Roots Of Symptomatic Plants Research Articles

Investigation, Identification and Pathogenicity Assessment of Leaf and Soil-Borne Fungal Diseases Causing Yield Reduction in Vegetables in Antalya

Vegetables significantly contribute to Antalya, Türkiye's economy. This study aimed to investigate and identify fungal pathogens causing leaf and soil-borne diseases in leafy vegetable crops. Surveys were conducted in 2021, focusing on morphological and microscopic diagnosis, as well as the prevalence rate of these diseases in five regions (Aksu, Serik, Muratpaşa, Kepez, and Korkuteli). Isolates were obtained from both the leaves and roots of symptomatic plants, and pathogenicity tests were conducted. Survey results showed that lectuca downy mildew was the most important disease, with a 40% plant infection rate and 16.9% disease severity in most parts of the region. In areas where parsley was cultivated, Fusarium sp. was identified as the predominant soil-borne pathogen at 23.82%, and Sclerotinia sclerotiorum was prevalent in lettuce at 14.28%. The lowest incidence of Albugo candida was observed in cress. In conclusion, this study provides crucial insights into the prevalence of fungal diseases and the specific pathogens responsible for damage to lettuce, parsley, dill, cress, mint, and basil cultivation in Antalya province. The incidence and prevalence of these fungal diseases were assessed through surveys in Antalya. In conclusion, this study provides important information on fungal disease agents and their prevalence in lettuce, parsley, dill, cress, mint and basil cultivated areas in Antalya province.

Soybean SDS in South Africa is Caused by Fusarium brasiliense and a Novel Undescribed Fusarium sp.

Soybean sudden death syndrome (SDS) was detected in South Africa for the first time during pathogen surveys conducted in 2013 to 2014. The primary objective of this study was to characterize the 16 slow-growing Fusarium strains that were isolated from the roots of symptomatic plants. Molecular phylogenetic analyses of a portion of translation elongation factor 1-α (TEF1) and the nuclear ribosomal intergenic spacer region (IGS rDNA) indicated that the etiological agents were Fusarium brasiliense and a novel, undescribed Fusarium sp. This is the first report of F. brasiliense outside of Brazil and Argentina and the novel Fusarium sp. causing soybean SDS. Koch's postulates were completed for both fusaria on seven soybean cultivars that are commercially available in South Africa. Results of the pathogenicity experiment revealed that the strains of F. brasiliense and Fusarium sp. differed in aggressiveness to soybean, as reflected in differences in foliar symptoms, root rot, and reduction in shoot length. Cell-free culture filtrates of the two soybean SDS pathogens from South Africa and two positive control strains of F. virguliforme from the United States induced typical SDS symptoms on susceptible soybean cultivars in a whole-seedling assay, indicating that they contained phytotoxins.

First Report of Fusarium cuneirostrum Causing Root Rot Disease in Dry Bean (Phaseolus vulgaris) in Canada.

Root rot is a major disease of dry bean and can cause significant yield reductions due to weakened root systems and poor plant stands. An in-depth study on root rot pathogen identification was conducted in 2011 in three commercial dry bean fields from the major production areas in Manitoba. Ten plants, sampled at each of four random sites within each field, were rated for disease severity. Twenty roots were processed for pathogen isolation and identification in the laboratory. Roots were cut into eight sections (~1 cm) and surface-sterilized in a laminar flow bench. Four root sections were placed on potato dextrose agar plates amended with 0.02% streptomycin sulfate (PDA-Strep) and four root sections were placed on peptone-pentachloronitrobenzene agar amended with 0.1% streptomycin sulfate and 0.012% neomycin sulfate. Afterward, 960 monosporic cultures were obtained representing 320 single spore isolates of potential root rot pathogens per commercial field. Common monosporic cultures from each field were subcultured on PDA-Strep and Spezieller Nährstoffarmer Agar (SNA) media. Based on morphological characteristics, 74 isolates were identified as Fusarium cuneirostrum (1). Colonies grew slowly on PDA-Strep with undulated margins, radial cream-grey mycelia, and conidia pustules with a cream-greyish pigmentation. Sporodochial conidia were falcate, mostly 5-septate, with a wedge shape and slightly protruding basal foot cell (56.3 to 71.8 × 4.6 to 6.2 μm on average). Species identity was confirmed for two isolates by sequencing the translation elongation factor 1 alpha (EF1-α) gene (2), the internal transcribed spacer (ITS) region (4), and the ribosomal intergenic spacer (IGS) (3) (GenBank Accession Nos. KF530848, KF530849, and KF025648 to 51). Sequence homology was compared using BLAST analysis and the FUSARIUM-ID database. The F. cuneirostrum isolates were deposited at the Canadian Collection of Fungal Cultures (DAOM 242540 and 242541). Pathogenicity screenings of two isolates was performed using sterilized seed of navy bean cv. Envoy. Seeds were germinated on moist filter paper for 3 days at 25°C and then inoculated by immersion in a prepared conidial suspension (2.5 × 105 conidia/ml) for 5 min. Seeds of the controls were immersed in sterile water. After inoculation, the germinated seeds were planted in 10-cm diameter pots, filled with sterile soilless mix (Sunshine #3). In the greenhouse, the experiment was arranged as a completely randomized design with three replicates with four germinated seeds per isolate, and was repeated twice. Disease assessment was performed 14 days after inoculation. Infected plants displayed dark brown lesions on the hypocotyl and primary root with a disease severity of 4 scored on a 0 to 5 scale. Fusarium cuneirostrum was re-isolated from roots of symptomatic plants. To our knowledge, this is the first report of F. cuneirostrum causing root rot of dry bean in Canada. It has been previously isolated from mung bean (Vigna radiata) in Ontario (1).

