Development of a mungbean (Vigna radiata) RFLP linkage map and its comparison with lablab (Lablab purpureus) revelas a high level of colinearity between the two genomes

A genetic linkage map of mungbean (Vigna radiata, 2n = 2x = 22) consisting of 255 RFLP loci was developed using a recombinant inbred population of 80 individuals. The population was derived from an intersubspecific cross between the cultivated mungbean variety 'Berken' and a wild mungbean genotype 'ACC 41' (V radiata subsp. sublobata). The total length of the map, which comprised 13 linkage groups, spanned 737.9 cM with an average distance between markers of 3.0 cM and a maximum distance between linked markers of 15.4 cM. The mungbean map was compared to a previously published map of lablab (Lablab purpureus, 2n = 2x = 24) using a common set of 65 RFLP probes. In contrast to some other comparative mapping studies among members of the Fabaceae, where a high level of chromosomal rearrangement has been observed, marker order between mungbean and lablab was found to be highly conserved. However, the two genomes have apparently accumulated a large number of duplications/deletions after they diverged.

Relationship between oviposition preference and offspring performance in Australian Helicoverpa armigera (Hubner) (Lepidoptera : Noctuidae)

We investigated the oviposition preference and larval performance of Helicoverpa armigera under laboratory conditions to determine if the oviposition preference of individual females on maize, cowpea and cotton correlates with offspring performance on the leaves of the same host plants. The host-plant preference hierarchy of females did not correlate with their offspring performance. Female moths chose host plants that contributed less to their offspring fitness. Plant effects accounted for the largest amount of variation in offspring performance, while the effects of female (family) was low. The offspring of most females (80%, n = 10) were broadly similar, but 20% (two out of 10), showed marked difference in their offspring performance across the host-plant species. Similarly, there was no relation between larval feeding preference and performance. However, like most laboratory experiments, our experi-mental design does not allow the evaluation of ecological factors (for example, natural enemies, host abundance, etc.) that can play an important role in larval performance in the field. Overall, the results highlight the importance of carrying out preference performance analysis on the individual or family level, rather than pooling individuals to obtain average population data.

Interspecific hybrids between the homothallic Phytophthora sojae and Phytophthora vignae

An interspecific cross was attempted between two homothallic species of Phytophthora, P. sojae and P. vignae. From 1640 single-oospore cultures isolated, DNA was extracted from 800, and two interspecific F-1 hybrids (F(1)1121 and F(1)1426) were putatively identified using RAPD markers. The true hybrid nature of these F-1 hybrids was confirmed using additional AFLP analysis. Single- zoospore cultures were generated for each F-1 hybrid and one single-zoospore culture of each was used in pathogenicity and virulence tests. Both F-1 hybrids were pathogenic to soybean and cowpea, causing symptoms including lesions, wilting and death of susceptible soybean and cowpea cultivars. However, the aggressiveness of the F-1 hybrids was reduced and was substantially more variable when compared with that of the parental isolates on their respective hosts. The F-1 hybrids were reisolated from infected seedlings and their hybrid nature confirmed using RAPD and AFLP analysis. These results provide a basis for further research aimed at obtaining an increased understanding of the genetics of host specificity in the Oomycetes.

Microbial biomass and soil chemical properties under different land use systems in northeastern Pará

The increase in agricultural production in the Brazilian Amazon region is mostly a result of the agricultural frontier expansion, into areas previously influenced by humans or of native vegetation. At the same time, burning is still used to clear areas in small-scale agricultural systems, leading to a loss of the soil productive capacity shortly after, forcing the opening of new areas. This study had the objective of evaluating the effect of soil preparation methods that involve plant residue shredding, left on the surface or incorporated to the soil, with or without chemical fertilization, on the soil chemical and biological properties. The experiment was conducted in 1995, in an experimental field of Yellow Latosol (Oxisol) of the Embrapa Amazônia Oriental, northeastern Pará (Brazil). The experiment was arranged in randomized blocks, in a 2x6 factorial design, with two management systems and six treatments evaluated twice. The management systems consisted of rice (Oriza sativa), followed by cowpea (Vigna unguiculata) with manioc (Manihot esculenta). In the first system the crops were planted in two consecutive cycles, followed by a three-year fallow period (natural regrowth); the second system consisted of one cultivation cycle and was left fallow for three years. The following treatments were applied to the secondary forest vegetation: slash and burn, fertilized with NPK (Q+NPK); slash and burn, without fertilizer NPK (Q-NPK); cutting and shredding, leaving the residues on the soil surface, fertilized with NPK (C+NPK); cutting and shredding, leaving residues on the soil surface, without fertilizer (C-NPK); cutting and shredding, with residue incorporation and fertilized with NPK (I+NPK); cutting and shredding, with residue incorporation and without NPK fertilizer (I-NPK). The soil was sampled in the rainier season (April 2006) and in the drier season (September 2006), in the 0-0.1 m layer. From each plot, 10 simple samples were collected in order to generate a composite sample. In the more intensive management system the contents of microbial C (Cmic) and microbial N (Nmic) were higher, while the C (Corg) level was higher in the less intensive system. The treatments with highest Cmic and Nmic levels were those with cutting, shredding and distribution of biomass on the soil surface. Under both management systems, the chemical characteristics were in ranges that classify the soil as little fertile, although P and K (in the rainy season) were higher in the less intensive management system.

