First Report of Brown Rot Caused by Monilinia fructicola on Apple in Italy

Monilinia fructicola (G. Wint.) Honey, the causal agent of brown rot, is one of the most important fungal pathogens of stone fruit but may also affect pome fruits. The pathogen is common in North America, Oceania, South America, and Asia. It is a quarantined pathogen in Europe (3), but was recently detected in apple from the Czech Republic, Germany, and Serbia (1,2,4). In January 2012, during a survey for fungal postharvest pathogens, stored apple (Malus domestica Borkh.) belonging to the cultivars Gala and Pink Lady showing brown rot symptoms were observed in the Emilia Romagna region, Italy. Typical decay spots were circular and brown, tending toward black. Decayed tissues remained firm, and numerous grayish pustules containing spores appeared on rotted areas. The pathogen was isolated on V8 juice agar and culture plates were incubated at 25°C in darkness for 5 days. A conidial suspension was spread on malt extract agar and single spores were selected. The colonies were morphologically identified as M. fructigena. Two colonies developing a gray mass of spores in concentric rings with the reverse side black were further studied by molecular tools. The colony margins were even and the conidia were one-celled, limoniform, hyaline, and 12.1 to 17.4 × 8.4 to 11.2 μm. The ribosomal ITS1-5.8S-ITS2 region was PCR-amplified from genomic DNA obtained from mycelium using primers ITS1 and ITS4. A BLAST search in GenBank revealed the highest similarity (99%) to M. fructicola sequences (GenBank Accession Nos. HQ893748.1 and FJ515894.1). Pathogenicity was confirmed using surface-sterilized mature ‘Gala’ apples, wounded with a sterile needle, and inoculated with an isolate conidial suspension (103 spores/ml). A 20 μl droplet was placed in the wound; control fruits received sterile water without conidia. After 5 days of incubation at 20°C in plastic containers with high humidity, typical symptoms of brown rot developed on inoculated fruits, while control fruits remained symptomless. The fungus isolated from inoculated fruit exhibited the same morphological and molecular features shown by the original isolates. To our knowledge, this is the first report of the fungus M. fructicola on apple in Italy. Further studies are necessary to determine geographic distribution, prevalence and economic importance of this quarantine organism in Italy. References: (1) J. Duchoslavovà et al. Plant Dis.91:907, 2007. (2) A. Grabke et al. Plant Dis. 95:772, 2011. (3) OEPP/EPPO. EPPO A2 list of pests recommended for regulation as quarantine pests. Version 2010-09. Retrieved from http://www.eppo.int/QUARANTINE/listA2.htm , 2010. (4) M. Vasic et al. Plant Dis. 96:456, 2012.

Download Full-text

First Report of Brown Rot of Apple Caused by Monilinia fructicola in Germany

Plant Disease ◽

10.1094/pdis-02-11-0113 ◽

2011 ◽

Vol 95 (6) ◽

pp. 772-772 ◽

Cited By ~ 14

Author(s):

A. Grabke ◽

M. J. Hu ◽

C. X. Luo ◽

P. K. Bryson ◽

G. Schnabel

Keyword(s):

Brown Rot ◽

The State ◽

Its2 Region ◽

Monilinia Fructicola ◽

Conidial Suspension ◽

First Report ◽

Plastic Bags ◽

Sterile Needle ◽

Average Size ◽

Equidistant Points

Monilinia fructicola (G. Wint.) Honey is a causal agent of brown rot of stone fruits but may also affect pome fruits. M. fructicola is common in North America, Oceania, and South America as well as in Asia, but it is listed as a quarantine pathogen in Europe (3). Since its first discovery in Europe in 2001 (France), it has been reported in Spain, Slovenia, Italy, and Switzerland. Recently, the fungus was also detected in orchards of blackberries and plums in the State of Baden-Württemberg, Germany (4). In July 2010, apples (Malus domestica Borkh.) of the cultivar Jonagold were found in a residential backyard in Fronhausen an der Lahn located in the State of Hessen, Germany with symptoms resembling brown rot caused by Monilinia species. Affected apples were at or near maturity with brown decay that had spread throughout the fruits. On the surface of the decaying apples was tan to white zones of sporulation. Upon isolation, the mycelium grew at a linear rate of 9.2 mm per day at 22°C on potato dextrose agar forming branched, monilioid chains of grayish colonies with concentric rings and little sporulation. The lemon-shaped spores had an average size of 14 × 9 μm, a shape and size consistent with M. fructicola. The ribosomal ITS1-5.8S-ITS2 region was PCR-amplified from genomic DNA obtained from mycelium using primers ITS1 and ITS4. A BLAST search in GenBank revealed highest similarity (99%) to M. fructicola sequences from isolates collected in China, Italy, and Slovenia (GenBank Accession Nos. FJ515894.1, FJ411109.1, GU967379.1). The M. fructicola sequence from the apple isolate was submitted to GenBank (Accession No. JF325841). The pathogen was also identified to the species level and confirmed to be M. fructicola using two novel PCR techniques based on cytochrome b sequences (1,2). Pathogenicity was confirmed by inoculating three surface-sterilized, mature apples cv. Gala with a conidial suspension (105 spores/ml) of the apple isolate. Fruit were stab inoculated at three equidistant points to a depth of 10 mm using a sterile needle. A 30-μl droplet was placed on each wound; control fruit received sterile water without conidia. After 5 days of incubation at room temperature in air-tight plastic bags, the inoculated fruits developed typical brown rot symptoms with sporulating areas (as described above). The developing spores on inoculated fruit were confirmed to be M. fructicola. All control fruits remained healthy. To our knowledge, this is the first report of M. fructicola on apple in Germany and more indication of further geographical spread of the quarantine disease in Germany. References: (1) J.-M. Hily et al. Pest Manag. Sci. Online publication. doi 10.1002/ps.2074, 2011. (2) S. Miessner and G. Stammler. J. Plant Dis. Prot. 117:162, 2010. (3) OEPP/EPPO. EPPO A2 list of pests recommended for regulation as quarantine pests. Version 2009-09. Retrieved from http://www.eppo.org/QUARANTINE/listA2.htm , September 22, 2010. (4) OEPP/EPPO. Reporting Service. No. 1, January 2010. Retrieved from http://archives.eppo.org/EPPOReporting/2010/Rse-1001.pdf , September 22, 2010.

