First Report of Root Rot of Pedunculate Oak and Other Forest Tree Species Caused by Phytophthora plurivora in the Czech Republic

From 2006 to 2008, several similar Phytophthora isolates were obtained from roots of mature Quercus robur and other tree species (Acer platanoides, Fraxinus excelsior, Q. rubra, and Tilia cordata) in forests and parks in several areas in the Czech Republic. The trees were characterized by chlorotic and reduced foliage, crown dieback, and reduced root hairs. Several isolates of Phytophthora were obtained from necrotic roots of these trees and identified as Phytophthora plurivora Jung & Burgess (1). Isolated colonies grown on V8A medium were radiate to slightly chrysanthemum shaped with limited aerial mycelium in the center. Optimum growth was at 25°C, minimum at 5°C and maximum at 32°C. Radial growth of colonies averaged 6.4 mm/day at 20°C. The isolates were homothallic and produced abundant smooth-walled, spherical oogonia (23.3 to 29.1 μm in diameter), oospores were nearly plerotic or plerotic (21.8 to 26.9 μm in diameter), and the oospore wall was 1.2 to 1.4 μm thick. Antheridia were usually paragynous and measured 8.4 to 12 × 6.5 to 8 μm, but amphigynous antheridia were occasionally observed. Noncaducous, semipapillate sporangia formed on simple or sympodial sporangiophores, were obpyriform, ovoid, ellipsoid or irregular in shape, and occasionally distorted with more than one apex. Sporangia dimensions were 33 to 65 × 24 to 33 μm; L/B ratio 1.2 to 1.6 (–2.1). Comparison of DNA sequences of internal transcribed spacer (ITS) regions of isolates (representative strain GenBank Accession No. FJ952382) confirmed the 100% identity of P. plurivora (1). The soil infestation test was conducted using a P. plurivora isolate acquired from roots of Q. robur and 20 3-year-old plants of Q. robur. Sterilized millet seeds colonized by pathogen with the method as described (2) were used as inoculation medium and added into sterilized peat substrate at the rate of 0.5% (vol/vol). The plants were cultivated in 5.8-liter pots in a greenhouse (20°C, 16-h/8-h photoperiod). After 4 months, the roots of all plants were washed, dried, and weighed. The root biomass of 20 infected plants was significantly reduced by approximately 25% on average compared with the control 20 plants (P < 0.05, t-test, Statistica 7.1). The pathogen was consistently reisolated from the roots of infected plants but not from control plants. Stem inoculation tests were conducted with 20 replicates in each group of 2-year-old plants of oak, maple, ash, and lime and isolates acquired from the hosts. On each seedling, a 5-mm-diameter bark plug was removed 5 cm above the collar. The inoculum (5-mm-diameter V8A agar plug with actively growing mycelium) was applied to the exposed substrate. The wounds were sealed with Parafilm. Stem necrosis developed in all cases after 1 to 2 weeks, whereas control plants remained healthy. The pathogen was successfully reisolated from necrotic stem tissues. To our knowledge, this is the first report of P. plurivora causing root rot on oak, maple, ash, and lime in the Czech Republic. On the basis of the host range and distribution of P. plurivora in the Czech Republic, it can be assumed that, as elsewhere in Europe (1), this pathogen is widespread and is a common cause of decline of many tree species. References: (1) T. Jung and T. I. Burgess. Persoonia 22:95, 2009. (2) C. Robin et al. Plant Pathol. 50:708, 2001.

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First Report of Phoma exigua var. populi Causing Canker of Twigs and Shoots of Poplar in the Czech Republic

Plant Disease ◽

10.1094/pdis-92-10-1473a ◽

2008 ◽

Vol 92 (10) ◽

pp. 1473-1473

Author(s):

K. Cerny ◽

M. Malinova ◽

M. Tomsovsky ◽

V. Strnadova ◽

V. Holub ◽

...

