Petiole Lesions Research Articles

First Report of Neopestalotiopsis spp. Causing Leaf Spot and Petiole Blight on Strawberry in New England.

In October 2023, a Connecticut grower contacted The Connecticut Agricultural Experiment Station about a field of strawberry plants (Fragaria × ananassa) (cv. Ruby June) showing symptoms of severe leaf spotting and visual wilting. Upon visiting the field, leaves had lesions with a diffuse black halo and a light brown center and wilting symptoms, which appeared driven by petiole lesions and presented as dark brown stripes with a reddish-purple halo. Symptoms were observed on 80 to 90% of plants within the block, nearly all of which (>90%) presented with both leaf spots and severe wilting. Diseased tissue was collected from 20 leaves and 25 petioles, sterilized in 0.6% NaOCL, and plated on potato dextrose agar. After hyphal tipping a morphologically identical fungus was isolated from 70% of leaves and 88% of petioles, which formed a dense white mycelial mat with moderate aerial mycelium and conidiomata that exuded dark brown conidial masses. The underside of the mycelial mat was yellowish. Conidia were fusoid, ellipsoid, straight to slightly curved, 4-septate with a single basal appendage and 2-5 apical, matching the description of species within the genus Neopestalotiopsis (Maharachchikumbura et al. 2014). The average conidia (n=74) length, not including appendages, was 29.9 ± 2.1 µm and the average width, at the widest point, was 7.5 ± 0.7 µm. Aerial hyphae were collected from two isolates, CT58-1 and CT62-2, and DNA was extracted for further molecular characterization. PCR was performed with primers targeting actin (ACT), β-tubulin (TUB2), and ITS prior to amplicon sequencing (Carbone and Kohn 1999; Hassan et al. 2018). Sequences were queried against the NCBI whole genome shotgun database, and aligned sequences from 13 species (including Neopestalotiopsis, Pestalotiopsis, and Pseudopestalotiopsis) were collected for each locus. Sequences were aligned, trimmed, and concatenated using Mega11, and IQ-TREE was employed for model selection (Nguyen et al. 2015; Tamura et al. 2021). A maximum-likelihood tree placed the isolates in a high-confidence cluster with Neopestalotiopsis rosae, confirming this placement of these isolates within the genus (CT58-1 Accession #: PP715979-89; PP707735). To confirm pathogenicity, CT58-1 was grown on autoclaved strawberry leaves to induce sporulation, and a suspension of 105 spores/ml was made. Five milliliters of this spore suspension was sprayed on six 6-week-old strawberries (cv. Jewel), and water was sprayed on the same number of control plants. Plants were at 100% humidity for two days and then kept in the greenhouse for 3 weeks to observe symptoms. Inoculated plants presented with identical leaf spot and petiole lesions to field samples and no visual symptoms were observed on control plants. New isolations were made from infected petioles, which produced morphologically identical spores to those described above, and ITS/ACT loci sequencing yielded sequences identical to those of CT58-1. Spore production and plant inoculations were repeated with this new isolate, and identical symptoms were observed. This is the first report of Neopestalotiopsis infecting strawberries in New England and given the high disease incidence in the initial infected field and relative lack of disease in a neighboring field, it is likely that this pathogen was introduced on bare root plants. As the plants were sourced from a nursery in Ontario, Canada, it is likely that the pathogen is capable of overwintering in the Northeastern United States.

Damping-off of Edible Aster Caused by Rhizoctonia solani AG-4

In August 2021, we surveyed diseases of wild vegetables grown in Taebaek, Gangwon Province, Korea. During the disease survey, we observed severe damping-off symptoms in young edible aster (<i>Aster scaber</i>) plants in a vinyl greenhouse investigated. The incidence of the disease in the plants ranged from 5% to 20%. Diseased plants of edible aster were collected from the vinyl greenhouse, and fungi were isolated from petiole lesions of the diseased plants. <i>Rhizoctonia</i> sp. was consistently isolated from the petiole lesions. We examined morphological characteristics and anastomosis groups of nine <i>Rhizoctonia</i> sp. isolates obtained from the petiole lesions. The examination results revealed that all the isolates corresponded to <i>Rhizoctonia solani</i> AG-4 based on the morphological characteristics and anastomosis test. Three isolates of <i>R. solani</i> AG-4 were tested for their pathogenicity on edible aster plants by artificial inoculation. Inoculation tests showed that the tested isolates caused damping-off symptoms on the inoculated plants. The induced symptoms were similar to those observed in the vinyl greenhouse investigated. Damping-off of edible aster caused by <i>R. solani</i> AG-4 is first reported in this study.

First Report of Cercospora rodmanii Causing Leaf and Petiole Lesions on Water Hyacinth in China.

