Detection of Varicosavirus and Ophiovirus in lettuce associated with lettuce big-vein symptoms in Brazil

During surveys undertaken from 1998 to 2003 in the major vegetable growing areas of the city of São Paulo green belt, lettuce (Lactuca sativa) and endive (Cichorium endivia) plants were observed, which showed chlorotic thickening of foliar veins, defective growth and, in some cases, failure to form complete heads. Biological and serological [DAS-Enzyme linked immunosorbent assay (Elisa)] tests together with electron microscope observations, revealed the presence of Lettuce big-vein virus and Mirafiori lettuce virus, in these plants both responsible for the lettuce big-vein syndrome.

First report of Fusarium oxysporum f. sp. lycopersici race 3 on tomato in Brazil

Fusarium wilt, caused by three races of Fusarium oxysporum f. sp. lycopersici, is one of the most important diseases of tomato (Lycopersicon esculentum). Races 1 and 2 are distributed worldwide whereas race 3 has a more limited geographic distribution with no report thus far in Brazil. Seven F. oxysporum isolates were obtained from wilted tomato plants of race 1 and 2-resistant hybrids 'Carmen' and 'Alambra' in Venda Nova do Imigrante (State of Espírito Santo), Brazil. Virulence assays were performed using a set of the race differential cultivars: 'Ponderosa' (susceptible to all races), 'IPA-5' (resistant to race 1), 'Floradade' (resistant to races 1 and 2) and 'BHRS-2,3' (resistant to race 3). All isolates were highly virulent to 'Ponderosa', 'IPA-5' and 'Floradade' and were able to infect only a few plants of 'BHRS-2,3'. An additional virulence test was conducted including the same set of cultivars plus Lycopersicon pennellii 'LA 716'. Identical results were obtained with L. pennellii displaying an extreme (immune-like) resistant response. These results indicated that all seven isolates could be classified as F. oxysporum f. sp. lycopersici race 3. This new Fusarium wilt might became an economically important disease since race 3-resistant cultivars adapted to Brazil are not yet available.

Citrus exocortis viroid and Hop Stunt viroid Doubly infecting grapevines in Brazil

Viroids, non-protein-coding small (246-401 nt) circular single-stranded RNAs with autonomous replication, are currently classified into two families. Within the family Pospiviroidae, Citrus exocortis viroid (CEVd) belongs to the genus Pospiviroid while Hop stunt viroid (HSVd) is the single member of the genus Hostuviroid. These pathogens are distributed worldwide and infect a large number of hosts. In Brazil, isolates of CEVd and HSVd have been detected in both citrus and grapevine. To characterize and study the genetic variability of these viroids, total RNA from leaves of grapevine Vitis vinifera 'Cabernet Sauvignon' and V. labrusca 'Niagara Rosada' from Bento Gonçalves, RS, was used as a template for RT-PCR amplification with specific primers for the five viroids described infecting grapevines [HSVd, CEVd, Grapevine yellow speckle viroid 1 (GYSVd-1), Grapevine yellow speckle viroid 2 (GYSVd-2) and Australian grapevine viroid (AGVd)]. Leaf samples of Citrus medica infected with CEVd from São Paulo were also analyzed. The resulting products were separated by agarose gel electrophoresis and DNA fragments of the expected size were eluted, cloned and sequenced. The grapevine samples analyzed were doubly infected by CEVd and HSVd. A phylogenetic analysis showed that the Brazilian grapevine HSVd variants clustered with other grapevine HSVd variants, forming a specific group separated from citrus variants, whereas the Brazilian CEVd variants clustered with other citrus and grapevine variants.

A new Ophiodothella species associated with leaf spots on Annona squamosa in Brazil

A new species of Ophiodothella, O. annonae (Phyllachoraceae, Ascomycetes) is described and illustrated. The fungus causes brown lesions on leaves of Annonae squamosa, which can fall off prematurely, and is characterized by fusiform, slightly curved, biguttulate ascospores with a conspicuous central concentration of cytoplasm. This is the first species of Ophiodotella found on leaves of Annonaceae in Brazil.

