDA) plates. Six isolates (FS1-FS6) were selected from Fusarium-like colonies obtained from the diseased tissue and pure cultures were obtained by single spore isolation. All the isolates produced off-white colonies (Figure 2a) on PDA after five to seven days incubation at 28°C. Pinkish to reddish pigmentation appeared on the bottom side of seven to ten-day-old cultures (Figure 2b). Macroconidia, microconidia and chlamydospores (Figure 3) were produced on carnation leaf agar after five to seven days incubation at 28°C. Microconidia were septate, hyaline, elongated ovoid or reniform, with a mean size of 7.23 × 3.41 µm (n = 25). Macroconidia were hyaline, crescent or curved, slightly tapered at the apex with three-five septa and measured 32.1 × 5.3 µm (mean, n = 25). Chlamydospores were light brownish, globose or round with a mean diameter of 4.3 µm (n = 25) and were produced either singly, in clusters on conidium or intercalary. The rDNA-ITS and RPB2 genes of FS1, FS2 and FS3 were amplified with ITS1/ITS4 (White et al., 1990) and RPB2F/RPB2R primer pairs (O'Donnell et al., 2013) and sequenced (GenBank Accession Nos. OR272193-OR272195 and OR582636-OR582638 respectively). The sequences of FS1, FS2 and FS3 isolates were identical and the isolates grouped with other isolates of F. equiseti in phylogenetic trees based on the internal transcribed spacer region and RNA polymerase II subunit 2 gene (Figure 4). A pathogenicity test was done on healthy six-month-old subabul clones grown in sterile soil in pots (50 cm diameter, 35 cm height). Plants were inoculated separately with 100 mL of a spore suspension of isolates FS1, FS2 or FS3 (106 conidia/mL). Sterile distilled water was used for the controls (n = 3). Assays were repeated three times in a glasshouse kept at 28°C. Twelve days post-inoculation, clones treated with each of the Fusarium isolates exhibited yellowing and wilting symptoms similar to those observed in the field, but control pots remained asymptomatic (Figure 5). Fusarium equiseti
{"title":"Wilt and gummosis disease of subabul caused by <i>Fusarium equiseti</i> ‐ a first record from India","authors":"P. Balanagouda, C. T. Ganesh, P. Kotari, R. Rathinavelu","doi":"10.1002/ndr2.12227","DOIUrl":"https://doi.org/10.1002/ndr2.12227","url":null,"abstract":"Subabul (Leucaena leucocephala) is a perennial, fast-growing, non-climbing tree originating from tropical America. The species is economically important for the paper and pulp industry supplying 35–40% wood required for these industries in India (Orwa et al., 2009; Global Invasive Species Database, 2023). In March 2022, more than 30% of three-year-old subabul trees exhibited wilting and oozing symptoms in many farmers’ plantations in Khammam, Telangana, India. The affected plants initially showed yellowing of the leaves and wilting symptoms (Figures 1a-b). Symptoms of gummosis were also observed subsequently (Figures 1c-e). Cross-sections of affected roots in nursery-grown trees revealed brownish discolouration and rotting. Diseased roots and bark sections were surface-sterilised in 75% ethanol for 15 seconds, followed by sodium hypochlorite (1%) for 45 seconds and rinsed three times in distilled water. After drying, samples were placed onto potato dextrose agar (PDA) plates. Six isolates (FS1-FS6) were selected from Fusarium-like colonies obtained from the diseased tissue and pure cultures were obtained by single spore isolation. All the isolates produced off-white colonies (Figure 2a) on PDA after five to seven days incubation at 28°C. Pinkish to reddish pigmentation appeared on the bottom side of seven to ten-day-old cultures (Figure 2b). Macroconidia, microconidia and chlamydospores (Figure 3) were produced on carnation leaf agar