Keyword(s):

Czech Republic ◽

Morphological Characteristics ◽

Aerial Mycelium ◽

First Record ◽

The Czech Republic ◽

Short Rotation ◽

Agar Plug ◽

Southern Moravia ◽

Host Tissues ◽

Populus Spp

During 2007 and the spring of 2008, a disease of poplars (Populus spp.) resembling the Dothichiza canker was found in plantations of fast-growing trees in central Bohemia and in southern Moravia where it was more abundant. The yellowish brown-to-brown, round or elongated cankers occurred on damaged shoots and twigs. Tissues directly under the bark were discolored and turned black. As the cankers enlarged, infected shoots and twigs died after several months. Small, black, gregarious pycnidia were observed under the bark or in lenticels after several weeks. The disease occurred on Populus nigra, P. × euroamericana cvs. Regenerata, Robusta, Brabantica, Spreewald, CZ-425/58, Blanc du Poitou, and Flaschlanden, and other Populus spp. Isolates of a species of Phoma were acquired by culturing damaged tissues on agar plates containing 3% oatmeal agar (OA) and 2% malt agar. Initial identification of the isolates was done by cultural and morphological characteristics (1). Colonies were floccose, aerial mycelium was olivaceous gray to gray, reverse olivaceous gray sometimes with darker tones at the margins or in the colony center, and NaOH reaction was negative. The growth rate was 42 to 56 in diameter after 7 days at 20°C on OA (optimum temperature for growth was 22°C with a minimum of 1°C and a maximum of 28 to 29°C). Pycnidia in culture scattered, were globose or subglobose, obviously with one nonpapillate ostiolum, olivaceous black or black, 120 to 370 μm in diameter, and conidial exudate was whitish. Phialides were globose to ampulliform and 3 to 7 × 3 to 6 μm. Conidia were hyaline, ellipsoidal, often guttulate, 3.1 to 7.8 × 1.9 to 3.1 μm, and L/B ratio 1.4:3.1. Septate conidia occurred only on natural substrate up to 10.6 × 3.9 μm. Morphological and cultural characteristics resembled those of P. exiqua var. populi Gruyter & P. Scheer (1). The internal transcribed spacer (ITS) sequence (GenBank Accession No EU562206) for the representative isolate (CCF No 3759) confirmed 100% identity to P. exigua. Pathogenicity was confirmed with 1-year-old P. nigra plants during a 2-month greenhouse experiment at 15 to 20°C. Fifteen replicate plants were wounded (5-mm diameter), inoculated with 5-mm OA plugs from actively growing colonies (isolate CCF No 3759), and sealed by Parafilming. An additional 15 control plants following wounding were inoculated with a sterile agar plug. After 3 to 4 weeks, yellowish or brownish necrotic lesions ranging from 1 to 1.5 cm long developed on all inoculated plants. The pathogen was successfully reisolated from lesions and the control plants were asymptomatic. P. exigua var. populi is considered an opportunistic poplar and willow pathogen (2) that becomes more important in winter (1). The pathogen potentially invades host tissues damaged by frost, sun scald, or weakened by excessive transpiration during sunny winter days. To our knowledge, this is the first record of the pathogen on poplars in the Czech Republic, which may have an economic impact on short-rotation coppice plantations. References: (1) J. de Gruyter and P. Scheer. J. Phytopathol. 146:411, 1998. (2) H. A. van der Aa et al. Persoonia 17:435, 2000.

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First Report of Phytophthora hedraiandra Causing Rhododendron Dieback and Root Rot of Common Beech in the Czech Republic

Plant Disease ◽

10.1094/pdis-04-14-0339-pdn ◽

2014 ◽

Vol 98 (10) ◽

pp. 1434-1434

Author(s):

M. Hejna ◽

K. Cerny ◽

L. Havrdova ◽

M. Mrazkova

Keyword(s):