First Report of Fusarium solani Causing Crown and Root Rot on Strawberry Crops in Southwestern Spain

Spain is the fourth largest strawberry (Fragaria × ananassa) producing country in the world. Since April 2010, stunted and dead strawberry plants have been detected in four strawberry fruit production fields in Huelva (southwestern Spain) affecting less than 1% of plants. Symptoms consisted of foliage wilt, plant stunting and drying, and death of older leaves. Internal vascular and cortical tissues of plant crowns showed an orange to brown discoloration. Crowns and roots of symptomatic plants were surface sterilized in 1% sodium hypochlorite for 2 min, rinsed in sterile distilled water for 2 min, and air-dried in a laminar flow cabinet. Small disinfested pieces were transferred to petri dishes containing potato dextrose agar (PDA) and incubated for 10 days at 25°C with a 12-h photoperiod. Cultures derived from single spores were obtained, and morphological characterization was performed by microscopic examination. White to pale cream colonies developed after 10 days of incubation. Unbranched monophialides with microconidia in false heads, micro- (0 to 3 septa) and macroconidia (5 to 7 septa) wide and robust in shape, and chlamydospores were consistent with descriptions of Fusarium solani (Martius) Appel & Wollenweber emend. Snyder & Hansen (2). In addition, the fungus was isolated from asymptomatic runner plants from nurseries by the same method described above, and from soil samples from six fruit-producing fields. Soil samples were analyzed by dilution plating on Fusarium-selective agar medium (1). Genomic DNA from three isolates (FPOST-81 from dead plant 'Sabrina,' TOR-11 from runner plant 'Camarosa,' and TOR-1 from soil) was obtained with a DNA extraction kit (Isolate Plant DNA MiniKit, Bioline). A portion of the translation elongation factor-1 alpha (EF-1α) gene was sequenced using EF-1/-2 primers (3) (GenBank Accession Nos. KF275032, KF275033, and KF275034). The sequence comparison revealed a 99 to 100% match with F. solani sequences in GenBank and Fusarium-ID databases. To confirm the pathogenicity of the fungi, runner strawberry plants 'Camarosa' were inoculated by dipping crowns and roots into a conidial suspension (106 to 107 conidia per ml) for 30 min (8 plants per F. solani isolate) or into sterile distilled water for the controls. Plants were potted in 13-cm diameter pots with peat and maintained at 25/18°C and 70/40% relative humidity (day/night) in a growth chamber with a daily 16-h photoperiod of fluorescent light. Three plants inoculated with isolates TOR-11 and FPOST-81, and four plants inoculated with isolate TOR-1, died within 10 days after inoculation. After 8 to 12 weeks, all of the remaining inoculated plants were stunted and developed symptoms similar to those observed in the field. Production of new feeder roots was lacking or scarce. Control plants remained healthy and formed feeder roots. All plants inoculated with isolates TOR-1 and FPOST-81, and 50% of plants inoculated with TOR-11, showed brown discoloration in the crown. F. solani was re-isolated from symptomatic plants at frequencies of 100% and 80 to 100% from root and crown tissues, respectively. Although F. solani has been reported as a pathogen in other crops, to our knowledge, this is the first report of the occurrence of F. solani causing disease in strawberry plants in Spain.

A New TaqMan Real-Time Polymerase Chain Reaction Assay for Quantification of Fusarium virguliforme in Soil.

The quantification of the soilborne pathogen Fusarium virguliforme inoculum in soil is important for epidemiological studies of soybean sudden death syndrome (SDS). Classical dilution plating methods to determine inoculum density in soil have yielded inconsistent results due to slow growth, variable colony morphology of the pathogen, and the presence of other fungi with similar phenotype. A TaqMan real-time polymerase chain reaction assay was developed based on sequences of the FvTox1 gene of F. virguliforme. The gene differed by four single-nucleotide proteins from the other SDS-causing species. Assay specificity was tested on 48 fungal isolates that varied in taxonomic relatedness. Assay sensitivity was appraised on 10-fold serial dilutions of genomic DNA, conidia suspensions, and soil spiked with conidia. Applicability of the assay was evaluated on field and greenhouse soil samples, and on roots of symptomatic plants. The assay detected only DNA sequences specific to F. virguliforme. The detection limit of the assay was 5 pg/μl, 1,000 conidia/ml, and 1,000 conidia/g soil for genomic DNA, conidial suspensions, and soil with conidia, respectively. The assay was specific to F. virguliforme and was used successfully to quantify inoculum density in soil and soybean roots. The assay can be used as a diagnostic tool for rapid screens of field and greenhouse soil, and for symptomatic and asymptomatic plants.

First Report of Hiemalis begonias Wilt Disease Caused by Fusarium foetens in Canada.