after five to seven days incubation at 28°C. Microconidia were septate, hyaline, elongated ovoid or reniform, with a mean size of 7.23 × 3.41 µm (n = 25). Macroconidia were hyaline, crescent or curved, slightly tapered at the apex with three-five septa and measured 32.1 × 5.3 µm (mean, n = 25). Chlamydospores were light brownish, globose or round with a mean diameter of 4.3 µm (n = 25) and were produced either singly, in clusters on conidium or intercalary. The rDNA-ITS and RPB2 genes of FS1, FS2 and FS3 were amplified with ITS1/ITS4 (White et al., 1990) and RPB2F/RPB2R primer pairs (O'Donnell et al., 2013) and sequenced (GenBank Accession Nos. OR272193-OR272195 and OR582636-OR582638 respectively). The sequences of FS1, FS2 and FS3 isolates were identical and the isolates grouped with other isolates of F. equiseti in phylogenetic trees based on the internal transcribed spacer region and RNA polymerase II subunit 2 gene (Figure 4). A pathogenicity test was done on healthy six-month-old subabul clones grown in sterile soil in pots (50 cm diameter, 35 cm height). Plants were inoculated separately with 100 mL of a spore suspension of isolates FS1, FS2 or FS3 (106 conidia/mL). Sterile distilled water was used for the controls (n = 3). Assays were repeated three times in a glasshouse kept at 28°C. Twelve days post-inoculation, clones treated with each of the Fusarium isolates exhibited yellowing and wilting symptoms similar to those observed in the field, but control pots remained asymptomatic (Figure 5). Fusarium equiseti","PeriodicalId":36931,"journal":{"name":"New Disease Reports","volume":"43 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"136198697","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
G. V. Benatar, E. Hartini, Y. Nurhayati, N. F. Ridwan
{"title":"First report of Diaporthe pseudophoenicicola causing stem end rot of mango in Indonesia","authors":"G. V. Benatar, E. Hartini, Y. Nurhayati, N. F. Ridwan","doi":"10.1002/ndr2.12237","DOIUrl":"https://doi.org/10.1002/ndr2.12237","url":null,"abstract":"","PeriodicalId":36931,"journal":{"name":"New Disease Reports","volume":"6 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139329129","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"First report of Pantoea ananatis causing leaf blight disease of pomegranate in India","authors":"N. Chathalingath, A. Gunasekar","doi":"10.1002/ndr2.12233","DOIUrl":"https://doi.org/10.1002/ndr2.12233","url":null,"abstract":"","PeriodicalId":36931,"journal":{"name":"New Disease Reports","volume":"13 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139329410","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
P. L. Babu, D. Singh, J. Rajender, N. Geat, R. Patidar
India produces 5.7 million tonnes of chilli (Capsicum annuum L.) from 100,000 hectares of cultivated plants annually (Directorate of Economics and Statistics, 2020). During June-October, 2022 and 2023, chilli cv. Pusa Jwala cultivated at the Indian Agricultural Research Institute, New Delhi developed irregular, necrotic lesions surrounded by a chlorotic halo and brown necrosis at the leaf tip followed by defoliation (Figure 1). Disease incidence was 52% over an area of c. 800 m2. Bacterial colonies isolated from ten infected leaf samples, collected randomly across three fields, were circular, 2–3 mm, convex and greyish to white after 48 hours at 28±1°C on nutrient agar. They were positive in 3% KOH and oxidase tests (Schaad et al., 2001). They produced a hypersensitive reaction in tobacco cv. BX61 two days after injection with a suspension containing 109 CFU/ml. One isolate and a positive control strain (Enterobacter cloacae subsp. dissolvens NAIMCC-B-01345 from NAIMCC, India) were positive for ONPG, urease, nitrate