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Article ; Online: Dothiorella sarmentorum Causing Canker and Branch Dieback of English Walnut in Maule Region, Chile

Iqbal, S. / Hernández, Y. / Lolas, M. / Elfar, K. / Eskalen, A. / Latorre, B. A. / Díaz, G. A.

Plant Disease. 2023 Apr. 03, v. 107, no. 4 p.1219-

2023

Abstract: English walnut (Juglans regia) cv. Chandler is the most cultivated tree nut in Chile, grown on 43,734 ha. In Maule Region, central Chile, English walnut plantings have expanded over an additional 7,000 ha in the last 5 years. During a routine orchard ... ...

Abstract	English walnut (Juglans regia) cv. Chandler is the most cultivated tree nut in Chile, grown on 43,734 ha. In Maule Region, central Chile, English walnut plantings have expanded over an additional 7,000 ha in the last 5 years. During a routine orchard survey in 2019, branch and twig dieback symptoms were observed in two commercial orchards in San Rafael (10 years old) and Longaví (12 years old) in the Maule Region, with incidences of 45 and 65% of affected trees, respectively. Symptomatic branch samples (n = 15) were collected from the two commercial orchards, transported to the laboratory in a cooler, surface sterilized in 96% ethanol for 3 s, and briefly flamed. A cross-section of symptomatic branches revealed brown to dark brown, wedge-shaped wood cankers. Small (5 mm) pieces of wood from the edge of cankered tissues were placed on potato dextrose agar (PDA, 2%) amended with 0.005% tetracycline, 0.01% streptomycin, and 0.1% Igepal CO-630 (PDAm) (Díaz and Latorre 2014) and incubated at 25°C for 5 days in the dark. Pure cultures were obtained by transferring a hyphal tip from growing colonies to fresh PDA media. Each fungal isolate was recovered from a single diseased branch (47%). Seven isolates (Dsar-1 to Dsar-7) developed dark to olive brown, fast-growing colonies with scarce aerial mycelium after 7 days at 25°C on PDA. These isolates showed a dark-olive color on the reverse side of Petri dishes and developed abundant, aggregated, and dark-brown pycnidia after 15 days at 25°C. Conidia were hyaline and aseptate, dark brown, one septate, with a brown wall, ovoid with a broadly rounded apex and truncated base (17.5–) 19.5 ± 1.2 (−22.0) × (7.6–) 8.9 ± 0.6 (−10.1) μm (n = 30). These isolates were tentatively identified morphologically as Dothiorella sp. (Phillips et al. 2005). Molecular identification was performed using ITS1/ITS4 and EF1-728F/EF1-986R primers (Dissanayake et al. 2015; White et al. 1990) of the internal transcribed spacer (ITS1-5.8S-ITS2) region and part of the translation elongation factor (EF1-α) genes, respectively. A MegaBLAST search in GenBank showed a 100% similarity to isolate CBS 115038, the ex-type of Dothiorella sarmentorum. The sequences were added to GenBank (OM161950 to OM161956 for ITS; OM177188 to OM177194 for EF1-α). Pathogenicity of two isolates (Dsar-2 and Dsar-7) was tested in the orchard on fresh pruning wounds on attached branches of English walnut trees cv. Chandler. A second pathogenicity test was done on fresh pruning wounds in 1-year-old rooted cuttings (n = 15) (40 cm of long) of English walnut cv. Chandler. Each pruning wound was inoculated with 40 μl of conidial suspension (10⁵ conidia/ml). Sterile distilled water was used as a control treatment. Both pathogenicity tests were repeated once. After 7 months for attached branches and 4 months for rooted plants, necrotic streaks with mean lengths of 81.3 and 44.5 mm, respectively, were observed below the inoculated pruning wounds. No necrotic streaks were observed in any of the control wounds. D. sarmentorum was 100% reisolated from symptomatic tissues of inoculated branches and molecularly identified (EF1-α), fulfilling Koch’s postulates. Recently, D. sarmentorum has been reported causing English walnut dieback in Spain (López-Moral et al. 2020). To our knowledge, this is the first report of D. sarmentorum causing canker and branch dieback of English walnut in Chile. Further studies are needed to know the impact and extent of canker and branch dieback of walnut in commercial orchards in the Maule Region, central Chile.
Keywords	Dothiorella sarmentorum ; Juglans regia ; color ; conidia ; culture media ; dieback ; ethanol ; fungi ; hyphae ; internal transcribed spacers ; mycelium ; olives ; orchards ; pathogenicity ; peptide elongation factors ; pycnidia ; streptomycin ; surveys ; tetracycline ; walnuts ; wood ; Chile ; Spain ; etiology ; tree nuts
Language	English
Dates of publication	2023-0403
Publishing place	The American Phytopathological Society
Document type	Article ; Online
ZDB-ID	754182-x
ISSN	0191-2917
ISSN	0191-2917
DOI	10.1094/PDIS-03-22-0636-PDN
Database	NAL-Catalogue (AGRICOLA)

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Article: First Report of Cryptovalsa ampelina and Eutypella leprosa Associated with Grapevine Trunk Diseases in Chile.