Hiemalis begonias Fotsch (Begonia × hiemalis), a member of the family Begoniaceae, is a commercially important potted flowering plant in Europe and North America. In the spring of 2010, stunted growth and dull green leaves of H. begonias were observed in a commercial greenhouse in southern Ontario, Canada. Symptoms began with dull green foliage, followed by wilted leaves, then the stem base became water soaked with vascular discoloration, and finally, large macroconidial masses of a fungus developed on the collapsed stems and veins. A fungus was consistently isolated from the leaves, stems, and roots of symptomatic plants. Single conidia were isolated from sporodochia and cultured on potato dextrose agar (PDA) and oatmeal agar (OA) for 7 days. Isolates exhibited strong pungent odors on PDA and OA and a brownish orange colony on OA and a light orange colony on PDA. Masses of light orange and hemispherical-shaped conidia and stromata formed on OA. Conidiophores formed from aerial mycelium producing ellipsoidal microconidia without septation. Sporodochia formed on agar surface producing three-septate, slightly curved macroconidia. The cultural and conidial characteristics of the isolates were similar to those of Fusarium foetens Schroers (4). Partial translation elongation factor 1-α (TEF) gene was amplified and sequenced with primers ef1 and ef2. A comparison of a partial sequence has been deposited in GenBank (Accession No. HM748968) and showed a 100% match with F. foetens (2). Inoculations with F. foetens isolates were performed by injecting a 100-μl suspension of 1 × 106 conidia/ml into stems of five healthy plants near the ground or soaking the soil of five healthy 6-week-old H. begonias cv. Golden Edith with 50 ml of suspension. Control plants were similarly injected with sterile water or sown in sterile soil. After 4 weeks, all inoculated plants developed dark, wilting leaves and collapsed stems and veins similar to those observed in the commercial greenhouse. F. foetens was reisolated from diseased plants, and identification was reconfirmed by conidial characteristics and TEF 1-α sequence. Control plants were healthy and symptom free. F. foetens has recently been described in association with a new disease of H. begonias in Europe (3) and the United States (1). F. foetens can cause major economic losses to farmers and marketers of H. begonias in Europe and the United States. To our knowledge, this is the first report of F. foetens causing wilt disease of H. begonias in Canada.

First Report of Crotalaria spectabilis Fasciation Associated with 'Candidatus Phytoplasma asteris' in India.

Crotalaria spectabilis Roth. (Fabaceae), commonly known as showy rattlebox, is an herbaceous legume mainly used as a green manure crop to improve soil properties and as a source of durable fiber. However, the plant is toxic to mammals and birds because of the presence of pyrrolidizine alkaloids. A native of India and the Malay Peninsula, the species has been introduced into other areas such as the United States and Pacific Islands where the plant is an invader of cultivated lands. Fasciated rattlebox plants were sighted in fields in New Delhi in February 2010, with approximately 99% of the examined plants symptomatic. Symptoms included flattening of stems in a descending gradient of severity from the apex to the base of each affected branch. Shoots showed longitudinal undulations bearing highly reduced leaves and uneven distribution of flowers and fruits. To identify the causal agent, 10 symptomatic plants and 8 asymptomatic plants (latter sampled from a field approximately 1.5 km away) were collected for nucleic acid analysis. Total genomic DNA was extracted from flattened stems as well as the roots of symptomatic plants, and from the same tissues of asymptomatic plants, and subjected to nested-PCR using phytoplasma 16S ribosomal DNA universal primer pair P1/P7, followed by R16F2/R2 (4). A known aster yellows-infected Catharanthus roseus plant was used as a control sample. Results depicted a characteristic phytoplasma amplicon of 1.25 kb in all samples from symptomatic plants and the control plant. No amplification was observed from asymptomatic plants. To obtain a full-length sequence, a representative amplicon was purified with the QIAquick gel extraction kit (QIAGEN, Valencia, CA), cloned into pGEM-T Easy vector (Promega, Madison, WI), and sequenced. Comparison of the 1,243-bp sequence (Genbank Accession No. HM137557) using BLAST analysis of the NCBI database showed 99% homology with sequences of members of 16SrI group phytoplasmas, i.e., 'Candidatus Phytoplasma asteris' such as Japanese spurge yellows (AB551736.1), Mulberry yellow dwarf (GQ249410.1), and Bamboo witches'-broom (FJ853161.1) phytoplasmas. The profiles of in vitro restriction fragment length polymorphism (RFLP) analysis obtained by digestion of the nested-PCR products with HhaI, KpnI, and AluI were similar to those of in silico RFLP analyses and coincided with the pattern of the 16SrI-B subgroup. Phylogenetic analysis of phytoplasma 16S rDNA sequences based on the maximum likelihood method using MEGA Version 4.1 also placed the Crotalaria fasciation (CF) phytoplasma within the 16SrI-B cluster. In India, C. tetragona plants infected with 16SrI phytoplasma (FJ185141) causing witches'-broom symptoms (1) showed 98% similarity with the CF phytoplasma. However, the results support previous molecular investigations associating 16SrI phytoplasma with fasciation of herbaceous plants, including Lilium spp. (2) and Asparagus officinalis (3). To our knowledge, C. spectabilis represents a new host that can be fasciated as a result of phytoplasma infection. Because of the weedy nature of C. spectabilis, this host could facilitate spread of the phytoplasma.

Morphological and molecular characterisation ofCampylocarponandCylindrocarponspp. associated with black foot disease of grapevines in Uruguay

Black foot disease of grapevines is a problem in most grape-producing regions of the world. This disease affects both young and mature plants, which show retarded growth and eventually die. Species of Cylindrocarpon have been identified as the causal agent of this disease. Fungal isolates were obtained from roots of symptomatic plants and plantlets from commercial vineyards in Uruguay. Morphological characteristics on malt extract agar were recorded, and DNA purified. Internal transcribed spacer (ITS) rDNA was amplified with primers ITS 4 and ITS 5 and the amplified fragments were sequenced. Phylogenetic analyses were done with sequences from Uruguay and sequences from GenBank using PAUP* (Phylogenetic Analysis Using Parsimony). Six phylogenetic groups were found that matched six previously known taxa. These groups were supported by morphological characteristics. Isolates were assignable to Campylocarpon and Cylindrocarpon species. Ca. pseudofasciculare, C. destructans var. crassum, C. liriodendri, C. macrodydimum, C. olidum var. crassum and C. pauciseptatum represent new records for Uruguay. This work is the first step towards a better understanding and management of black foot disease in local conditions.