reduction, Voges-Proskauer, aesculin hydrolysis, utilisation of ornithine, malonate, citrate, arabinose, xylose, trehalose, and glucose, and negative for phenylalanine deamination, H2S production, methyl red, indole, utilisation of lysine, adonitol, and lactose using a KB003 Hi25-Enterobacteriaceae identification kit (Hi Media, India). The isolate did not utilise the carbohydrates rhamnose, cellobiose, melibiose, saccharose, and raffinose in contrast to the positive control strain. The pathogenicity of the ten isolates was tested by syringe injection of a bacterial suspension (107 CFU/ml) into leaves of 30-day old chilli plants (cv. Pusa Jwala); sterile water was used as a negative control. Dark brown necrotic patches developed after two days in the injected region (Figure 2), followed by leaf decay and defoliation. No symptoms were observed on the negative control. DNA was extracted from bacteria re-isolated from inoculated plants using a modified CTAB protocol (Wang et al.,2010). PCR was done using primer pairs 27F/1492R and Hsp60-F/Hsp60-R which amplify the 16S rRNA and Hsp60 genes, respectively (Poussier et al., 2000; Hoffmann & Roggenkamp, 2003). Amplicons of the expected size were Sanger sequenced by Barcode Biosciences Pvt. Ltd (Bengaluru, India) and aligned manually. Sequences were deposited in Genbank with Accession Nos. OP897636 (16S rRNA) and OP918670 (Hsp60). An isolate (DLC-1) was also deposited in the Indian Type Culture Collection, New Delhi with accession number ITCCBY0001. A phylogenetic tree based on the concatenated sequences of both genes revealed that isolate DLC-1 clustered with two E. cloacae strains, SBP-8 and GGT036 from India and Republic of Korea, respectively (Figure 3). Enterobacter cloacae is a ubiquitous Gram-negative, facultatively anaerobic human pathogen, that has been reported to infect plants such as Allium cepa, Capsicum annuum and Manihot esculenta (García-González et al., 2018). Enterobacter cloa
印度每年从10万公顷的栽培植物中生产570万吨辣椒(Capsicum annuum L.)(经济和统计局，2020年)。在2022年6月至10月和2023年期间，辣椒cv。在新德里印度农业研究所种植的Pusa Jwala出现了不规则的坏死灶，周围有褪绿晕，叶尖呈褐色坏死，随后脱落(图1)。在约800平方米的面积上，疾病发病率为52%。随机采集的10个感染叶片样本，在28±1℃条件下放置48小时，菌落呈圆形、2-3 mm、凸状，呈灰白色。他们在3% KOH和氧化酶测试中呈阳性(Schaad等人，2001年)。它们对烟草产生过敏反应。BX61注射后2天，含109 CFU/ml的混悬液。一个分离株和一个阳性对照菌株(阴沟肠杆菌亚种)。溶解物NAIMCC- b -01345(来自印度NAIMCC)对ONPG、脲酶、硝酸盐还原、Voges-Proskauer、aesculin水解、鸟氨酸、丙二酸盐、柠檬酸盐、阿拉伯糖、木糖、海藻糖和葡萄糖的利用呈阳性，对苯丙氨酸脱胺、H2S生成、甲基红、吲哚、赖氨酸、甘油三醇和乳糖的利用呈阴性(使用KB003 Hi25-Enterobacteriaceae鉴定试剂盒(Hi Media，印度))呈阴性。与阳性对照菌株相比，该分离株不利用碳水化合物鼠李糖、纤维二糖、糖蜜二糖、蔗糖和棉子糖。将细菌悬浮液(107 CFU/ml)注射到30 d辣椒叶片中，检测10株菌株的致病性。天Jwala);无菌水作为阴性对照。两天后，注射区域出现深棕色坏死斑块(图2)，随后是叶片腐烂和落叶。阴性对照组未观察到症状。使用改良的CTAB协议从接种植物中重新分离的细菌中提取DNA (Wang et al.，2010)。采用分别扩增16S rRNA和Hsp60基因的引物对27F/1492R和Hsp60- f /Hsp60- r进行PCR (Poussier et al.， 2000;Hoffmann & Roggenkamp, 2003)。预期大小的扩增子由Barcode Biosciences pto . Ltd (Bengaluru, India)进行Sanger测序，并手动对齐。序列保存在Genbank中，登录号为OP897636 (16S rRNA)和OP918670 (Hsp60)。分离株(DLC-1)也保存在新德里的印度版式文化收藏馆，编号为ITCCBY0001。基于两个基因序列的系统发育树显示，分离株DLC-1分别与来自印度和韩国的两株阴沟肠杆菌SBP-8和GGT036聚集在一起(图3)。阴沟肠杆菌是一种普遍存在的革兰氏阴性、兼性厌氧人类病原体，据报道可感染洋葱、辣椒和马尼奥特·埃斯culenta等植物(García-González et al.， 2018)。最近报道了阴沟肠杆菌菌株SD4L和FQY013感染水稻和番茄植株(Cao et al.， 2020;Jin et al.， 2023)，表明该细菌具有广泛的宿主范围。据我们所知，这是印度和全球首次报道阴沟肠杆菌引起辣椒叶斑病、边缘坏死和落叶症状。需要进一步研究阴沟肠杆菌菌株对印度植物的存在和影响。