The rapid spread and colonization of water hyacinth (Eichhornia crassipes) leads to a series of serious environmental problems for water bodies, prompting microbiologists to develop effective mycoherbicides to alleviate the water hyacinth population (Julien et al. 2001). In September 2020, numerous leaftip diebacks and petiole rots of water hyacinth, with 40 to 50% incidence, were observed within an area of 2 ha (ca. 2 km) mat on Jinjiang River tributary, Fujian, China. Ten infected leaf samples were collected and symptomatic tissues were cut into small pieces, surface disinfected in 75% ethanol followed by 0.1% MgCl2 solution and placed on potato dextrose agar (PDA). Pure cultures (Isolates J1 and J5) were obtained and their colonies on PDA appeared as white villi with wrinkled surfaces and dense colorless mycelium on the upper surface, while they were dark olivaceous-gray at the bottom. Internal mycelium consisted of septate, branched, smooth hyphae. There lacked stromata. Conidiophores were solitary to 2 to 5 in loose fascicles, thick-walled, conically truncated, 40 to 80 × 5 to 8 µm. Conidiogenous cells integrated, terminal, 30 to 150 × 4 to 6 µm, slightly protuberant, apical and lateral. Conidia were solitary, hyaline, acicular to obclavate, slightly curved, acute at the apex, 50 to 80 ×3 to 6 µm, indistinctly 1 to 3 septate. Genomic DNA from the two isolates was extracted for PCR, and the internal transcribed spacers (ITS), calmodulin (CAL), translation elongation factor 1-alpha (TEF), actin (ACT), histone H3 (H3) and chitin synthase (CHS) were amplified and sequenced using the primer pairs ITS1/ITS4, CL1/CL2A, EF1Fd/EF1Rd, ACT1Fd/ACT1Rd, CYLH3F/CYLH3R and CHS-79F/CHS-345R (Weir et al. 2012; White et al. 1990), respectively. The ITS (MZ436974-MZ436975), CAL (MZ519385-MZ519386), TEF (OK340826-OK340827), ACT (OK340824-OK340825), H3 (OK340828-OK340829) and CHS (MZ519387-MZ519388) sequences were deposited in GenBank. A phylogenetic tree based on concatenated sequences of ITS-CAL-TEF-ACT-H3-CHS from the genus Cercospora was constructed using a maximum likelihood method, showing that the present isolates and Cercospora rodmanii formed a monophyletic group with 99% bootstrap support. Therefore, the fungus was identified as C. rodmanii (Groenewald et al. 2013; Nguanhom et al. 2015). To test Koch's postulates, petioles of a set of 20 water hyacinth seedlings of 30 to 40 days old were wounded using a sterile needle and then spray-inoculated with 20 μl of the spore suspension of each isolate at 106 CFU/ml. Another set of 20 seedlings inoculated with sterile distilled water served as the controls. Inoculated plants were kept in 50-liter plastic tanks and maintained in a greenhouse at room temperature 22 to 28°C with a relative humidity of 80 to 85%. After 7 to 20 days, lesions were observed on the inoculated leaves but not on the control leaves. The same fungus was reisolated and identified by microscopy and PCR-sequencing. The pathogenicity test was conducted twice. Cercospora rodmanii and C. piaropi have been reported on water hyacinth in America, Brazil, México, and Zambia (Charudattan et al. 1985; Montenegro-Calderón, 2011; Moran, 2015). To our knowledge, this is the first report of C. rodmanii causing leaf and petiole lesions on water hyacinth in China. This report will help identify indigenous plant pathogens in China and develop a novel bioherbicide strategy for control of water hyacinth.

Characterization and Incidence of Target Spot Lesions in Unifoliate Leaves, Petioles, and Stems of Soybean Cultivars

This study aimed to classify the different types of leaf lesions caused by target spot and quantify their incidence in plant tissues of soybean cultivars infected by Corynespora cassiicola isolates. The experiment was conducted in a randomized block design in an 8 × 8 factorial arrangement, consisting of eight C. cassiicola isolates and eight soybean cultivars. Soybean plants were inoculated by spraying fungal suspension on the leaflets at a concentration of 2 × 104 conidia mL-1. Target spot lesions were classified, assigning scores for each type of symptom observed. The incidence of lesions in plant tissues was evaluated 10 days after inoculation, recording the presence of lesions. Five patterns of lesions were observed, ranging from small (0.74 mm) to large (9.30 mm) necrotic spots. Symptoms capable of causing defoliation occurred in BMX Elite IPRO, BRS 284, BMX Garra IPRO, and Nidera 5909 RG when inoculated with ISO 1S, ISO 4S, and ISO 11S. The highest frequency of lesioned trefoils was verified in the upper and lower (3rd and 1st trefoil) strata of soybean plants. Lesions were detected in cotyledons, unifoliate leaves, petioles, and stems of plants from all cultivars evaluated in this study. The isolate ISO 4S caused higher incidence of lesions in petioles compared with ISO 2A and ISO 2S. Moreover, ISO 4S produced more lesions in the stems of BMX Potência RR and BMX Force RR than BMX Elite IPRO. The incidence of petiole lesions caused by C. cassiicola increased as the petiole insertion height into the main stem decreased.

