scholarly journals First Report of Collar and Root Rot Caused by Phytophthora nicotianae on Daphne odora in Italy

Plant Disease ◽  
2009 ◽  
Vol 93 (8) ◽  
pp. 848-848
Author(s):  
A. Garibaldi ◽  
D. Bertetti ◽  
M. L. Gullino

Daphne odora is becoming popular in gardens because of its variegated foliage and fragrant flowers in late winter and early spring. During October of 2008 in a commercial nursery near Maggiore Lake (Verbano-Cusio-Ossola Province) in northwestern Italy, plants of D. odora showed extensive chlorosis and root rot. Diseased plants eventually wilted and died, dropping leaves in some cases. Most frequently, wilted leaves persisted on stems. At the soil level, dark brown-to-black water-soaked lesions that coalesced often girdled the stem. All of the crown and root system was affected. Disease was widespread and severe with 70% of 2,500 potted plants being affected. A Phytophthora-like organism was isolated consistently on a medium selective for oomycetes (4) after disinfestation of lower stem and root pieces of D. odora for 1 min in a solution containing 1% NaOCl. Tissue fragments of 1 mm2 were excised from the margins of the lesions and plated. The pathogen was identified based on morphological and physiological features as Phytophthora nicotianae (= P. parasitica) (2). Sporangia were produced for identification by growing a pure culture in sterilized soil extract solution at neutral pH (obtained by shaking and then centrifuging 300 g of soil in 1 liter of distilled water). They were spherical to ovoid, papillate, and measured 39.2 to 54.5 × 31.7 to 41.7 μm (average 44.8 × 34.5 μm). Papillae measured 2.4 to 4.9 μm (average 3.7 μm). Chlamydospores were spherical with a diameter ranging from 15.8 to 36.1 μm (average 25.4 μm). The internal transcribed spacer (ITS) region of rDNA of a single isolate was amplified using primers ITS4/ITS6 and sequenced. BLAST analysis (1) of the 804-bp segment showed a 100% homology with the sequence of P. nicotianae EF140988. The nucleotide sequence has been assigned GenBank No. FJ843100. Pathogenicity of two isolates obtained from infected plants was confirmed by inoculating 12-month-old plants of D. odora. Both isolates were grown for 15 days on a mixture of 70:30 wheat/hemp kernels and then 80 g/liter of the inoculum was mixed into a substrate containing sphagnum peat moss/pumice/pine bark/clay (50:20:20:10 vol/vol). One plant per 3-liter pot was transplanted into the substrate and constituted the experimental unit. Three replicates were used for each isolate and noninoculated control treatment; the trial was repeated once. All plants were kept in a greenhouse at temperatures from 20 to 25°C. Plants inoculated with isolate no. 1 developed symptoms of chlorosis and root rot within 14 days and then a wilt rapidly followed. Isolate no. 2 was less aggressive causing the same symptoms within 20 days. Control plants remained symptomless. P. nicotianae consistently was reisolated from inoculated plants. Previously, P. nicotianae (= P. parasitica) has been reported in several states of the United States on D. odora (3). To our knowledge, this is the first report of P. nicotianae on D. odora in Italy. The economic importance of the disease is low because of the limited number of farms that grow this crop in Italy, although spread could increase as the popularity of plantings expand. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997 (2) D. C. Erwin and O. K. Ribeiro. Phytophthora Diseases Worldwide. The American Phytopathological Society, St Paul, MN, 1996. (3) D. F. Farr et al. Fungi on Plants and Products in the United States. The American Phytopathological Society, St Paul, MN, 1989. (4) H. Masago et al. Phytopathology, 67:425, 1977.