{"title":"First report of <i>Enterobacter cloacae</i> causing leaf spot of chilli pepper (<i>Capsicum annuum</i>) in India","authors":"P. L. Babu, D. Singh, J. Rajender, N. Geat, R. Patidar","doi":"10.1002/ndr2.12225","DOIUrl":"https://doi.org/10.1002/ndr2.12225","url":null,"abstract":"India produces 5.7 million tonnes of chilli (Capsicum annuum L.) from 100,000 hectares of cultivated plants annually (Directorate of Economics and Statistics, 2020). During June-October, 2022 and 2023, chilli cv. Pusa Jwala cultivated at the Indian Agricultural Research Institute, New Delhi developed irregular, necrotic lesions surrounded by a chlorotic halo and brown necrosis at the leaf tip followed by defoliation (Figure 1). Disease incidence was 52% over an area of c. 800 m2. Bacterial colonies isolated from ten infected leaf samples, collected randomly across three fields, were circular, 2–3 mm, convex and greyish to white after 48 hours at 28±1°C on nutrient agar. They were positive in 3% KOH and oxidase tests (Schaad et al., 2001). They produced a hypersensitive reaction in tobacco cv. BX61 two days after injection with a suspension containing 109 CFU/ml. One isolate and a positive control strain (Enterobacter cloacae subsp. dissolvens NAIMCC-B-01345 from NAIMCC, India) were positive for ONPG, urease, nitrate reduction, Voges-Proskauer, aesculin hydrolysis, utilisation of ornithine, malonate, citrate, arabinose, xylose, trehalose, and glucose, and negative for phenylalanine deamination, H2S production, methyl red, indole, utilisation of lysine, adonitol, and lactose using a KB003 Hi25-Enterobacteriaceae identification kit (Hi Media, India). The isolate did not utilise the carbohydrates rhamnose, cellobiose, melibiose, saccharose, and raffinose in contrast to the positive control strain. The pathogenicity of the ten isolates was tested by syringe injection of a bacterial suspension (107 CFU/ml) into leaves of 30-day old chilli plants (cv. Pusa Jwala); sterile water was used as a negative control. Dark brown necrotic patches developed after two days in the injected region (Figure 2), followed by leaf decay and defoliation. No symptoms were observed on the negative control. DNA was extracted from bacteria re-isolated from inoculated plants using a modified CTAB protocol (Wang et al.,2010). PCR was done using primer pairs 27F/1492R and Hsp60-F/Hsp60-R which amplify the 16S rRNA and Hsp60 genes, respectively (Poussier et al., 2000; Hoffmann & Roggenkamp, 2003). Amplicons of the expected size were Sanger sequenced by Barcode Biosciences Pvt. Ltd (Bengaluru, India) and aligned manually. Sequences were deposited in Genbank with Accession Nos. OP897636 (16S rRNA) and OP918670 (Hsp60). An isolate (DLC-1) was also deposited in the Indian Type Culture Collection, New Delhi with accession number ITCCBY0001. A phylogenetic tree based on the concatenated sequences of both genes revealed that isolate DLC-1 clustered with two E. cloacae strains, SBP-8 and GGT036 from India and Republic of Korea, respectively (Figure 3). Enterobacter cloacae is a ubiquitous Gram-negative, facultatively anaerobic human pathogen, that has been reported to infect plants such as Allium cepa, Capsicum annuum and Manihot esculenta (García-González et al., 2018). Enterobacter cloa","PeriodicalId":36931,"journal":{"name":"New Disease Reports","volume":"43 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"136128375","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
In autumn 2022, a tomato sample (Solanum lycopersicum, cv. Ekstasis F1 hybrid) from Aspropyrgos, Attiki, Greece, infected with Tomato brown rugose fruit virus (ToBRFV) was analysed with RNA-Seq. Total RNA was isolated from leaves with an RNeasy Plant Mini Kit (Qiagen, Germany) and was subjected to RNA-Seq in an Illumina Novaseq 6000 platform. The BLASTn annotation of the contigs obtained from the analysis of the 12 M, single-end, 100 bp reads with Geneious (v. 11.1.5), revealed the presence of ToBRFV, Southern tomato virus (genus Amalgavirus), a virus already described in Greece, and Tomato fruit blotch virus (ToFBV). ToFBV has been recently reported in Italy, Australia, Brazil, Spain, Portugal, Slovenia and Tunisia (Kitajima et al., 