scholarly journals First Report of Lasiodiplodia theobromae Associated with Stem Canker of Almond in California

Plant Disease ◽  
2013 ◽  
Vol 97 (7) ◽  
pp. 994-994 ◽  
Author(s):  
S. F. Chen ◽  
D. Morgan ◽  
R. H. Beede ◽  
T. J. Michailides
Keyword(s):  
Average Length ◽  
Stem Canker ◽  
Macrophomina Phaseolina ◽  
Disease Diagnosis ◽  
Type Specimens ◽  
Lasiodiplodia Theobromae ◽  
First Report ◽  
Agar Plug ◽  
Almond Trees ◽  
Negative Controls

California is a major almond (Prunus dulcis) producer in the world. In September 2012, 2-year-old almond trees from an orchard in Fresno Co. with stem cankers were submitted for disease diagnosis. In a survey of the orchard, 12 ha (1,500 Nonpareil and 1,800 Monterey almond trees) of 48 ha trees had been killed apparently due to a stem canker. The cankers developed above the graft union, were covered with amber sap, and often girdled the trunk. Isolations made from tissues at the canker margins onto acidified potato dextrose agar (PDA) yielded two fungi, Macrophomina phaseolina (Tassi) Goid and Lasiodiplodia theobromae (Pat.) Griffon & Maubl (1). M. phaseolina and L. theobromae were isolated from eight and two of 10 cankered trees, respectively. No mixed infections were found. M. phaseolina isolates were characterized by gray hyphae that turned black with developing microsclerotia. L. theobromae isolates were characterized by white, aerial mycelium that turned mouse gray after 5 days. Young conidia were ellipsoidal, thick walled, initially hyaline, granular, and nonseptate; aged conidia were brown, 1-septate with longitudinal striations in the wall. Identity was confirmed by analyses of the internal transcribed spacer (ITS), β-tubulin 2 (BT2), and the translation elongation factor 1-alpha (TEF-1α) gene regions. BLAST searches at GenBank showed a high identity with reference sequences of type specimens both for M. phaseolina (isolates 7E64 to 7E69: ITS, 100%; BT2, 99%; TEF-1α, 99%) and L. theobromae (isolates 7E86 to 7E88: ITS, 99%; BT2, 99%; TEF-1α, 100%). Sequences of three gene regions were deposited as GenBank accessions KC357271 to KC357279 (ITS); KC357280 to KC357288 (BT2); and KC357289 to KC357297 (TEF-1α). The pathogenicity of M. phaseolina and L. theobromae to P. dulcis cultivars Butte, Carmel, Nonpareil, and Padre was investigated in an orchard at KARE using four isolates of M. phaseolina (7E64, 7E65, 7E66, and 7E69) and two isolates of L. theobromae (7E86 and 7E88). Ten 2-year-old branches per isolate from 7-year-old trees were inoculated with each isolate in late September 2012, after removing the bark with a 7-mm cork borer and placing a 7-day-old 7-mm-diameter agar plug bearing mycelium of each isolate directly into the fresh wound, mycelium side down. Ten additional branches of each of the four cultivars were inoculated with sterile PDA plugs and served as negative controls. Three weeks after inoculation, the average lesion produced by M. phaseolina on Butte, Carmel, Nonpareil, and Padre was 53, 52, 41, and 37 mm in length, respectively. Lesions produced by L. theobromae were 191, 206, 194, and 103 mm in length on the four cultivars, respectively. No disease lesion, only wounds, were produced on negative controls. Lesions produced by both pathogens were longer (P < 0.05) than wounds on the controls (average length 10 mm on all cultivars). Both L. theobromae isolates killed branches of cultivars Butte, Carmel, and Nonpareil in 2 weeks. M. phaseolina and L. theobromae were reisolated from the inoculated branches, and no fungus was reisolated from controls. Based on pathogenicity results, L. theobromae is more virulent to almond branches than M. phaseolina. To our knowledge, this is the second report of M. phaseolina (2) and the first report of L. theobromae as pathogens of P. dulcis trees in California. References: (1) A. Alves et al. Fungal Diversity 28:1, 2008. (2) P. Inderbitzin et al. Mycologia 102:1350, 2010.

scholarly journals First Report of Lasiodiplodia theobromae Causing Stem Canker of Fraxinus americana

