First Report of Canker Disease Caused by Diplodia olivarum on Carob Tree in Italy

The evergreen carob tree (Ceratonia siliqua L., Fabaceae), also called locust, is widespread in the Mediterranean Region. Carob pods have been traditionally consumed as animal and human food and seeds are mainly used in the pharmaceutical and cosmetic industries. In July 2009, symptoms of canker, branch dieback, and foliage reddening were observed on carob trees in several natural areas in the province of Ragusa, Italy. Disease incidence ranged from 5 to 80% across different sites and for most areas it was nearly 15%. All affected trees showed dark necrotic tissue in the bark, cambium, and sapwood of the trunk and branches. Cankers often girdled the stem or branch, causing wilting and death of the portions beyond the canker. Black, subepidermal pycnidia developed in and erupted through the dead bark. Fragments of discolored wood were collected from 36 symptomatic carob trees (12 trees for each area), transferred onto potato dextrose agar (PDA), and incubated for 5 days at 21°C in the dark. Fungal colonies were consistently obtained from these diseased tissues. They initially were pale, becoming gray-green and finally black. After 30 days of incubation at room temperature in the natural light, colonies produced pycnidia identical to those observed in nature. A total of 500 conidia on 10 isolates were examined with a compound microscope. Conidia were initially hyaline, smooth, oblong to ovoid, both ends rounded, and aseptate; at maturity they were pale brown, one-septate, and measured 24 to 28 × 10 to 13.5 μm (means ± S.D. = 24.3 ± 1.4 × 12.1 ± 1 μm, L/W = 2.0 ± 0.18). The nucleotide sequences of the β-tubulin (GenBank Accession No. HQ660080) and TE-1α (No. HQ660078) genes and ITS-rDNA region (No. HM028640) for a representative isolate (IMI 390972) from carob showed 100, 100, and 98% similarity, respectively, when compared with the sequences HQ660079, EU392279, and EU392302, respectively, of the ex-type isolate of Diplodia olivarum (strain CBS 121887). On the basis of morphological and molecular characters, the fungus was identified as D. olivarum A.J.L. Phillips, Frisullo & Lazzizera; teleomorph unknown (1). Two-year-old trees were wounded with a scalpel through the full thickness of the bark along 1-cm longitudinal direction and inoculated by applying a 5-mm-diameter plug of mycelial (isolate IMI 390972) on PDA to the wound site. Three control seedlings were similarly wounded and plugs of sterile PDA applied. Plugs were held in place by Parafilm. The inoculated seedlings were maintained at 20 to 22°C and a 12-h light/dark cycle. Sixty days after inoculation, all inoculated trees showed leaf chlorosis, sunken, necrotic bark at the inoculation sites and finally pycnidia of D. olivarum. All treated seedlings were killed within 6 months from the inoculation. No symptoms were observed in the control plants. The pathogen was consistently reisolated from all the inoculated trees, but not from the control plants. D. olivarum has been found on rotting olive drupes in Apulia (southern Italy) and was first described as a new species in 2008 (1). This fungal species could be phenotypically misidentified as the closely related species D. mutila, which differs by having larger mean dimensions of conidia. To our knowledge, this is the first report of D. olivarum inducing canker and dieback on carob tree. Reference: (1) C. Lazzizera et al. Fungal Divers. 31:63, 2008.

Download Full-text

First Report of a Branch Dieback of Olive Trees in Tunisia Caused by a Phoma sp.

Plant Disease ◽

10.1094/pdis-94-5-0636c ◽

2010 ◽

Vol 94 (5) ◽

pp. 636-636 ◽

Cited By ~ 7

Author(s):

A. Rhouma ◽

M. A. Triki ◽

S. Krid ◽

J. J. Tuset ◽

M. Msallem

Keyword(s):