SOIL FUNGISTASIS AGAINST FUSARIUM GRAMINEARUM UNDER DIFFERENT CROP MANAGEMENT SYSTEMS

Soil management, in terms of tillage and cropping systems, strongly influences the biological properties of soil involved in the suppression of plant diseases. Fungistasis mediated by soil microbiota is an important component of disease-suppressive soils. We evaluated the influence of different management systems on fungistasis against Fusarium graminearum, the relationship of fungistasis to the bacterial profile of the soil, and the possible mechanisms involved in this process. Samples were taken from a long-term experiment set up in a Paleudult soil under conventional tillage or no-tillage management and three cropping systems: black oat (Avena strigose L.) + vetch (Vicia sativa L.)/maize (Zea mays L.) + cowpea (Vigna sinensis L.), black oat/maize, and vetch/maize. Soil fungistasis was evaluated in terms of reduction of radial growth of F. graminearum, and bacterial diversity was assessed using ribosomal intergenic spacer analysis (RISA). A total of 120 bacterial isolates were obtained and evaluated for antibiosis, and production of volatile compounds and siderophores. No-tillage soil samples showed the highest level of F. graminearum fungistasis by sharply reducing the development of this pathogen. Of the cropping systems tested, the vetch + black oat/maize + cowpea system showed the highest fungistasis and the oat/maize system showed the lowest. The management system also affected the genetic profile of the bacteria isolated, with the systems from fungistatic soils showing greater similarity. Although there was no clear relationship between soil management and the characteristics of the bacterial isolates, we may conclude that antibiosis and the production of siderophores were the main mechanisms accounting for fungistasis.

Microenxertia ex vitro para eliminação do vírus CABMV em maracujá-azedo

O objetivo deste trabalho foi avaliar diferentes meios de cultura, utilizados sobre o ponto da enxertia, na microenxertia ex vitro para a eliminação do Cowpea aphid-borne mosaic virus (CABMV), em plantas de maracujá-azedo (Passiflora edulis Sims f. flavicarpa Deg.). Ápices caulinares, provenientes de plantas infectadas, foram microenxertados em plântulas obtidas pela germinação de sementes em substrato comercial esterilizado. Foram conduzidos experimentos com a microenxertia realizada no hipocótilo e no epicótilo, e foram utilizados cinco meios de cultura, que diferiam na concentração de fitorreguladores, aplicados no local da enxertia. O índice médio de microenxertos com folha expandida foi de 27,22 e 32,22%, quando a microenxertia foi realizada no hipocótilo e no epicótilo, respectivamente. Na microenxertia realizada no hipocótilo, não houve efeito da aplicação de meios de cultura. Na microenxertia realizada no epicótilo, o meio MS acrescido de 0,1 mg L-1 de AIB e 1 mg L-1 de BAP proporcionou 53,3% de microenxertos com folha expandida, número superior aos demais tratamentos e maior desenvolvimento das brotações. A indexação realizada pelo teste ELISA indireto, 80 a 100 dias após a microenxertia, mostrou que 93% das plantas testadas não apresentavam vírus detectável.

Polymers as carriers for rhizobial inoculant formulations

The aim of this work was to evaluate the efficiency of carboxymethyl cellulose (CMC) and starch blends as carrier materials of rhizobial inoculants regarding their capacity to maintain viable cells and promote cowpea (Vigna unguiculata) nodulation. The experimental design adopted was completely randomized, with three replicates. Forty different compositions of carboxymethyl cellulose (CMC) with starch, compatibilized or not with different proportions of MgO or ZnO, were evaluated regarding their ability of maintaining rhizobial viable cells during the storage period of one month at room temperature, in an initial screening. Thereafter, selected inoculant carrier blends were evaluated regarding their ability to maintain viable rhizobial cells for a period of 165 days, and their performance as inoculant carriers was compared to a peat-based inoculant carrier under greenhouse conditions. Rhizobial cells were better maintained in blends containing 50-60% CMC. Compatibilizing agents did not increase survival of rhizobial cells for 30 days of storage. The cowpea nodulation of polymer blends was statistically the same of peat-based inoculants. CMC/starch polymer blends are efficient carriers to rhizobial inoculants for up to 165 days of storage, when compatibilized with MgO (1%).