Plant Disease ◽  
2010 ◽  
Vol 94 (7) ◽  
pp. 917-917
Author(s):  
A. Garibaldi ◽  
D. Bertetti ◽  
M. L. Gullino

Edgeworthia papyrifera, Oriental paperbush, is a deciduous flowering shrub becoming increasingly popular because of its clove-like perfumed flowers appearing in late winter-early spring. During August of 2009 in a commercial nursery close to Maggiore Lake (Verbano-Cusio-Ossola Province) in northwest Italy, 2-year-old plants of E. papyrifera showed extensive chlorosis and root rot. Twigs wilted and died, dropping leaves in some cases. Most frequently, wilted leaves persisted on stems. At the soil level, dark brown-to-black water-soaked lesions formed and coalesced, girdling the stem. All of the crown and root system was affected. Infected plants died within 14 days of the appearance of symptoms. Disease was widespread and severe, affecting 90 of the 100 plants present. After disinfestation for 1 min in a solution containing 1% NaOCl, rotting root and collar pieces of E. papyrifera consistently produced a Phytophthora-like organism when plated on a medium selective for oomycetes (3). The pathogen was identified morphologically as Phytophthora nicotianae (= P. parasitica) (2). On V8 agar, coenocytic hyphae, 4 to 8 μm in diameter, formed fluffy, aerial colonies and spherical, intercalary chlamydospores, 21.0 to 36.5 (average 26.7) μm in diameter. Colonies grew well at 35°C and stopped growing at 40°C. Sporangia were produced by growing a pure hyphal-tip culture in a diluted, sterilized soil-extract. Sporangia were borne singly, laterally attached to the sporangiophore, were noncaducous, spherical to ovoid, papillate, and measured 28.6 to 55.2 × 22.4 to 45.1 (average 42.4 × 34.6) μm, length/breadth ratio (1.1:1)-1.2:1-(1.3:1). Papillae measured 3.1 to 7.6 (average 4.6) μm. The internal transcribed spacer (ITS) region of rDNA of a single isolate was amplified with primers ITS4/ITS6 and sequenced. BLAST analysis (1) of the 839-bp segment showed 99% homology with the sequence of P. nicotianae (No. AJ854296). The sequence has been assigned the GenBank No. GU353341. Pathogenicity of isolates Edg.1 and Edg.2 obtained, respectively, from the root and collar of an infected plant was confirmed by inoculating 1-year-old plants of E. papyrifera. Both strains were grown for 15 days on a mixture of 70:30 wheat/hemp kernels, and 4 g/liter of the inoculum was mixed into a substrate containing sphagnum peat moss/pumice/pine bark/clay (50:20:20:10 vol/vol). One plant per 3-liter pot was transplanted into the substrate and constituted the experimental unit. Five plants were used for each test strain and noninoculated control treatment; the trial was repeated once. All plants were kept in a greenhouse at 25 to 28°C. Plants inoculated with Edg.1 and Edg.2 developed chlorosis and root rot 18 and 14 days after the inoculation, respectively, and wilt rapidly followed. Control plants remained symptomless. P. nicotianae was consistently reisolated from inoculated plants. To our knowledge, this is the first report of P. nicotianae on E. papyrifera in Italy as well as worldwide. The current economic importance of the disease is minor due to the limited number of farms that grow this crop in Italy, although spread could increase as the popularity of plantings expand. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997 (2) D. C. Erwin and O. K. Ribeiro. Phytophthora Diseases Worldwide. The American Phtytopathological Society, St Paul, MN, 1996. (3) H. Masago et al. Phytopathology 67:425, 1977.


Plant Disease ◽  
2004 ◽  
Vol 88 (8) ◽  
pp. 905-905
Author(s):  
A. Garibaldi ◽  
D. Bertetti ◽  
M. L. Gullino