Disease Incidence ◽

Olive Tree ◽

Conidial Suspension ◽

Internal Transcribed Spacer Region ◽

Canker Disease ◽

First Report ◽

Brown Discoloration ◽

Plastic Bag ◽

Olive Trees ◽

Branch Dieback

From 2007 to 2008, a new dieback of branches of olive trees was observed in several orchards in central and southern Tunisia. The appearance of this new syndrome coincided with warm temperatures and frequent rainfall from February to April 2007. Affected trees were observed in seven commercial orchards; disease incidence ranged from 1 to 9% and affected trees were randomly distributed in each orchard. Symptoms included abundant dead branches and wilted leaves remained attached. Distinct brown areas appeared on the bark of current-year shoots as well as on larger branches. Cankers on branches that were >2 years old were difficult to detect but were conspicuous in current-year shoots. To determine the etiology of this new syndrome, a study was carried out on samples of affected branches collected from 2007 to 2008 from different areas of the country. Unidentified species of Chaetomium and Phoma were isolated from the margins of the cankers. Koch's postulates were performed with one isolate each of a Chaetomium sp. and a Phoma sp on 2-year-old olive trees, cv. Chemlali, grown in 13-cm-diameter pots containing a sand/lime/peat mixture. Stems were inoculated by placing 10 μl of conidial suspension (106 conidia/ml) on 1-cm-long longitudinal stem wounds that had been made with a sterile scalpel. Control plants were wounded, but inoculum was replaced with sterile distilled water. Three sets of 10 plants each were wound inoculated with each of the fungi on the same day. Inoculated plants were covered with a polyethylene plastic bag to retain moisture and incubated for 2 months at 30°C with a 12-h photoperiod. After 45 days, only branches inoculated with the Phoma isolate showed brown discoloration areas at the inoculation sites. A Phoma sp. was recovered from necrotic bark from each of the 10 inoculated plants. Conidia were hyaline, unicellular, slightly ellipsoidal, and 4.8 to 6.3 × 1.8 to 2.2 μm. To confirm the identification, DNA extraction was done with hyphae collected from a 7-day-old culture on PDA after incubation at 25°C (1). Fungal primers ITS1 and ITS4 (3) were used for amplification. Purified amplicons were directly sequenced using the ITS1 and ITS4 primers for the internal transcribed spacer region of rDNA. A BLAST search of the GenBank database revealed 96% homology with Phoma sp. isolate (AJ972865.1) and 98% homology with Phoma medicaginis isolate (DQ026014.1). P. incompta has been reported as responsible for branch dieback of olive tree in Italy (2). To our knowledge, this is the first report of a canker disease of olive caused by a Phoma sp. in Tunisia. References: (1) S. R. Tendulkar et al. Biotechnol. Lett. 22:1941, 2003. (2) L. Tosi and A. Zazzerini. Petria 4:161, 1994. (3) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA, 1990.

Download Full-text

First Report of Botryosphaeria obtusa as Causal Agent of Olive Tree Branch Dieback in Tunisia

Plant Disease ◽

10.1094/pdis-01-12-0026-pdn ◽

2012 ◽

Vol 96 (6) ◽

pp. 905-905 ◽

Cited By ~ 4

Author(s):

M. Chattaoui ◽

A. Rhouma ◽

M. Msallem ◽

M. Pérez ◽

J. Moral ◽

...

Keyword(s):

Sodium Hypochlorite ◽

Olive Tree ◽

Fruit Rot ◽

Fluorescent Light ◽

Internal Transcribed Spacer Region ◽

Canker Disease ◽

First Report ◽

Botryosphaeria Obtusa ◽

Olive Trees ◽

Branch Dieback

A branch dieback of olive trees (Olea europaea L. cv. Manzanilla de Sevilla) was observed in 2010 in an orchard (50 ha), located in the Testour region of northern Tunisia. More than 50% of trees were severely damaged by the disease. Symptomatic trees presented dead branches and wilted leaves, which remained attached to the shoots, and the affected tissues appeared abnormally dark compared with the inner bark of healthy branches. Numerous pycnidia were observed on the surface of the infected branches. For diagnosis, symptomatic stems were collected and small pieces of discolored tissues were excised from lesion margins, surface sterilized in 0.5% sodium hypochlorite for 1 min, rinsed and dried on sterilized filter paper, then placed on acidified Difco potato dextrose agar plates (APDA; 2.5 ml of 25% lactic acid per liter). Plates were incubated at 25°C for 4 to 5 days, and hyphal tips from developing fungal colonies were transferred to PDA and placed under fluorescent light (12 h/day). A fastgrowing, pycnidia-producing fungus was consistently isolated, with conidia exuding onto the agar surface of 10-day-old cultures. On the basis of colony characteristics, isolates were identified as Botryosphaeria obtusa (3). Conidia were large, dark brown, aseptate, rounded at both ends or truncate at base, and 25 to 26.8 × 10.5 to 12.03 μm. Pathogenicity tests were performed on detached stems of cv. Manzanilla by 7-mm diameter mycelial plugs cut from actively growing cultures of the fungus. Stems (30 cm length) were cleaned, surface sterilized with sodium hypochlorite (0.25% for 2 min), and wounded with a sterilized scalpel. Mycelial disks were placed over wounds and wrapped with Parafilm to prevent desiccation. Control stems were mock inoculated with sterile agar plugs. Inoculated and control stems were placed in polyethylene boxes and incubated at 25°C. After 45 days, inoculated stems developed brown discoloration, and small dark pycnidia appeared on stem surfaces. Controls remained healthy. Koch's postulates were verified by isolating the fungus from symptomatic stems. To confirm the identification, DNA of one isolate was extracted and the fungal primers ITS1 and ITS4 (4) were used to amplify the internal transcribed spacer region of rDNA. Purified amplicons were sequenced and a BLAST search of the GenBank database revealed 99% homology with B. obtusa isolate HO166525.1. The anamorph of the fungus, Diplodia seriata, has been recognized as the cause of fruit rot of olive (1) and branch canker or dieback (2). To our knowledge, this is the first report of a canker disease of olive trees caused by B. obtusa in Tunisia. References: (1) J. Moral et al. Plant Dis. 92:311, 2008. (2) J. Moral et al. Phytopathology 100:1340, 2010. (3) A. Taylor et al. Australas. Plant Pathol. 34:187, 2005. (4) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. Academic Press, San Diego, CA, 1990.