Plant Disease ◽  
2019 ◽  
Vol 103 (12) ◽  
pp. 3276
Author(s):  
F. Chen ◽  
X. Zheng ◽  
X. Zhao ◽  
F. Chen
Keyword(s):  
Stem Canker ◽  
Fraxinus Americana ◽  
Lasiodiplodia Theobromae ◽  
First Report

scholarly journals First Report of Stem Canker of Goldenrain Tree Caused by Lasiodiplodia theobromae in Tennessee and the United States

Plant Disease ◽  
2020 ◽  
Vol 104 (11) ◽  
pp. 3062-3062
Author(s):  
F. Baysal-Gurel ◽  
F. A. Avin ◽  
Cansu Oksel ◽  
T. Simmons
Keyword(s):  
United States ◽  
Stem Canker ◽  
The United States ◽  
Lasiodiplodia Theobromae ◽  
First Report

First Report of Macrophomina phaseolina, Fusarium brachygibbosum, and Lasiodiplodia theobromae Causing Fungal Watermelon Vine Decline in Southwest and West-Central Florida

2021 ◽  
Author(s):  
Cristina Pisani ◽  
Scott Adkins ◽  
William W Turechek ◽  
Pragna C Patel ◽  
Erin Rosskopf
Keyword(s):  
Disease Severity ◽  
Fungal Pathogens ◽  
Disease Incidence ◽  
Macrophomina Phaseolina ◽  
Lasiodiplodia Theobromae ◽  
First Report ◽  
Central Florida ◽  
Untreated Check ◽  
Vine Decline ◽  
Yellowing Virus

Wilt and vine decline symptoms were observed on watermelon plants in Glades and Hardee Counties in Florida in spring 2017 that resembled viral watermelon vine decline caused by squash vein yellowing virus (SqVYV). When no SqVYV was detected, greenhouse studies and morphological and molecular analyses revealed three fungal pathogens, Macrophomina phaseolina, Fusarium brachygibbosum, and Lasiodiplodia theobromae, that were not previously reported on watermelon in Florida. A previously reported oomycete, Pythium spinosum, was also detected in some, but not all isolates, and but when applied independently, resulted in disease incidence that was comparable to the untreated check, ruling it out as a primary causal agent of the symptoms observed in the field. In one of three experiments, seedlings inoculated with a combination of Macrophomina phaseolina, Fusarium brachygibbosum, and Pythium spinosum suffered the highest disease severity based on AUDPC values. In another experiment, seedlings inoculated with F. brachygibbosum exhibited the most severe symptoms and rapid disease development following inoculation. When seeds were inoculated with either a single or a combination of the isolated fungi, those inoculated with L. theobromae resulted in seedlings with the greatest disease severity. This is the first report of these three fungal pathogens on watermelon in Florida.

scholarly journals First Report of Lasiodiplodia theobromae Associated with Stem Canker of Cassia fistula in Guangxi, South China

Plant Disease ◽  
2015 ◽  
Vol 99 (2) ◽  
pp. 288-288 ◽  
Author(s):  
T. J. Deng ◽  
Q. L. Li ◽  
X. L. Chen ◽  
S. P. Huang ◽  
T. X. Guo ◽  
...  
Keyword(s):  
Ornamental Plants ◽  
Morphological Characteristics ◽  
Its Region ◽  
Stem Canker ◽  
Morphological Identification ◽  
Vegetable Crops ◽  
Cassia Fistula ◽  
Lasiodiplodia Theobromae ◽  
First Report ◽  
Surface Sterilization