2023) and is associated with fruit blotch disease, although Koch's postulates have not yet been fulfilled, and the virus is not mechanically or seed transmitted. ToFBV is a member of the genus Blunervirus, and its genome consists of four single stranded, positive sense RNAs. The RNA 1 (5764 bp; GenBank Accession No. OQ473416) and RNA 2 (3618 bp; OQ473417) sequences of the Greek isolate showed the highest identity with those of the Tunisian isolate (99.7 and 99.6% nucleotide identity with MZ401001.1 and MZ401002.1, respectively). The RNA 3 (2829 bp; OQ473418) and RNA 4 (1905 bp; OQ473419) segments showed 99.4 and 99.7% nucleotide identity with the RNA 3 (NC_078394.1) and RNA 4 (NC_078393.1) of the Italian isolate, respectively. Finally, RT-PCR was used for the amplification of a 500 bp region of RNA 1 (Nakasu et al., 2022). The PCR product was Sanger sequenced and the resulting sequence was identical to that obtained from the RNA-Seq analysis. To further associate the virus with the fruit blotch symptoms, 11 tomato fruit samples (cv. Esperia F1 hybrid) from a greenhouse in Ierapetra (Crete, Greece) and a sample from field in Prokopi (Evia, Greece), collected in February and August 2023 respectively, and exhibiting fruit blotches (Figure 1), were tested for the presence of ToFBV. Sap inoculation on indicator plants (Chenopodium quinoa, Nicotiana benthamiana and N. tabacum cvs. Xanthi-nc and Turkish) did not induce any symptoms, suggesting the presence of a non-mechanically transmitted virus. Total RNA was extracted from all samples and analysed with RT-PCR for the presence of Tomato chlorosis virus (ToCV, Louro et al., 2000), Tomato infectious chlorosis virus (TICV, Vaira et al., 2002) and ToFBV. All samples tested positive for ToFBV and negative for ToCV and TICV. Moreover, in both cases plants were infested by tomato russet mite (Aculops lycopersici) which is the putative vector of the virus. To the best of our knowledge this is the first report of ToFBV infecting tomato in Greece. The fact that the virus was identified in three distinct regions of Greece, its association with fruit blotch disease together with the probability of mite transmission suggests that this emerging virus poses a risk for tomato cultivation.
{"title":"First report of <i>Tomato fruit blotch fruit virus</i> infecting tomato in Greece","authors":"D. Beris, A. Galeou, O. Kektsidou, C. Varveri","doi":"10.1002/ndr2.12219","DOIUrl":"https://doi.org/10.1002/ndr2.12219","url":null,"abstract":"In autumn 2022, a tomato sample (Solanum lycopersicum, cv. Ekstasis F1 hybrid) from Aspropyrgos, Attiki, Greece, infected with Tomato brown rugose fruit virus (ToBRFV) was analysed with RNA-Seq. Total RNA was isolated from leaves with an RNeasy Plant Mini Kit (Qiagen, Germany) and was subjected to RNA-Seq in an Illumina Novaseq 6000 platform. The BLASTn annotation of the contigs obtained from the analysis of the 12 M, single-end, 100 bp reads with Geneious (v. 11.1.5), revealed the presence of ToBRFV, Southern tomato virus (genus Amalgavirus), a virus already described in Greece, and Tomato fruit blotch virus (ToFBV). ToFBV has been recently reported in Italy, Australia, Brazil, Spain, Portugal, Slovenia and Tunisia (Kitajima et al., 2023) and is associated with fruit blotch disease, although Koch's postulates have not yet been fulfilled, and the virus is not mechanically or seed transmitted. ToFBV is a member of the genus Blunervirus, and its genome consists of four single stranded, positive sense RNAs. The RNA 1 (5764 bp; GenBank Accession No. OQ473416) and RNA 2 (3618 bp; OQ473417) sequences of the Greek isolate showed the highest identity with those of the Tunisian isolate (99.7 and 99.6% nucleotide identity with MZ401001.1 