Skimmia japonica, an evergreen flowering shrub, is becoming increasingly popular as a potted ornamental plant in northern Italy and represents 5% of acidophilous plant production; cv. Rubella accounts for 99% of production. During the spring of 2003, in many commercial nurseries located in northwestern Italy, plants of S. japonica cv. Rubella showed extensive chlorosis and root rot, and diseased plants eventually wilted and died without dropping leaves. The disease was widespread and severe, and in some nurseries, 40% of plants were affected. A Phytophthora-like organism was isolated consistently from infected lower stem and root pieces of S. japonica that had been disinfested for 1 min in 1% NaOCl and plated on a medium selective for oomycetes (2). The pathogen was identified based on morphological and physiological features as Phytophthora nicotianae (= P. parasitica [1]). The sporangia produced on V8 medium were ± spherical to obpyriform, obturbinate, papillate, and measured 33 to 94 × 25 to 48 μm (average 56.4 × 36.8 μm). Papillae measured 3.5 to 19 μm (average 7.8 μm). Chlamydospores were spherical with a diameter ranging from 26 to 32 μm (average 29.2 μm). Pathogenicity of four isolates obtained from infected plants was confirmed by inoculating 9-month-old plants of S. japonica cv. Rubella grown in 1-liter pots containing a substrate based on sphagnum peatmoss, pine bark, and clay (70-20-10% vol/vol/vol). Inocula, which consisted of 90-mm-diameter V8 agar disks per pot containing mycelium of each isolate, were introduced and mixed into the substrate in all pots before transplanting. One plant was transplanted into each pot and served as a replicate, and noninoculated plants served as controls. Eight replicates were used for each isolate and the control treatment, and the trial was repeated. All plants were kept outside at temperatures ranging from 16 to 38°C (average temperature 27°C). Inoculated plants developed symptoms of chlorosis, root rot, and wilt within 20 days, while control plants remained symptomless. P. nicotianae consistently was isolated from inoculated plants. Previously, P. nicotianae has been reported on S. japonica in Poland (3). To our knowledge, this is the first report of P. nicotianae on S. japonica in Italy. References: (1) D. C. Erwin and O. K. Ribeiro. Phytophthora Diseases Worldwide. The American Phytopathological Society, St Paul, MN, 1996. (2) H. Masago et al. Phytopathology, 67:425, 1977 (3) G. Szkuta and L. B. Orlikowski. Prog. Plant Prot. 42:808, 2002.


Plant Disease ◽  
2011 ◽  
Vol 95 (5) ◽  
pp. 614-614
Author(s):  
T. D. Miles ◽  
C. I. Woelk ◽  
A. Rojas ◽  
A. M. C. Schilder