Isolation and characterization of Pseudomonas syringae isolates affecting stone fruits and almond in Montenegro

In Montenegro, stone fruit species are grown on intensive and semi-intensive commercial plantations. However, almond production is mainly organized on family gardens and for household consumption. During two seasons (2017–2018), we surveyed apricot, peach, nectarine, sweet cherry, Japanese plum, and almond orchards for the presence of bacterial diseases at different geographical locations in Montenegro. From leaf, petiole and fruit lesions, branch or twig cankers, and necrotizing buds, a total of 29 isolates were obtained and subjected to identification based on their morphological, pathogenic, biochemical, and molecular characteristics. Pathogenicity of the isolates was confirmed by reproducing the symptoms on leaves, fruits, and twigs of the corresponding host plants. The biochemical tests indicated that the isolates belong to Pseudomonas syringae. However, isolates’ characterization showed variation in their phenotypic and molecular features. The presence of the syrB gene and ice nucleation activity grouped most of the isolates within pathovar syringae. The results of rep-PCR using the BOX primer revealed high genetic diversity of isolates. Multilocus sequence analysis (MLSA), using four housekeeping genes, showed that 27 isolates belong to the genomic species 1, P. syringae sensu stricto, corresponding to P. syringae phylogroup 2. However, isolates from the same phylogroup 2 did not form a monophyletic group. One strain isolated from apricot was most distinct and similar to members of genomic species 2, phylogroup 3. All tested isolates showed significant levels of resistance to copper sulfate and high level of sensitivity to streptomycin sulfate in vitro.

Comparison of Whole Plant and Detached Leaf Screening Techniques for Identifying Anthracnose Resistance in Strawberry Plants.

Anthracnose is a destructive disease of strawberry caused by several Colletotrichum species including C. acutatum, C. fragariae, and C. gloeosporioides. Identification of anthracnose resistant strawberry germplasm has commonly relied on inoculation of whole plants with isolates of these pathogens. In this study, whole plants and detached leaves from 81 germplasm lines were inoculated with a conidial suspension of isolates of C. acutatum, C. fragariae, and C. gloeosporioides, incubated in the dark at 30°C, 100% relative humidity, for 48 h, and assessed for disease severity based on symptoms on inoculated petioles and leaves. The correlation between the disease severity ratings of the whole plants rated 30 days after inoculation and detached leaves rated 5 days after inoculation was determined. Based on leaf symptoms and petiole lesions, the association between the whole plant leaf disease severity rating (DSR) and detached leaf DSR was positive (rp = 0.70), and the association between the whole plant DSR and the detached leaf DSR was also positive (rp = 0.66). Whole plant and detached leaf DSRs were used to assign each germplasm line to a resistance category, and a posthoc Tukey's test showed that the whole plant DSR means and the detached leaf DSR means for each resistance category differed significantly at p < 0.05. This research was used to develop a strawberry detached leaf assay which can reliably and quickly determine the degree of resistance of strawberry germplasm to anthracnose.

First Report of Colletotrichum acutatum sensu lato Causing Leaf Curling and Petiole Anthracnose on Celery (Apium graveolens) in Michigan.