Download Full-text

Seasonal disease: Foamy bark canker of Citrus maxima in the delta region of Tamil Nadu

INTERNATIONAL JOURNAL PLANT SCIENCES ◽

10.15740/has/ijps/16.1/7-14 ◽

2021 ◽

Vol 16 (1) ◽

pp. 7-14

Author(s):

G. Venkatesan ◽

P.S. Sharavanan

Keyword(s):

Tamil Nadu ◽

Infected Plant ◽

Fungal Species ◽

Delta Region ◽

Canker Disease ◽

Decay Branch ◽

Short Period ◽

Infected Area ◽

Citrus Maxima ◽

Branch Dieback

The Citrus maxima, commonly called pummelo, are a Rutaceae family. The Canker disease recently had an issue on Citrus species in the Delta region of Tamil Nadu. This disease is appeared by foamy oozes from the bark. The infected plant dies slowly in a short period. This study was identified the microorganism causes of foamy disease from bark and infected area. Totally 19 fungi were isolated. Among these 16 fungi were isolated from uninfected bark, 3 fungal species belonging to Ascomycetes, 2 fungal species belonging to Coelomycetes, and 10 species be classed Hyphomycetes and one sterile form, though 11 fungal species were isolated from infected bark foamy ooze, eight Hyphomycetes, one Oomycete, and two sterile forms were isolated. The RPO statistical analysis resulted, the bark fungi have been separated a group fungus from foamy fungi such a few fungi as the Fusarium, Phytophthora, and yeast have isolated in the foam. Also, the Jaccard’s similarity showed 42.105% and dissimilar among to the bark and foamy fungus. The plant was decay, branch dieback and tree death may induce by fungi also that the Canker disease on Citrus may be caused by Phytophthora fungal species.

Download Full-text

First Report of Fruit Rot and Associated Branch Dieback of Almond in California Caused by a Phomopsis Species Tentatively Identified as P. amygdali

Plant Disease ◽

10.1094/pdis.1999.83.11.1073c ◽

1999 ◽

Vol 83 (11) ◽

pp. 1073-1073 ◽

Cited By ~ 7

Author(s):

J. E. Adaskaveg ◽

H. Förster ◽

J. H. Connell

Keyword(s):

Disease Incidence ◽

Morphological Characteristics ◽

The United States ◽

Tree Canopy ◽

Fruit Rot ◽

Control Treatment ◽

Field Inoculation ◽

First Report ◽

Branch Dieback ◽

Summer Fruit

A fruit rot of almond (Prunus dulcis (Mill.) D. Webb.) was observed in an orchard in Durham, CA (Butte County), in June of 1998 after an unusually wet spring with a total precipitation of 17.2 cm for April and May. Disease incidence on fully developed fruit of almond cv. Sonora was nearly 90% in the lower tree canopy by July. Almond cv. Nonpareil grown in alternate rows in the same orchard was much less affected. Fruit symptoms included extensive grayish brown discolored and shriveled hulls, often associated with a clear gum secretion and shriveled kernels. Affected fruit frequently abscised. Leaf symptoms and branch dieback were not associated with the disease in 1998. In May of 1999, however, extensive twig dieback was observed on almond cv. Sonora in the same orchard. Isolations from more than 100 symptomatic fruit were conducted from 9 sampling sites in the 9-ha orchard. Based on morphological characteristics, the same fungus was isolated from 93% of the fruit. The fungus also was isolated consistently from samples exhibiting twig dieback. During a major disease survey conducted in 1998, the fungus was only incidentally isolated from almond fruit from other California orchards. Ascomata were not observed in vivo or in vitro. The fungus produced alpha and beta spores in pycnidia when cultured on potato dextrose agar. Spore measurements were obtained from 10 spores for each of 3 isolates obtained from fruit or twig dieback of almond cv. Sonora. Conidial dimensions of fruit and twig isolates were very similar. Based on spore sizes, with alpha spores measuring 5.3 to 7.5 (to 8) × 1.7 to 2.5 μm and beta spores measuring12.8 to 29.8 × 0.6 to 0.7 μm, the fungus was tentatively identified as Phomopsis amygdali (Del.) Tuset & Portilla (2). Previous reports on this fungus (2), however, indicated that beta spores are not produced in culture, and disease symptoms have not been observed on fruit. The fungus was morphologically different from other species of Phomopsis reported from almond and other Prunus species, including P. mali Roberts, P. padina (Sacc. & Roum.) Died., P. parabolica Petrak, P. perniciosa Grove, P. pruni (Ellis & Dearn.) Wehm., P. prunorum (Cooke) Grove, P. ribetejana Camara, and P. stipata (Lib.) Sutton (3). Field inoculation studies were performed in May of 1999 on almond cvs. Carmel and Mission. Almond fruit were wounded (2 × 2 × 2 mm) or left unwounded and were sprayed with water (control) or a suspension of alpha spores (105 spores per ml). Branches were bagged for 4 days to maintain high humidity. Fruit symptoms on cv. Carmel were observed after 4 weeks on wounded and nonwounded inoculated fruit, and P. amygdali was successfully reisolated from diseased tissue. No symptoms were observed in the control treatment for almond cv. Carmel or in any treatment for cv. Mission. This is the first report of P. amygdali causing a late spring and summer fruit rot and associated branch dieback of almond in North America (1). References: (1) D. F. Farr et al. 1989. Fungi on Plants and Plant Products in the United States. The American Phytopathological Society, St. Paul, MN. (2) J. J. Tuset and M. T. Portilla. Taxonomic status of Fusicoccum amygdali and Phomopsis amygdalina. Can. J. Bot. 67:1275, 1989. (3) F. A. Uecker. 1988. A World List of Phomopsis Names with Notes on Nomenclature, Morphology, and Biology. Mycologia Memoir No. 13. J. Cramer, Berlin.