In September 2009, ~40 declining blueberry plants (Vaccinium corymbosum L. ‘Jersey’) were observed in a poorly drained area of a 30-year-old field near Fennville, MI. The stunted bushes had yellow leaves and defoliation; others were completely dead. The grower reported that the bushes had been declining over several years. Root samples tested positive in a Phytophthora ELISA test (Agdia Inc., Elkhart IN). Twenty root pieces (2 cm long and 2 to 3 mm in diameter) were surface disinfested and plated on Rye A agar; five yielded fungal-like colonies that were subcultured on potato dextrose agar (PDA). One isolate was white and grew slowly (3 to 4 mm/day at 22 to 24°C). Three isolates were white and grew faster (10 to 12 mm/day at 22 to 24°C) in a chrysanthemal pattern. The fifth was a Fusarium sp. DNA of the white colonies was extracted and the internal transcribed spacer (ITS) region was sequenced using ITS1 (5′-TCCGTAGGTGAACCTGCGG-3′) and ITS4 (5′-TCCTCCGCTTATTGATATGC-3′) primers. The slow-growing morphotype had 99% identity to Phytophthora sp. isolate 92-209C (Accession No. EU106591) in GenBank but failed to induce symptoms in multiple inoculation tests. The fast-growing morphotype (Accession No. HQ398249) had 98% identity to Pythium sterilum UASWS0265 from declining alder stands in Poland (Accession No. DQ525089). Sequencing of the COXII gene using the FM66/FM58 primer set (3) yielded a product (Accession No. HQ721468) with 100% identity to P. sterilum GD32a from forest soil in Poland (Accession No. EF421185). Hyphae were hyaline, coenocytic, and 4 to 7 μm wide with some swellings at the tips (7 to 9 μm wide). No sporangia, oogonia, or antheridia were observed. Mycelium tested positive in the ELISA test described above. According to Agdia Inc., 10 of 19 tested Pythium spp. have shown similar cross reactivity. Pythium spp. are known to cause root rot of blueberries in Oregon (2). In British Columbia, P. sterilum was commonly isolated from roots of declining blueberry bushes (4). P. sterilum Belbahri & Lefort only reproduces asexually (1). Our isolate was similar but did not produce sporangia in water or on PDA, V8 juice agar, Rye A agar, or water agar. Roots of 10 2-month-old ‘Bluecrop’ cuttings were placed in an aqueous suspension of rinsed mycelium (0.1 g/ml) from 21-day-old cultures grown in V8 broth or in sterile deionized water (control). After 1 h, plants were potted in peat moss/perlite (2:1) or autoclaved sand (five each) and placed in a glasshouse at 25°C. After 7 days, inoculated plants in both soil types had wilted or collapsed with significant necrosis on the roots and primary shoot. Control plants showed no symptoms. In a similar experiment with 6-month-old plants in sand, symptoms appeared after 10 to 12 days. The pathogen was recovered from surface-disinfested root and stem sections of all inoculated plants but not control plants and its identity was confirmed by sequencing of the ITS region. To our knowledge, this is the first report of P. sterilum on blueberries in the United States. While this disease appears to be uncommon in Michigan, it is a potential cause of plant decline, the diagnosis of which may be complicated by cross reactivity in ELISA testing. References: (1) L. Belbahri et al. FEMS Microbiol. Lett. 255:209, 2006. (2) D. R. Bryla and R. G. Linderman. HortScience 43:260, 2008. (3) F. N. Martin. Mycologia 92:711, 2000. (4) S. Sabaratnam. BC Plant Health Fund Final Report. B.C. Retrieved from http://www.agf.gov.bc.ca/cropprot/phf_final_report.pdf , 2008.