Czech Republic ◽

Western Europe ◽

Aerial Mycelium ◽

Its Region ◽

Average Diameter ◽

The United States ◽

The Czech Republic ◽

First Report ◽

Common Beech ◽

Forest Nurseries

From 2010 to 2012, Phytophthora isolates were obtained from brownish diffusion leaf lesions usually up to 2 to 3 cm in diameter of Rhododendron caucasicum ‘Cheer,’ from withered twigs of Rhododendron sp. with blackish elongated lesions up to ~5 cm in length, and from rotten feeder roots of 2-year-old, chlorotic, wilting seedlings of Fagus sylvatica collected from ornamental and forest nurseries in three areas (central and eastern Bohemia and northern Moravia) in the Czech Republic. Isolates formed chrysanthemum-like to slightly stellate, appressed colonies with sparse aerial mycelium on V8 agar (V8A) plates at 20°C after 5 days, whereas on carrot agar (CA) plates the pattern was vague with no aerial mycelium. The cardinal growth temperatures were: min. 3°C, optimum 23 to 27°C, and max. 31°C. Radial growth was 5.7 to 6.6 mm/day at 20°C on V8A. The isolates were homothallic and produced colorless, smooth-walled, spherical oogonia with an average diameter 29.9 to 33.8 μm on CA. Oospores were aplerotic (avg. 26.4 to 29.3 μm), oospore wall was hyaline and averaged 1.3 to 1.7 μm thick, oospore wall index was 0.26 to 0.32. Paragynous or occasionally amphigynous antheridia averaged 13.4 to 15.0 × 10.9 to 12.5 μm (height × width). Sporangia were caducous, papillate, globose, spherical to ovoid, with short pedicels (avg. 2.1 to 2.6 μm) and averaged 30.9 to 41.5 × 25.5 to 30.6 μm, L:B ratio was 1.2 to 1.4. Chlamydospores were not observed. Morphological characters resembled those described for P. hedraiandra (1). The isolates were deposited in the collection of phytopathogenic oomycetes of RILOG Pruhonice and given accession nos. 450.11, 531.11, and 578.12. The isolates were sequenced for nuclear rDNA ITS region and partial Cox I gene. Obtained sequences were compared with sequences present in GenBank database using BLAST. The ITS sequences of all isolates (GenBank Accession Nos. KJ567081, 82, and 83) of overall length of 792 bp were identical to that of P. hedraiandra AY707987 (1). The Cox I sequences of overall length of 880 bp (KJ567084, 85, and 86) showed 99% homology (1 bp substitution) with AY769115 (1) and 100% identity with other Cox I sequences of P. hedraiandra, i.e., JN376067 (4) and EF174432 (3). Koch's postulates were confirmed by wound-inoculating of 3-year-old rhododendron and common beech plants (10 host plants per corresponding isolate). Rhododendron leaves were gently abraded near the midrib, whereas 5-mm-diameter bark plugs were removed from the beech collars. The inoculum (5-mm-diameter V8A plug with actively growing mycelium) was placed over wounds and sealed with Parafilm. Control plants were treated in the same manner with sterile agar plugs. Plants were maintained in a greenhouse at 22°C. All inoculated plants showed disease symptoms after 10 days of incubation; the lesions were up to 2 cm in rhododendron leaves and ~1 cm in beech collars. Control plants remained healthy. The pathogen was re-isolated from all infected plants. To our knowledge, this is the first report of P. hedraiandra in the Czech Republic. Besides it, the pathogen was found in southern and western Europe (Italy, Slovenia, Spain, the Netherlands) and in the United States (2). References: (1) A. W. A. M. de Cock and A. Lévesque. Stud. Mycol. 50:481, 2004. (2) D. F. Farr and A. Y. Rossman. Fungal Databases, Syst. Mycol. Microbiol. Lab., ARS, USDA. Retrieved from http://nt.ars-grin.gov/fungaldatabases/ , May 13, 2014. (4) E. Moralejo et al. Span. J. Agric. Res. 5:82, 2007. (2) X. Yang et al. Plant Dis. 96:915, 2012.

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First Report of Phytophthora Rot on Alders Caused by Phytophthora alni subsp. alni in Spain

Plant Disease ◽

10.1094/pdis-94-2-0273a ◽

2010 ◽

Vol 94 (2) ◽

pp. 273-273 ◽

Cited By ~ 5

Author(s):

C. Pintos Varela ◽

C. Rial Martínez ◽

J. P. Mansilla Vázquez ◽

O. Aguín Casal

Keyword(s):