In September 2010, celery plants with leaf cupping and petiole twisting were observed in commercial production fields located in Barry, Kent, Newago, and Van Buren Counties in Michigan. Long, elliptical lesions were observed on petioles but signs (mycelia, conidia, or acervuli) were not readily observed. Celery petioles were incubated in humid chambers (acrylic boxes with wet paper towels). After 24 h, conidia corresponding to the genus Colletotrichum were observed. Isolations were performed by excising pieces of celery tissue from the lesion margin and placing them on potato dextrose agar (PDA) amended with 30 ppm of rifampicin and 100 ppm of ampicillin. Plates were incubated at 21 ± 2°C under fluorescent light for 5 days. Fungal colony morphology was gray with salmon-colored masses of spores when viewed from above, and carmine when viewed from below. Isolates were single-spored and placed on 30% glycerol in -20°C, and cryoconservation media (20% glycerol, 0.04% yeast extract, 0.1% malt extract, 0.04% glucose, 0.02% K2HPO4) at -80°C. Conidia were 8.5 to 12.0 × 2.8 to 4.0 μm and straight fusiform in shape. Three isolates were confirmed as C. acutatum sensu lato based on sequences of the internal transcribed spacer (ITS) region of the nuclear ribosomal RNA and the 1-kb intron of the glutamine synthase gene (3), both with 100% similarity with Glomerella acutata sequences. Sequences were submitted to GenBank (Accession Nos. JQ951599 and JQ951600 for ITS and GS, respectively). Additionally, C. acutatum specific primer CaIntg was used in combination with the primer ITS4 on 54 isolates from symptomatic celery plants, obtaining the expected 490-pb fragment (1). Koch's postulates were completed by inoculating 4-week-old celery seedlings of cultivars Sabroso, Green Bay, and Dutchess using three plants per cultivar. Prior to inoculation, seedlings were incubated for 16 h in high relative humidity (≥95%) by enclosing the plants in humid chambers. Seven-day-old C. acutatum s. l. colonies were used to prepare the inoculum. Seedlings were spray-inoculated with a C. acutatum s. l. conidial suspension of 1 × 106 conidia/ml in double-distilled water plus Tween 0.01%. Two control seedlings per cultivar were sprayed with sterile, double-distilled water plus 0.01% Tween. Plants were enclosed in bags for 96 h post inoculation and incubated in a greenhouse at 27°C by day/20°C by night with a 16-h photoperiod. Leaf curling was observed on all inoculated plants of the three cultivars 4 days after inoculation (DAI). Petiole lesions were observed 14 to 21 DAI. Conidia were observed in lesions after incubation in high humidity at 21 ± 2°C for 24 to 72 h. Symptomatic tissue was excised and cultured onto PDA and resulted in C. acutatum colonies. Control plants remained symptomless. C. acutatum (4) and C. orbiculare (2) were reported to cause celery leaf curl in Australia in 1966 (2,4). To our knowledge, this is the first report of C. acutatum s. l. infecting celery in Michigan.

Inheritance of Phytophthora root rot resistance in red raspberry determined by generation means and molecular linkage analysis

Classical and molecular methodologies were used to determine the inheritance of Phytophthora root rot (PRR) resistance in red raspberry. The varieties 'Latham' and 'Titan,' resistant and susceptible, respectively, were used to create F(1), F(2), B(1), B(2), and S(1) populations for analysis. Generational means analysis was used to calculate the components of genetic variation and estimates of narrow and broad sense heritability for the plant disease index and the incidence of petiole lesions. The plant disease index showed additive genetic variation with additional significant interactions, but the incidence of petiole lesions was non-additive. A dominant, two-gene model was shown to be the best fit for the observed segregation ratios when classification for resistance was based on a combination of all criteria measured. Molecular linkage maps were generated from the segregating B(2) population. Linkage maps of both parents were constructed from amplified fragment length polymorphism (AFLP), Random amplified polymorphic DNA (RAPD), and uncharacterized resistant gene analog polymorphism (RGAP) markers with seven linkage groups each totaling 440 and 370 cM of genetic distance, respectively. An analysis of the distributional extremes of the B(2) population identified several RAPD markers clustered on two linkage groups associated with PRR resistance. QTL analysis identified two similar genomic regions on each map that explained significant percentages of phenotypic variation observed for the disease assessment criteria. Genetic mapping supports the dominant two-gene model developed from generational means analysis. The results reconcile conflicting reports on inheritance of PRR resistance, provide a basis for further investigation of durable resistance to Phytophthora caused diseases, and indicates that recurrent selection is the appropriate approach for the development of new resistant cultivars.

Incidence of leaf rot disease in relation to root (wilt) in young coconut palms and fungi in lamina and petiole lesions

Coconut leaf rot in association with root (wilt) is widespread in southern districts of Kerala, India. The disease complex has spread to northern districts of the state and also adjacent districts in Tamil Nadu. Where as the root (wilt) is a systemic infection (due to phytoplasma) the leaf rot is a foliar syndrome due to fungi (Colletotrichum gloeosporioides, Exserohilum rostratum, Fusarium spp.). Occurrence of leaf rot has been related to the incidence of root (wilt) and its appearance in young palms remained a matter of contention as to which disease precedes the other. Leaf rot lesions do occur on leaf petiole/mid-vein/mid-rib even as the disease lesions are known to be common on lamina (spindles). Investigations brought out that in majority of leaf rot affected young palms (2-5 years old) the flaccidity symptom of root (wilt) could be also discerned. In lesser number of affected palms, the other symptoms of root (wilt), yellowing and marginal necrosis, were also visible. As such, in 88.5% of leaf rot affected young palms one or the other symptom of root (wilt) has been recorded irrespective of season. C. gloeosporioides, E. rostratum and other fungi were found both in lamina and petiole lesions in different months. C. gloeosporioides was detected from petiole and lamina tissues in all the months, followed by E. rostratum, Fusarium spp. etc. C. gloeosporioides was isolated from these parts in more numbers and consistently during January-December. E. rostratum appeared erratically. Aggressiveness of C. gloeosporioides during months of wet season was confirmed. Fusarium spp. was isolated predominantly from these parts in dry months (January-May). Knowledge on occurrence of leaf rot in young palms in relation to root (wilt) and dynamics of leaf rot pathogens in leaf parts among months/seasons of year (in inoculums build-up, spread and dissemination) are important in the context of integrated management of root (wilt)-leaf rot complex.