Download Full-text

First Report of Branch Blight of Tree Peony Caused by Phoma glomerata in China

Plant Disease ◽

10.1094/pdis-12-12-1129-pdn ◽

2013 ◽

Vol 97 (8) ◽

pp. 1114-1114 ◽

Cited By ~ 4

Author(s):

D. Zhao ◽

Y. B. Kang

Keyword(s):

Disease Incidence ◽

Morphological Characteristics ◽

Aerial Mycelium ◽

Its2 Region ◽

Tree Peony ◽

Disease Symptoms ◽

First Report ◽

Plastic Bag ◽

Pure Cultures ◽

Branch Dieback

Tree peony (Paeonia suffruticosa Andrews) is a perennial woody deciduous shrub native to China and famous for its beautiful flowers. Starting in early autumn 2010, blighted branches of tree peony were detected in the International Peony Garden in Luoyang. The disease incidence was greater than 10% and disease symptoms included bulb atrophy and twig and branch dieback. Pycnidia were embedded within the bark of diseased branches. They were small, black, ostiolate, and measured 145 to 275 × 140 to 251 μm. Pycnoconidia were single-celled, hyaline or sandy beige, rounded to ellipsoidal, and 3.9 to 10.3 × 2.3 to 7.0 μm. Pure cultures were obtained by plating the pycnoconidia on potato dextrose agar (PDA). In culture, the fungus produced a circular, white to pink colony with pyknotic and linter shaped aerial mycelium. Numerous pycnidia, initially brown and dark at maturity, were embedded in the mycelium, especially in the center of the colony, with a few of them scattered in the edge. The morphological characteristics were consistent with Phoma (2). The ITS1-5.8S-ITS2 region of three isolates were PCR amplified and sequenced with primers ITS1 and ITS4. Sequences (GenBank Accession No. JX885584) showed 99% identity with reference isolates of Peyronellaea glomerata (Corda) Goid (AB470906.1 and HQ380779.1) and Phoma glomerata (Corda) Wollenw. & Hochapfel (EU098115.1). These two species are synonyms (1). To test pathogenicity, nine healthy branches of 3-year-old potted tree peony plants were wound-inoculated with a PDA disk containing pycnidia from an actively growing colony of P. glomerata. Three control branches were inoculated with sterile PDA disks. Each inoculated branch was wrapped in a plastic bag and maintained in a greenhouse at 25 to 28°C. After 3 days, brown patches appeared on inoculated branches and extended by up to 1 cm. Pycnidia identical to those observed in the field and in storage appeared on all inoculated branches 7 days after inoculation. Control branches did not show symptoms. The pathogen was reisolated from inoculated branches, fulfilling Koch's postulates. P. glomerata was reported as the causal agent of withering of flowers and young shoots of grapevines in Yugoslavia (3). To our knowledge, P. glomerata and Botryosphaeria dothidea have always been reported together, causing branch wilting or dieback. To our knowledge, this is the first report of branch blight of tree peony caused by P. glomerata in China. References: (1) M. M. Aveskamp et al. Mycol. Soc. Am. 101:363, 2009. (2) G. H. Boerema et al. Studies in Mycology, 3, 1973. (3) A. Šaric-Sabadoš et al. Atti Ist. bot. Univ. Pavia 18:101, 1960.