First Report of Crown Rot and Root Rot Caused by Cylindrocladium pauciramosum on Feijoa (Feijoa sellowiana) in Italy.

Feijoa sellowiana (O. Berg) O. Berg is native to South America and grown in Italy as an ornamental plant and for its fruit. During 2006, 2007, and 2008, a severe wilting was noticed on ∼ 10,000 potted 6-month to 3-year-old plants obtained from seeds and produced by three commercial nurseries in Catania Province (southern Italy). Wilting was always associated with root rot and brown discoloration at the base of the stem. Leaf spots or stem lesions were not observed. A Cylindrocladium sp. was isolated consistently from the crown, basal stem, and roots of symptomatic plants on potato dextrose agar (PDA). Ten isolates of a Cylindrocladium sp. were obtained from single-spore colonies made from a 14-day-old culture grown on PDA and subsequently subcultured on carnation leaf agar (CLA). Cultures were incubated at 25°C under near-UV light with a 12-h light/dark regimen and examined after 7 days (1). Only fungal structures occurring on the carnation leaf tissue were examined. Isolates were identified based on morphological traits and mating type responses (2,4). Isolates were mated with opposite mating tester strains of C. pauciramosum (DISTEF-G128 [MAT1-1] and DISTEF-G87 [MAT1-2]) and C. scoparium (DISTEF-GCs7 = CBS 120892 [MAT1-1] and DISTEF-GP0 = CBS 119669 [MAT1-2]) selected from an Italian collection. Plates were parafilmed, sealed in plastic bags, incubated in a chamber at 25 ± 1°C with a photoperiod of 16 h of light and 8 h of darkness, and examined weekly until perithecia developed. Successful crosses were determined after 2 months of incubation and were regarded as those isolate combinations that produced perithecia with viable ascospores. Conidia were observed to be 30 to 60 × 3.5 to 5.0 μm while conidiophores mostly had two to three series of branches and stipes terminated in obpyriform to broadly ellipsoidal vesicles, which is characteristic of C. pauciramosum C.L. Schoch and Crous (teleomorph Calonectria pauciramosa C.L. Schoch and Crous). Fertile perithecia containing viable ascospores were similar to Calonectria pauciramosa. They were obtained only from crosses of the examined isolates with mating type testers of C. pauciramosum. Six isolates (when paired with DISTEF-G128) and four isolates (when mated with DISTEF-G87) provided fertile progeny. No perithecia were obtained in pairings of the 10 isolates with tester strains of C. scoparium. One representative isolate was deposited at the Fungal Biodiversity Centre, Centraalbureau voor Schimmelcultures (DISTEF-AS4 = CBS-120618). Koch's postulates were fulfilled by inoculating 30 6-month-old seedlings of feijoa with a spore suspension (105 conidia per ml) of one isolate of the pathogen (DISTEF-AS4) grown on CLA for 14 days. The same number of seedlings was used as a control. Following inoculation, plants were maintained in a growth chamber at 25 ± 1°C and 90 to 95% relative humidity. Wilting, crown rot, and root rot symptoms identical to those observed in the nurseries appeared within 15 to 50 days. Symptoms were not observed on the control plants. C. pauciramosum was previously reported to cause leaf spots on feijoa (3). To our knowledge, this is the first report of wilting due to crown and root rot caused by C. pauciramosum in this host. The disease is a limiting factor for feijoa cultivation in Sicilian nurseries.

Pratylenchus neglectus, P. thornei, and Paratylenchus hamatus Nematodes Causing Yield Reduction to Dryland Peas and Lentils in Idaho.

In June 2006, stunted and chlorotic plants were observed in large patches in two 40.5-ha fields of dryland peas (Pisum sativum) in Latah County, Idaho, which resulted in 90 and 75% crop loss. In the same region, a 121.4-ha field of dryland lentils (Lens culinaris) also had plants showing poor growth, wilting, and yellowing in large patches, which resulted in 40% crop loss. Two species of lesion nematodes (Pratylenchus neglectus and P. thornei) and one species of pin nematode (Paratylenchus hamatus) were extracted from rhizosphere soil and the roots of symptomatic plants from these fields. In a subsequent survey of seven dryland pea fields, under cv. Columbian, in Latah and Nez Perce counties and one dryland pea field, under cv. Small Sieve, in Latah County, plant samples had means of 551 and 2,178 mixed species of lesion nematodes per gram of dry root, respectively. Plant samples from 12 lentil fields in Latah County, six planted with cv. Red Chief and six with cv. Pardina, had means of 279 and 987 mixed species of lesion nematodes per gram of dry root, respectively. Soil samples from the same fields had a mean of 628 and 671 pin nematodes, Paratylenchus hamatus, per 250 cm3 soil for Red Chief and Pardina, respectively. Lentils cv. Pardina and peas cv. Columbian were planted separately in six pots, five seeds per pot containing 250 g of infested soil brought from the field to the greenhouse. Fumigated sandy loam soil was used as control. These assays were repeated three times. In addition, peas and lentils were planted to pots infested singly with each of the three nematode species. For this assay, nematodes were extracted from field soil, surface sterilized, and used to infest 250 g of fumigated sandy loam soil at two nematodes per gram of soil. Six plants per nematode species and an uninoculated control were used in the greenhouse assays, which were repeated three times. Nematodes in all of the assays reduced plant growth in comparison with controls; an average of 50 to 70% reduction in plant height was noted. The lesion nematode populations increased in all pots. The greenhouse assays verified the negative impact of these nematodes on growth of dryland peas cvs. Columbian and Small Sieve and lentils cvs. Red Chief and Pardina. P. neglectus, P. thornei, and Paratylenchus spp. previously have been reported from the semi-arid Pacific Northwest (1). However, to our knowledge, this is the first report attributing plant growth and yield reduction of certain cultivars of lentils and peas to these two species of lesion nematodes and pin nematodes, identified to species level as Paratylenchus hamatus. Reference: (1) R. W. Smiley et al. J. Nematol. 36:54, 2004.