Díaz, G A / Prehn, D / Latorre, B A

Plant disease

2019 Volume 95, Issue 4, Page(s) 490

Abstract: Trunk diseases (TD) of grapevines (Vitis vinifera L.) have increased considerably in Chile with an incidence of more than 25% found in ≥7-year-old vineyards. Only species of Botryosphaeriaceae, Phaeoacremonium, and Phaeomoniella were associated with TD ... ...

Abstract	Trunk diseases (TD) of grapevines (Vitis vinifera L.) have increased considerably in Chile with an incidence of more than 25% found in ≥7-year-old vineyards. Only species of Botryosphaeriaceae, Phaeoacremonium, and Phaeomoniella were associated with TD in Chile (1,2). From 2009 to 2010, isolations were made from the grapevines 'Cabernet Sauvignon', 'Carmenere', 'Flame Seedless', and 'Pinot Noir' collected in central Chile (33°27' to 34°39'S, 71°17' to 71°33'W). These grapevines showed cankers and vascular necrosis of trunks, arms, and spurs along with a general decline and dieback. Isolations were performed in potato dextrose agar (PDA) plus 0.005% tetracycline, 0.01% streptomycin, and 0.1% Igepal CO-630 (Sigma-Aldrich, St. Louis, MO), for 14 days at 20°C. On the basis of colony morphology and conidia production, two Libertella-like species were obtained in 26 (7.8%) of 335 trunk samples. On the basis of the internal transcribed spacer region (ITS4 and ITS5) of rDNA, Cryptovalsa ampelina (Nitschke) Fuckel (GenBank Accession Nos. HQ694976 and HQ694977), and Eutypella leprosa (Persoon) Berlese (HQ694974 and HQ694975) were identified, showing 98 to 100% similarity with the sequences of C. ampelina (GQ293913) and E. leprosa (AJ302463.1). C. ampelina produced white-to-creamy, smooth colonies with a creamy underside. Colonies of E. leprosa were white-to-gray with a white underside. Orange conidial masses were exuded after 30 days at 20°C. Conidia on PDA (n = 20) were unicellular, hyaline, filiform, slightly curved, and (19.8) 23.4 ± 2.6 (28.3) × (1.1) 1.4 ± 0.2 (1.8) μm and (19.2) 23.9 ± 3.0 (27.6) × (1.0) 1.2 ± 0.1 (1.5) μm for E. leprosa and C. ampelina, respectively. Stromatic perithecia of C. ampelina, embedded in the bark, were observed in dead pruning residues of infected vines (4). Pathogenicity tests were conducted with two isolates of each species, on 30-day-old 'Carmenere', rooted in vitro (n = 12), that were inoculated by placing a 5-mm agar plug on the surface of the propagation medium. Additionally, 15 cm long pieces (n = 5) of 1-year-old canes from 'Carmenere', 'Chardonnay', and 'Red Globe' were inoculated by placing a 5-mm agar plug underneath a cut aseptically made in each cane. An equal number of noninoculated plants and canes, but treated with sterile agar plugs, were used as controls. Leaf number (LN), shoot length (SL), and root length (RL) were assessed on plants in vitro after 28 days at 20°C. The extent of vascular discoloration (VD) obtained in canes was determined after 45 days in humid chambers at 20°C. One-way analysis of variance was performed and mean differences were studied by Tukey's test. C. ampelina and E. leprosa significantly (P < 0.05) reduced the LN, SL, and RL relative to the control plants. They also caused a VD of 10.1, 11.6, and 9.8 mm and 11.2, 13.4, and 10.0 mm in 'Carmenere', 'Chardonnay', and 'Red Globe', respectively. No symptoms were observed on the control canes. C. ampelina (100%) and E. leprosa (75%) were reisolated from inoculated plants and canes. To our knowledge, this is the first report of C. ampelina and E. leprosa on grapevines in Chile. However, their relative importance as causal agent of trunk disease remains to be determined. C. ampelina and E. leprosa have been associated with grapevine cankers in the United States and Spain (3,4). References: (1) J. Auger et al. Plant Dis. 88:1285, 2004. (2) J. Auger et al. Plant Dis. 88:1286, 2004. (3) M. T. Martin et al. Plant Dis. 93:545, 2009. (4) F. P. Trouillas et al. Mycologia 102:319, 2010.
Language	English
Publishing date	2019-02-12
Publishing country	United States
Document type	Journal Article
ZDB-ID	754182-x
ISSN	0191-2917
ISSN	0191-2917
DOI	10.1094/PDIS-12-10-0919
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Article: First Report of Seimatosporium botan Associated with Trunk Disease of Grapevine (Vitis vinifera) in Chile.

Díaz, G A / Elfar, K / Latorre, B A

Plant disease

2019 Volume 96, Issue 11, Page(s) 1696

Abstract: Grapevines are planted on 180,000 ha in Chile. In 2010 and 2011, necrotic lesions and hard texture were observed on woody tissue on 10-year-old vines of cvs. Cabernet Sauvignon, Carménère, Moscatel de Alejandría, and Pedro Jimenez in Ovalle (lat. 30°58' ... ...