Download Full-text

First Report of Asiatic Brown Rot Caused by Monilinia polystroma on Peach in Italy

Plant Disease ◽

10.1094/pdis-05-14-0551-pdn ◽

2014 ◽

Vol 98 (11) ◽

pp. 1585-1585 ◽

Cited By ~ 13

Author(s):

C. Martini ◽

A. Lantos ◽

A. Di Francesco ◽

M. Guidarelli ◽

S. D'Aquino ◽

...

Keyword(s):

Czech Republic ◽

Brown Rot ◽

Fruit Trees ◽

Economic Losses ◽

Its2 Region ◽

Malt Extract ◽

Conidial Suspension ◽

The Czech Republic ◽

First Report ◽

Plant Pathol

Monilinia spp. are well-known pathogens causing brown rot of fruit trees in many fruit production areas worldwide. In Italy, three Monilinia species are particularly significant with regard to fruit trees, causing blossom and twig blight and brown rot in fruits: Monilinia laxa (Aderhold and Ruhland) Honey, M. fructicola (Winter) Honey, and M. fructigena (Aderhold and Ruhland). In 2009, a new species, M. polystroma, was distinguished from M. fructigena based on morphological and molecular characteristics in Europe (3). M. polystroma is not known to occur in Italy and to date has been reported from the Czech Republic (1), Hungary (3), Poland (4), Serbia (5), and Switzerland (2). In July 2013, during a survey for fungal postharvest pathogens, stored peaches (Prunus persica (L.) Batsch) belonging to different cultivars showing brown rot symptoms were observed in the Emilia Romagna and Sardinia regions of Italy. Typical decay spots were circular and brown, tending toward black, and 5% of peaches presented a large number of yellowish or buff-colored stromata and firm decayed tissues, the symptoms originated by M. polystroma. The pathogen was isolated on V8 agar (V8A) and culture plates were incubated at 25°C in darkness for 5 days. A conidial suspension was spread on malt extract agar (MEA) and single spores were selected. M. polystroma colonies grown on potato dexstrose agar (PDA) were yellowish in color. Irregular black stromatal crusts occurred on the edges of the colonies after 10 to 12 days of incubation and on the margin was present sporogenous tissue slightly elevated above the colony surface, color buff/pale luteous (1). The conidia were one-celled, ovoid or limoniform, smooth and hyaline, and 12 to 20 × 8 to 12 μm in distilled water when grown on V8A at 22°C. The ribosomal ITS1-5.8S-ITS2 region was PCR-amplified from genomic DNA obtained from mycelium using primers ITS1 and ITS4. A BLAST search in GenBank revealed the highest similarity (99%) to M. polystroma sequences (GenBank Accession No. GU067539). Pathogenicity was confirmed using surface-sterilized mature ‘Red Heaven’ peaches. The fruits were wounded (2 × 2 × 2 mm) twice with a sterile needle and inoculated with 2-mm plugs of 7-day-old mycelia from fungal colony margins. The sample unit was represented by 10 fruits. Control fruits were inoculated with PDA. After 7 days of incubation at 20°C in plastic containers with high humidity, typical symptoms of brown rot developed on both the wounds of all inoculated fruits, while control fruits remained symptomless. By the 14th day, all fruits had rotted and the yellowish exogenous stromata appeared on the surface of infected peaches. The fungus isolated from inoculated fruit exhibited the same morphological and molecular features of the original isolates; the molecular analysis performed using the primers by Petroczy (3) confirmed the result of the PCR with ITS1 and ITS4 primers. To our knowledge, this is the first report of M. polystroma on peach in Italy. This is relevant because the new pathogen could spread into other European countries that are main peach producers (such as Spain), causing economic losses. Bringing it to the attention of the scientific community allows the arrangement of research studies for assessing potential resistances with a significant impact on disease control management. Further studies are necessary to determine geographic distribution, prevalence, and economic importance of this organism in Italy. References: (1) EPPO Reporting Service. 2011/134: First reports of Monilinia polystroma in Hungary and the Czech Republic. No. 6, 2011. (2) M. Hilber-Bodmer et al. Plant Dis. 96:146, 2012. (3) M. Petroczy and L. Palkovics. Eur. J. Plant Pathol. 125:343, 2009. (4) A. Poniatowska et al. Eur. J. Plant Pathol. 135:855, 2013. (5) M. Vasic et al. Plant Dis. 97:145, 2013.