and MZ401002.1, respectively). The RNA 3 (2829 bp; OQ473418) and RNA 4 (1905 bp; OQ473419) segments showed 99.4 and 99.7% nucleotide identity with the RNA 3 (NC_078394.1) and RNA 4 (NC_078393.1) of the Italian isolate, respectively. Finally, RT-PCR was used for the amplification of a 500 bp region of RNA 1 (Nakasu et al., 2022). The PCR product was Sanger sequenced and the resulting sequence was identical to that obtained from the RNA-Seq analysis. To further associate the virus with the fruit blotch symptoms, 11 tomato fruit samples (cv. Esperia F1 hybrid) from a greenhouse in Ierapetra (Crete, Greece) and a sample from field in Prokopi (Evia, Greece), collected in February and August 2023 respectively, and exhibiting fruit blotches (Figure 1), were tested for the presence of ToFBV. Sap inoculation on indicator plants (Chenopodium quinoa, Nicotiana benthamiana and N. tabacum cvs. Xanthi-nc and Turkish) did not induce any symptoms, suggesting the presence of a non-mechanically transmitted virus. Total RNA was extracted from all samples and analysed with RT-PCR for the presence of Tomato chlorosis virus (ToCV, Louro et al., 2000), Tomato infectious chlorosis virus (TICV, Vaira et al., 2002) and ToFBV. All samples tested positive for ToFBV and negative for ToCV and TICV. Moreover, in both cases plants were infested by tomato russet mite (Aculops lycopersici) which is the putative vector of the virus. To the best of our knowledge this is the first report of ToFBV infecting tomato in Greece. The fact that the virus was identified in three distinct regions of Greece, its association with fruit blotch disease together with the probability of mite transmission suggests that this emerging virus poses a risk for tomato cultivation.","PeriodicalId":36931,"journal":{"name":"New Disease Reports","volume":"80 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135762637","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
A. Farhaoui, A. Tahiri, N. Radouane, M. Khadiri, S. Amiri, N. El Alami, R. Lahlali
{"title":"First report of Clonostachys rosea causing sugar beet root rot in Morocco","authors":"A. Farhaoui, A. Tahiri, N. Radouane, M. Khadiri, S. Amiri, N. El Alami, R. Lahlali","doi":"10.1002/ndr2.12235","DOIUrl":"https://doi.org/10.1002/ndr2.12235","url":null,"abstract":"","PeriodicalId":36931,"journal":{"name":"New Disease Reports","volume":"22 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139329596","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
{"title":"First report of leaf spot disease of Tabernaemontana divaricata caused by Colletotrichum coffeanum in India","authors":"S. Saha, C. Mili, A. Sarma, K. Tayung","doi":"10.1002/ndr2.12232","DOIUrl":"https://doi.org/10.1002/ndr2.12232","url":null,"abstract":"","PeriodicalId":36931,"journal":{"name":"New Disease Reports","volume":"3 1","pages":""},"PeriodicalIF":0.0,"publicationDate":"2023-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"139329946","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
Robusta coffee (Coffea canephora) is the most produced coffee species in the Philippines, accounting for 70% of the country's total coffee production (Philippine Statistics Authority, 2023). Despite being considered more disease-resistant than C. arabica, a number of diseases have been reported infecting robusta coffee (Cao et al., 2014). In November 2022, necrotic leaf spots (Figure 1) were observed on the leaves of five-year-old robusta coffee trees in New Bantangan, Columbio, Sultan Kudarat, Philippines (6.3427 N; 124.5924 E). The leaf spots initially started as tiny circular or irregular brown spots that gradually enlarged and turned darker. The disease incidence was approximately 85% over an area of 1 hectare with 25% of