Abstract	Grapevines are planted on 180,000 ha in Chile. In 2010 and 2011, necrotic lesions and hard texture were observed on woody tissue on 10-year-old vines of cvs. Cabernet Sauvignon, Carménère, Moscatel de Alejandría, and Pedro Jimenez in Ovalle (lat. 30°58' S) and Cauquenes (lat. 35°58' S). Symptoms were on 10 to 25% of the arm cross sections, resembling symptoms caused by Botryosphaeriaceae (4). Prevalence of 5% was estimated visually in Ovalle (n = 920 grapevines) and Cauquenes (n = 350 grapevines). Small pieces (3 mm) of necrotic tissues from the margins of lesions in cordons (n = 32) were surface sterilized (96% ethanol, 15 s), and plated on acidified PDA plus 0.5 ml/liter of 92% lactic acid, 0.005% tetracycline, 0.01% streptomycin, and 0.1% Igepal CO-630 (Sigma-Aldrich, St. Louis, MO) (APDA). The plates were incubated at 20°C for 14 days. Isolates (n = 12) were obtained from the yellow to dark green slimy colonies with white irregular margins, staining brown the underside of APDA plates. Black acervuli and ellipsoid to fusiform conidia were obtained. Conidia were triple septated, with hyaline upper and bottom cells and brown middle cells (n = 30) of 17.7 ± 1.2 × 5.8 ± 0.8 μm. A basal conidial appendage (6.2 ± 1.0 μm) was always obtained, but conidia having appendages at both ends also were observed. Morphologically, these isolates were identified as Seimatosporium botan Sat. Hatak. & Y. Harada (2). The identification of isolates sei-302 and sei-316 was confirmed by amplifying and sequencing the region ITS1-5.8S-ITS2 of rDNA using ITS4 and ITS5 primers (GenBank Accession Nos. JN088482 and JN088483). BLAST analyses showed 100% similarity with S. botan (Accession No. HM067840) (2). Pathogenicity tests were conducted with isolates sei-302 and sei-316 on detached green shoots (GS) and on rooted 2-year-old vines 'Carménère.' Rooted vines were inoculated at the base of canes and trunks. Inoculations were performed by placing a mycelial agar plug taken from APDA on a wound aseptically made with a cork borer. Wounds were sealed with Parafilm to avoid a rapid dehydration. The inoculated GS were incubated for 2 weeks in a moisture chamber (relative humidity >80%) at 20°C. Inoculated 2-year-old vines were placed in a lath-house for 7 and 15 months for canes and trunk inoculation, respectively. An equal number of GS and vines were inoculated with sterile agar plugs and left as controls. Necrotic lesions with mean of 23.7 ± 2.5 mm on GS, 50.5 ± 3.4 mm on canes, and 41.9 ± 2.3 mm on trunks developed. No significant difference (P < 0.05) was obtained in lesion length between S. botan isolates. After 7 months, 40% of inoculated canes had died. No symptoms were observed in GS controls and rooted control vines treated with sterile agar plugs. S. botan was reisolated from 93 to 100% of the inoculated samples. Previously, S. botan was reported as pathogenic in Paeonia suffruticosa (1), and Seimatosporium sp. was isolated from V. vinifera in California, but their pathogenicity was not demonstrated (3). To our knowledge, this is the first report of pathogenic isolates of S. botan associated with trunk disease of grapevines. These results contribute to the knowledge of the trunk disease of grapevines worldwide. References: (1) Y. Duan et al. Plant Dis. 95:226, 2011. (2) S. Hatakeyama et al. Mycoscience 45:106, 2004. (3) Z. Morales et al. Phytopathol. Mediterr. 49:109, 2010. (4) J. R. Úrbez-Torres. Phytopathol. Mediterr. 50:S5, 2011.
Language	English
Publishing date	2019-02-06
Publishing country	United States
Document type	Journal Article
ZDB-ID	754182-x
ISSN	0191-2917
ISSN	0191-2917
DOI	10.1094/PDIS-05-12-0478-PDN
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article ; Online: Potencial relacionado con eventos cognitivos P300 en el diagnóstico y clasificación del trastorno neurocognitivo debido a enfermedad de Alzheimer posible.

Montoya-Pedrón, A / Ocaña-Montoya, C M / Bolaño-Díaz, G A

Revista de neurologia

2020 Volume 71, Issue 1, Page(s) 11–18

Abstract: Introduction: The value of the P300 cognitive event-related potential in the diagnosis of Alzheimer subtype neurocognitive disorders is still incipient. Recent studies suggest that combining it with neuropsychological tests by cognitive domains would ... ...