A novel postharvest rot of okra pods caused by Rhizoctonia solani in Brazil

Okra pods with unusual brown lesions and rot were collected in a local supermarket in Brasília DF. The objective of this paper was to characterize the causal agent, to fulfill Koch's postulates and to determine some conditions conducive to disease. The pathogen was identified as Rhizoctonia solani based on morphological characteristics which fitted the fungus description, such as pale to brown hyphae, with nearly right-angled side branches constricted at the base, hyphal cells 6-10 µm wide with a septum near the base. Five isolates were obtained from infected pods and identified as AG 1-IB anastomosis group. Wounded or unwounded okra pods cv. Santa Cruz 47 were inoculated with mycelium disks of R. solani and kept in humid chambers at 12 ºC or 25 ºC. After seven days at 25 ºC, both wounded and unwounded pods were completely rotted and brown, while those kept at 12 ºC showed small lesions ranging from 0.6 to 1.0 mm only in wounded pods. The pathogen was able to grow in different materials used for assembling crates and packs of horticultural products, such as pinewood, corrugated carton, plastic, Styrofoam and newspaper sheets when kept in humid chambers (24 ºC, 96 % RH). The disease occurrence can be related to careless handling practices and to the transmission of R. solani propagules by infected plant debris or soil particles. This is the first report of Rhizoctonia solani causing postharvest rot in okra pods in Brazil.

Variability of the coat protein gene of Grapevine leafroll-associated virus 3 in Brazil

Leafroll is an economically important disease affecting grapevines (Vitis spp.). Nine serologically distinct viruses, Grapevine leafroll-associated virus-1 through 9, are associated with this disease. The present study describes the coat protein gene sequence of four GLRaV-3 isolates occurring in the São Francisco River basin, Northeastern Brazil. The viral RNA was extracted from GLRaV-3 ELISA-positive plants and the complete coat protein gene was amplified by RT-PCR. Sequences were generated automatically and compared to the complete coat protein sequence from North American (NY1) and Chinese (Dawanhong Nº2 and SL10) GLRaV-3 isolates. The four studied isolates, named Pet-1 through 4, showed deduced amino acid identities of 98-100% (Pet-1 through 3) and 95% (Pet-4) with North American and Chinese isolates. A total of seventeen amino acid substitutions was detected among the four characterized isolates in comparison to the NY1, Dawanhong No.2 and SL10 sequences. The results indicated the existence of natural variation among GLRaV-3 isolates from grapevines, also demonstrating a lack of correlation between sequence data and geographic origin. This variability should be considered when selecting regions of the viral genome targeted for reliable and consistent virus molecular detection.

Powdery mildew of Ruta graveolens in Brazil caused by Oidiopsis haplophylli

Oidiopsis haplophylli is described causing powdery mildew on Ruta graveolens for the first time in Brazil (Viçosa, MG). The fungus causes yellowing of infected leaves accompanied by the presence of a whitish mycelial colony abaxially. The following combination of characters typical of this species was observed on the specimens collected in Viçosa: mycelium hypophyllous, hemiendophytic (partly external and partly internal), entering the leaves through the stomata; conidiophores hypophyllous, produced from the internal mycelium emerging through the stomata, cylindrical, hyaline, smooth; conidia, isolate, dimorphic - primary conidia lanceolate, 66.5-91.5 x 11.0-20.0 mm l/w ratio 3.5-6.2, secondary conidia cylindrical with rounded ends, 57.0-81.5 x 13.5-20.0 mm, l/w ratio 3.1 - 5.3, aseptate, hyaline, smooth.

Powdery mildew of ornamental species caused by Oidiopsis haplophylli in Brazil

Oidiopsis haplophylli (syn. Oidiopsis sicula) was identified as the causal agent of powdery mildew diseases occurring on five ornamental species in Brazil. This disease was observed in plastic house-grown lisianthus (Eustoma grandiflorum: Gentianaceae), in nasturtium (Tropaeolum majus: Tropaeolaceae) cultivated under open field conditions and in greenhouse-grown calla lily (Zantedeschia aethiopica: Araceae), impatiens (Impatiens balsamina: Balsaminaceae) and balloon plant (Asclepias physocarpa: Asclepiadaceae). Typical disease symptoms consisted of chlorotic areas on the upper leaf surface corresponding to a fungal colony in the abaxial surface. With the disease progression, these chlorotic areas eventually turned to necrotic (brown) lesions. Fungi morphology on all hosts was similar to that described for the imperfect stage of Leveillula taurica (O. haplophylli). The Koch's postulates were fulfilled by inoculating symptom-free plants via leaf-to-leaf contact with fungal colonies. Additional inoculations using an isolate of O. haplophylli from sweet pepper (Capsicum annuum) demonstrated that it is pathogenic to all five species belonging to distinct botanical families, indicating lack of host specialization. This is the first formal report of a powdery mildew disease on lisianthus, calla lilly, impatiens and nasturtium in Brazil. It is, to our knowledge, the first report of O. haplophyllii infecting A. physocarpa, extending the host range of this atypical powdery mildew-inducing fungus. This disease might become important on these ornamental crops especially in protected cultivation and also under field conditions in hot and dry areas of Brazil.