Download Full-text

First Report of Leaf Blight on Duying Caused by Phyllosticta anacardiacearum in China

Plant Disease ◽

10.1094/pdis-93-5-0546b ◽

2009 ◽

Vol 93 (5) ◽

pp. 546-546 ◽

Cited By ~ 2

Author(s):

B. G. Lou ◽

Y. D. Xu ◽

C. Sun ◽

X. M. Lou

Keyword(s):

Southern China ◽

Disease Incidence ◽

Its Region ◽

Culture Collection ◽

Leaf Blight ◽

Its Sequences ◽

Dead Leaf ◽

Conidial Suspension ◽

First Report ◽

Branch Dieback

Duying (Elaeocarpus glabripetalus Merr.; Elaeocarpaceae) is widely cultivated as an ornamental tree of commercial importance in southern China. From 2003 to 2008, severe outbreaks of Duying leaf blight occurred in the Hangzhou area, Zhejiang Province. Disease incidence was greater than 20% and mainly infected young leaves and shoots in the spring and autumn. Severely infected leaves and shoots died and eventually led to branch dieback. The overall growth decline of affected trees occurs over 4 to 6 years before tree death. Infection symptoms are characterized by grayish, round, semicircular- or irregular-shaped spots (5 mm to 5 cm long) with dark brown borders and the appearance of black, granular pycnidia within the dead leaf tissues. The primary infection zones are commonly observed on the leaf margins and apices, are brown, up to 2 mm in diameter, and often surrounded by a yellow zone. Pycnidia were globose and 122 to 127 μm (average 123.5 μm) in diameter. A fungus was consistently isolated from symptomatic tissues on potato dextrose agar (PDA). Ash-black pycnidia appeared on PDA after 10 days. Ascospores developed on modified PDA (1 liter of PDA + 20 g of Duying leaves) after 18 days. Conidiogenous cells were cylindrical to obpyriform. The hyaline conidia were obovoid and guttulate, 10 to 13 × 6 to 8 μm (average 11.5 × 7.5 μm), and usually surrounded by a mucilaginous sheath with a hyaline apical appendage that was 5 to 8 μm long. Pseudothecia were solitary and subglobose with long necks. Asci were 45 to 70 × 7.5 to 12 μm (average 62.5 × 10.8 μm). Ascospores were 12 to 13 × 4 to 5 μm with rounded apices and hyaline, mucilaginous, apical caps. The fungus was morphologically identified as Phyllosticta anacardiacearum van der Aa (teleomorph Guignardia mangiferae A. J. Roy). This identification was also confirmed by the China General Microbiological Culture Collection Center (CGMCC). Six representative fungal isolates were identified by sequencing the internal transcribed spacer (ITS) region of the rDNA and comparing the sequences with those in GenBank using BLAST searches. The ITS sequences of six cultures (GenBank Accession Nos. EU821356–EU821361) showed 100% identity with the ITS sequences of an isolate of a Phyllosticta sp. (GenBank Accession No. AF532314) (2) and G. mangiferae (GenBank Accession No. AY277717) (1). To fulfill Koch's postulates, a conidial suspension (106 conidia per ml) collected from PDA cultures (isolate phy01) was used to spray inoculate leaves of potted 3-year-old Duying trees. Inoculated trees were kept for 48 h under a polyethylene sheet cover and grown at 10 to 15°C in a greenhouse. A total of 30 leaves of five healthy trees were inoculated with the pathogen. In addition, five 3-year-old trees were sprayed with sterile water to serve as uninoculated controls. After 10 to 14 days, inoculated leaves showed infection symptoms resembling those observed on Duying trees naturally infected with P. anacardiacearum. The pathogen was reisolated from the margins of necrotic tissues, but not from controls. To our knowledge, this is the first report of leaf blight on E. glabripetalus caused by P. anacardiacearum in China. Reference: (1) F. R. Katia et al. Mycol. Res. 108:45, 2004. (2) A. K. Pandey et al. Mycol. Res. 107:439, 2003.

Download Full-text

First Report on the Occurrence of a New Pathotype, 714, of Plasmopara halstedii (Sunflower Downy Mildew) in Hungary

Plant Disease ◽

10.1094/pdis-02-14-0142-pdn ◽

2014 ◽

Vol 98 (11) ◽

pp. 1580-1580 ◽

Cited By ~ 4

Author(s):

R. Bán ◽

A. Kovács ◽

K. Körösi ◽

M. Perczel ◽

Gy. Turóczi

Keyword(s):