Download Full-text

First Report of Monilinia fructicola Causing Brown Rot on Stored Japanese Plum Fruit in Chile

Plant Disease ◽

10.1094/pdis-06-13-0647-pdn ◽

2014 ◽

Vol 98 (1) ◽

pp. 160-160 ◽

Cited By ~ 3

Author(s):

B. A. Latorre ◽

G. A. Díaz ◽

A. L. Valencia ◽

P. Naranjo ◽

E. E. Ferrada ◽

...

Keyword(s):

San Francisco ◽

Brown Rot ◽

Soft Rot ◽

The United States ◽

Its2 Region ◽

Monilinia Fructicola ◽

Conidial Suspension ◽

First Report ◽

Japanese Plum ◽

Official Control

In autumn 2013, fruit of Japanese plum (Prunus salicina) cvs. Angelino and Black Kat developed an unusual brown and soft rot after 2 months in cold storage (0°C) on nearly 1% of the fruit. Fruit showed small, circular, light brown spots that eventually destroyed the entire fruit. Small sporodochia appeared on the fruit surface. Fruit was harvested from orchards located near San Francisco de Mostazal (33°59′ S, 70°41′ W), Chile. Small pieces of diseased tissue were selected from margins of lesions of surface disinfected (96% ethanol) fruit (n = 7) and placed on acidified potato dextrose agar (PDA) plates for 5 days at 20°C. Light brown colonies with even margins and concentric rings of spores were obtained. The conidia of five isolates were one-celled, hyaline, lemon-shaped, (min. 10.7) 14.9 ± 1.5 (max. 18.6) × (min. 8.1) 9.4 ± 0.8 (max. 10.8) μm (n = 30), and borne in branched monilioid chains. This fungus was identified as Monilinia fructicola (G. Winter) Honey (1). Identification was confirmed by amplifying and sequencing the ribosomal ITS1-5.8S-ITS2 region using ITS1 and ITS4 primers (3). BLAST analysis of Chilean plum isolates (GenBank Accession Nos. KF148610 and KF148611) were 99 to 100% identical to isolates of M. fructicola originating from the United States (DQ314727 and HQ846966, respectively) and 100% identical to the first Chilean isolate (JN001480) found in nectarines originating from California at the supermarkets in Santiago in June 2009. Koch's postulates were fulfilled by reproducing brown rot symptoms on mature wounded Japanese plums cv. Angelino (n = 8) inoculated with 10 μl of a conidial suspension (105 conidia/ml) or with a mycelium plug (5-mm diameter). After 2 days in humid chambers (>80% relative humidity) at 25°C, all inoculated fruit developed brown rot symptoms with necrotic lesion means of 15.8 and 21.5 mm in diameter in fruit inoculated with conidia and mycelium, respectively. Non-inoculated control fruit remained healthy. Re-isolations were performed on PDA and the presence of M. fructicola was morphologically confirmed in 100% of the symptomatic fruits. To our knowledge, this is the first report demonstrating the presence of M. fructicola causing brown rot in stored Japanese plums in Chile after its first interception in 2009 in Chile, suggesting that this pathogen has been established in the field. Currently, M. fructicola is a quarantine organism under official control, restricted to Prunus orchards between Santiago and Nancagua in central Chile (2). References: (1) EPPO. EPPO Bull. 39:337, 2009. (2) Servicio Agrícola y Ganadero, SAG, Ministerio de Agricultura, Gobierno de Chile. www.sag.cl , accessed 15 November 2013. (3) T. J. White et al. Page 315 in: PCR Protocols: A Guide to Methods and Applications. M. A. Innis et al., eds. Academic Press, NY, 1990.