Dna Sequences ◽

Mitochondrial Gene ◽

Aerial Mycelium ◽

Selective Medium ◽

Mt Dna ◽

First Report ◽

Inner Bark ◽

Agar Plug ◽

Plant Pathol ◽

Phytophthora Rot

Phytophthora alni, a soil- and waterborne pathogen, causes aggressive root and collar rot on riparian alder populations (1,2,4). The disease has been described from several European countries with a destructive impact in Great Britain (1,2). All European alder species and the red alder (Alnus rubra) are highly susceptible. P. alni has multiple variants that have been placed in three subspecies: P. alni subsp. alni, P. alni subsp. uniformis, and P. alni subsp. multiformis (1). In July 2009, a survey of symptoms of Phytophthora rot from A. glutinosa at 20 riparian stands along the Avia River in Galicia (northwest Spain) was conducted. Affected trees showed symptoms of Phytophthora rot including abnormally small, sparse, and yellowish foliage, dieback in the canopy, necroses of the inner bark and cambium, and bleeding cankers on the trunks (2,4). Phytophthora spp. were baited from saturated rhizosphere soil and watercourses using oak leaflets (4). Roots and tissue from fresh active inner bark lesions were transferred to selective medium V8-PARPH agar (4) and incubated for 7 days at 22°C in the dark. A Phytophthora sp. was isolated, transferred to carrot agar (CA), and incubated in the dark. Colonies were appressed, often irregular in outline, and with limited aerial mycelium (1). Growth on CA occurred from 4 to 31°C with optimum growth at 23 to 25°C. Chlamydospores were not observed. Ellipsoid, nonpapillate, noncaducous sporangia had a length/breadth average ratio of 1.4. Nesting and extended internal proliferation occurred. Oogonia, antheridia, and oospores were abundantly produced in a single culture. Oogonia with tapered stalks were spherical (mature oogonia 38 to 50 μm in diameter) and some had ornamented walls or bullate protuberances (1,2). Antheridia were large, amphigynous, and predominantly two-celled (23 to 37 × 16 to 23 μm). Oospores were plerotic. Distorted comma-shaped or smaller oogonia and aborted oospores were observed (1). Amplification of DNA was accomplished by using sequence-characterized amplification region-PCR primers (3). The amplicon sizes obtained were identical to P. alni subsp. alni (3). Internal transcribed spacer (ITS)-DNA and nadh1 mitochondrial gene were also amplified. DNA sequences of ITS and mt-DNA regions were deposited in GenBank (Nos. GU108602 and GU108603). Comparison of the sequences showed 100% homology with P. alni subsp. alni (GenBank Nos. FJ746679 and DQ202490). P. alni subsp. alni was recovered from trees at 3 of 20 riparian alder stands with symptoms. Pathogenicity of one representative isolate was confirmed by inoculating 10 3-year-old A. glutinosa seedlings grown in pots. One shallow cut was made into the bark at the collar level. A colonized agar plug, from the margin of an actively growing colony of P. alni subsp. alni, was inserted beneath the flap that was sealed with Parafilm. Five controls seedlings received only sterile CA agar plugs. Plants were incubated at 24°C and 95% humidity for 30 days. On inoculated plants, necroses progressed bidirectionally from the wound, and dead leaves and wilting of shoots were observed. P. alni subsp. alni was recovered from inoculated seedlings, but not from controls. To our knowledge, this is the first report of Phytophthora rot on alder caused by P. alni subsp. alni in Spain. References: (1) C. M. Brasier et al. Mycol. Res. 108:1172, 2004. (2) J. Gibbs et al. For. Comm. Bull. 126, 2003 (3) R. Ioos et al. Eur. J. Plant Pathol. 112:323, 2005. (4) T. Jung et al. Plant Pathol. 53:197, 2004.

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First Report of Leaf Spot, Shoot Blight, and Stem and Collar Canker of Rhododendron spp. Caused by Phytophthora citricola in the Czech Republic

Plant Disease ◽

10.1094/pdis-91-11-1515b ◽

2007 ◽

Vol 91 (11) ◽

pp. 1515-1515

Author(s):

M. Mrazkova ◽

K. Cerny ◽

S. Gabrielova ◽

M. Tomsovsky

Keyword(s):