Cassia fistula, a member of the Fabaceae, known as the golden shower tree, is native to South Asia. It is now distributed worldwide and is popular as an ornamental plant as well as being used in herbal medicine. In October 2013, symptoms of stem canker were observed on C. fistula in a nursery (108°38′ E, 22°87′ N) in Nanning, Guangxi, China. The symptoms began as small brown lesions, which enlarged over several months to long, striped, slightly sunken lesions, 1 to 9 cm in width and 16 to 135 cm in length. The conspicuous cankers had vertical cracks outlining the canker and evenly spaced horizontal cracks, eventually resulting in whole plants dying back. The cankers were found on 90% of six-year-old plants in this nursery and were also observed in other plantings. On potato dextrose agar (PDA), isolates with similar morphological characteristics were consistently recovered from symptomatic plant tissues after surface sterilization in 75% ethanol for 30 sec and then in 0.1% mercuric chloride for 2 min. Over 100 conidia were examined from three isolates and were found to be elliptical and hyaline when immature, becoming dark brown, one-septate, and longitudinally striate when mature and ranging from 20 to 31 × 11 to 16 μm (average 25.5 × 13.6 μm). The rDNA internal transcribed spacer (ITS) region of isolate LC-1 was sequenced (GenBank Accession No. KM387285), and it showed 100% identity to Lasiodiplodia theobromae (Pat.) Griffon & Maubl. (GenBank KC964548), confirming the morphological identification (2) as L. theobromae (also known as Botryosphaeria rhodina (Cooke) Arx). A culture of this isolate has been preserved in the Guangxi Academy of Agricultural Sciences fungal collection. The pathogenicity of the isolate was tested on healthy twigs and branches of C. fistula trees in a field setting at Guangxi Agricultural Vocational-Technical College, Nanning, Guangxi, in June and August 2014. For each treatment, five green twigs and five 2-year-old branches were used. Five adjacent needle punctures were made on each branch with a sterilized needle. A mycelial plug was then placed on the wound of each branch and wrapped with Parafilm. Control twigs were treated with sterile PDA plugs. One week later, typical lesions were observed on the inoculated branches, with symptoms becoming more extensive after two weeks, but no symptoms were seen on the controls. Koch's postulates were fulfilled by re-isolation of L. theobromae from diseased branches. L. theobromae is recognized as an important wood pathogen and has been reported to cause cankers, dieback, and fruit and root rots in over 500 different hosts, including perennial fruit and nut trees, vegetable crops, and ornamental plants (2). The fungus has been reported on C. fistula in India since the 1970s (1); however, to our knowledge, this is the first report of L. theobromae infecting C. fistula in China. References: (1) R. S. Mathur. The Coelomycetes of India. Bishen Singh Mahendra Pal Singh, Delhi, India, 1979. (2) J. R. Úrbez-Torres et al. Plant Dis. 92:519, 2008.

scholarly journals First Report of Dragon Fruit Stem Canker Caused by Lasiodiplodia theobromae in Bangladesh

Plant Disease ◽  
2019 ◽  
Vol 103 (10) ◽  
pp. 2686 ◽  
Author(s):  
P. S. Briste ◽  
M. A. H. B. Bhuiyan ◽  
A. M. Akanda ◽  
O. Hassan ◽  
N. U. Mahmud ◽  
...  
Keyword(s):  
Stem Canker ◽  
Lasiodiplodia Theobromae ◽  
First Report ◽  
Dragon Fruit

scholarly journals First Report of Lasiodiplodia Fruit Rot of Jackfruit in Taiwan

Plant Disease ◽  
2008 ◽  
Vol 92 (7) ◽  
pp. 1137-1137 ◽  
Author(s):  
H. F. Ni ◽  
R. S. Chen ◽  
S. F. Chang ◽  
H. R. Yang
Keyword(s):  
Early Stage ◽  
Plant Diseases ◽  
Fruit Rot ◽  
Commonwealth Mycological Institute ◽  
Tropical Fruit ◽  
Lasiodiplodia Theobromae ◽  
First Report ◽  
Median Septum ◽  
Agar Plug ◽  
Diseased Fruit

Jackfruit (Artocarpus heterophyllus Lam.) is a tropical fruit that is native to India. Five diseases, including Rhizopus fruit rot and anthracnose fruit rot, have been recorded in Taiwan (2). In 2003, brown lesions were observed on mature or harvested fruits at the Chiayi Agricultural Experiment Branch. The disease caused fruits to collapse and was easily distinguished from anthracnose and Rhizopus fruit rot. In the field, Rhizopus fruit rot was characterized by black flocci sporangia and mycelia covering the flowers and young fruits. Lasiodiplodia fruit rot often occurred on mature or wounded fruit and diseased fruit were covered with gray or black flat mycelia under humid conditions. In the early stage of Lasiodiplodia fruit rot, tiny yellow-brown lesions appeared on the peel. The lesions could rapidly expand to 10 cm in diameter within 5 days and became dark brown with a light margin. The rot symptoms progressed quickly from the peel surface into the sarcocarps that eventually turned black and soft. A fungus was isolated from the margin of the lesions and cultured on acidified potato dextrose agar (PDA) (pH 3.8). The morphology of the fungus was similar to Lasiodiplodia theobromae (Pat.) Griff. & Maubl. (synonym Botryodiplodia theobromae Pat.), which causes the stem-end rot of mango, papaya, and banana in Taiwan. The fungus grew well and produced pycnidia and conidia on PDA. Young conidia were ovate, hyaline, and thin walled without septa. Mature conidia (20 to 28 × 12 to 15 μm) were dark brown and thick walled with one median septum and longitudinal striations. The internal transcribed spacer (ITS) sequence of ribosomal DNA of this fungus was submitted to GenBank (Accession No. EU 407235) and showed 100% sequence identity with that of Botryosphaeria rhodina (anamorph Lasiodiplodia theobromae; GenBank Accession No. DQ458890). On the basis of morphological and molecular criteria, the fungus was identified as L. theobromae (1). Three healthy jackfruit fruits were wounded and inoculated with 2 × 2 mm mycelial agar plugs of the fungus from a monoconidial culture. A sterile agar plug was placed on the wounded site as a control. The fruits were kept in a box to maintain high humidity for 2 days at room temperature. Brown lesions were observed on all inoculated sites 6 days post infection. The pathogen was reisolated from the lesions of inoculated fruits, fulfilling Koch's postulate. The experiment was repeated twice. To our knowledge, this is the first report of L. theobromae causing fruit rot of jackfruit in Taiwan. References: (1) B. C. Sutton. The Coelomycetes. Commonwealth Mycological Institute, Kew, UK, 1980. (2) Y. P. Tsai, ed. List of Plant Diseases in Taiwan. 4th ed. Taiwan Phytopathological Society, 2002.