Download Full-text

First Report of Brown Rot Caused by Monilinia fructicola on Stored Apple in Serbia

Plant Disease ◽

10.1094/pdis-06-11-0531 ◽

2012 ◽

Vol 96 (3) ◽

pp. 456-456 ◽

Cited By ~ 10

Author(s):

M. Vasić ◽

N. Duduk ◽

M. M. Ivanović ◽

A. Obradović ◽

M. S. Ivanović

Keyword(s):

Online Publication ◽

Slovak Republic ◽

Brown Rot ◽

Pathogenic Fungi ◽

Its2 Region ◽

Morphological Identification ◽

Monilinia Fructicola ◽

First Report ◽

Molecular Features ◽

Quarantine Organism

Monilinia fructicola (G. Winter) Honey is a causal agent of brown rot of stone fruits, occasionally affecting pome fruits as well. The pathogen is commonly present in North and South America, Oceania, and Asia, but listed as a quarantine organism in Europe (4). After its first discovery in France in 2001, its occurrence has been reported in Germany, Hungary, Italy, Poland, Romania, Slovenia, Spain, Switzerland, Austria, and the Slovak Republic (1). In February 2011, during a survey for fungal postharvest pathogens in cold storage conditions, apple fruits (Malus domestica Borkh.) grown and stored in the Grocka Region, Serbia, were collected. All pathogens from symptomatic fruits were isolated on potato dextrose agar (PDA). One isolate from apple fruit cv. Golden Delicious with brown rot symptoms was identified as M. fructicola based on morphological and molecular characters. Colonies cultivated on PDA at 22°C in darkness were colorless, but later became grayish, developing mass of spores in concentric rings. Colony margins were even. Conidia were one-celled, limoniform, hyaline, measured 12.19 to 17.37 (mean 13.8) × 8.62 to 11.43 μm (mean 9.9), and were produced in branched monilioid chains (3). Morphological identification was confirmed by PCR (2) using genomic DNA extracted from the mycelium of pure culture, and an amplified product of 535 bp, specific for the species M. fructicola, was obtained. Sequence of the ribosomal (internal transcribed spacer) ITS1-5.8S-ITS2 region was obtained using primers ITS1 and ITS4 and deposited in GenBank (Accession No. JN176564). Control fruits were inoculated with sterile PDA plugs. After 3 days of incubation in plastic containers with high humidity at room temperature, typical symptoms of brown rot developed on inoculated fruits, while control fruits remained symptomless. The isolate recovered from symptomatic fruits showed the same morphological and molecular features of the original isolate. To our knowledge, this is the first report of M. fructicola in Serbia. Further studies are necessary for estimation of economic importance and geographic distribution of this quarantine organism in Serbia. References: (1) R. Baker et al. European Food Safety Authority. Online publication. www.efsa.europa.eu/efsajournal . EFSA J. 9(4):2119, 2011. (2) M.-J. Côté et al. Plant Dis. 88:1219, 2004. (3) J. E. M. Mordue. CMI Descriptions of Pathogenic Fungi and Bacteria. No. 616, 1979. (4) OEPP/EPPO. EPPO A2 List of Pests Recommended for Regulation as Quarantine Pests. Online publication. Version 2010-09. Retrieved from http://www.eppo.org/QUARANTINE/listA2.htm , June 27, 2011.

Download Full-text

First Report of Brown Rot Caused by Monilinia fructicola Affecting Peach Orchards in Slovenia

Plant Disease ◽

10.1094/pdis-94-9-1166a ◽

2010 ◽

Vol 94 (9) ◽

pp. 1166-1166 ◽

Cited By ~ 15

Author(s):

A. Munda ◽

M. Viršček Marn

Keyword(s):