Czech Republic ◽

Morphological Characteristics ◽

Aerial Mycelium ◽

Culture Collection ◽

The Czech Republic ◽

First Report ◽

Public Gardens ◽

Phytophthora Citricola ◽

Plastic Bag ◽

Shoot Blight

During the summer and autumn of 2006, a disease of rhododendron plants (Ericaceae) was found in nurseries and public gardens in several areas of the Czech Republic. Leaves of damaged plants showed dark brown-to-black lesions extending along the mid-rib and commonly spreading to petioles and shoots. The infected shoots turned black and died. The cankers on branches, stems, and collars were characterized by reddish, brownish, or blackish discoloration. The disease was identified on Rhododendron catawbiense, R. repens, and other Rhododendron spp. After plating pieces of symptomatic tissue on PARPNH medium (2), several isolates of a homothallic Phytophthora sp. were acquired. Ten representative isolates of the pathogen were cultivated on V8A plates and examined for cultural and morphological characteristics. Colonies had a stellate pattern of growth with sparse aerial mycelium at 20°C; optimum temperature for growth was 25 to 28°C, minimum was 4°C, and maximum was 33°C. Radial growth was 14 mm per day at 20°C on V8A. The isolates produced terminal, spherical, smooth-walled oogonia, which were 19 to 37 μm in diameter. Oospores were plerotic (17 to 32 μm) with walls 2 to 4 μm thick; antheridia were paragynous. Single, terminal, noncaducous, semipapillate sporangia were formed on simple (occasionally sympodial) sporangiophores in nonsterile soil filtrate. The sporangia (28 to 61 × 24 to 35 μm, L:B ratio 1.5) were mostly obpyriform, rarely obovoid, or ovoid-ellipsoid. Morphological and cultural characters resembled those described for Phytophthora citricola Sawada (1). The ITS sequences of the rDNA of the two representative isolates (GenBank Accession Nos. EF194772 and EF194773) showed 100% homology to P. citricola sequences obtained from GenBank, thus the identity was confirmed as P. citricola. Both specimens were deposited in CCF (Culture Collection of Fungi, Charles University, Prague, Czech Republic). To confirm the pathogenicity of isolates, Koch's postulates were tested using 40 3-year-old potted rhododendron (R. catawbiense and R. repens) plants and the two P. citricola strains deposited in CCF. Surfaces of attached healthy leaves were disinfected with 95% ethanol and gently abraded with a sterile scalpel near the mid-rib. Agar plugs from the margin of a 5-day-old colony grown on carrot agar were placed on leaf surfaces and also inserted under flaps of stem tissues made with a sterile scalpel. The leaves and stems were then sealed with Parafilm. Control plants were treated in the same manner with sterile agar plugs. All plants were watered with deionized water, covered with a plastic bag, and maintained in a greenhouse at 21°C for 6 weeks. All inoculated plants exhibited necrotic lesions on leaves and stems around the points of inoculation after 4 days, whereas the control plants remained healthy. The pathogen was consistently reisolated from symptomatic plants. P. citricola is well known as a pathogen of rhododendron (1), but to our knowledge, this is the first report of P. citricola on Rhododendron sp. in the Czech Republic. P. citricola has been found at five different locations and in the most frequently isolated Phytophthora spp. from rhododendron in the Czech Republic. References: (1) D. C. Erwin and O. K. Ribeiro. Phytophthora Diseases Worldwide. The American Phytopathological Society. St. Paul, MN, 1996. (2) T. Jung et al. Eur. J. For. Pathol. 26:253, 1996.

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First report of Cherry virus F infecting Japanese plum in Korea

Plant Disease ◽

10.1094/pdis-08-20-1725-pdn ◽

2020 ◽

Author(s):

Yeonhwa Jo ◽

Hoseong Choi ◽

Jin Kyong Cho ◽

Won Kyong Cho

Keyword(s):

