Differential responses of nematode communities to soybean genotypes resistant and susceptible to Heterodera glycines race 3

Heterodera glycines and Helicotylenchus dihystera were the two most abundant plant-parasitic nematodes found in two H. glycines race 3-infested fields, Chapadão do Céu, MS and Campo Alegre, MG. These fields had been planted with resistant (R) and susceptible (S) plants to cyst nematodes. In the first field, soybean (Glycine max) FT-Cristalina (S) was susceptible to H. glycines but resistant to H. dihystera, while GOBR93 122243 (R) was just the opposite. In the second field, M-Soy 8400 (R) was more resistant to the spiral nematode than M-Soy8411 (S), but the resistance to the cyst nematode was not different between the two genotypes. The total abundance of nematodes was not different between the susceptible and resistant plants in the two fields, suggesting that H. dihystera and/or bacterial feeders and other trophic groups replaced the reduced abundance of the cyst nematodes in resistant plants. Bacterial feeders acted as a compensatory factor to plant-parasitic nematodes in ecological function. The populations of fungal feeders were higher in GOBR93 122243 (R) than in susceptible FT-Cristalina (S) in Chapadão do Céu, but lower in M-Soy 8400 (R) than in M-Soy 8411 (S) in Campo Alegre. This is being attributed to the different periods of soil samplings that were made at the florescent period in the first field, and at the final growing cycle in the second field. Only four nematodes, H. glycines, H. dihystera, Acrobeles sp. and Panagrolaimus sp. dominated the nematode resistant community GOBR93 122243 (R) in Chapadão do Céu, but dominance was shared by ten genera in Campo Alegre, which explains why the five diversity indexes (S, d, Ds, H' and T) were higher in resistant plants than in susceptible plants in field two.

Injuries caused by citrus leafminer (Phyllocnistis citrella) exacerbate citrus canker (Xanthomonas axonopodis pv. citri) infection

After the introduction of citrus leafminer in São Paulo State, an increase in the number of new plants infected with citrus canker has been observed. The interaction between these two organisms is known, but there is no information about how the leafminer damage intensifies citrus canker incidence and severity. The objectives of this paper were to evaluate the effects of leafminer damage in citrus canker infection and its influence on the monocyclic components of the disease on Citrus limonia. Higher incidence of diseased plants, AUDPC (area under the disease progress curve), disease severity and shorter incubation periods were observed in plants inoculated after insect infestation. These factors explain the association found between the higher citrus canker intensity and the damage caused by the insect and show, albeit partially, the consequences of these changes in the spread of the pathogen under natural conditions of infection.

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.

Diversidade de Phytophthora parasitica isolados de Citrus usando seqüências de nucleotídeos da região ITS-5.8S rDNA

Realizou-se estudo para caracterização e verificação da diversidade genética de Phytophthora parasitica, agente causador da gomose dos citros. Quatorze isolados de Phytophthora parasitica, provenientes do Estado de São Paulo, foram seqüenciados a partir das regiões internas transcritas (ITS1 e ITS2) do gene 5.8S. Obtiveram-se seqüências de 812 pb a 860 pb que foram comparadas com seqüências de outras espécies de Phytophthora spp depositadas no NCBI. Foram feitos estudos filogenéticos, utilizando-se o método "neighbor-joining" com 1000 "bootstrap" e construído o dendrograma mais representativo. Obtiveram-se os resultados de 98,88% a 100% de similaridade genética entre os 14 isolados paulistas, e 99,5% a 98,8% entre estes e a seqüência de P. nicotianae (gi| 8927482) obtida do GenBank NCBI.

The citrus leprosis pathosystem

Citrus leprosis is considered the main viral disease for the Brazilian citrus production, particularly for the State of São Paulo, due to the high costs spent for the chemical control of its vector, the tenuipalpid mite Brevipalpus phoenicis. In addition, its global importance has significantly increased in the last years, with the dissemination of the virus to new countries in South and Central America. In Brazil, despite its economical importance and occurrence for more than seven decades, the most significant advances towards understanding the pathosystem interactions have been obtained only in the last ten years. This review focuses on various aspects of the disease, beginning with a historical view, its main characteristics, alternatives for its control, its increasing economical importance in Brazil and abroad, and the new data on the search for understanding the interactions amongst the mite vector, the virus, and the plant host.