Title translation	P300 cognitive event-related potentials in the diagnosis and classification of possible Alzheimer-type neurocognitive disorders.
Abstract	Introduction: The value of the P300 cognitive event-related potential in the diagnosis of Alzheimer subtype neurocognitive disorders is still incipient. Recent studies suggest that combining it with neuropsychological tests by cognitive domains would allow an objective and early characterisation of the cognitive impairment in its initial stages. Aims: To characterise the electrophysiological patterns in the P300 potential that obtain a discriminatory value for the diagnosis and classification of the neurocognitive disorders with a possible Alzheimer-type aetiology. Subjects and methods: This study examines 39 patients classified, according to the DSM-5, with possible Alzheimer-type minor and major neurocognitive disorders, aged between 50 and 85 years, and 53 control subjects with normal cognitive functions. The P300 potential is registered in the auditory mode, oddball paradigm and centroparietal recording. Results: P300 latency is significantly prolonged in subjects with neurocognitive disorder; there are significant differences in the mean values and confidence intervals between healthy controls and patients. No significant differences are obtained in P300 latency between groups with minor and major neurocognitive disorder. The mean amplitude value decreases in neurocognitive disorder, and the P300 amplitude logarithm of the control groups and those with minor and major neurocognitive disorder reaches significantly different mean values and confidence intervals. Conclusions: The parameters quantified in the P300 potential can be used as complementary biomarkers to classify the presence and level of cognitive dysfunction with a possible Alzheimer-type aetiology.
MeSH term(s)	Aged ; Aged, 80 and over ; Alzheimer Disease/diagnosis ; Alzheimer Disease/physiopathology ; Cognitive Dysfunction/diagnosis ; Cognitive Dysfunction/physiopathology ; Confidence Intervals ; Early Diagnosis ; Event-Related Potentials, P300 ; Female ; Humans ; Male ; Middle Aged ; Neurocognitive Disorders/classification ; Neurocognitive Disorders/diagnosis ; Neurocognitive Disorders/physiopathology ; Neuropsychological Tests ; Reaction Time
Language	Spanish
Publishing date	2020-06-24
Publishing country	Spain
Document type	Comparative Study ; Journal Article
ZDB-ID	1468278-3
ISSN	1576-6578 ; 0210-0010
ISSN (online)	1576-6578
ISSN	0210-0010
DOI	10.33588/rn.7101.2019341
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: First Report of Cordon Dieback of Kiwifruits Caused by Diaporthe ambigua and D. australafricana in Chile

Díaz, G. A / B. A. Latorre

Plant disease. 2018 Feb., v. 102, no. 2

2018

Abstract: A cordon dieback of kiwifruit (Actinidia deliciosa) cultivar Hayward was observed in 16 orchards in Chile (Quillota to Linares) in 2013 to 2015. Prevalence varied greatly from 5 to 75%. Symptomatic plants of >9 years showed short internodes, weak shoots, ...

Abstract	A cordon dieback of kiwifruit (Actinidia deliciosa) cultivar Hayward was observed in 16 orchards in Chile (Quillota to Linares) in 2013 to 2015. Prevalence varied greatly from 5 to 75%. Symptomatic plants of >9 years showed short internodes, weak shoots, small and chlorotic leaves, and cane and cordon dieback. Brown, irregular, hard cankers were observed in cross sections of diseased cordons. Cordon samples (n = 80) were surface disinfected (96% ethanol for 10 s and flamed for 8 s), and pieces (2 to 3 cm in length) from the edges of cankered tissues were plated on potato dextrose agar (PDA) plus 0.1% Igepal detergent (octylphenoxy poly[ethyleneoxy]ethanol, branched) (Sigma-Aldrich) for 7 days at 20°C (Díaz and Latorre 2014). Isolates (n = 47) developing white to creamy fast-growing colonies with black-globose pycnidia were obtained after 30 days. All isolates produced hyaline, one-cell, biguttulate, and ellipsoidal α conidia of 5.3 to 7.6 × 2.4 to 3.1 μm and were identified as Diaporthe (Phomopsis) spp. (Gomes et al. 2013). To determine the species identity, six isolates were sequenced using PCR amplification of the rDNA internal transcribed spacer (ITS4/ITS5) and translation elongation factor 1-α gene (EF728/EF968) (Gomes et al. 2013). ITS and EF1-α sequences of three isolates (GenBank accession nos. KU679325 to KU679327 and KU695196 to KU695198, respectively) were 100% identical to the ex-epitype (CBS 114015) of Diaporthe ambigua Nitschke (GenBank accession nos. KC343010 and KC343736 for ITS and EF1-α, respectively). ITS and EF1-α sequences of the other three isolates (GenBank accession nos. KU679315 to KU679317 and KU695186 to KU695188, respectively) were 100% identical to the ex-type (CBS 111886) of Diaporthe australafricana Crous & Van Niekerk (GenBank accession nos. KC343038 and KC343764 for ITS and EF1-α, respectively). Pathogenicity tests were conducted with D. ambigua, isolate Dam KM-5, and D. australafricana, isolate Daus KM-13, on canes (n = 16) and 3-year-old trunks (n = 10) of Hayward kiwifruit in winter months. Canes were pruned and immediately inoculated with 20 μl of conidial suspension (106 α-conidia/ml). Trunk inoculations were performed at the trunk base with a 5 mm mycelial plug, taken from 7-day-old colonies on PDA, and placed aseptically in wounds made with a corker borer. An equal number of canes inoculated with sterile water and trunks inoculated with sterile agar plugs served as controls. On canes, necrotic lesions of 66 and 94 mm in length were caused by D. ambigua and D. australafricana, respectively, 9 months after inoculation. Trunk necrotic lesions of 51 and 62 mm were obtained with D. ambigua and D. australafricana, respectively, 12 months after inoculation. Reisolations were successful from inoculated tissues, and isolates were reidentified molecularly, completing Koch’s postulates. Noninoculated controls remained symptomless. To our knowledge, this is the first report of cordon dieback caused by D. ambigua and D. australafricana in kiwifruit worldwide. Previously, D. ambigua and D. australafricana have been reported causing blueberry stem canker and kiwifruit rots in Chile (Díaz et al. 2017; Elfar et al. 2013). Previously, D. actinidiae was associated to kiwifruit dieback in Chile (Palma and Piontelli 2000). The understanding of the pathogens associated to kiwifruit cane dieback can help to improve management practices to prevent severe damages.
Keywords	Actinidia deliciosa ; agar ; blueberries ; canes ; conidia ; cultivars ; culture media ; detergents ; Diaporthe ; dieback ; ethanol ; genes ; internal transcribed spacers ; internodes ; kiwifruit ; leaves ; mycelium ; orchards ; pathogenicity ; pathogens ; peptide elongation factors ; polymerase chain reaction ; pycnidia ; ribosomal DNA ; shoots ; stem cankers ; tissues ; translation (genetics) ; winter ; Chile
Language	English
Dates of publication	2018-02
Size	p. 446.
Publishing place	Plant Disease
Document type	Article
ZDB-ID	754182-x
ISSN	0191-2917
ISSN	0191-2917
DOI	10.1094/PDIS-07-17-1105-PDN
Database	NAL-Catalogue (AGRICOLA)