Plant Disease ◽  
2001 ◽  
Vol 85 (4) ◽  
pp. 445-445
Author(s):  
E. Ilieva ◽  
W. A. Man in 't Veld ◽  
B. F. Wessels-Berk ◽  
R. P. Baayen

Limonium (statice or sea-lavendar, family Plumbaginaceae) is grown in the Netherlands as a perennial (Limonium sinense) or annual (Limonium sinuatum) crop. Plants have tufted leaves and numerous clustered flowers of different colors and are used for flower arrangements. In August 2000, we received diseased plants of L. sinense cv. Diamond and L. sinuatum. Disease symptoms consisted of leaf wilting followed by plant collapse. The base of the leaves showed progressive necrotic areas that later turned dark brown to black. The cortex of the stem and roots was water-soaked and dark brown to black. Longitudinal sections of stems and roots of diseased plants displayed discoloration of tissues. Rotted root tissue was brown with a characteristic black margin. Rotted vascular tissues and other stem parts were also dark brown. Pith parenchyma turned gray-brown and had a firm, wet rot. In plants with advanced disease symptoms, a cavity in the stem parenchyma was observed. Isolations were made from sections of symptomatic leaves, stems and roots of both Limonium species on cherry and water agar (WA), followed by incubation at 20°C. Phytophthora sp. was isolated consistently from the base of leaves, stems, and roots of diseased plants and identification of isolates was based on morphological characteristics and by isozyme analysis (3). Observations of colony morphology and growth at 35°C were made on V8 agar. Mating type was determined in dual cultures with mating type A2 (P. nicotianae, P 1923 [4]) and A1 (P. nicotianae, PD98/8/10402). Sporangial features were observed from liquid cultures of the isolates (autoclaved soil-extract or sterile distilled water). All isolates formed colonies consisting of loose, fluffy aerial mycelia. Sporangia and chlamydospores were present in all fungal isolates and all isolates were able to grow at 35°C. Few sporangia were produced on solid media (WA and V8 juice agar), but were abundant in liquid cultures. Sporangia were borne singly or in simple sympodial sporangiophores (3 to 4 sporangia), and were ovoid/spherical, obturbinate with rounded base and had prominent papillae (some had two papillae). Sporangia measured 40 to 64 × 24 to 56 μm, (average 50.4 × 38.4 μm) and had an average length:breath ratio of 1.3:1. Chlamydospores were terminal and intercalary and measured 18 to 44 μm (average 31.6 μm). Hyphal swellings with hyphal outgrowths were present. Isolates of the fungus were heterothallic and produced oogonia and oospores rapidly and abundantly on V8 agar at 22°C only with the A1 mating type of P. nicotianae. We concluded that all isolates from Limonium had the A2 compatibility type. Antheridia were amphigynous. Oogonia were spherical and ranged from 20 to 30 μm, (average 27.5 μm). Oospores ranged from 18 to 27 μm, (average 23.1 μm). The observed characteristics are similar to those described for P. nicotianae. Isozyme analysis, using the dimeric enzymes malic enzyme (EC 1.1.1.40) and malate dehydrogenase (EC 1.1.1.37), revealed the presence of the Mdhp100 allele and the Mdh-2100 allele. Both alleles are characteristic for P. nicotianae (3). Based on morphological features and isozyme genotyping, isolates of Phytophthora from diseased Limonium plants could be assigned to P. nicotianae van Breda de haan (1). A report from Florida associated Phytophthora sp. with root rot of Limonium plants (2) but did not identify the species. According to the multi-decade records at the Netherlands Plant Protection Service (unpublished data) Phytophthora has never been observed on Limonium before. This is the first report of P. nicotianae associated with root rot and basal rot of Limonium plants in Europe. References: (1) D. C. Erwin and O. K. Ribeiro. 1996. Phytophthora Diseases Worldwide. American Phytopathological Society, St. Paul, MN. (2) D. F. Farr et al. 1989. Fungi on Plants and Plant Products in the United States. American Phytopathological Society, St. Paul, MN. (3) W. A. Man in 't Veld et al. Phytopathology 88:922–929, 1998. (4) P. Oudemans and M. D. Coffey. Mycol. Res. 95:1025–1046, 1991.


Plant Disease ◽  
2012 ◽  
Vol 96 (9) ◽  
pp. 1381-1381
Author(s):  
A. Garibaldi ◽  
D. Bertetti ◽  
A. Poli ◽  
L. Bizioli ◽  
M. L. Gullino