Prunus Persica ◽

Brown Rot ◽

Fruit Rot ◽

Morphological Identification ◽

Monilinia Fructicola ◽

Conidial Suspension ◽

Stone Fruits ◽

The European Union ◽

First Report ◽

Representative Isolate

Monilinia fructicola, the causal agent of brown rot, is a destructive fungal pathogen that affects mainly stone fruits (Prunoideae). It causes fruit rot, blossom wilt, twig blight, and canker formation and is common in North and South America, Australia, and New Zealand. M. fructicola is listed as a quarantine pathogen in the European Union and was absent from this region until 2001 when it was detected in France. In August 2009, mature peaches (Prunus persica cv. Royal Glory) with brown rot were found in a 5-year-old orchard in Goriška, western Slovenia. Symptoms included fruit lesions and mummified fruits. Lesions were brown, round, rapidly extending, and covered with abundant gray-to-buff conidial tufts. The pathogen was isolated in pure culture and identified based on morphological and molecular characters. Colonies on potato dextrose agar (PDA) incubated at 25°C in darkness had an average daily growth rate of 7.7 mm. They were initially colorless and later they were light gray with black stromatal plates and dense, hazel sporogenous mycelium. Colony margins were even. Sporulation was abundant and usually developed in distinct concentric zones. Limoniform conidia, produced in branched chains, measured 10.1 to 17.7 μm (mean = 12.1 μm) × 6.2 to 8.6 μm (mean = 7.3 μm) on PDA. Germinating conidia produced single germ tubes whose mean length ranged from 251 to 415 μm. Microconidia were abundant, globose, and 3 μm in diameter. Morphological characters resembled those described for M. fructicola (1). Morphological identification was confirmed by amplifying genomic DNA of isolates with M. fructicola species-specific primers (2–4). Sequence of the internal transcribed spacer (ITS) region (spanning ITS1 and ITS 2 plus 5.8 rDNA) of a representative isolate was generated using primers ITS1 and ITS4 and deposited in GenBank (Accession No. GU967379). BLAST analysis of the 516-bp PCR product revealed 100% identity with several sequences deposited for M. fructicola in NCBI GenBank. Pathogenicity was tested by inoculating five mature surface-sterilized peaches with 10 μl of a conidial suspension (104 conidia ml–1) obtained from one representative isolate. Sterile distilled water was used as a control. Peaches were wounded prior to inoculation. After 5 days of incubation at room temperature and 100% relative humidity, typical brown rot symptoms developed around the inoculation point, while controls showed no symptoms. M. fructicola was reisolated from lesion margins. Peach and nectarine orchards in a 5-km radius from the outbreak site were surveyed in September 2009 and M. fructicola was confirmed on mummified fruits from seven orchards. The pathogen was not detected in orchards from other regions of the country, where only the two endemic species M. laxa and M. fructigena were present. To our knowledge, this is the first report of M. fructicola associated with brown rot of stone fruits in Slovenia. References: (1) L. R. Batra. Page 106 in: World Species of Monilinia (Fungi): Their Ecology, Biosystematics and Control. J. Cramer, Berlin, 1991. (2) M.-J. Côté et al. Plant Dis. 88:1219, 2004. (3) K. J. D. Hughes et al. EPPO Bull. 30:507, 2000. (4) R. Ioos and P. Frey. Eur. J. Plant Pathol. 106:373, 2000.

Download Full-text

First Report of Phaeoacremonium mortoniae Associated with Grapevine Decline in Iran

Plant Disease ◽

10.1094/pdis-05-11-0361 ◽

2011 ◽

Vol 95 (8) ◽

pp. 1034-1034 ◽

Cited By ~ 6

Author(s):

H. Mohammadi

Keyword(s):

Fungal Pathogens ◽

Vascular Tissue ◽

Sodium Hypochlorite Solution ◽

Malt Extract ◽

Fars Province ◽

Distilled Water ◽

Conidial Suspension ◽

Single Spore ◽

First Report ◽

Grapevine Decline

In July 2009, a survey was conducted in individually owned rooted vineyards in Iran to determine fungal pathogens associated with grapevine decline. Symptoms of grapevine decline such as slow dieback, stunted growth, small chlorotic leaves, and reduced foliage were observed on 7-year-old grapevines (cv. Askari) in Bavanat (Fars Province, southwestern Iran). Internal wood symptoms such as black spots and dark brown-to-black vascular streaking were observed in cross and longitudinal sections of stems and trunks. Wood samples were collected from symptomatic trunks and cordons. The bark of each fragment was removed and 10 thin cross sections (2 to 3 mm thick) were cut from symptomatic vascular tissue of the samples. These disks were immersed in 1.5% sodium hypochlorite solution for 4 min, washed thrice with sterile distilled water, and plated onto malt extract agar (MEA) supplemented with 100 mg liter–1 of streptomycin sulfate. Plates were incubated at 25°C in darkness. All colonies were transferred to potato dextrose agar (PDA) and incubated at 25°C. Five isolates of a Phaeoacremonium sp. were obtained. Single-spore isolates were transferred to PDA, MEA, and oatmeal agar (OA) media and incubated at 25°C for 8 to 16 days in the dark (2). Colonies reached a radius of 9.5 to 12 mm after 8 days of incubation. Colonies were flat and yellowish white on PDA and OA and white-to-pale gray after 16 days of incubation on MEA. Conidiophores were short and unbranched, 14 to 38.5 (23.5) μm long, and often ending in a single terminal phialide. Phialides were terminal or lateral and mostly monophialidic. Conidia were hyaline, oblong to ellipsoidal or reniform, 2 to 6.5 (4.9) μm long, and 1.1 to 1.7 (1.4) μm wide. On the basis of these characteristics, the isolates were identified as Phaeoacremonium mortoniae (1,2). Additionally, identity of the PMH1 isolate was confirmed by sequencing a fragment of the -tubulin gene with primers T1 and Bt2b (GenBank Accession No. JF831449). The sequence of this isolate was identical to the sequence of P. mortoniae (GenBank Accession No. HM116767). Pathogenicity tests were conducted on 2-month-old grapevine seedlings of cv. Askari by watering the roots with 25 ml of a conidial suspension (107 conidia ml–1) harvested from 21-day-old cultures grown on MEA. Controls were inoculated with 25 ml of sterile distilled water. Fifteen replicates were used for each isolate with an equal number of noninoculated plants. All plants were grown under greenhouse conditions (25 to 30°C). Two months after inoculation, inoculated seedlings showed reduced growth, chlorotic leaves, epinasty, severe defoliation, and finally wilting, while control seedlings remained healthy. The fungus was reisolated from internal tissues of the stems of inoculated seedlings. To my knowledge, this is the first report of P. mortoniae causing grapevine decline in Iran. References: (1) M. Groenewald et al. Mycol. Res. 105:651, 2001. (2) L. Mostert et al. Stud. Mycol. 54:1, 2006.