Assessing the Resistance of Red Raspberry (Rubus idaeus L.) Genotypes to Phytophthora fragariae var. rubi in Hydroponic Culture

A hydroponic method was developed and tested for screening red raspberry genotypes for resistance to Phytophthora fragariae var. rubi, the most common causal agent of Phytophthora root rot in raspberry. Plants of `Titan' and `Encore' exhibited typical disease symptoms, with the latter developing significantly smaller stem lesions and fewer petiole lesions. The resistant cultivar, `Latham', regenerated healthy root tissue from the crown and older-order roots after initial infection and necrosis of young roots and exhibited no other symptoms beyond minor leaf chlorosis. This component of the resistance reaction has not been documented previously. A segregating F1 population from the cross of `Latham' × `Titan' had a survival rate of 56% with 42% classified as resistant, exhibited minimal symptoms, and produced varying amounts of healthy root tissue. This screening method allows multiple observations of all plant tissues, including roots, under repeatable and definable growth chamber conditions. It should be useful for classifying the phenotype of individuals in segregating red raspberry populations to investigate the inheritance of Phytophthora root rot resistance using molecular markers.

First Report of Anthracnose of Gaultheria procumbens Caused by Colletotrichum gloeosporioides.

In 2001, a stem canker and dieback associated with a Colletotrichum sp. was observed on Gaultheria procumbens L. (wintergreen) seedlings grown from seed in soilless potting mix at four commercial nurseries in coastal British Columbia, Canada. Black, water-soaked lesions developed into tan cankers girdling stems and petioles, and a black rot extended into the base of leaves. In 2002, a Colletotrichum sp. was isolated from similar stem cankers from wintergreen plants. One flat of healthy, 13-week-old seedlings of G. procumbens (one- to three-leaf stage; approximately 380 seedlings per flat) was inoculated with a mycelial and spore suspension of one of the Colletotrichum sp. isolates at 108 CFU/ml using a hand atomizer. The flat was covered with black plastic for 24 h after inoculation and maintained in a growth chamber at 22°C with overhead watering. After 7 days, 98% of seedlings in the inoculated flat were necrotic with abundant sporulation of the Colletotrichum fungus on stem and petiole lesions. Plants in noninoculated flats remained healthy. Reisolation of the fungus from symptomatic tissues confirmed the causal agent to be Colletotrichum gloeosporioides (Penz.) Penz. & Sacc. in Penz, based on morphology (1,2) and polymerase chain reaction (PCR) using the species-specific primer CgInt (3). Cylindrical, hyaline, aseptate conidia measuring 16 to 20 × 4 to 6 μm were produced in acervuli with setae. Perithecia of the teleomorph, Glomerella cingulata, observed on the plants and in culture, measured 100 to 128 μm in diameter and contained acsi measuring 56 to 60 × 8 to 10 μm. Ascospores were hyaline, unicellular, slightly curved, and measured 18 to 20 × 4 to 6 μm. Subsequently, a single-spore isolate of the fungus was used to inoculate 21-week-old seedlings in the greenhouse as described above. Four inoculated and four noninoculated half-flats containing an average of 160 seedlings each were randomized on the greenhouse bench and maintained at 22°C with daily overhead misting. Stem and petiole cankers appeared on 65.1% of inoculated seedlings at 10 days after inoculation (mean of four inoculated half-flats); increasing to 90.5% at 14 days. At 14 days, 5.9% of seedlings in four noninoculated half-flats were also symptomatic, possibly from spore contamination as a result of overhead watering. To our knowledge, this is the first confirmation of the causal agent of wintergreen seedling death in seed propagation nurseries in British Columbia.

First Report of the Necrotic Strain of Potato virus Y (PVYN) on Potatoes in the Northwestern United States