Evaluation of Phytophthora ramorum in Nursery Crop Tissue Culture Propagation

Phytophthora ramorum, cause of Ramorum blight on numerous woody ornamental shrubs, is a regulated pathogen in the US and internationally. Currently, nurseries are inspected to detect infected plants; however, many plants are propagated by tissue culture nurseries and the behavior of P. ramorum in this system is unknown. Pathogen growth and sporulation in propagation vessels containing different multiplication and rooting media, with a range of plants and without plants, was evaluated with regard to pathogen visibility and induction of disease symptoms. Within 2 weeks, the pathogen colonies were visible to the naked eye on all 26 multiplication media and on 9 of 11 rooting media tested (without plants). The appearance of colonies on different media was variable and no sporangia but occasional chlamydospores were produced. The pathogen growth was very visible on multiplication media containing susceptible plants, inoculated plants exhibiting obvious discoloration and mortality. The pathogen was reisolated from terminal shoot tissue and roots of symptomatic plants. Variability occurred in susceptibility of different cultivars of a plant species, in virulence of the two isolates of the pathogen, and in recovery from shoot tissue. We conclude that fungal growth on the media, with or without plants, and symptoms of disease were apparent enough that contaminated vessels would be destroyed. Accepted for publication 9 April 2007. Published 22 August 2007.

First Record of Crown and Root Rot Caused by Cylindrocladium pauciramosum on California Lilac in Italy.

California lilacs, or Ceanothus, are evergreen, drought tolerant, colorful ornamental shrubs belonging to the Rhamnaceae family. Ceanothus thyrsiflorus var. repens is the most common species grown in Italy. In October 2005, a severe wilting was noticed on approximately 3,000 potted, 6-month-old plants obtained from cuttings produced by a commercial nursery in Lecco Province (northern Italy). Wilting was always associated with root rot and brown discoloration at the base of the stem. No leaf spots or stem lesions were detected. A Cylindrocladium sp. was isolated consistently from crown, basal stems, and roots of symptomatic plants with potato dextrose agar. Although many crown rots are caused by Phytophthora spp., this pathogen was not found associated with rotten roots and crown plating small pieces of diseased root tissue and crowns onto selective medium. A random selection of five Cylindrocladium sp. isolates was obtained from the infected crown and basal stem. Subsequently, they were identified on carnation leaf agar (CLA) as Cylindrocladium pauciramosum C.L. Schoch & Crous on the basis of their obpyriform to broadly ellipsoidal terminal vesicles, conidiophore branching pattern, conidium, and perithecial morphology, as well as mating type with tester strains of C. pauciramosum selected isolates (1,4). All single-conidial isolates were mated with tester strains DISTEF-G87 (MAT1-1) and DISTEF-G128 (MAT1-2) of C. pauciramosum on CLA, which confirmed both mating types to be present. Two of the isolates were deposited at Fungal Biodiversity Centre, Centraalbureau voor Schimmelcultures (CBS-120145 and CBS-120146). Pathogenicity tests were performed by macerating CLA cultures of C. pauciramosum, obtained from a single-spore isolate and incorporating them on the soil surface of 20 8-month-old C. thyrsiflorus var. repens potted plants. The same number of plants was used as the control. Following inoculation, plants were well irrigated and kept in a growth chamber at 25 ± 1°C. All inoculated plants developed crown rot, basal stem rot, and root rot 25 days after inoculation. Uninoculated control plants remained healthy. C. pauciramosum was reisolated from the artificially inoculated plants. C. pauciramosum causes considerable damage to the ornamental industry in Italy, where projects have been carried out since the first record of the fungus in Europe (3). C. pauciramosum was previously detected on Ceanothus sp. in the United Kingdom (2), where foliar and stem lesions were described. To our knowledge, this is the first record of the pathogen on C. thyrsiflorus var. repens and the first report of wilting due to crown and basal stem rot.

First Report of Sudden Death Syndrome of Soybean Caused by Fusarium solani f. sp. glycines in Nebraska.

During August of 2004, soybean (Glycine max (L.) Merr.) plants exhibiting symptoms typical of sudden death syndrome (SDS) caused by Fusarium solani (Mart.) Sacc. f. sp. glycines (= Fusarium virguliforme Akoi, O'Donnell, Homma, & Lattanzi) (1) were observed in Nemaha and Pierce counties in eastern Nebraska. Leaf symptoms ranged from small chlorotic spots to prominent interveinal necrosis on plants at R5-R6 growth stages. Taproots of symptomatic plants were plated on potato dextrose agar (PDA) amended with hymexazol, ampicillin, and rifampicin (HAR). Resulting fungal isolates grew slowly and developed masses of blue macroconidia, characteristic of F. solani f. sp. glycines. Sorghum seed infested with the isolates were placed 1.5 cm below soybean seeds of the susceptible cv. Sloan planted in clay pots (3). Noninfested sorghum seed and sorghum seed infested with F. oxysporum were controls. Plants were maintained for 32 days at 27.5 ± 2.5°C in the greenhouse. Small cholorotic spots were observed on leaves of F. solani f. sp. glycines-inoculated plants within 21 days followed by the development of interveinal chlorosis. Roots of symptomatic plants were plated on PDA with HAR and F. solani f. sp. glycines was recovered. Identification of the fungal cultures was further confirmed as F. solani f. sp. glycines by a real-time quantitative polymerase chain reaction (qPCR) assay described by Gao et al. (2). During 2005, SDS symptoms were also reported in early planted soybeans from Jefferson and Seward counties and the presence of SDS was confirmed by qPCR. The confirmation of SDS at multiple locations suggests that the pathogen is widely distributed in the eastern one-third of Nebraska. SDS could be a serious threat to soybean production in this area since spring weather conditions favor SDS infection and many producers plant soybean early in cool soils.