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Article: Occurrence of Botrytis prunorum Causing Calyx-End Rot in European Pear Fruits During Cold Storage in Chile

Ferrada, E. E / Briceño, E. X / Díaz, G. A / Lolas, M / Naranjo, P

Plant disease. 2020 Feb., v. 104, no. 2

2020

Abstract: The European pear tree (Pyrus communis L.) is an important fruit species, with 8,217 ha concentrated in central Chile. Previously, only Botrytis cinerea has been described causing calyx-end rot in pears in Chile. During the 2016 to 2017 postharvest ... ...

Abstract	The European pear tree (Pyrus communis L.) is an important fruit species, with 8,217 ha concentrated in central Chile. Previously, only Botrytis cinerea has been described causing calyx-end rot in pears in Chile. During the 2016 to 2017 postharvest season, pears with symptoms of calyx-end rot were observed from a commercial packinghouse in Curicó, Maule Region, after 60 days at 0°C. The rot prevalence registered during cold storage was 3, 2, and 7% for cultivars Beurré Bosc, Forelle, and Packham’s Triumph, respectively. The symptoms started in the pear calyx end as a light brown to dark brown lesion with soft watery decay toward the endocarp. To isolate the causal agent, symptomatic pear fruits (n = 72) were surface disinfected with 75% ethanol, and then small fragments (5-mm length) from rotten internal tissue were placed on Petri dishes containing PDA acidified with 0.5 ml/liter of 92% lactic acid (APDA) and incubated for 7 days at 20°C. Fifty-one isolates of Botrytis were obtained and classified according to conidia production as high sporulating (HS) and low sporulating (LS) on pea agar medium (PAM) containing per liter 160 g of pea (liquefied for 2 min in 1 liter of distilled water, adjusted to pH 6.0 using HCl), 5 g of sucrose, and 25 g of agar. The HS isolates (n = 38) were morphologically and molecularly identified as Botrytis cinerea Pers. The LS isolates (n = 13) produced white to yellowish colonies, floccose, tufted, and scarce sporulation, with a mean of 0.6 and 7.0 conidia/cm2 after 7 days at 20°C on APDA and PAM media, respectively. Conidia were ovoid, hyaline, smooth, and (5.9) 11.2 ± 2.0 (14.7) × (5.3) 6.3 ± 0.6 (8.1) μm (n = 40). Conidiophores were septate, dark, and constricted at the base, hyaline and branched at the apex, and shorter than would correspond to B. cinerea. The molecular identification of six representative isolates was made by analysis of glyceraldehyde-3-phosphate dehydrogenase (G3PDH), heat-shock protein 60 (HSP60), and DNA-dependent RNA polymerase subunit II (RPB2) genes (Staats et al. 2005). BLAST search showed 99 to 100% identity with isolate type Bpru8 of GenBank accessions KP339979 (G3PDH), KP339993 (HSP60), and KP339986 (RPB2) of Botrytis prunorum (Ferrada et al. 2016). Sequences of isolates BEP-10-1 to BEP-10-6 were deposited in GenBank (MN136239 to MN136244, MN136245 to MN136250, and MN136251 to MN136256 for G3PDH, HSP60, and RPB2, respectively). Pathogenicity testing was performed using three isolates (BEP-10-1 to BEP-10-3) in ripe pears (n = 75 fruits), cultivar Beurré Bosc. An isolate of B. cinerea (BAP-4) was included in this test. Fruits were surface disinfected (75% ethanol, 2 min), wounded with a sterile hypodermic syringe in the calyx-end area, and inoculated with 50 μl of a conidial suspension (106 conidia/ml). The control fruits were inoculated with distilled water. After 10 days at 20°C in a humid chamber (>85% relative humidity), all inoculated fruits with B. prunorum developed brown, watery soft rot in the calyx-end area with lesions from 15.7 to 38.6 mm. Fruits inoculated with B. cinerea showed a lesion from 37.8 to 41.9 mm. B. prunorum isolates had a significantly lower virulence than B. cinerea (P < 0.05). Control fruit remained healthy. The pathogen was reisolated from diseased fruit and molecularly identified as B prunorum. To our knowledge, this is the first report of B. prunorum causing pear calyx-end rot in Chile and worldwide. Recently, B. prunorum was described causing kiwifruit rot in Chile (Elfar et al. 2017). These results confirm the occurrence of B. prunorum, with an isolation frequency of 25% of the total Botrytis isolates obtained during cold storage in Chile.
Keywords	Botrytis cinerea ; calyx ; cold storage ; conidia ; conidiophores ; cultivars ; culture media ; DNA-directed RNA polymerase ; endocarp ; ethanol ; fruit trees ; genes ; glyceraldehyde-3-phosphate dehydrogenase ; heat shock proteins ; kiwifruit ; lactic acid ; packing houses ; pathogens ; pears ; Pyrus communis ; relative humidity ; sporulation ; syringes ; virulence ; Chile
Language	English
Dates of publication	2020-02
Publishing place	Plant Disease
Document type	Article
ZDB-ID	754182-x
ISSN	0191-2917
ISSN	0191-2917
DOI	10.1094/PDIS-07-19-1556-PDN
Database	NAL-Catalogue (AGRICOLA)