scholarly journals First Report of Cankers and Dieback Caused by Neofusicoccum mediterraneum and Diplodia corticola on Grapevine in Spain

Plant Disease ◽  
2011 ◽  
Vol 95 (10) ◽  
pp. 1315-1315 ◽  
Author(s):  
C. Pintos Varela ◽  
V. Redondo Fernández ◽  
O. Aguín Casal ◽  
J. P. Mansilla Vázquez
Keyword(s):  
Average Length ◽  
Aerial Mycelium ◽  
Its Region ◽  
Single Spore ◽  
Mycelium Growth ◽  
First Report ◽  
Tubulin Genes ◽  
Agar Plug

In November 2010, four grapevine plants of cv. Crimson from a vineyard located in Sevilla (south Spain) revealed trunk cankers. Several pathogens were isolated, including Cylindrocarpon liriodendri (2), Phaeoacremonium aleophilum (2), Pleurostomophora richardsiae, Neofusicoccum parvum, and Botryosphaeria dothidea (2). Among Botryosphaeriaceae fungi isolated on potato dextrose agar (PDA) were two types that did not fit the above mentioned species. Isolates of type 1 produced an abundant, gray mycelium with a diurnal zonation that gradually became dark olivaceous. Mycelium growth occurred from 5 to 37°C with an optimum at 28°C. Conidia were hyaline, fusiform, aseptate, thin walled, but gradually became obscured and septate with age, and measured (18.4-) 21.4 (-24.3) × (4.2-) 5.5 (-7.2) μm with a length/width (L/W) ratio of 4.0 ± 0.5 (n = 100). Isolates of type 1 were identified as N. mediterraneum (3). Single-spore cultures of type 2 developed a whitish, dense, aerial mycelium and remained white up to 10 days on PDA and darkened to gray thereafter. Mycelium growth occurred from 3 to 37°C with an optimum at 29 to 30°C. Conidia were hyaline, aseptate, thick walled, oblong to cylindrical, sometimes becoming light brown and one or two septate after discharge, and measured (24.6-) 30.2 (-42.8) × (10.9-) 14.3 (-18.6) μm with a L/W ratio of 2.1 ± 0.2 (n =100). Isolates of type 2 were identified as Diplodia corticola (1). Nucleotide sequences of the ribosomal internal transcribed spacer (ITS) region and the -tubulin genes were used to confirm the identifications through BLAST searches in GenBank. Comparison of the sequences of types 1 and 2 showed 99 to 100% homology with N. mediterraneum (HM443604 (4) and GU251836) and D. corticola (AY268421 (1) and EU673117), respectively. Representative sequences of N. mediterraneum (JF949757 and JF949756) and D. corticola (JF949758 and JF949759) were deposited in GenBank. The pathogenicity of one representative isolate of each of N. mediterraneum and D. corticola was confirmed by inoculating 10 detached grapevine canes (averaging 12 mm in diameter and 30 cm long) per isolate. A shallow wound was made with a scalpel on the internodes. A colonized 6-mm agar plug, from the margin of an actively growing colony, was inserted in every wound and sealed with Parafilm. Ten grapevine canes controls received only sterile PDA agar plugs. Canes were maintained at 25°C and 70% humidity. After 5 weeks, all inoculated canes developed cankers and pycnidia around the inoculation site. Vascular necroses that developed on the inoculated canes were an average of 28.6 mm for N. mediterraneum and 27.7 mm for D. corticola. One-way analysis of variance and Tukey's test confirmed significant differences in the extent of vascular necroses. The average necroses length in the inoculated canes was significantly greater (P < 0.05) than the average length of discoloration induced by the simulated inoculation process in the control. Both pathogens were reisolated from all inoculated plants but not from controls. To our knowledge, this is the first report of N. mediterraneum and D. corticola as pathogens on grapevine in Spain. References: (1) A. Alves et al. Mycologia 96:603, 2004. (2) A. Aroca and D. Gramaje et al. Eur. J. Plant. Pathol. 126:165, 2010. (3) P. W. Crous et al. Fungal Planet. No. 19, 2007. (4) F. P. Trouillas et al. Plant. Dis. 94:1267, 2010.