the leaves on affected plants having disease symptoms. Twenty diseased leaves were collected, one each from 20 plants, and 3 mm discs were cut from the advancing margin of the infection adjoining healthy tissues. The discs were soaked immediately in a 10% solution of NaOCl for one minute, followed by three rinses in sterile distilled water, and blot dried on aseptic, dry tissue paper inside a laminar flow hood. The tissues were then placed equidistantly on potato dextrose agar (PDA) medium and incubated at 28 ±1°C. The active mycelial tip was transferred and incubated for seven days. The colonies produced white aerial mycelium with black conidial masses as they aged (Figure 2). Microscopic examination showed four-celled spindle-shaped conidia (n = 20), measuring 24.42-31.08 × 6.66 μm, and basal appendages 28.86-33.30 μm long (Figure 3). The fungal isolate was deposited in the fungal repository of the Plant Pathology Laboratory of the University of Southern Mindanao Research and Development Center (P3LSS01). To further ascertain its identity, genomic DNA was extracted from a representative seven-day-old isolate (001) grown in PDA broth, using a Zymo Quick-DNA™ Fungal/Bacterial Miniprep Kit (Zymo Research, USA). The rDNA ITS region of the representative isolate was amplified and sequenced using universal primers ITS4/ITS5 (White et al., 1990). The sequence was deposited in Genbank (Accession No. OR125548). A BLASTn search revealed that the isolate had 99.61% identity to isolate FAFU03 (MH470257.1). On the basis of the morphological and molecular characters, the fungus was identified as Pseudopestalotiopsis theae. A pathogenicity test was performed thrice on detached healthy leaves of robusta coffee. Ten wounded (pin-pricked) and ten unwounded leaves were sprayed with 30 μl spore suspension (107 spores/ml) taken from a seven-day-old pure culture of the pathogen (isolate P3LSS01). Sterile distilled water was sprayed onto leaves as a control. The leaves were incubated at 26 ±1°C and the development of the infection and symptoms were observed regularly over a seven-day period. The first symptoms, small brown spots, were observed on wounded leaves after three days and these enlarged to 5–6 mm in diameter after seven days. Small bro
罗布斯塔咖啡(Coffea canephora)是菲律宾产量最大的咖啡品种，占该国咖啡总产量的70%(菲律宾统计局，2023年)。尽管罗布斯塔咖啡被认为比阿拉比卡咖啡更抗病，但据报道，许多疾病感染了罗布斯塔咖啡(Cao et al.， 2014)。2022年11月，在菲律宾Sultan Kudarat (6.3427 N;[124.5924 E] .叶斑最初是小的圆形或不规则的棕色斑点，逐渐扩大并变暗。在1公顷的面积上，该病发病率约为85%，受影响植物上25%的叶子有疾病症状。采集20株病叶20片，每片1片，从感染边缘邻近健康组织处切下3 mm的圆盘。膜片立即在10%的NaOCl溶液中浸泡1分钟，然后在无菌蒸馏水中冲洗三次，然后在层流罩内的无菌干燥纸巾上吸干。将组织等距置于马铃薯葡萄糖琼脂(PDA)培养基上，28±1℃孵育。将活性菌丝尖端转移培养7天。菌落衰老后产生白色气生菌丝，分生孢子团呈黑色(图2)。显微镜检查显示4细胞梭形分生孢子(n = 20)，尺寸为24.42 ~ 31.08 × 6.66 μm，基部附属物长28.86 ~ 33.30 μm(图3)。该真菌分离物保存在南棉兰老大学研究与发展中心植物病理学实验室真菌库(P3LSS01)。为了进一步确定其身份，使用Zymo Quick-DNA™真菌/细菌迷你试剂盒(Zymo Research, USA)从PDA培养液中培养的具有代表性的7日龄分离物(001)中提取基因组DNA。使用通用引物ITS4/ITS5扩增代表性分离物的rDNA ITS区域并进行测序(White et al.， 1990)。该序列已存入Genbank(登录号:OR125548)。BLASTn检索结果显示，分离物与FAFU03 (MH470257.1)的同源性为99.61%。根据形态和分子特征鉴定该真菌为假estestalotiopsis theae。对罗布斯塔咖啡分离的健康叶片进行了三次致病性试验。从病原菌(分离株P3LSS01) 7 d的纯培养液中提取孢子悬浮液30 μl(107孢子/ml)，喷洒10片伤叶(针刺叶)和10片未伤叶。在叶片上喷洒无菌蒸馏水作为对照。将叶片在26±1°C下孵育，并在7天内定期观察感染和症状的发展。第一个症状是3天后在受伤的叶片上观察到小的棕色斑点，7天后这些斑点直径扩大到5-6毫米。7天后，未受伤的测试样品才开始出现小的棕色斑点。对照叶片未出现任何症状(图4)。尽管最初与茶叶有关，但也发现它会感染世界温带和热带地区的几种重要作物(Maharachchikumbura et al.， 2014)。这是菲律宾首次报告的茶杆菌感染，也是全球首次报告的罗布斯塔咖啡感染。这种病原体的影响尚不清楚，但可能被证明是咖啡的一种重要的经济疾病。与其他重要作物的交叉感染研究也很重要，因为在菲律宾，咖啡通常与其他作物间作。需要研究这种病原体的管理方法，以保障菲律宾和全世界的咖啡生产。作者对PhilCafe和World Coffee Research对本研究的财政支持表示感谢。还要感谢Tamie C. Solpot博士，他允许我们在病原体分离和分子鉴定过程中使用植物病理学研究实验室。