Kalmia latifolia L., common name Mountain Laurel, is an evergreen shrub that is becoming increasingly popular in gardens. It is also grown as a potted plant for its round flowers that range from light pink to white and occur in clusters in late spring. During July 2011, 3-year-old plants of K. latifolia ‘Olympic Fire’ showed extensive chlorosis and root rot on several commercial nurseries close to Maggiore Lake (Verbano-Cusio-Ossola Province) in northwestern Italy. Twigs wilted and died and leaves dropped, although in some cases, wilted leaves persisted on stems. The whole root system was affected with vascular tissues on the lower stem exhibiting brown discoloration, followed by plant death. The disease was severe and widespread, affecting 5% of approximately 3,500 plants. Tissue fragments of 1 mm2 were excised from the margins of the lesions and plated. A Phytophthora-like organism was consistently isolated on an oomycete-selective medium (BNPRA + HMI at 25 μg/ml) (4) after disinfesting root pieces for 1 min in a 1% NaOCl solution. The pathogen was identified based on morphological and physiological features as Phytophthora cinnamomi (2). Oogonia didn't form in single culture. On V8 agar, the microorganism was characterized by coenocytic coralloid hyphae, with spherical lateral and terminal swellings 23 to 46 (average 34) μm in diameter (n = 50), single or in clusters, and produced spherical, terminal chlamydospores 35 to 47 (average 40) μm in diameter (n = 50). No sporangia were produced after growing pure cultures in sterilized soil extracts nor were they produced on V8 agar. The internal transcribed spacer (ITS) region of the rDNA of a single isolate was amplified using the primers ITS1/ITS4 and sequenced. BLAST analysis (1) of the 898-bp segment showed a 99% homology with the sequence of P. cinnamomi (GU799638). The nucleotide sequence has been assigned the GenBank Accession No. JQ951607. Pathogenicity of one isolate obtained from infected plants was confirmed by inoculating 18-month-old plants of K. latifolia ‘Olympic Fire’. The isolate was grown for 50 days in a mixture of 70:30 wheat/hemp kernels and then mixed into a substrate containing sphagnum peat moss/pumice/pine bark/clay (50:20:20:10 v/v) at a rate of 3 g/l. One plant per 2-l pot was transplanted into the substrate and constituted the experimental unit. Five plants were inoculated. Noninoculated plants represented the control treatment and the trial was repeated once. All plants were kept in a greenhouse at 24 to 27°C. Two of five plants inoculated developed symptoms of chlorosis, wilting, and root rot after 70 days and remaining plants after about 80 days. P. cinnamomi was reisolated consistently from inoculated plants but not from controls that remained symptomless. To our knowledge, this is the first report of P. cinnamomi on K. latifolia in Italy and in Europe. The disease has been reported in the United States (3). The economic importance of the disease is uncertain because of the limited number of nurseries that grow this crop in Italy, although its importance could increase as the popularity of K. latifolia increases. References: (1) S. F. Altschul et al. Nucleic Acids Res. 25:3389, 1997 (2) D. C. Erwin and O. K. Ribeiro. Phytophthora Diseases Worldwide. American Phytopathological Society, St Paul, MN, 1996. (3) L. F. Grand. North Carolina Agric. Res. Serv. Techn. Bull. 240, 1985. (4) H. Masago et al. Phytopathology 67:425, 1977.


Plant Disease ◽  
2004 ◽  
Vol 88 (6) ◽  
pp. 681-681
Author(s):  
A. Garibaldi ◽  
D. Bertetti ◽  
M. L. Gullino

Aquilegia flabellata Sieb. and Zucc. (columbine) is a perennial garden species belonging to the family Ranunculaceae. During the summer of 2003, a severe outbreak of a previously unknown powdery mildew was observed in several gardens near Biella (northern Italy). Upper surfaces of leaves were covered with a white mycelium and conidia, and as the disease progressed infected leaves turned yellow and died. Foot cell was cylindric and appressorium lobed. Conidia were hyaline, ellipsoid, and measured 31.2 to 47.5 × 14.4 to 33 μm (average 38.6 × 21.6 μm). Fibrosin bodies were not present. Cleistothecia were globose, brown, had simple appendages, ranged from 82 to 127 (average 105) μm in diameter, and contained one to two asci. Ascocarp appendages measured five to eight times the ascocarp diameter. Asci were cylindrical (ovoidal) and measured 45.3 to 58.2 × 30.4 to 40.2 μm. Ascospores (three to four per ascus) were ellipsoid or cylindrical and measured 28.3 to 31.0 × 14.0 to 15.0 μ;m. On the basis of its morphology, the pathogen was identified as Erysiphe aquilegiae var. aquilegiae (1). Pathogenicity was confirmed by gently pressing diseased leaves onto leaves of five, healthy A. flabellata plants. Five noninoculated plants served as controls. Inoculated and noninoculated plants were maintained in a garden where temperatures ranged between 20 and 30°C. After 10 days, typical powdery mildew symptoms developed on inoculated plants. Noninoculated plants did not show symptoms. To our knowledge, this is the first report of the presence of powdery mildew on Aquilegia flabellata in Italy. E. communis (Wallr.) Link and E. polygoni DC. were reported on several species of Aquilegia in the United States (2), while E. aquilegiae var. aquilegiae was previously observed on A. flabellata in Japan and the former Union of Soviet Socialist Republics (3). Specimens of this disease are available at the DIVAPRA Collection at the University of Torino. References: (1) U. Braun. Nova Hedwigia, 89:700, 1987. (2) D. F. Farr et al. Fungi on Plants and Plant Products in the United States. The American Phytopathological Society, St Paul, MN, 1989. (3) K. Hirata. Host Range and Geographical Distribution of the Powdery Mildews. Faculty of Agriculture, Niigata University, 1966.