First Report of Pythium Root Rot of Rau Ram (Polygonum odoratum).

Polygonum odoratum (= Persicaria odorata), known as rau ram or sang hum, is native to southeastern Asia and is a common herb in Vietnamese cuisine (1). It has been studied most extensively for its aromatic compound content (2). In Florida, rau ram commonly is grown hydroponically in greenhouses using large, cement beds with recirculated water. The plants form dense mats from which new growth is trimmed for market. During January of 2002, a severe dieback was observed in one production house in Saint Lucie County, FL. Plants with less severe symptoms were yellowed and stunted. Roots of symptomatic plants were largely decayed with root symptoms beginning as a tip necrosis. The cortex of severely affected roots slipped off easily, leaving a stringy vascular system. Plating of symptomatic tissue from 20 randomly selected plant samples was performed with multiple general and selective media including potato dextrose agar, corn meal agar with pimaricin, ampicillin, rifampicin, and pentachloronitrobenzene (PARP) (3). All colonies produced were identified as Pythium helicoides Drechsler on the basis of sporangial, oogonial, and antheridial characteristics (4). Isolates had proliferous, obovoid, papillate sporangia, and were homothallic with smooth-walled oogonia and thick-walled, aplerotic oospores. Multiple antheridial attachments per oogonium were common with the antheridium attached along its entire length. Pathogenicity tests were conducted using P. odoratum plants grown from commercial transplants. Two tests were performed. Each test was conducted using eight inoculated and eight control plants. In the first test, plants were maintained in 10-cm pots immersed in sterilized pond water for the duration of the test. Plants were inoculated with five 7- × 70-mm sections of freshly growing mycelial culture per plant using 10-day-old cultures of Pythium helicoides grown on water agar. Chlorosis was observed at approximately 2 months after inoculation. Root necrosis was observed in inoculated plants approximately 5 months after inoculation. This test was performed in the greenhouse with temperatures ranging from 20 to 30°C. The second test was performed in growth chambers at 35 to 40°C. Plants were maintained in 10-cm pots immersed in Hoagland's solution and were inoculated with four 6-mm plugs per plant. Symptoms were observed on inoculated plants at this temperature within 1 week of inoculation. No chlorosis or root decay was observed in noninoculated, immersed plants. The pathogen was reisolated from inoculated, symptomatic tissue. To our knowledge, this is the first report of root rot of P. odoratum caused by Pythium helicoides.

Crown and Root Infection of Lisianthus Caused by Fusarium solani in South Africa.

Cultivation of lisianthus (Eustoma grandiflorum (Raf.) Shinn.) is a minor industry in South Africa, with only a few growers producing the crop commercially. Commercial production at a location in Gauteng Province is hampered by rotting of the crowns and roots of plants that result in mortality of as much as 22% of the plants. At advanced stages of infection, the crowns of affected plants characteristically are covered with masses of fusoid, curved hyalophragmospores. Crowns and roots of symptomatic plants that were submitted by the grower in January 2003 were surface disinfested by immersing for 2 min in a 3% solution of sodium hypochlorite, and segments excised from the plant tissue were plated on potato dextrose agar supplemented with 50 mg l-1 of rifampicin. Fusarium solani (Mart.) Appel & Wollenw. (1), was consistently and exclusively isolated from the segments. Teleomorph Nectria haematococca Berk. & Broome, commonly developed in culture after incubation for 4 to 6 weeks, although no sexual structures were observed on infected plants. A spore suspension containing 104 micro- and macroconidia ml-1 was prepared for each of two single-conidial isolates of F. solani. Using a 0.8-mm-diameter hypodermic needle, 100 μl of each suspension was injected subepi-dermally into the crown of each of three 1-month-old disease-free lisian-thus plantlets (cv. Texas Blue Bell) growing in 500-ml plastic pots filled with sterilized vermiculite. In addition, each suspension was incorporated at 2% (vol/vol) into three pots with sterile vermiculite, and a plantlet was planted in each pot. Control plantlets were treated similarly, but with sterile distilled water. All inoculated plantlets developed crown rot and wilted within 2 weeks while maintained at 28°C in a greenhouse, regardless of mode of inoculation, and F. solani was readily reisolated from their crowns and roots. Control plantlets remained symptomless and did not yield F. solani. Crown and root infection of lisianthus by F. solani has been described (2,3), but to our knowledge, this is the first report of the disease in South Africa.