Czech Republic ◽

High Throughput Sequencing ◽

Prunus Avium ◽

De Novo ◽

Protein Database ◽

Specific Primers ◽

The Czech Republic ◽

First Report ◽

Japanese Plum ◽

Leaf Spots

Cherry virus F (CVF) is a tentative member of the genus Fabavirus in the family Secoviridae, consisting of two RNA segments (Koloniuk et al. 2018). To date, CVF has been documented in only sweet cherry (Prunus avium) in the Czech Republic (Koloniuk et al. 2018), Canada, and Greece. In May 2014, we collected leaf samples from four symptomatic (leaf spots and dapple fruits) and two asymptomatic Japanese plum cultivars (Sun and Gadam) grown in an orchard in Hoengseong, South Korea, to identify viruses and viroids infecting plum trees. Total RNA from individual plum trees was extracted using two commercial kits: Fruit-mate for RNA Purification Kit (Takara, Shiga, Japan) and RNeasy Plant Mini Kit (Qiagen, Hilden, Germany). We generated six mRNA libraries from the six different plum cultivars for RNA-sequencing using the TruSeq RNA Library Preparation Kit v2 (Illumina, CA, U.S.A.) as described previously (Jo et al. 2017). The mRNA libraries were paired-end (2 X 100 bp) sequenced with a HiSeq 2000 system (Macrogen, Seoul, Korea). The raw sequence reads were de novo assembled by Trinity program v. 2.8.6, with default parameters (Haas et al. 2013). The assembled contigs were subjected to BLASTX search against the non-redundant protein database in NCBI. Of the two asymptomatic cultivars, the transcriptome of asymptomatic plum cv. Gadam contained five contigs specific to CVF. Two and three contigs were specific to CVF RNA1 (2,571 reads, coverage 42.15%) and RNA2 (2,025 reads, coverage 53.04%), respectively. The size of these five contigs ranged from 241 to 5,986 bp. Contigs of 5,986 and 3,867 bp in length, referred to as CVF isolate Gadam RNA1 (GenBank MN896996) and RNA2 (GenBank MN896995), respectively, were subjected to BLASTP search against NCBI’s non-redundant protein database. The results showed that the polyprotein sequences of RNA1 and RNA2 shared 95.3% and 93.11% amino acid identities with isolates SwC-H_1a from the Czech Republic (GenBank acc. no. AWB36326) and Stac-3B_c8 from Canada (AZZ10055), respectively. To confirm the infection of CVF in cv. Gadam, RT-PCR was conducted using CVF RNA1-specific primers designed based on the CVF reference genome sequences (MH998210 and MH998216), including 5’-CCACCAAATAGGCAAGAGGTCAC-3’ (position 3190–3212) and 5’-CACAATCACCATCAATGGTCTCTGC-3’ (position 3742–3766), and CVF RNA2-specific primers, including 5’-CTGCTTTATGATGCTAGACATCAAGATG-3’ (position 1015–1042) and 5’-ACAATAGGCATGCTCATCTCAACCTC-3’ (position 1594–1619). We amplified 577-bp RNA1-specific and 605-bp RNA2-specific amplicons that were cloned and then performed Sanger sequencing. Sequencing of the cloned amplicons for isolate Gadam RNA1 (GenBank MN896993) and RNA2 (GenBank MN896994) revealed values of 99.48% and 99.17% nucleotide identity to that of RNA1 and RNA2 determined by high-throughput sequencing, respectively. Additionally, we tested five plants for each of the six plum cultivars grown in the same orchard. The detection of CVF was carried out through PCR using the primers and protocol described above. Of the 30 trees, CVF was detected in three trees of cv. Gadam by both primer pairs. To our knowledge, this is the first report of CVF infecting Japanese plum and the first report of the virus in Korea. However, its prevalence in other Prunus species, including apricot, European plum, and peach, should be further elucidated.

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First Report of Colletotrichum acutatum on Tomato and Apple Fruits in the Czech Republic

Plant Disease ◽

10.1094/pdis-10-11-0849-pdn ◽

2012 ◽

Vol 96 (5) ◽

pp. 769-769 ◽

Cited By ~ 4

Author(s):

J. Víchová ◽

B. Staňková ◽

R. Pokorný

Keyword(s):

Czech Republic ◽

Tomato Fruit ◽

Colletotrichum Acutatum ◽

Distilled Water ◽

Species Determination ◽

Specific Primers ◽

The Czech Republic ◽

First Report ◽

Pcr Products ◽

Apple Fruits

Apple (Malus domestica Borkh.) is a fruit traditionally grown in the Czech Republic, and tomatoes (Solanum lycopersicum Mill.), too, are widely raised in this region. Colletotrichum acutatum J. H. Simmonds is a polyphagous fungal plant pathogen. Earlier, this pathogen caused disease on strawberry in the Czech Republic (2), and now it has become an important pathogen on safflower (4). During the 2010 harvest, anthracnose symptoms were noticed on the fruits of apple and tomato. Infected apples fruits (localities Velká Bíteš and Znojmo) and tomatoes (localities Velká Bíteš and Žabčice) were collected. Typical symptoms on fruit surfaces were round, brown, shriveled and sunken spots, 1.2 to 2.0 cm, with orange conidial masses appearing on the spots. A fungus was isolated from each host on potato dextrose agar and cultured at 25 ± 2°C for 10 days. Mycelium was superficial, partly immersed, and white to gray with occurrence of orange conidial masses. Conidia of the tomato and apple isolates were colorless and fusiform. The size of conidia from the apple and tomato isolates, respectively, ranged from 11 to 15 × 2.5 to 3.5 μm and 11 to 16 × 2.5 to 4 μm. Morphological characteristics suggested that the isolated fungi was a Colletotrichum sp. To fulfill Koch's postulates, healthy tomato and apple fruits were disinfected with 3% sodium hypochlorite for 2 min and rinsed in sterile distilled water. Fruits were pinpricked with a sterile needle and 10 μl of a spore suspension (1 × 105 conidia ml–1) was inoculated by pipetting into the wound. Control fruits were treated with sterile distilled water. The fruits were transferred to a growth cabinet and maintained at a temperature of 25 ± 2°C, relative humidity of 70 ± 5%, and a photoperiod of 12 h. Similar disease symptoms as in the collected fruits were observed on tomato fruits at 7 days and apple fruits at 20 days after inoculation, while no symptoms appeared on control fruits. The pathogen was reisolated from infected fruits. Species determination of the isolates was confirmed by PCR. Specific primers designed in region ITS1, the 5.8S RNA gene, and region ITS2 of the pathogen DNA were selected. Specific primers CaInt2 and ITS4 were used to identify C. acutatum (3), and primers CgInt and ITS4 were used to determine C. gloeosporioides isolate CCM 177 (1), which was used as a control. Our isolates yielded PCR products (490 bp) only with primers designed for C. acutatum. The C. gloeosporioides isolate yielded a PCR product (450 bp) only with CgInt and ITS4 primers. PCR products were sequenced and identified with the BLAST program. The sequence of the tomato fruit isolate (Accession No. JN676199) and apple fruit isolate (Accession No. JN676198) matched with 100% similarity to the C. acutatum sequences in GenBank. The control isolate of C. gloeosporioides matched 100% to sequences AJ749682 and AJ749692. To our knowledge, this is the first report of C. acutatum on tomato and apple fruits in the Czech Republic. This pathogen can endanger the production and storage of apples and tomatoes in this region. References: (1) P. R. Mills et al. FEMS Microbiol. Lett. 98:137, 1992. (2) D. Novotný et al. Plant Dis. 91:1516, 2007. (3) S. Sreenivasaprasad et al. Plant Pathol. 45:650, 1996. (4) J. Víchová et al. Plant Dis. 95:79, 2011.