{"title":"First report of <i>Pseudopestalotiopsis theae</i> causing leaf spot of robusta coffee in the Philippines","authors":"N. P. D. N. Sumaya, P. II M. Caluban, B. T. Borja","doi":"10.1002/ndr2.12218","DOIUrl":"https://doi.org/10.1002/ndr2.12218","url":null,"abstract":"Robusta coffee (Coffea canephora) is the most produced coffee species in the Philippines, accounting for 70% of the country's total coffee production (Philippine Statistics Authority, 2023). Despite being considered more disease-resistant than C. arabica, a number of diseases have been reported infecting robusta coffee (Cao et al., 2014). In November 2022, necrotic leaf spots (Figure 1) were observed on the leaves of five-year-old robusta coffee trees in New Bantangan, Columbio, Sultan Kudarat, Philippines (6.3427 N; 124.5924 E). The leaf spots initially started as tiny circular or irregular brown spots that gradually enlarged and turned darker. The disease incidence was approximately 85% over an area of 1 hectare with 25% of the leaves on affected plants having disease symptoms. Twenty diseased leaves were collected, one each from 20 plants, and 3 mm discs were cut from the advancing margin of the infection adjoining healthy tissues. The discs were soaked immediately in a 10% solution of NaOCl for one minute, followed by three rinses in sterile distilled water, and blot dried on aseptic, dry tissue paper inside a laminar flow hood. The tissues were then placed equidistantly on potato dextrose agar (PDA) medium and incubated at 28 ±1°C. The active mycelial tip was transferred and incubated for seven days. The colonies produced white aerial mycelium with black conidial masses as they aged (Figure 2). Microscopic examination showed four-celled spindle-shaped conidia (n = 20), measuring 24.42-31.08 × 6.66 μm, and basal appendages 28.86-33.30 μm long (Figure 3). The fungal isolate was deposited in the fungal repository of the Plant Pathology Laboratory of the University of Southern Mindanao Research and Development Center (P3LSS01). To further ascertain its identity, genomic DNA was extracted from a representative seven-day-old isolate (001) grown in PDA broth, using a Zymo Quick-DNA™ Fungal/Bacterial Miniprep Kit (Zymo Research, USA). The rDNA ITS region of the representative isolate was amplified and sequenced using universal primers ITS4/ITS5 (White et al., 1990). The sequence was deposited in Genbank (Accession No. OR125548). A BLASTn search revealed that the isolate had 99.61% identity to isolate FAFU03 (MH470257.1). On the basis of the morphological and molecular characters, the fungus was identified as Pseudopestalotiopsis theae. A pathogenicity test was performed thrice on detached healthy leaves of robusta coffee. Ten wounded (pin-pricked) and ten unwounded leaves were sprayed with 30 μl spore suspension (107 spores/ml) taken from a seven-day-old pure culture of the pathogen (isolate P3LSS01). Sterile distilled water was sprayed onto leaves as a control. The leaves were incubated at 26 ±1°C and the development of the infection and symptoms were observed regularly over a seven-day period. The first symptoms, small brown spots, were observed on wounded leaves after three days and these enlarged to 5–6 mm in diameter after seven days. Small bro","PeriodicalId":36931,"journal":{"name":"New Disease Reports","volume":"100 1","pages":"0"},"PeriodicalIF":0.0,"publicationDate":"2023-10-01","publicationTypes":"Journal Article","fieldsOfStudy":null,"isOpenAccess":false,"openAccessPdf":"","citationCount":null,"resultStr":null,"platform":"Semanticscholar","paperid":"135656077","PeriodicalName":null,"FirstCategoryId":null,"ListUrlMain":null,"RegionNum":0,"RegionCategory":"","ArticlePicture":[],"TitleCN":null,"AbstractTextCN":null,"PMCID":"","EPubDate":null,"PubModel":null,"JCR":null,"JCRName":null,"Score":null,"Total":0}
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