scholarly journals First Report of Coniella granati as a Causal Agent of Pomegranate Crown Rot in Southern Italy

Plant Disease ◽  
2016 ◽  
Vol 100 (7) ◽  
pp. 1498-1498 ◽  
Author(s):  
S. Pollastro ◽  
C. Dongiovanni ◽  
D. Gerin ◽  
P. Pollastro ◽  
G. Fumarola ◽  
...  
Plant Disease ◽  
2014 ◽  
Vol 98 (7) ◽  
pp. 995-995 ◽  
Author(s):  
F. De Curtis ◽  
D. Palmieri ◽  
D. Vitullo ◽  
G. Lima

Epidemiological investigations in representative chickpea (Cicer arietinum L.) fields in southern Italy (Larino, Campobasso, 41°50′45″ N, 14°55′28″ E) identified severe withering (25 to 51%) of plants during flowering. Diseased plants showed a reduced total root biomass associated with less vigorous and chlorotic foliage. Browning and necrosis of subcortical and xylematic tissues of the crown and main roots were observed in affected plants. Symptomatic root and stem portions from 50 plants were sampled, surface disinfected with a sodium hypochlorite water solution (2% v/v for 2 min), rinsed with sterile distilled water, and placed in petri dishes containing potato dextrose agar with streptomycin sulfate (200 mg/l) and incubated at 25°C for 10 days. The most frequent fungal colony isolated showed macro- and microscopic characters specific of the genus Fusarium (3), with falcate and three-septate macroconidia (24.0 to 43.8 μm long) and microconidia (6.8 to 10.4 μm long) with zero or one septa. The ribosomal DNA of the fungal isolate processed by PCR using the ITS1F/ITS4 primers (2) produced an amplicon of 545 bp (ENA, Accession No. HG423346). A BLAST search with the amplified sequence in the database of the International Mycological Association ( www.mycobank.org ) revealed 99% identity with F. oxysporum sequences. Additional molecular analysis using the specific primers Foc0-12/Foc0-12rf for F. oxysporum f.sp. ciceris (Foc) produced an amplicon only in the chickpea virulent strain Foc-7952, race 0 (1) used as control; furthermore, PCR amplification for the Pisatin Demetylase gene by using the specific primers PDAF2a and PDAR3a (4) yielded the expected amplicon only for the new isolate, whereas no amplification was obtained with the control strain Foc-7952. Pathogenicity assays were carried out to complete Koch's postulates. To this aim, horticultural peat was infested with a conidial suspension (1 × 104 conidia/g of soil) from the new fungal pathogen, dispensed in plastic pots, and sown with surface sterilized seeds of chickpea (cv. Real, ISEA, Italy). Uncontaminated peat was used as control. For both treatments, 3 replicates of 10 seeds were used and experiments repeated twice. The plastic pots were kept in a growth chamber (28°C; 70% RH; 15/9 h light/dark) where the first disease symptoms on plants appeared 20 days after sowing. At the end of the experiments, all plants inoculated with the new isolate showed a high disease severity (98%), whereas non-inoculated plants remained healthy. The seedlings from infested soil demonstrated the same symptoms previously observed in the field, and after re-isolation, the causal agent demonstrated the same morphological features of the isolate used for inoculation. Pathogenicity tests were performed on pea, faba bean, melon, and tomato by using three cultivars for each crop. The results demonstrated high virulence of the new isolate of F. oxysporum f.sp. pisi (Fop) on both chickpea and pea with seed germination reduction, rot on main and secondary roots and cotyledonary leaves, and root biomass reduction and foliage chlorosis. No symptoms were observed on other inoculated vegetal species. Collectively, data of our investigation allow us to affirm that this is the first report of Fop as a new pathogen of chickpea. This result has great economic importance since it enables specific monitoring and management plans for this new disease caused by Fop on chickpea, a key crop for human and animal nutrition. References: (1) M. M. Jiménez-Gasco and R. M. Jiménez-Díaz, Phytopathology 93:201, 2003. (2) I. Larena et al. J. Biotechnol. 75:187, 1999. (3) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell Publishing, Ames, IA, 2006. (4) N. A. Milani et al. Fungal Genet. Biol. 933:942, 2012.