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The First Report of Downy Mildew Caused by Peronospora belbahrii on Sweet Basil in Greenhouses in the Czech Republic

Plant Disease ◽

10.1094/pdis-07-13-0747-pdn ◽

2014 ◽

Vol 98 (11) ◽

pp. 1579-1579 ◽

Cited By ~ 3

Author(s):

I. Šafránková ◽

L. Holková

Keyword(s):

Czech Republic ◽

Downy Mildew ◽

Width Ratio ◽

Specific Sequence ◽

The Czech Republic ◽

First Report ◽

Sweet Basil ◽

Potted Plants ◽

Length Width ◽

Necrotic Spots

Sweet basil (Ocimum basilicum L.) is an aromatic plant that is cultivated as a pot plant in greenhouses or in fields in the Czech Republic. The plants are intended for direct consumption or for drying. In April of 2012, the first large chlorotic from the middle necrotic spots occurred gradually on leaves of pot plants O. basilicum cv. Genovese in greenhouses in Central Bohemia. The characteristic gray to brown furry growth of downy mildew appeared on abaxial surfaces of leaves in the place of chlorotic spots within 3 to 4 days. The infested leaves fell off in the late stages of pathogenesis. The infestation gradually manifested itself in ever-younger plants and in July, cotyledons and possibly the first true leaves were already heavily infected and damaged and these plants rapidly died. The plant damage reached 80 to 100%, so it was necessary to stop growing the plants in the greenhouse at the end of July. The causal agent was isolated and identified as Peronospora belbahrii Thines by means of morphological and molecular characters (2,3). Conidiophores were hyaline, straight, monopodial, 280 to 460 μm, branched three to five times, ended with two slightly curved branchlets with a single conidia on each branchled tip. The longer branchlets measured 13 to 24 μm (average 18.2 μm), the shorter one 4 to 15 μm (average 9.7 μm). Conidia were rounded or slightly ovoid, from brownish to dark brownish, measured 22 to 31 × 20 to 28 μm (length/width ratio 1.2). A pathogen-specific sequence was detected with the help of the pathogen ITS rDNA specific primers in symptomatic leaves (1). DNA from plant tissues was isolated using the DNeasy plant Mini Kit (Qiagen, Germany) following the standard protocol. PCR was performed using KAPA2G Robust HotStar kit (Kapa Biosystems, United States) according to the conditions recommended in Belbahri et al. (1). The specific products were visualized by electrophoresis through 1.5% agarose gels. Leaves of 20-day-old potted plants O. basilicum ‘Genovese’ were inoculated by spraying with 5 × 105 conidia/ml of the pathogen. Each pot contained 10 plants. Sterilized distilled water was applied to control plants. Plants were covered with polyethylene bags during the entire incubation period to maintain high humidity, and kept at a temperature of 22 to 24°C. Typical disease symptoms appeared on leaves 5 to 9 days after inoculation. Control plants were symptomless. P. belbahrii was re-isolated from the lesions of inoculated plants, thus fulfilling Koch's postulates. Downy mildew on sweet basil was reported in countries in Africa, Europe, and South and North America (4). To our knowledge, this is the first report of downy mildew on sweet basil in the Czech Republic. References: (1) L. Belbahri et al. Mycol. Res. 109:1276, 2005. (2) Y.-J. Choi et al. Mycol. Res. 113:1340, 2009. (3) M. Thines et al. Mycol. Res. 113:532, 2009. (4) C. A. Wyenandt et al. HortScience 45:1416, 2010.