Characterization of Citrus tristeza virus isolates from grapefruit (Citrus paradisi Macf.) accessions of Citrus Active Germplasm Bank

Citrus tristeza virus (CTV) isolates from 35 grapefruit accessions belonging to Citrus Active Germplasm Bank of the "Instituto Agronômico de Campinas" located at the "Centro APTA Citros Sylvio Moreira", Cordeirópolis, São Paulo state, Brazil, were characterized and evaluated through symptoms in the trees, biological indexing, immunological diagnosis with different monoclonal antibodies and SSCP analysis (single-strand conformation polymorphism) of the coat protein gene. Symptomatology indicated that, in general, the group of plants with smaller canopy volume and severe stem pitting differed significantly from the group that presented greater vegetative development and mild to moderate stem pitting. However, the isolates from most of the accessions induced mild reaction on Mexican lime. The serological evaluation through the DAS-ELISA using monoclonal antibodies did not reveal any association between virus titer in the plant tissue and symptoms. The reaction with different monoclonal antibodies and the distinct electrophoresis patterns obtained through SSCP showed that there is a high degree of diversity among the isolates that infect these grapefruit accessions. High complexity within the same isolate was also observed in the SSCP profiles. This finding indicates that the CTV isolates from these plants are a complex mixture of CTV haplotypes. Similar SSCP banding patterns were observed among some plants with strong stem pitting symptoms, and among some plants with weak or moderate stem pitting symptoms.

Variabilidade biológica de isolados do Citrus leprosis virus (CiLV) oriundos de cultivares de laranjeira

A leprose, causada pelo Citrus leprosis virus (CiLV), é uma das principais doenças presentes em pomares cítricos fluminenses. O objetivo deste trabalho foi comparar o quadro sintomatológico desenvolvido por isolados de CiLV obtidos de cultivares comerciais de laranjeira (Lima, Pêra e Seleta), inoculados mecanicamente em Chenopodium amaranticolor, em três diluições. Após cinco a sete dias da inoculação foram observadas lesões necróticas, com pequeno halo clorótico quando observadas contra a luz. O maior número de lesões, nas três diluições, foi obtido do isolado de 'Seleta', seguido por 'Pêra' e 'Lima'. A melhor diluição utilizada para a observação das lesões foi de 1:10. Os resultados demonstram uma possível variabilidade biológica entre os isolados virais e/ou uma menor ou maior replicação viral, dependendo da cultivar, indicando um possível mecanismo de resistência da planta ao vírus.

Management of crotalaria and pigeon pea for control of yam nematode diseases

Management of plant-parasitic nematodes with the use of nematicides has not been recommended for small farmers that grow yam in the Northeastern region of Brazil, due to its high cost and residue toxicity. The use of plants with antagonistic effect to nematodes and green manure which improves soil chemical, physical and biological characteristics can be a viable and low cost alternative to control parasitic nematodes. This work aimed to evaluate the effect of crotalaria (Crotalaria juncea) and pigeon pea (Cajanus cajan) plants on the control of yam nematodes. Three experiments were carried out. The first was conducted under in vitro conditions to evaluate the nematostatic and nematicide effect of extracts from fresh and dry matter of the above ground parts of crotalaria, pigeon pea, and the combination of both. The second experiment was carried out under greenhouse conditions to evaluate the effect of soil amendment with crotalaria, pigeon pea, and the combination of both in the infectivity of Scutellonema bradys, using tomato plants as the host plant. The third experiment was conducted under field conditions to evaluate the effect of crotalaria, pigeon pea, and the combination of both, cultivated between yam planting rows and incorporated to soil surface, on yam nematodes. The aqueous extract obtained form fresh matter of crotalaria had a nematicide effect of 100% for S. bradys. Extracts from dry matter of both crotalaria and pigeon pea did not have any nematicide effect, but had a nematostatic effect. Incorporation of crotalaria to soil inhibited infectivity of S. bradys in tomato seedlings. These results showed that planting crotalaria alone or in combination with pigeon pea, between the yam planting rows, is an efficient method for controlling S. bradys and Rotylenchulus reniformis associated with yams. Crotalaria can be used for controlling these plant-parasitic nematodes in soil.

Caracterização biológica e molecular de isolados do Citrus tristeza virus com potencial para utilização em programas de pré-imunização

O experimento teve como objetivo caracterizar biológica e molecularmente os isolados de CTV, obtidos de árvores de laranja Pêra em pomares das regiões Norte e Noroeste do Paraná, visando selecionar isolados fracos do Citrus tristeza virus com capacidade protetiva para pré-imunização. Os resultados apontaram sintomas fracos a moderados de caneluras e revelaram similaridade genética da maioria dos isolados analisados e o isolado fraco controle, sugerindo que as plantas selecionadas estão infectadas por haplótipos fracos de CTV, com potencial para serem utilizadas como plantas matrizes.