More than 50 isolates of Potato virus Y (PVY) with characteristics of strains that cause tobacco veinal necrosis (PVYN) were obtained from potatoes (Solanum tuberosum L.) grown in the northwestern United States. These isolates are being characterized at the biological and molecular levels. Isolate RR1 was obtained from leaves of potato cv. Ranger Russet showing distinct mottling and leaf deformity, which is in contrast to the leaf-drop and necrosis usually observed with ordinary strains of PVY (PVYO) in this variety. Isolate AL1 was obtained from tubers of potato cv. Alturas showing distinct internal light brown rings and blotches. When RR1 and AL1 were transmitted to tobacco (Nicotiana tabacum L. cvs. Samsun NN and 423), they caused systemic veinal necrosis, including stem and petiole lesions typical of PVYN strains (2). Symptoms induced by RR1 and AL1 on tobacco appeared 9 to 11 days after inoculation, whereas some other isolates caused delayed veinal necrosis. All isolates that produced veinal necrosis on tobacco were detectable with PVY polyclonal antisera. Potato virus X was not detected by enzyme-linked immunosorbent assay in tobacco plants showing veinal necrosis. Some isolates, including AL1, failed to react in serological tests using PVYN-specific monoclonal antibodies obtained from three commercial sources. Other isolates, including RR1, were detectable with these monoclonal antibodies. Reverse transcription-polymerase chain reaction (RT-PCR) products obtained with primers specific for the coat protein (CP) open reading frame (ORF) were cloned and sequenced. AL1 possesses a CP more closely related to PVYO type isolates, which would account for its failure to react with PVYN monoclonal antibodies. In this regard, AL1 is similar to the PVYN-Wilga isolate (1). Other isolates that are detectable with the PVYN monoclonal antibodies possess a CP more consistent with N strains of the virus. Results of RT-PCR tests using primers derived from the P1 ORF sequence (3), and the restriction enzyme analysis and sequencing of the RT-PCR products, all suggest that AL1 and RR1 are related to European-type members of PVY tuber necrotic (NTN) or N strains. However, other isolates under investigation appear to be more closely related to previously reported North American NTN types (3). The symptomatology of these viruses on tobacco and potato, and the serological and molecular data clearly show that at least two distinct variants of PVYN have been found for the first time in a major potato production area of the United States, and pose a potential threat to the potato industry.

Caracterização patogênica de Colletotrichum acutatum e C. fragariae associados à antracnose do morangueiro

A antracnose é uma das principais doenças do morangueiro morangueiro (Fragaria X ananassa) e na sua sintomatologia pode estar envolvida mais de uma espécie de Colletotrichum. Neste trabalho objetivou-se caracterizar a especialização patogênica de Colletotrichum acutatum e C. fragariae em frutos, pecíolos, rizomas e inflorescências, utilizando-se inoculação artificial em várias cultivares. Em frutos, C. acutatum provocou lesões maiores que C. fragariae, enquanto que em pecíolos e rizomas C. fragariae foi mais patogênico. A cultivar Reiko foi mais suscetível às lesões dos pecíolos, enquanto 'IAC- Princesa Isabel', 'Sequoia', 'IAC-Guarani' e 'Chandler' foram mais resistentes. Os sintomas de podridão de rizoma foram menos severos em 'IAC-Princesa Isabel' e 'Sequoia' e apenas para essas duas cvs. foi observada semelhança na patogenicidade de C. acutatum e C. fragariae. Para as demais, C. fragariae foi sempre mais patogênico. Nas inflorescências houve maior severidade de sintomas quando se inoculou C. acutatum e, dentro de cada cultivar, confirmou-se esta constatação, exceto para 'IAC-Princesa Isabel', para a qual os dois fungos foram igualmente patogênicos. Evidenciou-se uma maior especialização patogênica de C. acutatum em frutos e inflorescências e de C. fragariae em pecíolos e rizomas. Porém, quando em interação com as cultivares, os dois patógenos apresentaram diferentes graus de patogenicidade, podendo causar sintomas de intensidade semelhante no mesmo órgão da planta.

First Report of Strawberry Anthracnose (Colletotrichum acutatum) in Strawberry Fields in New York.

Strawberry plants with red to black, sunken, fusoid lesions on runners and leaf petioles were found in several first-year plantings in grower's fields in western New York in 2000. Affected cultivars included Honeoye, Jewel, and Primetime. Sections of petiole were excised from lesion margins and plated on potato dextrose agar (PDA) amended with chloramphenicol, streptomycin sulfate, and tetracycline hydrochloride at 100 μg/ml. The fungus, Colletotrichum acutatum (J.H. Simmonds), was consistently isolated and identified based on conidia morphology and its growth rate in culture relative to reference cultures of C. acutatum, C. gloeosporoides, and C. fragariae (1,2). The average size of conidia produced on PDA was 15.2 × 5 μm. For each of six isolates, three plants each of six-week-old Honeoye and Kent were spray inoculated with a conidial suspension (106 conidia per ml), and petioles, leaves, and crowns were stabbed with a sterile pin following inoculation. After 14 days at 20 to 25°C, petioles and leaves on all inoculated plants developed lesions consistent with those seen in the field. The fungus was readily reisolated from leaf and petiole lesions. Subsequent inoculations on detached fruit under the same conditions yielded circular, sunken, dry lesions that produced a salmon-colored, slimy, spore mass typical of C. acutatum. This is the second report of strawberry anthracnose in the northeastern United States (3) and to our knowledge, the first report in New York.