Download Full-text

First Report of Peach Brown Rot Caused by Monilinia fructicola in Central and Western China

Plant Disease ◽

10.1094/pdis-03-13-0310-pdn ◽

2013 ◽

Vol 97 (9) ◽

pp. 1255-1255 ◽

Cited By ~ 2

Author(s):

L. F. Yin ◽

S. N. Chen ◽

N. N. Yuan ◽

L. X. Zhai ◽

G. Q. Li ◽

...

Keyword(s):

Prunus Persica ◽

Direct Sequencing ◽

Eastern China ◽

Morphological Characteristics ◽

Brown Rot ◽

Its Sequences ◽

Western China ◽

Its2 Region ◽

Monilinia Fructicola ◽

First Report

Brown rot of peach (Prunus persica) in China has been reported to be caused by at least three Monilinia species (1). In the present study, peaches with symptoms resembling brown rot caused by Monilinia species were collected from commercial orchards in the northwestern province of Gansu in August 2010, the southwestern province of Yunnan in July 2011, and in the central province of Hubei in July 2012. Affected fruit showed the typical symptoms of brown rot with zones of sporulation. Fungal isolates were single-spored and cultured on potato dextrose agar (PDA). Colonies showed grayness with concentric rings of sporulation after incubation at 25°C in the dark. Mean mycelial growth of isolates YHC11-1a and YHC11-2a from Yunnan, GTC10-1a and GTC10-2a from Gansu, and HWC12-14a and HWC12-23a from Hubei, was 4.6 ± 0.4 and 7.5 ± 0.7 cm after 3 and 5 days incubation, respectively. Conidia were lemon shaped and formed in branched monilioid chains, and the mean size was 9.3 (6.7 to 11.5) × 12.5 (7.9 to 17.8) μm, which was consistent with the characteristics of Monilinia fructicola (1,2). The species identification was confirmed by sequencing of the ribosomal ITS sequences. The ribosomal ITS1-5.8S-ITS2 region was amplified from each of the six isolates using primers ITS1 and ITS4 (3). Results indicated that the ITS sequences of these isolates were identical and showed the highest similarity (100%) with M. fructicola ITS sequences from isolates collected in China (GenBank Accession Nos. HQ893748, FJ515894, and AM887528), Slovenia (GU967379), Italy (FJ411109), and Spain (EF207423). The pathogen was also confirmed to be M. fructicola based on the detection of an M. fructicola- specific band (534 bp) using a PCR-based molecular tool developed for distinguishing Chinese Monilinia species affecting peach (1). Pathogenicity was tested on surface-sterilized, mature peaches (Shui Mi Tao) with representative isolates. Fruits were holed at three equidistant positions to a depth of 5 mm using a sterile cork borer. Mycelial plugs (5 mm in diameter) from the periphery of a 4-day-old colony of each isolate were placed upside down into each hole, control fruits received water agar. After 3 days of incubation at 22°C in a moist chamber, inoculated fruits developed typical brown rot symptoms while control fruits remained healthy. Pathogens from the inoculated fruit were confirmed to be M. fructicola based on morphological characteristics. To our knowledge, this is the first report of occurrence of M. fructicola in Gansu, Yunnan, and Hubei provinces, thousands of kilometers away from eastern China where occurrence of peach brown rot caused by M. fructicola has been confirmed (2,4). The results indicated the further geographical spread of the M. fructicola in China. References: (1) M. J. Hu et al. Plos One 6(9):e24990, 2011. (2) M. J. Hu et al. Plant Dis. 95:225, 2011. (3) T. J. White et al. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics. Academic Press, San Diego, 1990. (4) X. Q. Zhu et al. Plant Pathol. 54:575, 2005.

Download Full-text

First Report of Brown Rot Caused by Monilia polystroma on Apple in Serbia

Plant Disease ◽

10.1094/pdis-07-12-0670-pdn ◽

2013 ◽

Vol 97 (1) ◽

pp. 145-145 ◽

Cited By ~ 9

Author(s):

M. Vasić ◽

N. Duduk ◽

M. S. Ivanović

Keyword(s):