Downy Mildew ◽

Resistance Gene ◽

Disease Incidence ◽

Signs And Symptoms ◽

Weather Conditions ◽

Plasmopara Halstedii ◽

First Report ◽

Severe Stunting ◽

Plastic Bags ◽

Leaf Chlorosis

Downy mildew of sunflower, caused by Plasmopara halstedii (Farlow) Berlese et de Toni, is an economically important disease in Hungary and much of Europe. The known pathotypes (races) of the pathogen influence the resistance genes (Pl genes) incorporated into new sunflower hybrids to manage the disease. There are at least 36 pathotypes of P. halstedii worldwide (3), but the number of races is increasing rapidly. In 2010, race 704 was identified in Hungary for the first time (2). Race 704 has been reported to confer virulence on Pl6, a broad spectrum resistance gene that is widely used in sunflower hybrids. This has coincided with a significant increase in disease severity since 2010 in the country. Our objectives are to continuously monitor this pathogen and identify pathotypes of P. halstedii. Because of the unfavorable weather conditions for downy mildew in 2013, samples were collected at a single site (Kunszentmárton, South Hungary) in the beginning of July from NK Neoma sunflower hybrids. Disease incidence (early and late primary infection) was as high as 40%. Systemically mildewed plants showed severe stunting and leaf chlorosis, signs and symptoms consistent with downy mildew. P. halstedii was identified microscopically. Examination of isolates was carried out using a set of sunflower differential lines based on the internationally standardized method for race identification of P. halstedii (1). Inoculum of the isolates was increased on a susceptible cultivar (cv. Iregi szürke csíkos) and tested by inoculating 3-day-old seedlings of sunflower differential lines. Inoculated seedlings were planted in trays in glasshouse. After 8 to 9 days, seedlings were sprayed with distilled water, covered with black plastic bags, and left overnight to induce sporulation. Disease incidence was determined by examining cotyledons at 9 days after inoculation for sporulation and true leaves on 12 to 13 days after inoculation for secondary symptoms, such as leaf chlorosis and stunting (1). While several differential lines showed no typical susceptible/resistant reactions, i.e., the infection was much lower than 100%, it was concluded that the isolates were mixtures of different P. halstedii pathotypes. To obtain single isolates, we collected zoosporangia from the differential lines in question separately, and then inoculated the seedlings of the same genotype and a uniformly susceptible line. A single isolate caused as high as 100% infection on HA-335, containing resistance gene Pl6. Subsequent evaluation of this isolate with the entire differential set resulted in an aggregate virulence phenotype of 714. As resistance gene Pl6 is incorporated to the majority of sunflower hybrids grown in Hungary, pathotypes virulent on this gene, such as 704 and 714, are likely to spread. This underscores the need to prove the resistance to these races in the newly registered hybrids and for further research to identify P. halstedii pathotypes. It is also important to establish the identity of this new pathotype by already discovered 714 pathotypes in other countries like France and Italy and to discover the real conditions of local evolving of new pathogens. To our knowledge, this is the first report of pathotype 714 of P. halstedii in both Hungary and Central Europe. References: (1) T. J. Gulya et al. Helia 14:11, 1991. (2) K. Rudolf et al. Növényvédelem 47:279, 2011. (3) F. Virányi and O. Spring. Eur. J. Plant Pathol. 129:207, 2011.

Download Full-text

First Report of Broad Bean Canker Caused by Pestalotiopsis disseminata in India

Plant Disease ◽

10.1094/pdis.2001.85.2.229d ◽

2001 ◽

Vol 85 (2) ◽

pp. 229-229

Author(s):

N. Iboton Singh ◽

R. K. Tombisana Devi

Keyword(s):

Broad Bean ◽

Agricultural Research ◽

Disease Incidence ◽

Indian Agricultural Research Institute ◽

Culture Collection ◽

New Delhi ◽

Canker Disease ◽

First Report ◽

Plastic Bags ◽

Initial Symptoms

A canker disease of broad bean (Vicia faba L.) pods was observed annually from February to April every year since 1995. Disease incidence ranged from 45 to 100% in commercial fields and experimental plots at the Central Agricultural University, Imphal, Manipur. Initial symptoms on young pods appeared as bright yellow, scattered spots 1 to 2 mm in diameter that later became raised, black, and corky within 12 days. As the disease progressed, the lesions coalesced and formed large cankers, which resulted in unmarketable pods. Surface-sterilized symptomatic tissues were plated on potato-dextrose agar (PDA) and the fungus that grew out was identified as Pestalotiopsis disseminata. One culture was deposited at the Indian Type Culture Collection, Division of Plant Pathology, Indian Agricultural Research Institute, New Delhi. Pathogenicity of the fungus was determined by pipetting two drops of a 3 × 103 conidia/ml spore suspension on each of five healthy young pods and repeating on each of 10 plants. A similar five pods inoculated with sterile water served as controls. Inoculated broad bean plants were covered with plastic bags for 48 h and placed into the open under shade having a temperature range of 25 to 28°C. Corky lesions similar to those originally observed developed within 16 days on inoculated pods. Symptoms did not appear on control pods. Necrotic tissues from artificially induced broad bean cankers were plated on PDA and consistently yielded P. disseminata. This is the first report of P. disseminata on broad bean in India.