Differences in symptom development in subterranean clover infected with Kabatiella caulivora Race 1 and Race 2 are related to host resistance

Clover scorch (Kabatiella caulivora) is a severe fungal disease of Trifolium spp. contributing to the collapse of pasture swards across southern Australia during warm, humid spring weather. Host plant responses associated with resistance to the disease were determined in 2 cultivars of subterranean clover (T. subterraneum) separately inoculated with K. caulivora Race 1 or Race 2. Germination of conidia of both races reached a maximum 5 days post-inoculation on cv. Woogenellup (susceptible to both races) and 4 days post-inoculation on cv. Daliak (resistant to Race 1 but susceptible to Race 2). Germ tube growth of Race 1 was inhibited on cv. Daliak and the percentage of conidia penetrating leaf surfaces was lowest on this race–cultivar combination. Susceptibility was characterised by large petiole lesions, with invasive hyphae extending through the mesophyll tissue into the pith and then through the phloem tissue of vascular bundles, eventually causing the petioles to collapse. Resistance was characterised by small, black lesions with invasive hyphae extending no further than the fourth layer of mesophyll cells. A suberin-based material was observed beneath infected mesophyll cells in the incompatible interaction, beyond which no further growth of hyphae occurred. Race 2 caused a faster rate of host tissue necrosis than Race 1 and also the breakdown of starch grains in uninvaded petiole tissues. Starch grains in plants infected with Race 1 were evenly distributed in uninvaded tissue. Sporulation was rarely observed in the incompatible interaction but was common in compatible interactions within 15 days post-inoculation. These responses to K. caulivora can now be used as a breeding tool in evaluating and selecting improved resistance to clover scorch disease among breeding lines of subterranean clover.

Bacterial Leaf Spot Diseases of Leafy Crucifers in Oklahoma Caused by Pathovars of Xanthomonas campestris.

Fields of kale, spinach mustard, and turnip were severely damaged by bacterial leaf spots during 1994 to 1996. Symptoms included circular to angular necrotic lesions with yellow halos and water-soaking on the abaxial leaf surface. Yellow, mucoid strains isolated from leaf spots were identified as Xanthomonas campestris using Biolog. Four strains caused black lesions on stems of cabbage seedlings in an excised cotyledon assay, leaf spots and sunken dark lesions on petioles of spray-inoculated crucifers, and leaf spots on spray-inoculated tomato. These strains were classified as X. campestris pv. armoraciae. Most other strains from leafy crucifers and all strains from a cabbage field caused black rot in the cotyledon assay and in spray-inoculations. Many of these strains also caused leaf spots on collard and kale but not stem and petiole lesions. The strains causing black rot were classified as X. campestris pv. campestris. Cluster analysis of Biolog profiles yielded a small group that contained local strains of both pathovars, and a large group comprised of reference and local strains of each pathovar, and some local, nonpathogenic strains. Five fingerprint groups were identified by rep-polymerase chain reaction using the BOXA1R primer. Local and reference strains of each pathovar occurred in two of the groups. Two pathovars of X. campestris are involved in the leaf spot diseases. Both pathovars were recovered within several fields, and also were recovered along with Pseudomonas syringae pv. maculicola. This is the first report of Xanthomonas leaf spot caused by X. campestris pv. armoraciae in Oklahoma.

First Report of Anthracnose of Lentil Incited by Colletotrichum truncatum in Bulgaria.

In June 1992 and 1995, anthracnose of lentil (Lens culinaris Medik.) incited by Colletotrichum truncatum (Schwein.) Andrus & W. D. Moore was widespread in field trials at the Institute for Wheat and Sunflower 'Dobroudja' near General Toshevo in northeastern Bulgaria. Lesions on the leaves, stems, and pods were usually white to grayish on younger plants, often turning brown as plants matured. Severe infection usually resulted in dieback and/or death of plants. Acervuli containing spores and dark setae were observed within lesions, and conidia from the acervuli produced pure cultures of C. truncatum. Conidia were hyaline, onecelled, falcate to nearly straight with a prominent clear area in the center of highly granular cytoplasm, and measured 17.6 to 19.8 × 4.4 μm. C. truncatum was seed-borne in naturally infected lentil cv. Tadjikskaya 95 at low frequencies (<2%). Koch's postulates were fulfilled by inoculating the foliage of lentil cvs. Brewer and Pardina and reisolating the fungus from stem and petiole lesions. In pathogenicity tests, three isolates of C. truncatum from the foliage and seeds of lentil caused severe symptoms on inoculated lentil cvs. Brewer and Pardina, similar to those observed on diseased lentils in Bulgaria. The fungus also caused moderate symptoms on inoculated faba bean (Vicia faba L.) and pea (Pisum sativum L.), and light symptoms on inoculated chickpea (Cicer arietinum L.). In 1995, 258 USDA Plant Introduction (PI) accessions from the USDA lentil core collection were screened in replicated trials in northeastern Bulgaria and disease symptoms were observed in >90% of the lines. Anthracnose severity ranged from light to severe. A few accessions appeared to have acceptable levels of resistance to the disease. These included accessions from Iran (PI 431714 and 431717) and Spain (PI 533693). Also that year, C. truncatum was isolated from stem lesions of naturally infected bitter vetch (Vicia ervilia (L.) Willd.) at the Institute for Wheat and Sunflower 'Dobroudja'. The disease in Bulgaria appears to be identical to one causing anthracnose of lentil in Canada (1) and the United States (2). This is the first report of C. truncatum causing anthracnose of lentil in Bulgaria.