Brown Rot ◽

Apple Fruit ◽

Molecular Characteristics ◽

Its2 Region ◽

Morphological Identification ◽

Japanese Species ◽

First Report ◽

Control Fruit ◽

Molecular Features ◽

Monilia Polystroma

Monilia polystroma van Leeuwen is a new Japanese species, similar to M. fructigena but distinguishable based on morphological and molecular characteristics (3). After its first discovery on apple in Japan, occurance of M. polystroma in Europe has been reported in Hungary, the Czech Republic, and Switzerland (2,3,4). In October 2011, during a survey for apple fungal pathogens in the Bela Crkva district, 15 apple fruit (Malus domestica Borkh.) cv. Golden Delicious were collected. Two isolates of Monilinia polystroma were obtained from apple fruit showing brown rot, covered with small yellowish sporodohia. The pathogen was identified as M. polystroma based on morphological and molecular features (1,3). Upon isolation, colonies cultivated on PDA were white to grayish and the mycelium grew 8.85 mm per day at 22 ± 1°C in 12-h light/12-h dark regime. After 6 to 8 days of incubation, black stromatal plates were observed on the reverse sides of the inoculated petri dishes. Conidia were one-celled, limoniform, hyaline, 14.7 to 21.88 μm (16.2 mean) × 7.85 to 12.92 μm (10.8 mean), and were produced in branched monilioid chains on inoculated apple fruit. Morphological identification was confirmed by PCR (1) using genomic DNA extracted from the mycelium of pure cultures, and amplified products of 425 bp in length, specific for M. polystroma were amplified as expected with primers MO368-5 and MO368-8R. For one isolate, the ribosomal ITS1-5.8S-ITS2 region was obtained, using primers ITS1 and ITS4, and deposited in GenBank (Accession No JX315717). The sequence was 498 bp in length and showed 100% identity with sequences deposited for M. polystroma in NCBI GenBank (JN128835, AM937114, GU067539). Pathogenicity was confirmed by wound-inoculating five surface-sterilized, mature apple fruit with mycelium plugs (5 mm in diameter) of both isolates grown on PDA. Control fruit were inoculated with sterile PDA plugs. After 3 days of incubation in plastic containers, under high humidity (RH 90 to 95%) at 22 ± 1°C, typical symptoms of brown rot developed on inoculated fruit, while control fruit remained symptomless. Isolates recovered from symptomatic fruit showed the same morphological and molecular characteristics as original isolates. To the best of our knowledge, this is the first report of M. polystroma in Serbia. Further studies are necessary to estimate the economic importance and geographic distribution of this organism in Serbia. References: (1) M.-J. Côté et al. Plant Dis. 88:1219, 2004. (2) M. Hilber-Bodmer et al. Plant Dis. 96: 146, 2012. (3) G. C. M. van Leeuwen et al. Mycol. Res. 106: 444, 2002. (4) OEPP/EPPO Reporting Service. Retrieved from http://archives.eppo.int/EPPOReporting/2011/Rse-1106.pdf

Download Full-text

First Report of Brown Rot of Stone Fruit Caused by Monilinia fructicola in Italy

Plant Disease ◽

10.1094/pdis-93-6-0668a ◽

2009 ◽

Vol 93 (6) ◽

pp. 668-668 ◽

Cited By ~ 39

Author(s):

C. Pellegrino ◽

M. L. Gullino ◽

A. Garibaldi ◽

D. Spadaro

Keyword(s):

Fungal Pathogens ◽

Brown Rot ◽

Stone Fruit ◽

Fruit Rot ◽

Brown Spot ◽

Morphological Identification ◽

Monilinia Fructicola ◽

Stone Fruits ◽

First Report ◽

Sequence Characterized Amplified Region

Monilinia fructicola, causal agent of brown rot, is one of the most important fungal pathogens of stone fruit. M. fructicola is a quarantined pathogen in Europe. During the summer of 2008 in 15 orchards located in Piedmont (northern Italy), 12,500 stone fruits (cherries, apricots, peaches, nectarines, and plums) were stored in cold chambers at 4 and 6°C and monitored for 8 weeks for the presence of Monilinia spp. M. fructicola was detected on 0.5% of nectarines (cvs. Sweet Red and Orion) that originated from two orchards in Lagnasco. Symptoms appeared on the fruit during storage, starting 3 weeks after harvest. Fruit rot lesions were brown, sunken, and covered with grayish tufts. The majority of infected fruit became dry and mummified. Brown rot symptoms were similar to those caused by endemic M. fructigena and M. laxa. Symptoms began with a small, circular, brown spot, and the rot spread rapidly. At the same time, numerous, small, grayish stromata developed. Finally, the whole surface of the fruit was covered by conidial tufts. Tissues were excised from diseased stone fruits and cultured on potato dextrose agar (PDA) amended with 25 μg of streptomycin per liter. The isolates produced abundant mycelium on PDA at 20 ± 2°C. Colonies were initially gray, but after sporulation, they became hazel, showing concentric rings (sporulation is sparse in M. laxa or M. fructigena). Conidia were one-celled, ellipsoid, hyaline, 15.2 × 10.1 μm, and produced in branched monilioid chains (2). Preliminary morphological identification of fungi resembling M. fructicola was confirmed by PCR using genomic DNA extracted from the mycelia of pure cultures. The DNA was amplified with a common reverse primer and three species-specific forward primers (3) obtained from a sequence characterized amplified region and a product of 535 bp, diagnostic for the species M. fructicola, was obtained. BLAST analysis of the amplified sequence (GenBank Accession No. FI569728) showed 96% similarity to the sequence of a M. fructicola isolated from Canada (GenBank Accession No. AF506700), 15% similarity to M. laxa ATCC11790 (GenBank Accession No. AF506702), and 35% similarity to a M. fructigena sequence isolated in Italy (GenBank Accession No. AF506701). Moreover, two sequences obtained through the amplification of ribosomal region ITS1-5.8S-ITS2, showing 100% similarity to the same ribosomal sequence of M. fructicola, were deposited in GenBank (Accession Nos. FJ411109 and FJ411110). The pathogen was detected on some mummified fruit from the same orchards in November of 2008. Pathogenicity was tested by spraying 103 conidia/ml on 10 surface-sterilized artificially wounded nectarines per strain of M. fructicola. After 5 days of incubation at 20 ± 2°C, typical, brown, rot symptoms developed on inoculated fruit. M. fructicola was reisolated from the inoculated fruit on PDA. Symptoms did not appear on control fruit. To our knowledge, this is the first report of M. fructicola in Italy. Its occurrence in Europe has been reported sporadically in Austria and France, and in 2006, it was detected in Hungary and Switzerland on peaches and nectarines imported from Italy and Spain (1,4). References: (1) E. Bosshard et al. Plant Dis. 90:1554, 2006. (2) R. J. W. Byrde and H. J. Willetts. The Brown Rot Fungi of Fruit: Their Biology and Control. Pergamon Press, Oxford, 1977. (3) M. J. Coté et al. Plant Dis. 88:1219, 2004. (4) M. Petròczy and L. Palkovics. Plant Dis. 90:375, 2006.