Use of botanical extracts, cassava wastewater and nematicide for the control of root-knot nematode on carrot

Aqueous extracts of several plant species have shown promising in controlling root-knot nematode, Meloidogyne incognita (Kofoid & White), one of the most limiting agents for carrot cultivation. The current study evaluated the effect of aqueous extracts from seven botanical species applied to 40, 50, 60, 70 and 80 days after sowing 'Nantes' carrots in soil infested with root-knot nematode. Three other treatments included cassava wastewater, distilled water (control), which were applied in the same periods of the extracts application, in addition to carbofuran 50G (80Kg/ha), which was applied once at 60 days after carrot sowing. Evaluations were performed at 90 days after inoculation to determine shoot and root fresh weight, as well as the diameter and the length of principal roots and the number of galls on primary and secondary roots. Plants treated with cassava wastewater, extracts of Ricinus communis L. seeds, Crotalaria juncea L. seeds, R. communis leaves + branches + fruits, Chenopodium ambrosioides L. leaves + branches + inflorescences and Azadirachta indica A. Juss. seeds showed the highest rates of total weight (root + shoot) and shoot weight. The extract of R. communis leaves + branches + fruits provides the highest total root weight and principal root diameter. Cassava wastewater and extracts of R. communis seeds provided the highest principal root weight. The extract of R. communis seeds and cassava wastewater can be considered promising for the alternative control of M. incognita.

Potencial do flavedo (epicarpo) de Citrus aurantifolia cv. Tahiti no controle do bolor verde e da antracnose em citros

O Brasil é considerado o maior produtor de citros e o maior exportador de suco de laranja. Doenças de pós-colheita representam uma grande perda para a citricultura, sendo que para a exportação de frutos são rígidas as exigências com relação a isenção de resíduos químicos nos mesmos. Patógenos de importância em pós-colheita de citros incluem o Penicillium digitatum, agente causal do bolor-verde e o Colletotrichum gloeosporioides, agente causal da antracnose. Dada a importância econômica que representam estas doenças dos frutos cítricos, tanto em termos de comprometimento da qualidade e dificuldade de controle, a busca de alternativas adicionais que possam viabilizar a capacidade produtiva e garantir a obtenção de frutos com excelentes padrões de qualidade torna-se imprescindível. Portanto, estudou-se os efeitos dos extratos aquosos do flavedo de Citrus aurantifolia var. Tahiti, Lentinula edodes, Agaricus subrufescens (syn. Agaricus brasiliensis), albedo de Citrus sinensis var. Valência e do ácido jasmônico no controle póscolheita do bolor verde e da antracnose e na indução de resistência em frutos de laranjeira Valência (Citrus sinensis). Foi possível observar que o extrato aquoso do flavedo (C. aurantifolia) apresentou efeito inibitório sobre os patógenos, quando tratados em pós-colheita, em função da redução dos sintomas e esporulação. Porém, os extratos de albedo (C. sinensis), L. edodes, A. subrufescens e o ácido jasmônico não apresentaram efeitos sobre P. digitatum e C. gloeosporioides.

Nematicidal effect of volatile organic compounds (VOCs) on the plant-parasitic nematode Meloidogyne javanica

Previous studies have demonstrated that volatile organic compounds (VOCs), produced by the yeast Saccharomyces cerevisiae, were able to inhibit the development of phytopathogenic fungi. In this context, the nematicidal potential of the synthetic mixture of VOCs, constituted of alcohols and esters, was evaluated for the control of the root-knot nematode Meloidogyne javanica, which causes losses to crops of high economic value. The fumigation of substrate containing second-stage juveniles with VOCs exhibited nematicidal effect higher than 30% for the lowest concentration tested (33.3 µL g-1 substrate), whereas at 66.6 and 133.3 µL g-1 substrate, the nematode mortality was 100%. The present results stimulate other studies on VOCs for nematode management.

Avaliação da resistência de variedades de Citrus spp. à Xanthomonas citri subsp. citri na região Noroeste Paranaense, em condições de campo

A identificação de cultivares com determinados níveis de resistência ao cancro cítrico, com características agronômicas e comerciais desejáveis, assume grande importância para o manejo desta doença. A avaliação de genótipos promissores pode ser fetuada em condições controladas ou em campo. O trabalho teve como objetivo avaliar genótipos de laranja doce (Citrus sinensis), tangerinas (Citrus reticulata) e híbridos quanto à resistência ao cancro cítrico, em diferentes épocas de avaliação. O experimento foi conduzido em pomar experimental no município de Maringá, Paraná. Para a determinação da severidade média da doença, foram utilizadas escalas diagramáticas, sendo avaliadas quatro plantas por bloco e amostrados ramos na porção mediana de cada planta. Além disso, foi calculada a Área Abaixo da Curva de Progresso da Severidade (AACPS). De acordo com os dados obtidos, ocorreram diferenças significativas na severidade e na AACPS, ao longo das avaliações, sendo que a severidade variou de 0,25% a 4,88% e a AACPS de 49,80 a 742,65. As variedades cítricas Tang. Ponkan, Rosehaugh Nartjee, Harris, De wildt, Dekopon, Lee, África do Sul, Tarocco#23, Solid Scarlet, Satsuma Okitsu SPA 29, Cristalina, Shamouti e Pêra Olimpia se destacaram e apresentaram as menores médias de severidade, sendo essas classificadas como resistentes. Os resultados sugerem que esses genótipos podem ser importantes ferramentas nos programas de melhoramento de genótipos de citros, visando resistência a Xanthomonas citri subsp. citri.