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Economic Impact of Douglas-fir (Pseudotsuga menziesii [Mirb.] Franco) production in the Czech Republic

Journal of Forest Science ◽

10.17221/27/2014-jfs ◽

2014 ◽

Vol 60 (No. 7) ◽

pp. 297-306 ◽

Cited By ~ 5

Author(s):

K. Pulkrab ◽

M. Sloup ◽

M. Zeman

Keyword(s):

Czech Republic ◽

Economic Impact ◽

Pseudotsuga Menziesii ◽

Tree Species ◽

Economic Effect ◽

Douglas Fir ◽

European Countries ◽

Natural Conditions ◽

The Czech Republic ◽

Forest Production

The article addresses the issues of Douglas-fir (Pseudotsuga menziesii [Mirb.] Franco) production in the Czech Republic (CR). Our analysis shows that the tree species can occupy 149,616–163,713 ha in the CR (with respect to ecological limits set by the Czech legislation). The potential economic effect expressed by the gross yield of forest production might be higher by 27–30 million EUR·yr–1.  The results of the analysis support the forest owners’ interest to extend Douglas-fir production in the CR, similarly like it has been extended systematically in all European countries where natural conditions allow.

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Testing of germination of spruce, pine and larch seed after 10 years from collection

Journal of Forest Science ◽

10.17221/74/2013-jfs ◽

2014 ◽

Vol 60 (No. 12) ◽

pp. 540-543

Author(s):

I. Tomášková ◽

J. Vítámvás ◽

J. Korecký

Keyword(s):

Czech Republic ◽

Picea Abies ◽

Pinus Sylvestris ◽

Tree Species ◽

Abiotic Factors ◽

Forest Tree ◽

Germination Capacity ◽

The Czech Republic ◽

Germination Energy ◽

Seed Ageing

:Germination capacity and germination energy are usually the most frequently used quantitative parameters of forest tree seed. With seed ageing both parameters decreased and the rate of the collapse is given by tree species, age of tree and its seed and biotic and abiotic factors. Relatively little attention has been paid to the age of seed. As it was found, the longevity of the main tree species remained relatively high, and spruce (Picea abies [L.] Karsten and pines (Pinus sylvestris L.) from the investigated areas across the Czech Republic maintained minimally one third of germination capacity or germination energy during the 10 years with the exception of larch (Larix decidua Mill.) where germination capacity decreased almost to zero after 10 years. Although the germination energy and germination capacity decreased significantly, it is possible to use the seed in the case of shortage of the seed of better quality.  

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Occurrence and distribution of mating types A1 and A2 of Phytophthora infestans (Mont.) de Bary in the Czech Republic

Plant Protection Science ◽

10.17221/2697-pps ◽

2010 ◽

Vol 42 (No. 2) ◽

pp. 41-48 ◽

Cited By ~ 7

Author(s):

J. Mazáková ◽

V. Táborský ◽

M. Zouhar ◽

P. Ryšánek ◽

E. Hausvater ◽

...

Keyword(s):

Czech Republic ◽

Phytophthora Infestans ◽

Mating Type ◽

Mating Types ◽

Pcr Markers ◽

The Czech Republic ◽

First Report

A total of 199 Phytophthora infestans isolates were obtained from leaves, tubers and fruits of infected crops of potato and tomato in different regions of the Czech Republic in 2003, 2004 and 2005. They were analysed for mating type using the conventional pairing assay and PCR markers; 107 isolates were of A1 and 92 of A2 mating type. No self-fertile isolate was found. Our study is the first report of the presence and distribution of the A2 mating type of P. infestans in the Czech Republic. The co-existence of the two mating types may enable the pathogen to reproduce sexually, thus enhancing the diversity of its population countrywide.

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