Plant Disease ◽  
2007 ◽  
Vol 91 (11) ◽  
pp. 1519-1519
Author(s):  
E. Lahoz ◽  
R. Caiazzo ◽  
A. Carella ◽  
E. Cozzolino

In each of two fields of buckwheat (Fagopyrum esculentum L.) grown in Benevento Province (southern Italy), 60 to 70% of the plants developed severe root and crown rot. Symptoms included irregular, water-soaked spots on stems that were eventually covered with cottony mycelia as the lesions enlarged. Black sclerotia usually developed within the mycelium. The fungus was isolated on potato dextrose agar and 2% water agar. On the basis of colony morphology, including the production of black sclerotia (1), the fungus was identified as Sclerotinia sclerotiorum (Lib.) De Bary. The identity of the fungus was confirmed by near exact identity of internal transcribed spacer sequences (99%) with two isolates of S. sclerotiorum in GenBank (Accession Nos. Z73800 and Z73799). Pathogenicity of the fungus on buckwheat was evaluated by transplanting 20 20-day-old healthy plants in a mixture of soil and fungal inoculum (0.5% of wet millet seeds colonized by four isolates of S. sclerotiorum). Lesions on crowns and roots developed after 12 days and sclerotia appeared approximately 20 days later. No symptoms developed on noninoculated plants. Reisolation from inoculated plants yielded colonies of S. sclerotiorum. To our knowledge, this is the first report of S. sclerotiorum on buckwheat in Italy. The high incidence and severity of the disease may be limiting factors in the development of buckwheat as an alternative crop of tobacco in southern Italy. Reference: (1) J. E. M. Mordue and P. Holliday. Sclerotinia sclerotiorum. No. 513 in: Descriptions of Pathogenic Fungi and Bacteria. CMI, Kew, Surrey, UK, 1976.


Plant Disease ◽  
2013 ◽  
Vol 97 (7) ◽  
pp. 996-996 ◽  
Author(s):  
S. Sánchez ◽  
M. Gambardella ◽  
J. L. Henríquez ◽  
I. Díaz

In recent years, an increase of collapsed and dead strawberry plants has been observed in several fields in central Chile, specifically in San Pedro, Melipilla, an important area for strawberry cultivation in the country. To determine the causal agent of the disease and the extent of the problem, 25 sample sites of 1 ha each, distributed in different San Pedro zones, were surveyed at the end of the 2011 season (from December 2010 to February 2011). Cross sections of the crowns of symptomatic strawberry plants showed necrotic tissue and brown-red to dark brown areas on the vascular ring. Samples of the affected crowns were superficially disinfested and plated on potato dextrose agar with 200 μg/ml of streptomycin sulfate. Dark gray colonies were observed after 7 days of incubation at 24°C. Pure cultures of the pathogen showed aerial mycelium and abundant dark oblong sclerotia. Fifty sclerotia were measured, averaging 120 × 74 μm. Twenty-one isolates were identified molecularly utilizing the species specific primers MpKFI and MpKRI (2) that yielded a 350-bp fragment. The amplified DNA fragments were sequenced and BLAST analysis showed a 99% nucleotide sequence identity with Macrophomina phaseolina (GeneBank Accession No JX535007.1). Both morphological and molecular analyses confirmed that the isolated species corresponded to M. phaseolina, causal agent of crown and root rot in strawberry. Four representative isolates were selected to conduct pathogenicity tests. Inoculum was prepared by incubating the pathogen for 28 days at 20°C in sterilized oat seeds. Pots of 1.5 liters were filled with a mixed substrate of peat and perlite (2:1), amended with inoculated oats at 9 g per liter of substrate. ‘Camarosa’ strawberry plants were planted and grown in a glasshouse for 1 month. Six replicated plants per isolate and six plants growing on non-inoculated substrate were left as controls. Ninety-five percent of the inoculated plants showed wilt and collapse symptoms 22 days after transplant, whereas no symptoms were observed in the control plants. M. phaseolina was reisolated from the crowns of symptomatic plants, fulfilling Koch's postulates. The pathogen was isolated from symptomatic strawberry plants in 14 of the 25 sampled sites. Although M. phaseolina was described previously on other crops in Chile, to our knowledge, this is the first report of M. phaseolina causing crown rot of strawberry. The disease has been recently reported in Spain, the United States, and Argentina (1,3,4). References: (1) M. Avilés et al. Plant Pathol. 57:382, 2008. (2) B. Babu et al. Mycologia 99:797, 2007. (3) O. Baino et al. Plant Dis. 95:1477, 2011. (4) S. Koike. Plant Dis. 92:1253, 2008.