scholarly journals First Report of Lasiodiplodia theobromae Causing Stem Canker of Jatropha curcas in Malaysia

Plant Disease ◽  
2012 ◽  
Vol 96 (5) ◽  
pp. 767-767 ◽  
Author(s):  
R. Sulaiman ◽  
S. S. Thanarajoo ◽  
J. Kadir ◽  
G. Vadamalai
Keyword(s):  
Jatropha Curcas ◽  
Disease Incidence ◽  
Stem Canker ◽  
Molecular Characteristics ◽  
Width Ratio ◽  
Commonwealth Mycological Institute ◽  
Lasiodiplodia Theobromae ◽  
First Report ◽  
Wide Range ◽  
Vascular Discoloration

Physic nut (Jatropha curcas L.) is an important biofuel crop worldwide. Although it has been reported to be resistant to pests and diseases (1), stem cankers have been observed on this plant at several locations in Peninsular Malaysia since early February 2008. Necrotic lesions on branches appear as scars with vascular discoloration in the tissue below the lesion. The affected area is brownish and sunken in appearance. Disease incidence of these symptomatic nonwoody plants can reach up to 80% in a plantation. Forty-eight samples of symptomatic branches collected from six locations (University Farm, Setiu, Gemenceh, Pulau Carey, Port Dickson, and Kuala Selangor) were surface sterilized in 10% bleach, rinsed twice with sterile distilled water, air dried on filter paper, and plated on water agar. After 4 days, fungal colonies on the agar were transferred to potato dextrose agar (PDA) and incubated at 25°C. Twenty-seven single-spore fungal cultures obtained from all locations produced white, aerial mycelium that became dull gray after a week in culture. Pycnidia from 30-day-old pure cultures produced dark brown, oval conidia that were two celled, thin walled, and oval shape with longitudinal striations. The average size of the conidia was 23.63 × 12.72 μm with a length/width ratio of 1.86. On the basis of conidial morphology, these cultures were identified as Lasiodiplodia theobromae. To confirm the identity of the isolates, the internal transcribed spacer (ITS) region was amplified with ITS1/ITS4 primers and sequenced. The sequences were deposited in GenBank (Accession Nos. HM466951, HM466953, HM466957, GU228527, HM466959, and GU219983). Sequences from the 27 isolates were 99 to 100% identical to two L. theobromae accessions in GenBank (Nos. HM008598 and HM999905). Hence, both morphological and molecular characteristics confirmed the isolates as L. theobromae. Pathogenicity tests were performed in the glasshouse with 2-month-old J. curcas seedlings. Each plant was wound inoculated by removing the bark on a branch to a depth of 2 mm with a 10-mm cork borer. Inoculation was conducted by inserting a 10-mm-diameter PDA plug of mycelium into the wound and wrapping the inoculation site with wetted, cotton wool and Parafilm. Control plants were treated with plugs of sterile PDA. Each isolate had four replicates and two controls. After 6 days of incubation, all inoculated plants produced sunken, necrotic lesions with vascular discoloration. Leaves were wilted and yellow above the point of inoculation on branches. The control plants remained symptomless. The pathogen was successfully reisolated from lesions on inoculated branches. L. theobromae has been reported to cause cankers and dieback in a wide range of hosts and is common in tropical and subtropical regions of the world (2,3). To our knowledge, this is the first report of stem canker associated with L. theobromae on J. curcas in Malaysia. References: (1) S. Chitra and S. K. Dhyani. Curr. Sci. 91:162, 2006. (2) S. Mohali et al. For. Pathol. 35:385, 2005. (3) E. Punithalingam. Page 519 in: CMI Descriptions of Pathogenic Fungi and Bacteria. Commonwealth Mycological Institute, Kew, Surrey, UK. 1976.