Download Full-text

First Report of Fruit Rot on Plum Caused by Monilinia fructicola at Alcalá del Río (Seville), Southwestern Spain

Plant Disease ◽

10.1094/pdis-11-11-0965 ◽

2012 ◽

Vol 96 (4) ◽

pp. 590-590 ◽

Cited By ~ 1

Author(s):

F. T. Arroyo ◽

M. Camacho ◽

A. Daza

Keyword(s):

Fungal Pathogens ◽

Brown Rot ◽

Fruit Rot ◽

Morphological Data ◽

Monilinia Fructicola ◽

First Report ◽

Sequence Characterized Amplified Region ◽

Agar Plug ◽

Pure Cultures ◽

Del Rio

Monilinia fructicola, causal agent of brown rot, is one of the most important fungal pathogens of stone fruit. In the summer of 2011, Japanese plum fruit of ‘Larry Ann’ (Prunus salicina Lindl) showing symptoms of fruit rot disease were detected and collected from trees in an experimental field at Alcalá del Río (Seville), southwestern Spain. Fruit rot lesions were brown, sunken, and covered with grayish brown tufts or pustules. The majority of infected fruit became dry and mummified on the trees after 30 days. Symptoms were similar to those caused by three Monilinia species, M. laxa, M. fructigena, and M. fructicola (2). Pieces of infected tissue, previously disinfested in 0.6% NaOCl, were placed on potato dextrose agar (PDA) amended with 50 μg of streptomycin per liter and incubated at 22°C with a 12-h photoperiod for 15 days. The isolates produced abundant, grayish white mycelium, which after sporulation became hazel in color, and colonies displayed concentric rings. Colonies produced scarce conidia, which were arranged in branched, monilioid chains. Conidia were one celled, hyaline, ellipsoid to lemon shaped, and measured 15.42 ± 1.91 × 8.02 ± 0.9 μm. The morphological data and growth rates match the description of M. fructicola (Winter) Honey (2–4). Fungal identification was confirmed by PCR using genomic DNA extracted from the mycelia of pure cultures. The DNA was amplified with a common reverse primer and three specific forward primers obtained from a sequence-characterized, amplified region that distinguishes between M. fructicola, M. fructigena, and M. laxa. The size of the amplified fragment (a product of 535 bp) fit with the one described for M. fructicola (2). To confirm the pathogenicity of the isolate, mature ‘Larry Ann’ and ‘Sungold’ plum fruits (six fruits per cultivar) were inoculated by placing an agar plug from the edge of an actively growing colony on PDA directly on the fruit surface. After 5 days of incubation, typical brown rot symptoms developed on inoculated fruit and the fungus was successfully reisolated, thus fulfilling Koch's postulates. No symptoms appeared on control fruit. To our knowledge, this is the first report of M. fructicola on plums in southwestern Spain. M. fructicola is a quarantined pathogen in Europe and has been reported on imported apricot and nectarine (1) and peach in several European countries (3,4). References: (1) E. Bosshard et al. Plant Dis. 90:1554, 2006. (2) M. J. Côté. Plant Dis. 88:1219, 2004. (3) A. De Cal and I. Gell. Plant Dis. 93:763, 2009. (4). C. Pellegrino et al. Plant Dis. 93:668, 2009.

Download Full-text