Plant Disease ◽  
2020 ◽  
pp. PDIS-05-20-1045
Author(s):  
H.-Y. Wu ◽  
C.-Y. Tsai ◽  
Y.-M. Wu ◽  
H.-A. Ariyawansa ◽  
C.-L. Chung ◽  
...  
Keyword(s):  

Plant Disease ◽  
2014 ◽  
Vol 98 (5) ◽  
pp. 698-698 ◽  
Author(s):  
Y. Tomitaka ◽  
T. Usugi ◽  
R. Kozuka ◽  
S. Tsuda

In 2009, some commercially grown tomato (Solanum lycopersicum) plants in Chiba Prefecture, Japan, exhibited mosaic symptoms. Ten plants from a total of about 72,000 cultivated plants in the greenhouses showed such symptoms. To identify the causal agent, sap from leaves of the diseased plants was inoculated into Chenopodium quinoa and Nicotiana benthamiana plants. Local necrotic lesions appeared on inoculated leaves of C. quinoa, but no systemic infection was observed. Systemic mosaic symptoms were observed on the N. benthamiana plants inoculated. Single local lesion isolation was performed three times using C. quinoa to obtain a reference isolate for further characterization. N. benthamiana was used for propagation of the isolate. Sap from infected leaves of N. benthamiana was mechanically inoculated into three individual S. lycopersicum cv. Momotaro. Symptoms appearing on inoculated tomatoes were indistinguishable from those of diseased tomato plants found initially in the greenhouse. Flexuous, filamentous particles, ~750 nm long, were observed by electron microscopy in the sap of the tomato plants inoculated with the isolate, indicating that the infecting virus may belong to the family Potyviridae. To determine genomic sequence of the virus, RT-PCR was performed. Total RNA was extracted from the tomato leaves experimentally infected with the isolate using an RNeasy Plant Mini kit (QIAGEN, Hilden, Germany). RT-PCR was performed by using a set of universal, degenerate primers for Potyviruses as previously reported (2). Amplicons (~1,500 bp) generated by RT-PCR were extracted from the gels using the QIAquick Gel Extraction kit (QIAGEN) and cloned into pCR-BluntII TOPO (Invitrogen, San Diego, CA). DNA sequences of three individual clones were determined using a combination of plasmid and virus-specific primers, showing that identity among three clones was 99.8%. A consensus nucleotide sequence of the isolate was deposited in GenBank (AB823816). BLASTn analysis of the nucleotide sequence determined showed 99% identity with a partial sequence in the NIb/coat protein (CP) region of Colombian datura virus (CDV) tobacco isolate (JQ801448). Comparison of the amino acid sequence predicted for the CP with previously reported sequences for CDV (AY621656, AJ237923, EU571230, AM113759, AM113754, and AM113761) showed 97 to 100% identity range. Subsequently, CDV infection in both the original and experimentally inoculated plants was confirmed by RT-PCR using CDV-specific primers (CDVv and CDVvc; [1]), and, hence, the causal agent of the tomato disease observed in greenhouse tomatoes was proved to be CDV. The first case of CDV on tomato was reported in Netherlands (3), indicating that CDV was transmitted by aphids from CDV-infected Brugmansia plants cultivated in the same greenhouse. We carefully investigated whether Brugmansia plants naturally grew around the greenhouses, but we could not find them inside or in proximity to the greenhouses. Therefore, sources of CDV inoculum in Japan are still unclear. This is the first report of a mosaic disease caused by CDV on commercially cultivated S. lycopersicum in Japan. References: (1) D. O. Chellemi et al. Plant Dis. 95:755, 2011. (2) J. Chen et al. Arch. Virol. 146:757, 2001. (3) J. Th. J. Verhoeven et al. Eur. J. Plant. Pathol. 102:895, 1996.


