In June of 2009, sweet pepper (Capsicum annuum cvs. Elisa and Prador) plants exhibiting interveinal chlorosis, some necrosis, and mild upward leaf curling on the intermediate leaves were found in three protected crops in the municipality of São Miguel Arcanjo, São Paulo state, Brazil. Incidence of symptomatic plants varied from 70 to 100%. Abundant whitefly adults were seen in all crops. Initially, total DNA was separately extracted from seven symptomatic plants and submitted to a PCR reaction using the universal primer pairs PAL1v1978/PAR1c496 and PBL1v2040/PCRc1 for begomovirus (3). The results were negative. The same samples were also analyzed for infection with Tomato infectious chlorosis virus (TICV) and Tomato chlorosis virus (ToCV) (genus Crinivirus, family Closteroviridae). Total RNA was extracted separately from leaves of each symptomatic plant and used for one-step reverse transcription (RT)-PCR using the HS-11/HS-12 primer pair, which amplifies a fragment of 587 bp from the highly conserved region of the heat shock protein (HSP-70) homolog gene reported for TICV and ToCV (1). The RT-PCR product was subsequently tested by nested-PCR for single detection of TICV and ToCV using primer pairs TIC-3/TIC-4 and ToC-5/ToC-6, respectively (1). Only one fragment of approximately 463 bp was amplified from the five plants with the primer pair specific for ToCV. No amplification was obtained with the primers specific for TICV. Four purified amplicons of 463 bp were directly sequenced in both directions. Sequence comparisons of the 419-bp consensus sequence, encompassing nucleotides 750 and 1,167 of the HSP-70 homolog gene, revealed 98% identity with the reported sequences of tomato infecting isolates of ToCV from Brazil (GenBank Accession No. EU868927) and the United States (GenBank Accession No. AY903448). Virus-free adults of Bemisia tabaci biotype B were confined on symptomatic pepper leaves for a 48-h acquisition access period. Twenty adults were transferred to one plant of sweet pepper cv. Magda for a 24-h inoculation access period. The sweet pepper plant exhibited the original symptoms on the leaves 67 days after inoculation under greenhouse conditions. Infection by ToCV was confirmed by RT-PCR. The susceptibility of sweet pepper plants to ToCV was previously reported in Spain (2), whereas in the United States, this species was experimentally found as nonhost for this virus (4). Further studies are needed to better understand the variable susceptibility of sweet pepper to ToCV. References: (1) C. I. Dovas et al. Plant Dis. 86:1345, 2002. (2) G. Lozano et al. Plant Dis. 88:224, 2004. (3) M. R. Rojas et al. Plant Dis. 77:340, 1993. (4) W. M. Wintermantel et al. Plant Dis. 90:814, 2006.

Doze isolados de Pythium foram obtidos de raízes de alface cultivada em sistemas hidropônicos comerciais, apresentando ou não sintomas de apodrecimento. Três desses isolados foram identificados como Pythium helicoides Drechsler (H1, H2 e H3), cinco como pertencentes ao grupo F (F1 a F5) e quatro ao grupo T (T1 a T4) de Pythium. A identificação das espécies foi realizada baseando-se nas características morfológicas. O efeito da temperatura (10, 15, 18, 21, 24, 27, 30, 37 e 40ºC) sobre o crescimento micelial foi determinado para todos os isolados. As temperaturas mínima e máxima, estimadas pela função beta generalizada, variaram de 3,5 a 10ºC e de 40 a 40,7ºC, respectivamente. A temperatura ótima foi de 24 a 37ºC para P. helicoides, de 25 a 35ºC para o isolado F4 e de 21 a 30ºC para os demais isolados. A patogenicidade e a agressividade dos isolados foram avaliadas, inoculando-se sementes de alface cv. Verônica, semeadas em ágar-água, a 21 e 30ºC. A 30ºC, os isolados de P. helicoides foram notadamente os mais agressivos, ocasionando 100 % de mortalidade das sementes logo após sua germinação. A 21ºC, todos os isolados induziram subdesenvolvimento de plântulas, acompanhado ou não de necrose dos tecidos radiculares. Trata-se do primeiro relato de P. helicoides para o Brasil e a primeira referência mundial da espécie em hidroponia.