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Article: Severe Outbreak of Dry Core Rot in Apple Fruits cv. Fuji Caused by Kalmusia variispora During Preharvest in Maule Region, Chile

Gutierrez, M. / Pinheiro, C. / Duarte, J. / Sumonte, C. / Ferrada, E. E. / Elfar, K. / Eskalen, A. / Lolas, M. / Galdós, L. / Díaz, G. A.

Plant disease. 2022 Oct. 03, v. 106, no. 10

2022

Abstract: Apple (Malus × domestica) is an important fruit crop in Chile, with a cultivated area of 32,313 ha concentrated (63%) in the Maule Region (35°25′ S). Unusual core rot on ‘Fuji’ apples was observed at harvest in a commercial orchard in Curicó, Maule ... ...

Abstract	Apple (Malus × domestica) is an important fruit crop in Chile, with a cultivated area of 32,313 ha concentrated (63%) in the Maule Region (35°25′ S). Unusual core rot on ‘Fuji’ apples was observed at harvest in a commercial orchard in Curicó, Maule Region, with an incidence of from 22 to 35% in 2018 and 2019. In 2017, an incidence of 30% was estimated on Fuji fruits destined to the Asian market. Internal decay symptoms consisted of dry, corky, light to dark-brown tissue, initially within the seed locules. In moderate to severe cases, the necrotic lesion progresses deeper into the mesoderm. External symptoms were quite subtle, and, typically, the disease goes unnoticed. However, infected fruit ripen earlier. Small pieces (2 to 3 mm) from the internal lesion margin of symptomatic apples (n = 50) were placed on potato dextrose agar (2%) and incubated at 20°C for 10 days. Pure cultures (n = 41) were obtained and transferred to malt extract agar (MEA) (2%). Colonies on MEA produced an even to slightly undulating buff margin with white woolly aerial mycelium, changing to gray to olivaceous with age. On the underside, colonies were umber and buff in the center and margin, respectively. After 10 days, numerous densely aggregated dark-brown mature pycnidia were observed. Aseptate conidia were subglobose to cylindrical, straight, sometimes curved and rounded at both ends, initially hyaline to pale olive, thin, smooth walled, and with mean dimensions of (2.9–) 3.4 (–4.4) × (1.5–) 1.8 (2.2) µm (n = 50). Based on morphology, the fungus was identified as Kalmusia variispora (Verkley et al. 2014). The internal transcribed space (ITS), portion of β-tubulin (TUB), and large subunits of the nuclear ribosomal RNA (LSU) loci were used for molecular identification, using primers ITS4/ITS5, Bt2a/Bt2b, and LR0R/LR5, respectively (Ariyawansa et al. 2014). BLAST searches indicated 100% identity with K. variispora (ex-type CBS 121517). The maximum parsimony phylogenetic analysis placed Chilean isolates in the K. variispora clade. Sequences were deposited in GenBank (OL711706 to OL711709, OL739499 to OL739502, and OL711710 to OL711713 for ITS, TUB, and LSU, respectively). Pathogenicity tests were conducted using four K. variispora isolates. Fuji apples (n = 20) were surface disinfested (75% ethanol, 30 s) and then wounded and inoculated with conidial suspension (50 μl of 10⁶ conidia/ml) deposited in the middle and into the core region using a sterile fine-tipped micropipette. Additionally, 20 1-year-dormant rooted cuttings of Fuji and ‘Cripps Pink’ were pruned and immediately inoculated on the pruning wound. An equal number of apples and rooted cuttings treated with sterile water were used as controls. The experiments were repeated once. All inoculated fruits developed lateral lesions (22 to 37 mm) and dry core rot (18 to 36 mm) symptoms identical to those described in the original outbreak after 20 days at 20°C in a commercial packing box. The inoculated cuttings produced canker lesions of 10 to 21 mm in length, and dieback symptoms were observed after 3 months. No symptoms were observed on the negative controls. Koch’s postulates were fulfilled by 100% reisolating K. variispora. Previously, Alternaria spp. have been reported as the primary pathogen associated with moldy core and dry core rot of apples worldwide (McLeod et al. 2014) and in Chile (Elfar et al. 2018). However, Kalmusia spp. have been associated with dry core rot in apples (McLeod et al. 2014) and have been isolated from canker symptoms on apples in Chile (Díaz et al. 2022). To our knowledge, this is the first report of a severe outbreak of K. variispora causing dry core rot in apples in Chile and worldwide.
Keywords	Alternaria ; Malus domestica ; conidia ; cultivation area ; culture media ; dieback ; ethanol ; fruits ; markets ; mesoderm ; molds (fungi) ; mycelium ; olives ; orchards ; pathogenicity ; pathogens ; phylogeny ; pycnidia ; ribosomal RNA ; Chile
Language	English
Dates of publication	2022-1003
Publishing place	The American Phytopathological Society
Document type	Article
ZDB-ID	754182-x
ISSN	0191-2917
ISSN	0191-2917
DOI	10.1094/PDIS-12-21-2776-PDN
Database	NAL-Catalogue (AGRICOLA)