scholarly journals First Report of Canker Disease Caused by Neofusicoccum parvum and N. australe on Blueberry Bushes in Spain

Plant Disease ◽  
2013 ◽  
Vol 97 (8) ◽  
pp. 1112-1112 ◽  
Author(s):  
S. Castillo ◽  
C. Borrero ◽  
R. Castaño ◽  
A. Rodríguez ◽  
M. Avilés
Keyword(s):  
Phylogenetic Analyses ◽  
Elongation Factor ◽  
Stem Canker ◽  
Healthy Plant ◽  
Neofusicoccum Parvum ◽  
First Report ◽  
Progress Curve ◽  
Agar Plug ◽  
Southern Highbush ◽  
Pathogenicity Assay

A field survey conducted in September 2009 at five plantations of six different cultivars of southern highbush blueberries (Vaccinium spp.) in Huelva, Spain, yielded 35 diseased plants. Diseased plants exhibited red-brown cankers and stem dieback. Blueberry cultivation in Huelva rose from 290 ha in 2007 to 777 ha in 2012, and the increase of these symptoms is of concern to producers. Stem pieces cut from the edge of lesions on infected plants were surface-disinfected with 5% sodium hypochlorite and cultured on potato dextrose agar (PDA). Based on colony characteristics on PDA, 18 colonies (one each from 18 different plants) were identified as Botryosphaeria spp. Species identities were confirmed by analysis of nucleotide sequences of the internal transcribed spacer (ITS), rDNA, and elongation factor 1-alpha (EF1-α) sequences, using ITS1-ITS4 (3) and EF728f-EF986r (2) as primer pairs, respectively. BLAST searches of GenBank showed a high similarity of the isolate sequences to the reference sequences. Molecular results confirmed these species as Neofusicoccum parvum, N. australe, and B. dothidea. N. parvum was the most prevalent (on 34% of the plants analyzed), followed by N. australe and B. dothidea (9% each). In phylogenetic analyses, isolates that clustered in the same group belonged to the same species with a high homogeneity index (>99%). One representative isolate of each species was selected for a pathogenicity assay. Amplified sequences from each selected isolate were deposited in GenBank with the following accession numbers: N. parvum, KC556958 (ITS) and KC556961 (EF); N. australe, KC556959 (ITS) and KC556962 (EF); and B. dothidea, KC556960 (ITS) and KC556963 (EF). The pathogenicity assay of these three isolates was conducted using two cultivars of southern highbush blueberry, ‘Misty’ and ‘Star.’ The isolates were cultured on acidified PDA at 25°C for 5 days. Stems of the plants were wounded at a height of 10 cm with a drill (5 mm diameter and ~4 mm deep). Six replicates per cultivar were inoculated per isolate by placing a colonized agar plug (4 to 5 mm diameter) in the hole and wrapping the stem with Parafilm. Plants treated identically with sterile agar plugs were used as controls. The plants were then maintained at 100% relative humidity for 2 h. This trial was conducted in a growth chamber at 28°C (night) and 30°C (day) with a 14-h photoperiod for 3 months. Disease was measured on a six-point scale: 0 = healthy plant; 1 = plant with a canker smaller than 3.5 cm; 2 = plant with a canker bigger than 3.5 cm; 3 = plant with one dry shoot; 4 = plant with some dry shoots; 5 = dead plant. At the end of the trial, disease was expressed as area under the disease progress curve. The results showed the N. parvum isolate to be the most aggressive, followed by the N. australe isolate. Espinoza et al. (1) also found that N. parvum showed more aggressiveness than N. australe on blueberries in Chile. B. dothidea was not pathogenic and behaved similarly to the controls (P < 0.05). Each pathogen was reisolated from all the inoculated plants, fulfilling Koch's postulates. To our knowledge, this is the first report of isolates of these pathogens, N. parvum and N. australe, causing stem canker and dieback on blueberry bushes in Spain. References: (1) J. G. Espinoza et al. Plant Dis. 93:1187, 2009. (2) A. J. L. Phillips et al. Mycol. 97:513, 2005. (3) T. J. White et al. Pages 315-322 in: PCR Protocols: a Guide to Methods and Amplifications. M. A. Innis et al., eds. Academic Press, San Diego, CA. 1990.

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