First Report of Root and Crown Rot of Sage Caused by Phytophthora cryptogea in Italy

Sages are cultivated as aromatic and ornamental plants in Italy and represent the common name of certain species of Salvia and Phlomis (family Lamiaceae). In Sicily (southern Italy) during the summer of 2001, ≈40% of 1,400 2-year-old landscape plants of S. leucantha Cav. (Mexican bush sage or velvet sage) showed symptoms of stunting, chlorosis, and gradual dieback or sudden wilt, which are associated with root and crown rot. Plants were supplied by a commercial nursery, transplanted from pots in the spring, and irrigated using a trickle system. Phytophthora was isolated consistently from roots and basal stems of symptomatic plants on a BNPRAH medium (2). The species was identified as P. cryptogea Pethybr. & Laff., primarily on the basis of morphological and cultural characteristics. Five representative single-hypha isolates were characterized. On potato dextrose agar, they formed colonies with a slight petaloid pattern. Cardinal temperatures for mycelium growth were 2°C, minimum; 25°C, optimum; and 30 to 35°C, maximum. Hyphal swellings were abundant in aqueous culture. Sporangia were obpyriform, persistent, nonpapillate, and proliferous (2). All isolates were the A1 mating type and formed oogonia, amphigynous antheridia, and oospores in dual cultures with reference isolates of the A2 mating type of P. cryptogea and P. drechsleri. Identification was confirmed by electrophoresis of mycelium proteins on a polyacrylamide slab gel (1). Electrophoretic patterns of total soluble proteins from the sage isolates were identical or very similar to those from 10 reference isolates of P. cryptogea from various hosts, including isolate IMI 180615 (ex-type isolate). Conversely, the electrophoretic pattern of the isolates of P. cryptogea from sage was clearly distinct from those from reference isolates of other species included in Waterhouse's taxonomic group VI. Esterase (EC 3.1.1.2.) zymograms of the sage isolates corresponded to those of isolates of P. cryptogea included in electrophoretic group 2 (1). The pathogenicity of a representative isolate of P. cryptogea from sage was tested in the greenhouse using 4-month-old plants of Mexican bush sage. Inoculum was produced on a mixture of vermiculite and autoclaved oat seeds (4) and mixed with steam-sterilized sandy loam soil at a concentration of 4% (vol/vol). Plants were transplanted in pots (12 cm diameter) filled with infested soil; control plants were grown in pots containing noninfested soil. After transplanting, all pots were placed in shallow trays filled with water for 24 h to saturate the soil. All plants grown in infested soil showed extensive root necrosis and dieback ≈30 days after transplanting, and P. cryptogea was reisolated from roots of symptomatic plants. Control plants did not develop symptoms. Root and crown rot of sage caused by P. cryptogea has been reported previously in California (3). To our knowledge, this is the first report of P. cryptogea on sage in Italy. Root rot caused by P. cryptogea may be a potential problem for commercial cultivation of sage as no serious disease of this plant has been reported in Italy so far.

Phytophthora palmivora a New Pathogen of Lavender in Italy.

Root rot caused by Phytophthora nicotianae is considered the most serious disease of lavender in commercial cultivations in Italy. In summer 2001, in the Gela area (Sicily), ≈60% of 34,000 2-year-old landscape shrubs of English lavender (L. angustifolia) grown in a clay loam soil showed symptoms of dieback associated with root rot. Plants had been transplanted from pots in May and watered using a trickle irrigation system. A species of Phytophthora was isolated consistently from roots of symptomatic plants using potato dextrose agar (PDA) containing benomyl, nystatin, pentachloronitrobenzene, rifampicin, ampicillin, and hymexazol. The species was identified as P. palmivora on the basis of morphological and cultural characters. Ten representative single-zoospore isolates were characterized. On agar media, the isolates produced elliptical to ovoid, papillate sporangia, with a mean length/breadth ratio of 1.8. Sporangia, produced on sporangiophores forming simple sympodia (as many as 20 sporangia per sympodium), were caducous with a short pedicel (mean pedicel length = 5 μm) and a conspicuous basal plug. In addition to typical sporangia, all isolates produced sporocysts, i.e., subglobose, nonpapillate sporangia (2). The minimum temperature for mycelium growth on PDA was 10°C, the optimum was 27°C, and the maximum was 35°C. All isolates were A1 mating type. Antheridia were amphyginous. The identification was confirmed by electrophoresis of mycelial proteins on a polyacrylamide slab gel. Electrophoretic banding patterns of total soluble proteins and eight isozymes of the isolates from lavender were identical to those of a reference isolate of P. palmivora from olive (1). Conversely, the electrophoretic phenotype of the isolates from lavender was distinct from those of reference isolates of other species, including P. cactorum, P. capsici, P. citrophthora, P. nicotianae, and P. tropicalis. The pathogenicity of a representative isolate of P. palmivora from lavender was tested in the greenhouse using 6-month-old plants of English lavender, Rosea, a commercial cultivar very susceptible to root rot caused by P. nicotianae (3). Inoculum was produced on a mixture of vermiculite and autoclaved oat seeds (4) and mixed with soil (sand/lime/peat 1:1:1) at a concentration of 4% (vol/vol). Plants were transplanted to pots filled with infested soil. Control plants were grown in pots containing noninfested soil. After transplanting, all pots were flooded for 24 h by plugging the drain hole. Three months after transplanting all plants grown in pots containing infested soil showed extensive root necrosis and dieback symptoms. Control plants remained healthy. P. palmivora was recovered from roots of symptomatic plants. To our knowledge, this is the first report from Italy of P. palmivora on lavender. Root rot caused by P. palmivora may be a potential problem for commercial cultivation of lavender.

First Report of Beet soil-borne virus on Sugar Beet in Iran.