A alface é a folhosa de maior importância no Brasil. O presente trabalho visou determinar a reação de cultivares de alface à murchadeira provocada pelo fungo Thielaviopsis basicola, na fase juvenil. Um experimento foi conduzido em casa-de-vegetação em delineamento inteiramente casualizado, em esquema fatorial 37 x 2 (cultivares, com e sem inoculação), com três repetições. Mudas com 30 dias foram transplantadas para bandejas de 128 células preenchidas com 1/3 de substrato colonizado com 7,5 x 10(5) conídios/g de substrato. Logo após o transplante, inoculou-se 3 mL de suspensão de esporos de concentração 2 x 10(6) conídios/mL, próximo ao colo de cada planta. A reação do hospedeiro ao patógeno e sua avaliação foi realizada utilizando escala de nota de 1 (ausência de sintomas) a 5 (mais de 90% das raízes afetadas), com base na severidade da doença. Cultivares do tipo crespa e batavia foram todas resistentes. Cultivares do tipo americana e lisa apresentaram variação inter-varietal quanto à reação da hospedeira ao patógeno.

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Root infections in hydroponic lettuce are frequent and mostly caused by Pythium species, which are extremely well adapted to aquatic environments. The present study aimed to evaluate in vitro the pathogenic potential of Pythium middletonii and Pythium dissotocum in four cultivars of lettuce, Elisa (smooth), Vera (curly), Mimosa (mimosa) and Tainá (American). Lettuce seeds from each cultivar were superficially disinfected, pre-germinated for 24 hours and placed on the water agar medium surface. Then, a dish containing mycelium from Pythium isolates was placed in the center of each plate. Control consisted of plates containing lettuce seeds only. Experimental design was completely randomized, with five replicates, each one represented by a Petri dish. Hypocotyl and radical length besides surviving seedlings percen tage after ten days of in cuba tion were evaluated. pathogenicity of the isolates was investigated at the ideal temperature for the lettuce (20ºC), and at the ideal temperatures for the growth of the isolates, 23ºC for P. middletonii and 27ºC for P. dissotocum. At 20ºC, P. dissotocum had higher pathogenicity on lettuce cultivars than P. middletonii, significantly decreasing the length of hypocotyls, especially radicles, of most cultivars. For P. dissotocum, 27ºC was the most suitable temperature for the specimen growth; however, it led to the lowest percentage of surviving seedlings among all cultivars, with the lowest percentage (54%) detected for Vera. Among the cultivars, Mimosa presented higher percentage of survivor seedlings and higher length of radicles in relation to the control, and thus was considered less susceptible to the pathogen. P. dissotocum presented higher mycelium growth and was more pathogenic than P. middletonii in all experiments.experimento foi conduzido na temperatura ideal de crescimento da alface (20ºC), e nas ideais para o crescimento dos isolados, 23ºC para P. middletonii e 27ºC para P. dissotocum. A 20ºC, P. dissotocum foi mais patogênico que P. middletonii, reduzindo significativamente o comprimento dos hipocótilos e, principalmente, das radículas, de praticamente todas as cultivares. Na temperatura de 27ºC, P. dissotocum foi responsável pela mais baixa porcentagem de plântulas sobreviventes entre as cultivares, sendo mais patogênico em Vera (54% de sobrevivência). Dentre as cultivares analisadas, a Mimosa mostrou tendência de menor suscetibilidade a este patógeno, apresentando a maior porcentagem de plântulas sobreviventes e o maior comprimento das radículas em relação ao controle. P. dissotocum apresentou maior crescimento micelial e foi mais patogênico que P. middletonii em todos os experimentos realizados.

This contribution describes biological and molecular features of an isolate of the chrysanthemum stem necrosis orthotospovirus (CSNV) found naturally infecting sweet pepper plants (Capsicum annuum L.). High-throughput sequencing resulted in three contig sequences of 2948, 4829 and 8959 nucleotides for CSNV-Ca small, medium and large RNAs, respectively. They shared 98.4%, 99.3% and 99.1% nucleotide identities, respectively, with the corresponding nucleotide sequences of a CSNV isolate from Slovenia. The virus was mechanically transmitted to sweet pepper and some additional test plants, and RT-PCR confirmed the infection with specific primers for CSNV. Additionally, transmission electron microscopic analysis of ultrathin sections prepared from symptomatic sweet pepper fruit tissue showed orthotospoviruses-like particles in elements of the endoplasmic reticulum. Studies are needed to evaluate the occurrence of CSNV infecting sweet pepper in commercial crops, to assess yield damage and develop recommendations for management.