Download Full-text

First Report of Coffee Canker Disease Caused by Ceratocystis fimbriata in China

Plant Disease ◽

10.1094/pdis-08-21-1808-pdn ◽

2022 ◽

Author(s):

Kecheng Xu ◽

Ruiqi Zhang ◽

Haixia Lu ◽

Jinglong Zhang ◽

Jing Yang ◽

...

Keyword(s):

Disease Incidence ◽

Morphological Characteristics ◽

Molecular Characteristics ◽

Outer Cell ◽

Leaf Margin ◽

Malt Extract ◽

Distilled Water ◽

Ceratocystis Fimbriata ◽

Canker Disease ◽

First Report

Coffee (Coffea arabica L.) is one of the most important agricultural commodities in the world market. As an important cash crop in China, coffee is cultivated mainly in Yunnan and Hainan provinces. During October 2013 and September 2020, coffee trees showing typical dieback and wilt symptoms were found in Nanping town (N 22° 38', E 101° 0'), Pu’er, and Puwen town (N 22° 32', E 101° 4'), Xishuangbanna in Yunnan province, China. Symptomatic trees initially exhibited yellowing of foliage, expanding in size along the leaf margin, then became blighted and dry, and the internal xylem in main stem discolored brown to black. Infected trees eventually developed dieback and wilt. Disease incidence ranged from 10% to 22% and 25% to 40% of crown symptoms in the affected coffee trees. In extreme cases, 50% out of 380 trees were affected. The stems of coffee trees with typical symptoms were collected, and then the diseased tissues were surface disinfected with 75% ethanol for 30 s and 0.1% mercuric chloride (HgCl2) solution for 2 min, rinsed three times with sterile distilled water, plated onto potato dextrose agar (PDA) medium, and incubated at 25°C. After 6 days, fungal mycelium was observed growing from the tissue. Three isolates (C3-1, C3-2, and C3-2-1) were obtained by picking spore masses from the apices of perithecia and transferring them to malt extract agar (MEA) medium and incubated at 25°C for 10 days to observe the cultural features. In culture, colonies reaching 65 mm within 10 days, mycelium initially white, then becoming light blue-green. After 6 days of formation, perithecia were black, globose (123.8 - 173.4 μm × 138.2 - 180.6 μm), and showed a long black neck (414.2 - 650.0 μm). Ascospores with outer cell wall forming a brim, hat-shaped, accumulating in a mucilaginous mass at the tips of ostiolar hyphae (4.3 μm × 6.0 μm). Cylindrical endoconidia (14.1 - 45.2 μm × 3.5 - 5.7 μm) were hyaline. Chain of barrel-shaped conidia (6.6 - 10.2 μm × 6.8 - 8.8 μm) were found. Aleuroconidia (10.8 - 16.9 μm × 9.1 - 13.0 μm) were olive-brown, ovoid or obpyriform, and smooth. Morphological characteristics of the fungus were consistent with the description of Ceratocystis fimbriata Ellis & Halst. (Engelbrecht and Harrington 2005). The three isolates were used for molecular identification, and their genomic DNA was extracted using the chelex-100 method (Xu et al. 2020). The internal transcribed spacer (ITS) region of rDNA was sequenced using the procedures of Thorpe et al. (2005). Analysis of the ITS sequence data (GenBank accessions KY580836, KJ511480, and KJ511479) showed that the isolates were 100% homologous to isolates of C. fimbriata from Punica granatum, Camellia sinensis, and Cucumis sativus in China (GenBank accessions KY580891, KY580870, and MH535909, respectively) by BLAST analysis. Neighbor-joining (NJ) phylogenetic analysis was performed using MEGA 6.06 based on the ITS sequences. The three isolates were clustered on the same clade with other C. fimbriata isolates with a high bootstrap value (90%). Therefore, the fungus was identified as C. fimbriata based on both morphological and molecular characteristics. Pathogenicity of the three isolates was tested by inoculating one-year-old pot grown coffee seedlings (C. arabica) through drenching the loams with 30 ml spore suspension (1 × 106 spores/ml). Control plants were inoculated with 30 ml of sterile distilled water. The trees were kept in a controlled greenhouse at 25°C and watered weekly. One month after inoculation, all inoculated plants produced typical dieback and wilt symptoms, whereas the control trees showed no symptoms. The same fungus was isolated from the inoculated trees on PDA and identified as C. fimbriata according to the methods described above, and no fungal growth was observed in the controls, thus fulfilling the Koch's postulates. Coffee canker disease caused by C. fimbriata has been reported in Indonesia and Colombia (Marin et al. 2003). To our knowledge, this is the first report of C. fimbriata causing canker disease of coffee trees in China.