Sugar beet is a main field crop in Iran and is cultivated in 186,000 ha. During the summer of 2001, sugar beet (Beta vulgaris) plants with pale, often upright, narrow, and rolled leaves were collected from the six main beet cultivation provinces of Iran (Fars, Ghazvin, Kermanshah, Khorasan, Semnan, and Isfahan). Roots of symptomatic plants were small, often with constriction, and exhibited warty outgrowth, proliferation of fibrous roots, and vascular necrosis. Beet soil-borne virus (BSBV) and Beet necrotic yellow vein (BNYVV, genus Benyvirus) were detected in sugar beet root samples by tissue-blot immunoassay (TBIA) using BSBV- and BNYVV-specific monoclonal antibodies (As-0576.1 and As-0799.1/CG6-F4, respectively; DSMZ Plant Virus Collection, Germany). Root extracts of sugar beet plants infected with BSBV, were infective by mechanical inoculation to Chenopodium quinoa causing necrotic ring spots. BSBV was detected in inoculated plants by TBIA. Laboratory tests using TBIA on 2,387 randomly collected samples showed that BSBV was present in 406 plants (17%) and BNYVV was present in 1,347 plants (56.43%). BSBV resembles BNYVV, the causal agent of sugar beet rhizomania, morphologically and in its transmission by Polymyxa betae (1). BNYVV has been reported previously from Iran (2). To our knowledge, this is the first report of BSBV occurring on sugar beet in Iran.

First Report of Phytophthora palmivora as a Pathogen of Olive in Italy.

Olive (Olea europea L.) is an economically important crop in Italy and is planted on about 1 million ha. The Apulia, Calabria, and Sicily regions of Southern Italy account for about 70% of the production. Many new plantations have been established during the last 10 years. In summer 1999, 1- to 2-year-old olive trees (cv. Carolea) with decline symptoms were observed in new plantations in Catanzaro Province (Calabria). The symptoms associated with the root rot were leaf chlorosis, defoliation, wilting, twig dieback, and eventual plant collapse. In some cases, more than 40% of the trees were affected. A Phytophthora sp. was isolated consistently from rotted rootlets of diseased trees using a selective medium (2). Single-zoospore isolates were obtained from the colonies. The species isolated from olive roots was identified as P. palmivora (E. Butler) E. Butler on the basis of morphological and cultural characters according to Erwin and Ribeiro (1). All isolates produced papillate sporangia, which were elliptical to ovoid, caducous (mean pedicel length = 5 µm), with a length-breadth ratio of 1.8. In addition, some isolates produced subglobose, non-papillate sporangia with a length-breadth ratio ranging from 1.2 to 1.5. Electrophoresis of mycelial proteins on polyacrylamide gels confirmed that all isolates were pure cultures and that they all belonged to the same species. The electrophoretic banding patterns of total soluble mycelial proteins and eight isozymes (alkaline phosphatase, esterase, fumarase, NAD-glucose dehydrogenase, glucose-6-phosphate dehydrogenase, lactate dehydrogenase, malate dehydrogenase, and superoxide dismutase) of all olive isolates were identical to those of two strains of P. palmivora from parlor palm and from pittosporum. Conversely, they were clearly distinct from the electrophoretic banding patterns of reference isolates of P. cactorum (Lebert & Cohn) Schroter, P. capsici Leonian, P. citrophthora (R. E. Sm. & E. H. Sm.) Leonian, and P. nicotianae van Breda de Haan. All isolates of P. palmivora from olive were of the A1 mating type. The pathogenicity of four P. palmivora isolates from olive, two producing only typical and two producing both types of sporangia, was tested in the greenhouse at 18 to 25°C, using 20 1-year-old rooted cuttings of olive cv. Carolea for each isolate. Twenty noninoculated cuttings were used as a control. The inoculum for pathogenicity tests was produced on autoclaved rice grains moistened with V-8 juice. Cuttings were transplanted to pots filled with soil infested with inoculum at 2% (wt/vol). Control plants were grown in pots containing a mixture of soil and 2% autoclaved rice. After transplanting, all pots were flooded once a week for 24 h by plugging the drain hole of the pot. One month after planting, all the plants in infested soil had died and no difference in virulence was observed among the isolates. Control plants remained healthy. P. palmivora was reisolated from the roots of symptomatic plants. Pathogenicity tests were repeated twice with similar results. In a survey of nurseries in Southern Italy, P. palmivora was recorded frequently from roots of young olive trees suggesting that infections originated from nurseries. This is the first report from Italy of P. palmivora on olive. This species has been described recently as a pathogen of olive in Spain (3).

First Report of Root and Collar Rot by Sclerotium rolfsii on Apple Trees in Italy.

Sclerotium rolfsii Sacc. (teleomorph Athelia rolfsii (Curzi) Tu & Kimbrough) is a polyphagous, soilborne plant pathogen. In summer 1998, a sudden death of 2-year-old apple trees (Malus domestica Borkh.) cv. Royal Gala grafted on M9 rootstock was observed in an orchard near Rome, Italy. Symptoms were stunted vegetation, leaf chlorosis, and root and collar rot. A fungus identified as S. rolfsii was observed producing sclerotia and whitish mycelial strands on root and collar bark. Isolations from roots and at the margin of subcortical necrosis on the collar consistently yielded S. rolfsii colonies on potato dextrose agar (PDA); sclerotia developed within 7 days. Koch's postulates were fulfilled by inoculating 10 1-year-old apple tree cv. M9 rootstocks, grown in 3.5-liter pots, with an S. rolfsii isolate grown for 1 week on PDA at 25°C. One ground plate per plant was used, placed around collar and main roots. Five control plants were treated with PDA only. Rootstocks were kept in the greenhouse at 26 ± 2°C. Within 2 months, 70% of inoculated plants died, with marked necrosis girdling the collar. The other inoculated plants showed a general decline, with widespread necrosis on collars and main roots. Control plants remained healthy. S. rolfsii was reisolated from collars and roots of symptomatic plants. S. rolfsii has been recorded on apple trees in the U.S., India, China, and Israel. In Italy, it is destructive on several crops, and was recently recorded on walnut (1). This first outbreak of S. rolfsii on apple in Italy may have been favored by exceptionally warm late spring and summer temperatures. Reference: (1) A. Belisario and L. Corazza. Plant Dis. 80:824, 1996.