Plant Disease ◽  
2006 ◽  
Vol 90 (8) ◽  
pp. 1109-1109 ◽  
Author(s):  
A. Garibaldi ◽  
G. Gilardi ◽  
M. L. Gullino

Lamb's lettuce or corn salad (Valerianella olitoria) is increasingly grown in Italy and used primarily in the preparation of mixed processed salad. In the fall of 2005, plants of lamb's lettuce, cv Trophy, exhibiting a basal rot were observed in some commercial greenhouses near Bergamo in northern Italy. The crown of diseased plants showed extensive necrosis, progressing to the basal leaves, with plants eventually dying. The first symptoms, consisting of water-soaked zonate lesions on basal leaves, were observed on 30-day-old plants during the month of October when temperatures ranged between 15 and 22°C. Disease was uniformly distributed in the greenhouses, progressed rapidly in circles, and 50% of the plants were affected. Diseased tissue was disinfested for 1 min in 1% NaOCl and plated on potato dextrose agar amended with 100 μg/liter of streptomycin sulfate. A fungus with the morphological characteristics of Rhizoctonia solani was consistently and readily isolated and maintained in pure culture after single-hyphal tipping (3). The five isolates of R. solani, obtained from affected plants successfully anastomosed with tester isolate AG 4, no. RT 31, received from R. Nicoletti of the Istituto Sperimentale per il Tabacco, Scafati, Italy (2). The hyphal diameter at the point of anastomosis was reduced, and cell death of adjacent cells occurred (1). Pairings were also made with AG 1, 2, 3, 5, 7, and 11 with no anastomoses observed between the five isolates and testers. For pathogenicity tests, the inoculum of R. solani (no. Rh. Vale 1) was grown on autoclaved wheat kernels at 25°C for 10 days. Plants of cv. Trophy were grown in 10-liter containers (20 × 50 cm, 15 plants per container) on a steam disinfested substrate (equal volume of peat and sand). Inoculations were made on 20-day-old plants by placing 2 g of infected wheat kernels at each corner of the container with 3 cm as the distance to the nearest plant. Plants inoculated with clean wheat kernels served as controls. Three replicates (containers) were used. Plants were maintained at 25°C in a growth chamber programmed for 12 h of irradiation at a relative humidity of 80%. The first symptoms, consisting of water-soaked lesions on the basal leaves, developed 5 days after inoculation with crown rot and plant kill in 2 weeks. Control plants remained healthy. R. solani was consistently reisolated from infected plants. The pathogenicity test was carried out twice with similar results. This is, to our knowledge, the first report of R. solani on lamb's lettuce in Italy as well as worldwide. The isolates were deposited at the AGROINNOVA fungal collection. The disease continues to spread in other greenhouses in northern Italy. References: (1) D. Carling. Rhizoctonia Species: Pages 37–47 in: Taxonomy, Molecular Biology, Ecology, Pathology and Disease Control. B. Sneh et al., eds. Kluwer Academic Publishers, the Netherlands, 1996. (2) J. Parmeter et al. Phytopathology, 59:1270, 1969. (3) B. Sneh et al. Identification of Rhizoctonia Species. The American Phytopathological Society, St. Paul, MN, 1996.