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Article: First Report of Neofusicoccum australe Associated with Botryosphaeria Canker of Grapevine in Chile.

Besoain, X / Torres, C / Díaz, G A / Latorre, B A

Plant disease

2019 Volume 97, Issue 1, Page(s) 143

Abstract: A survey of trunk diseases was conducted in 2010 in vineyards (n = 14) in central Chile (latitude 33°51' to 36°30'), specifically of Vitis vinifera 'Cabernet Sauvignon,' which is the main wine-grape cultivar (38,806 ha) in Chile. The following symptoms ... ...

Abstract	A survey of trunk diseases was conducted in 2010 in vineyards (n = 14) in central Chile (latitude 33°51' to 36°30'), specifically of Vitis vinifera 'Cabernet Sauvignon,' which is the main wine-grape cultivar (38,806 ha) in Chile. The following symptoms of trunk disease were observed in 5- to 19-year-old grapevines: short internodes, dead spurs, dead cordons (arms), and shoot dieback. Upon cutting into cordons and trunks of symptomatic vines, brown, V-shaped cankers of hard consistency were observed. A total of 56 wood cankers were collected, and small pieces of symptomatic wood (approximately 4 mm in diameter) taken from the canker margin were surface disinfected (75% ethanol, 30 s) and placed on acidified PDA (0.5 ml of 96% lactic acid per liter; APDA), which was incubated for 4 to 7 days at 24°C. Colonies, tentatively identified as a species within the Botryosphaeriaceae based on the presence of whitish-to-gray aerial mycelium and exhibiting rapid growth (4 to 5 cm colony diameter in 48 h), were hyphal-tip purified to APDA for identification. Colonies produced globose, black pycnidia with unicellular, hyaline, ellipsoidal, densely granulate, externally smooth, and thin-walled conidia of 17.0 ± 0.7 ± 6.7 ± 0.4 μm (n = 20). A yellow pigmentation was observed at the center of 48-h colonies on APDA. Morphologically, these isolates were identified as Neofusicoccum australe (Slippers, Crous & M.J. Wingfield) Crous, Slippers & A.J.L. Phillips (2,3). BLASTn searches of the ITS rDNA region, amplified with PCR primers ITS4/ITS5 (532 bp), and a 400-bp section of the beta-tubulin subunit 2 gene amplified with primers Bt2a and Bt2b of N. australe (GenBank Accession No. JX290091 and JX679868, respectively) revealed 99% similarity with the ITS and beta-tubulin sequences of N. australe reference strains EF638778 and HQ392761, respectively. Pathogenicity tests were conducted using N. australe isolate Vid1559 on 2-year-old Cabernet Sauvignon plants (n = 4), which were inoculated by wounding the woody stem with a scalpel approximately 1 cm below the most basal bud, placing an 8-mm mycelial plug taken from a 7-day culture into the wound, and then sealing the wound with Parafilm. Non-inoculated controls (n = 4) were 'mock' inoculated with sterile agar plugs. After 3 months under field conditions, during spring and summer, the woody stems were examined for vascular discoloration (VD), characteristic of a wood canker. Inoculated plants had stems with light-brown, necrotic VD with a mean length of 15.2 cm, measured from the inoculation point. No VD was observed on the controls. N. australe was reisolated from 100% of the inoculated plants, completing Koch's postulates. Of 14 vineyards surveyed, 8% were infected with N. australe. N. australe is known as a trunk pathogen of grape (4), and other species of Botryosphaeriaceae have been associated with grapevine trunk disease in Chile (1). To our knowledge, this is the first report of N. australe causing Botryosphaeria canker of grape in Chile, where the pathogen is previously reported on blueberry (2). References: (1) G. A. Díaz et al. Plant Dis. 95:1032, 2011. (2) J. G. Espinoza et al. Plant Dis. 92:1407, 2008. (3) Slippers et al. Mycologia 96:1030, 2004. (4) J. R. Úrbez-Torres Phytopathol. Mediterr. 50:S5, 2011.
Language	English
Publishing date	2019-02-05
Publishing country	United States
Document type	Journal Article
ZDB-ID	754182-x
ISSN	0191-2917
ISSN	0191-2917
DOI	10.1094/PDIS-07-12-0652-PDN
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: A Severe Outbreak of Charcoal Rot in Cantaloupe Melon Caused by Macrophomina phaseolina in Chile.