First Report of Sclerotinia sclerotiorum on Coneflower.

Purple coneflower (Echinacea purpurea (L.) Moench; Asteraceae), a perennial herb originating from North America, is used as a garden ornamental and is grown commercially for use in medicinal preparations as an immunostimulant. In October 1996, a previously undescribed stem rot disease was observed in a research plot of 6-month-old echinacea plants at Brooks. Seedlings had been raised in small rockwool cubes (2 × 2 × 5 cm3) in a greenhouse, then transplanted into the field in early June. By late August, dead and dying plants were observed throughout the stand. They had dark brown to black stem lesions above and at the soil level and dead leaves with bleached petiole lesions that extended ca. 15 cm above the axil. Diseased stems and petioles often disintegrated, leaving only fibrous tissues intact. Roots were rotted and black. Superficial white mycelium developed over the basal part of affected stems. Black, oblong to irregular-shaped sclerotia, 5.1 to 17.6 mm in size, formed externally on the crown areas after plant death. Sclerotinia sclerotiorum (Lib.) de Bary (1) was isolated from the diseased plants. Five isolates were selected to fulfill Koch's postulates with 3-month-old echinacea seedlings grown in 12-cm pots of soilless mix. Sclerotia from wilted, field-grown echinacea plants were transferred onto potato dextrose agar medium for 2 days at 20°C. Agar disks were cut with a 1-cm cork borer and two plugs containing sclerotial and mycelial tissues were inserted into the soilless mix 0.5 cm deep and 0.5 cm from the opposite sides of stems of test plants. Inoculated plants were enclosed in transparent plastic bags for 5 days and incubated in a growth chamber at 15/18°C (night/day) with a 12-h photoperiod. One to four lower leaves per plant wilted within 1 week after inoculation and aerial mycelia appeared on the petioles. Infected leaves quickly withered, dried, and dropped off the plant after the bags were removed. Plants often died 3 weeks after inoculation and S. sclerotiorum was reisolated from infected crown tissues. This disease was also found on 3-year-old plants of E. pallida (Nutt.) Nutt. var. angustifolia (DC.) Cronq. in Vernon, British Columbia, Canada, in May 1997. This is the first report of sclerotinia stem rot on Echinacea spp., a disease that could have a significant impact on the longevity and productivity of this crop in the field and greenhouse. Reference: (1) L. H. Purdy. Phytopathology 69:875, 1979.

Leaf Spot and Stem Blight of Sweetpotato Caused byAlternaria bataticola:A New Record to South America

A severe leaf spot and stem blight disease was observed on several sweetpotato genotypes at an experimental field and in commercial plantings of the cultivar Brazlandia Roxa in the Brasilia-DF area. An Alternaria sp. was consistently isolated from the lesions. Pathogenicity tests were done under greenhouse conditions (air temperature, 18-40 C) using a spore suspension adjusted to 5 × 10 4 conidia per milliliter. The same Alternaria sp. was reisolated from leaf and petiole lesions. Even though identified as A. bataticola, the Brazilian isolates completely lacked the forked beaks in the conidia that are characteristic of this species. This is apparently the first report of A. bataticola in South America, extending the geographic range of this pathogen to the New World

Weeds as hosts of Mycocentrospora acerina

SummaryWeeds were collected from carrot fields and some other sites at 27 localities in seven counties in Norway during August and September of 1986 and 1987. Leaf and petiole lesions were microscopically examined for Mycocentrospora acerina. Viola arvensis and Viola tricolor were commonly observed with the pathogen. The fungus was also registered on Fumaria officinalis, Galium aparine, Matricaria inodora, Matricaria matricarioides, Ranunculus repens, Senecio vulgaris and Stachys palustris. In inoculation experiments in the greenhouse, M. acerina infected carrot leaves and 13 of 16 species of weeds. The symptoms on the various susceptible plants showed some variation, but were essentially of the same pattern. The investigation indicates that weeds are common hosts of M. acerina. A list of 23 host weed species is given.