Plant Disease ◽  
2021 ◽  
Author(s):  
Samara A. Oliveira ◽  
Daniel M. Dlugos ◽  
Paula Agudelo ◽  
Steven N. Jeffers

Root-knot nematodes (RKNs), Meloidogyne spp., are some of the most economically important pathogens of cultivated plants. Meloidogyne javanica is one of the most destructive RKN species and is well known for its broad host range and the severe damage it causes to plant roots (Perry et al. 2009). In Feb 2018, four mature dead and dying hybrid lavender plants (Lavandula ×intermedia ‘Phenomenal’) were collected in Edgefield County, South Carolina, and suspected of having Phytophthora root and crown rot (Dlugos and Jeffers 2018). Greenhouse-grown plants had been transplanted in Dec 2016 and Jan 2017 into a sandy loam soil on a site that had been fallow or in pasture for over 30 years. Some plants began to turn gray and die in summer 2017, and approximately 40% of 1230 plants were symptomatic or dead by Feb 2018. Phytophthora spp. were not isolated from the collected plants but were isolated from plants collected on subsequent visits. Instead, all four plants had small, smooth galls on the roots. Lavender roots were examined microscopically (30-70×), and egg masses of RKNs were observed on the galls. Mature, sedentary RKN females were handpicked from galled roots, and perineal patterns of 10 specimens were examined and identified as M. javanica. Juveniles and eggs were extracted from lavender roots by the method of Coolen and D’herde (1972). To confirm species identification, DNA was extracted from 10 individual juveniles, and a PCR assay was conducted using species-specific primers for M. javanica, Fjav/Rjav (Zijlstra et al. 2000). A single amplicon was produced with the expected size of approximately 720 bp, which confirmed identity as M. javanica. To determine pathogenicity, M. javanica from lavender roots were inoculated onto susceptible tomato plants for multiplication, and severe gall symptoms occurred on tomato roots 60 days later. Nematodes were extracted from tomato roots and inoculated onto healthy, rooted cuttings of ‘Phenomenal’ lavender plants growing in pots of soilless medium in a greenhouse. Plants were inoculated with 0, 1000, 2000, 5000, or 10000 eggs and juveniles of M. javanica. Five single-plant replicates were used for each treatment, and plants were randomized on a greenhouse bench. Plants were assessed 60 days after inoculation, and nematodes were extracted from roots and counted. The reproduction factor was 0, 43.8, 40.9, 9.1, 7.7, and 2.6 for initial nematode populations 0, 1000, 2000, 5000, and 10000, respectively, which confirmed pathogenicity (Hussey and Janssen 2002). Meloidogyne javanica also was recovered in Mar 2018 from galled roots on a ‘Munstead’ (L. angustifolia) lavender plant from Kentucky (provided by the Univ. of Kentucky Plant Disease Diagnostic Laboratories), and an unidentified species of Meloidogyne was isolated in Aug 2020 from a ‘Phenomenal’ plant grown in Florida. COI mtDNA sequences from the SC (MZ542457) and KY (MZ542458) populations were submitted to Genbank. M. javanica previously was found associated with field-grown lavender (hybrid and L. angustifolia) in Brazil, but pathogenicity was not studied (Pauletti and Echeverrigaray 2002). To our knowledge, this is the first report of M. javanica pathogenic to L. ×intermedia in the USA, and the first time RKNs have been proven to be pathogenic to Lavandula spp. following Koch’s Postulates. Further studies are needed to investigate the geographic distribution of M. javanica on lavender and the potential threat this nematode poses to lavender production in the USA.


Plant Disease ◽  
2017 ◽  
Vol 101 (6) ◽  
pp. 1059 ◽  
Author(s):  
R. Petry ◽  
M. E. N. Fonseca ◽  
L. S. Boiteux ◽  
A. Reis

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