Jacob, C J / Krarup, C / Díaz, G A / Latorre, B A

Plant disease

2019 Volume 97, Issue 1, Page(s) 141

Abstract: A severe outbreak of charcoal rot was observed in cantaloupe melon (Cucumis melo L.) in the summer of 2011 to 2012 in Curacaví Valley, Chile. Prior to harvest, of 72 plants per cultivar, charcoal rot prevalence varied from 32% to 82% in cvs. Colima, ... ...

Abstract	A severe outbreak of charcoal rot was observed in cantaloupe melon (Cucumis melo L.) in the summer of 2011 to 2012 in Curacaví Valley, Chile. Prior to harvest, of 72 plants per cultivar, charcoal rot prevalence varied from 32% to 82% in cvs. Colima, Charantias, Navigator, Origami, Otero, and Samoa. Symptoms were wilting and leaf browning associated with water-soaked lesions at the base of the crown with amber to dark brown exudates. Lesions dried out progressively, turned tan, and cracked. Affected plants declined and died before harvest. Reddish fruit decay was observed. Symptomatic stem and root samples (n = 97) were collected, surface disinfected (96% ethanol, 30 s), plated on PDA acidified with 0.5 ml/liter of 92% lactic acid (APDA), and incubated at 20 ± 1°C. A white, fast-growing mycelium was obtained that turned gray to black after 7 days due to the presence of spherical to oblong black microsclerotia 136 ± 52 μm (n = 80) in diameter. On the basis of colony morphology and microsclerotia, 57 isolates (59%), obtained from 97 melon samples, were tentatively identified as Macrophomina phaseolina (Tassi) Goid. (2,3). The morphological identification of four isolates M1HB-B, M2CO-B, M3CH-R, and M4OT-B (GenBank Accession Nos. JX203630, JX203631, JX203632, and JX203633) was confirmed by sequencing of the internal transcribed spacer region (ITS1-5.8S-ITS2) of rDNA, using primers ITS4 and ITS5, with >99% similarity with the sequences of M. phaseolina (GenBank Accession No. HQ660592) (4). Pathogenicity tests were conducted with isolates M1HB-B, M2CO-B, M3CH-R, and M4OT-B on melon fruits cvs. Colima, Origami, Charantias, and Diva. Four mature melon fruits per cultivar per isolate were surface disinfected with 0.5% sodium hypochlorite for 2 min before inserting a mycelium plug (19 mm
Language	English
Publishing date	2019-02-05
Publishing country	United States
Document type	Journal Article
ZDB-ID	754182-x
ISSN	0191-2917
ISSN	0191-2917
DOI	10.1094/PDIS-06-12-0588-PDN
Database	MEDical Literature Analysis and Retrieval System OnLINE

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Article: First Report of Blossom Blight Caused by Sclerotinia sclerotiorum on Japanese Plum, Nectarine, and Sweet Cherry Orchards in Chile.

Ferrada, E E / Díaz, G A / Zoffoli, J P / Latorre, B A

Plant disease

2019 Volume 98, Issue 5, Page(s) 695

Abstract: Blossom blight of Japanese plum (Prunus salicina), nectarine (P. persica var. nectarina), and sweet cherry (P. avium) was observed in commercial orchards in central Chile in 2012. Disease prevalence of 8% and 1% were estimated in 2012 and 2013, ... ...

Abstract	Blossom blight of Japanese plum (Prunus salicina), nectarine (P. persica var. nectarina), and sweet cherry (P. avium) was observed in commercial orchards in central Chile in 2012. Disease prevalence of 8% and 1% were estimated in 2012 and 2013, respectively. Early symptoms appeared as small pale-brown necrotic lesions on the petals that eventually affected the entire flowers. White and cottony fungal colonies were consistently isolated on potato dextrose agar acidified with 0.5 ml/liter of 92% lactic acid (APDA), incubated for 5 days at 20°C. Black spherical to elongated sclerotia of 2.5 to 4.2 × 2.8 to 5.3 mm (n = 60) were formed on APDA. This fungus was tentatively identified as Sclerotinia sclerotiorum (Lib.) de Bary. The identity of the fungus was confirmed by BLAST analysis of the internal transcribed spacer (ITS) region (GenBank Accession Nos. KF148604 to KF148609) of rDNA, amplified with PCR primers ITS1/ITS4 (3), demonstrating a 99 to 100% similarity with the reference S. sclerotiorum strains (EU082466 and JX307092). The pathogenicity was studied in detached flowers of 'Larry Ann' Japanese plum, 'Summer Bright' nectarine, and 'Bing' sweet cherry that were inoculated with a mycelial suspension (10
Language	English
Publishing date	2019-02-01
Publishing country	United States
Document type	Journal Article
ZDB-ID	754182-x
ISSN	0191-2917
ISSN	0191-2917
DOI	10.1094/PDIS-10-13-1045-PDN
Database	MEDical Literature Analysis and Retrieval System OnLINE

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