Download Full-text

First Report of Fusarium avenaceum Causing Canker Disease on Kiwi Tree in China

Plant Disease ◽

10.1094/pdis-11-21-2459-pdn ◽

2022 ◽

Author(s):

Qing Sun ◽

Yongjing Xie ◽

Tangmin Chen ◽

Jianping Zhang ◽

Pedro Laborda ◽

...

Keyword(s):

Fungal Species ◽

Kiwi Fruit ◽

Canker Disease ◽

Molecular Phylogenetic Tree ◽

Three Samples ◽

Molecular Phylogenetic ◽

Symptomatic Tissue ◽

White Mycelium ◽

Yellow Pigmentation ◽

Old Trees

In May 2021, canker symptoms were detected on ‘Xuxiang’ kiwi trees in southwestern Shaanxi (Hanzhong municipality; 107.27° E, 33.23° N) in China. Seven-year-old trees exhibited black necrotic lesions and cracked areas in the trunk (Figure 1). The symptoms were observed in approximately 10% of the trees in 6 orchards (31 ha in total). Application of commercial fungicides did not control the advancement of the pathogen, and infected trees were removed to control the spread. Three samples, approximately 1 cm2 in size, of symptomatic tissue were collected and surface sterilized in 2% NaOCl for 1 min, and washed with sterile ddH2O. Four isolates showing white mycelium with yellow pigmentation were obtained after 4 days of incubation on PDA, containing chloramphenicol (50 µg/mL), at 28 ºC. The pathogen was isolated from all collected samples. ITS, EF1-α, TUB2, RPB1 and RPB2 genes were amplified using ITS1/ITS4, EF1-728F/EF1-986R, T1/T22, RPB1-5F/RPB1-8R and RPB2-5F/RPB2-7cR (strain NJC06), or RPB2-c7F/RPB2-11aR (strains NJC07 and NJC08), primers, respectively. Two isolates shared the same sequences (strain NJC08). Obtained sequences were submitted to GenBank under accession numbers MZ669205 and OL347898-OL347899 (ITS), OL439731-OL439733 (EF1-α), OL439734-OL439736 (TUB2), OL439737-OL439739 (RPB1), and OL439740-OL439742 (RPB2). The sequences shared >99% (ITS; F. avenaceum CBS 128538, MH864972), >99% (EF1-α; F. avenaceum 55-2, MN473124), 100% (TUB2; F. avenaceum SICAUCC 18-0001, MK253102), >98% (RPB1; F. avenaceum NRRL 26911, MG282372), and >98% (RPB2; F. avenaceum SICAUCC 18-0001, MK396098; or F. avenaceum FRC R-09495, CQ915486) homology to multiple F. avenaceum strains. Molecular phylogenetic tree (Figure 2) was constructed using MEGA7 with Fusarium strains found causing rot in various hosts (Wang et al. 2015), and other fungal species, such as Cadophora nalorum, Diaporthe ambigua, D. australafricana, and Neofusicoccum parvum, which were reported to cause cordon dieback on kiwi tree in Chile (Diaz et al. 2021). Microscope observations after cultivation of all isolates on barley-honey-tryptone medium (Song et al. 2020) showed the presence of septate mycelium, fusiform microconidia (8-15 µm in length, containing between 0 and 3 septa; n = 77) and chlamydospores (n = 21), and agree with the morphology of F. avenaceum (Zhao et al. 2020). To confirm pathogenicity, a sterilized spatula was used to make wounds (3 mm diameter, 1 mm depth) on the trunk of 3-months-old ‘Xuxiang’ kiwi trees. Solutions containing 1 × 106 spores/mL (20 µL) of the isolates were injected in the wounds. Sterile ddH2O was used for the control experiment. Inoculated plants were maintained in a growth chamber at 28 °C and 80% relative humidity for 4 days. The pathogen was recovered from the canker lesions, which were similar to those observed in the orchards, and its identity was confirmed by sequence analysis. The pathogen only infected wounded trees, and probably invaded the orchards during the pruning in February 2021. F. avenaceum was reported to cause canker on almond tree (Stack et al. 2020), stem rot on Anthoxanthum aristatum and Polygonatum cyrtonema (Pieczul et al. 2018; Xu et al. 2019), and root rot on carrot, Coptis chinensis and wheat (Le Moullec-Rieu et al. 2020; Mei et al. 2020; Ozer et al. 2020). Recently, F. avenaceum was found causing fruit blotch in kiwi fruit in Anhui (China) (Zhao et al. 2020). Here, F. avenaceum was found causing canker disease in kiwi tree, demonstrating the host and tissue promiscuity of this pathogen. Kiwi is an important crop in China with nearly 1.5 million tons produced in 2019. This report will help to better understand the pathogens reducing kiwi production in China.

Download Full-text