Plant Disease ◽  
2011 ◽  
Vol 95 (10) ◽  
pp. 1316-1316 ◽  
Author(s):  
M. M. Díaz Arias ◽  
G. P. Munkvold ◽  
L. F. Leandro

Fusarium spp. are widespread soilborne pathogens that cause important soybean diseases such as damping-off, root rot, Fusarium wilt, and sudden death syndrome. At least 12 species of Fusarium, including F. proliferatum, have been associated with soybean roots, but their relative aggressiveness as root rot pathogens is not known and pathogenicity has not been established for all reported species (2). In collaboration with 12 Iowa State University extension specialists, soybean roots were arbitrarily sampled from three fields in each of 98 Iowa counties from 2007 to 2009. Ten plants were collected from each field at V2-V3 and R3-R4 growth stages (2). Typical symptoms of Fusarium root rot (2) were observed. Symptomatic and asymptomatic root pieces were superficially sterilized in 0.5% NaOCl for 2 min, rinsed three times in sterile distilled water, and placed onto a Fusarium selective medium. Fusarium colonies were transferred to carnation leaf agar (CLA) and potato dextrose agar and later identified to species based on cultural and morphological characteristics. Of 1,230 Fusarium isolates identified, 50 were recognized as F. proliferatum based on morphological characteristics (3). F. proliferatum isolates produced abundant, aerial, white mycelium and a violet-to-dark purple pigmentation characteristic of Fusarium section Liseola. On CLA, microconidia were abundant, single celled, oval, and in chains on monophialides and polyphialides (3). Species identity was confirmed for two isolates by sequencing of the elongation factor (EF1-α) gene using the ef1 and ef2 primers (1). Identities of the resulting sequences (~680 bp) were confirmed by BLAST analysis and the FUSARIUM-ID database. Analysis resulted in a 99% match for five accessions of F. proliferatum (e.g., FD01389 and FD01858). To complete Koch's postulates, four F. proliferatum isolates were tested for pathogenicity on soybean in a greenhouse. Soybean seeds of cv. AG2306 were planted in cones (150 ml) in autoclaved soil infested with each isolate; Fusarium inoculum was applied by mixing an infested cornmeal/sand mix with soil prior to planting (4). Noninoculated control plants were grown in autoclaved soil amended with a sterile cornmeal/sand mix. Soil temperature was maintained at 18 ± 1°C by placing cones in water baths. The experiment was a completely randomized design with five replicates (single plant in a cone) per isolate and was repeated three times. Root rot severity (visually scored on a percentage scale), shoot dry weight, and root dry weight were assessed at the V3 soybean growth stage. All F. proliferatum isolates tested were pathogenic. Plants inoculated with these isolates were significantly different from the control plants in root rot severity (P = 0.001) and shoot (P = 0.023) and root (P = 0.013) dry weight. Infected plants showed dark brown lesions in the root system as well as decay of the entire taproot. F. proliferatum was reisolated from symptomatic root tissue of infected plants but not from similar tissues of control plants. To our knowledge, this is the first report of F. proliferatum causing root rot on soybean in the United States. References: (1) D. M. Geiser et al. Eur. J. Plant Pathol. 110:473, 2004. (2) G. L. Hartman et al. Compendium of Soybean Diseases. 4th ed. The American Phytopathologic Society, St. Paul, MN, 1999. (3) J. F. Leslie and B. A. Summerell. The Fusarium Laboratory Manual. Blackwell Publishing, Oxford, UK, 2006. (4) G. P. Munkvold and J. K. O'Mara. Plant Dis. 86:143, 2002.


Plant Disease ◽  
2011 ◽  
Vol 95 (9) ◽  
pp. 1187-1187
Author(s):  
J. J. Sadowsky ◽  
T. D. Miles ◽  
A. M. C. Schilder

Necrotic stems and leaves were observed on 2- to 4-month-old, rooted microshoot plants (Vaccinium corymbosum L. ‘Liberty’ and ‘Bluecrop’, V. angustifolium Aiton ‘Putte’, and V. corymbosum × V. angustifolium ‘Polaris’) in a Michigan greenhouse in 2008 and 2009. As the disease progressed, leaves fell off and 80 to 100% of the plants died in some cases. Root rot symptoms were also observed. A fungus was isolated from stem lesions. On potato dextrose agar (PDA), cultures first appeared light tan to orange, then rusty brown and zonate with irregular margins. Chains of orange-brown chlamydospores were abundant in the medium. Macroconidiophores were penicillately branched and had a stipe extension of 220 to 275 × 2.5 μm with a narrowly clavate vesicle, 3 to 4 μm wide at the tip. Conidia were hyaline and cylindrical with rounded ends, (1-)3-septate, 48 to 73 × 5 to 7 (average 60 × 5.5) μm and were held together in parallel clusters. Perithecia were globose to subglobose, yellow, 290 to 320 μm high, and 255 to 295 μm in diameter. Ascospores were hyaline, 2- to 3-septate, guttulate, fusoid with rounded ends, slightly curved, and 30 to 88 × 5 to 7.5 (average 57 × 5.3) μm. On the basis of morphology, the fungus was identified as Calonectria colhounii Peerally (anamorph Cylindrocladium colhounii Peerally) (1,2). The internal transcribed spacer region (ITS1 and ITS2) of the ribosomal DNA and the β-tubulin gene were sequenced (GenBank Accession Nos. HQ909028 and JF826867, respectively) and compared with existing sequences using BLASTn. The ITS sequence shared 99% maximum identity with that of Ca. colhounii CBS 293.79 (GQ280565) from Java, Indonesia, and the β-tubulin sequence shared 97% maximum identity with that of Ca. colhounii CBS 114036 (DQ190560) isolated from leaf spots on Rhododendron sp. in North Carolina. The isolate was submitted to the Centraalbureau voor Schimmelcultures in the Netherlands (CBS 129628). To confirm pathogenicity, 5 ml of a conidial suspension (1 × 105/ml) were applied as a foliar spray or soil drench to four healthy ‘Bluecrop’ plants each in 10-cm plastic pots. Two water-sprayed and two water-drenched plants served as controls. Plants were misted intermittently for 2 days after inoculation. After 7 days at 25 ± 3°C, drench-inoculated plants developed necrotic, sporulating stem lesions at the soil line, while spray-inoculated plants showed reddish brown leaf and stem lesions. At 28 days, three drench-inoculated and one spray-inoculated plant had died, while others showed stem necrosis and wilting. No symptoms were observed on control plants. Fungal colonies reisolated from surface-disinfested symptomatic stem, leaf, and root segments appeared identical to the original isolate. Cy. colhounii was reported to cause a leaf spot on blueberry plants in nurseries in China (3), while Ca. crotalariae (Loos) D.K. Bell & Sobers (= Ca. ilicicola Boedijn & Reitsma) causes stem and root rot of blueberries in North Carolina (4). To our knowledge, this is the first report of Ca. colhounii causing a disease of blueberry in Michigan or the United States. Because of its destructive potential, this pathogen may pose a significant threat in blueberry nurseries. References: (1) P. W. Crous. Taxonomy and Pathology of Cylindrocladium (Calonectria) and Allied Genera. The American Phytopathological Society, St. Paul, MN, 2002. (2) L. Lombard et al. Stud. Mycol. 66:31, 2010. (3) Y. S. Luan et al. Plant Dis. 90:1553, 2006. (4) R. D. Milholland. Phytopathology 64:831, 1974.


Plant Disease ◽  
2018 ◽  
Vol 102 (10) ◽  
pp. 2035 ◽  
Author(s):  
J. L. Jacobs ◽  
K. Oudman ◽  
H. Sang ◽  
M. I. Chilvers

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