Production of policlonal antisera specific to plant viruses by rabbit oral immunization

Serological techniques are of great importance for plant virus identification and characterization. The major limiting factor for using these techniques for plant virus identification is the requirement of a good virus purified preparation to be used in immunizing animals for antiserum production. In the present study, two New Zealand rabbits were orally immunized with extracts from cowpea (Vigna unguiculata) plants systemically infected with Cowpea severe mosaic virus (CPSMV) and with extracts from papaya (Carica papaya) infected with Papaya lethal yellowing virus (PLYV). The leaf extracts were prepared in saline solution 0.15 M in the rate of 1:1 (w/v) and clarified by a centrifugation of 10,000 g for 10 min. The clarified extracts containing the viruses were orally administered to the New Zealand rabbits in two series of five daily doses of 1.0 ml each. The obtained policlonal antisera were shown to be very specific to their respective viruses in double immunodiffusion and indirect ELISA. These seem to be the first antisera specific for plant virus obtained by rabbit oral immunization. The results open up some possibilities for producing antisera to plant viruses of difficult purification. It is a simple, fast and inexpensive method for production of antisera for plant viruses when compared to the traditional techniques that involve rabbit injections with purified virus preparations.

Análise molecular de segmento do RNA2 de comovirus isolados de soja no estado do Paraná

Nas áreas produtoras de feijão (Phaseolus vulgaris) do Estado do Paraná observa-se anualmente a ocorrência do vírus do mosaico em desenho do feijoeiro (Bean rugose mosaic virus, BRMV), principalmente em infecções mistas com o vírus do mosaico dourado do feijoeiro (Bean golden mosaic virus, BGMV), acarretando maior severidade de sintomas e causando perdas na produção. Recentemente constatou-se a presença do vírus do mosaico severo do caupi (Cowpea severe mosaic virus, CPSMV) associado a sintomas de queima do broto em plantações de soja (Glycine max) na região de Londrina, sendo este um fato novo no Estado. Neste trabalho, parte do RNA2 de dois comovirus isolados de soja no Paraná foram clonados e sequenciados, sendo 600 pares de bases (pb) do BRMV-PR e 594 pb do CPSMV-PR. Posteriormente, as seqüências correspondentes de aminoácidos foram comparadas com seis seqüências de vírus do gênero Comovirus depositadas no GenBank. Com base nestes dados observou-se que o segmento do RNA2 do isolado CPSMV-PR apresentou homologia de 85% com parte de uma seqüência já conhecida do RNA2 do CPSMV, enquanto que o segmento do RNA2 do isolado BRMV-PR apresentou homologia de 39% com o CPSMV, e de 44% com o Bean pod mottle virus (BPMV). Este trabalho apresenta pela primeira vez dados de sequenciamento parcial do BRMV, o que poderá contribuir para sua completa caracterização molecular e para o estabelecimento de estratégias para obtenção de plantas resistentes ao vírus.

Genotypic diversity among brazilian isolates of Macrophomina phaseolina revealed by RAPD

Macrophomina phaseolina has been considered one of the most prevalent soybean (Glycine max) pathogens in Brazil. No genetic resistance has been determined in soybean and very little is known about the genetic diversity of this pathogen in tropical and sub-tropical regions. Fifty-five isolates from soybean roots were collected in different regions and analyzed through RAPD for genetic diversity. The UPGMA cluster analysis for 74 loci scored permitted identification of three divergent groups with an average similarity of 99%, 92% and 88%, respectively. The three groups corresponded to 5.45%, 59.95% and 34.6%, respectively of all isolates used. A single plant had three different haplotypes, while 10.9% of the analyzed plants had two different haplotypes. In another study the genetic similarity was evaluated among isolates from different hosts [soybean, sorghum (Sorghum bicolor), sunflower (Helianthus annuus), cowpea (Vigna unguiculata), corn (Zea mays) and wheat (Triticum aestivum)] as well as two soil samples from native areas. Results showed that more divergent isolates originated from areas with a single crop. Isolates from areas with crop rotation were less divergent, showing high similarity values and consequently formed the largest group. Amplification of the ITS region using primers ITS1 and ITS4 produced only one DNA fragment of 620 bp. None of the isolates were differentiated through PCR-RFLP. Our results demonstrated genetic variability among Brazilian isolates of M. phaseolina and showed that one single root can harbor more than one haplotype. Moreover, cultivation with crop rotation tends to induce less specialization of the pathogen isolates. Knowledge of this variation may be useful in screening soybean genotypes for resistance to charcoal rot.

Análise filogenética de potyvírus causando endurecimento dos frutos do maracujazeiro no Nordeste do Brasil

Amostras foliares de plantas de maracujá-amarelo (Passiflora edulis f. flavicarpa) com sintomas típicos de endurecimento dos frutos foram coletadas nos estados de Pernambuco, Paraíba e Sergipe. A infecção viral foi comprovada por meio de teste sorológico e gama de hospedeiros. Os seis isolados virais obtidos foram capazes de infetar várias espécies testadas, porém apresentando diferenças na intensidade dos sintomas induzidos nessas hospedeiras. Teste de ELISA indireto demonstrou que os isolados obtidos a partir de plantas de maracujá são sorologicamente relacionados entre si e com o potyvírus Cowpea aphid-borne mosaic virus (CABMV). A seqüência de aminoácidos da proteína capsidial foi determinada para os seis isolados. A comparação dessas seqüências com as de outros potyvírus indicou uma identidade máxima com isolados de CABMV (86 a 94%). A identidade com isolados de Passionfruit woodiness virus (PWV) foi de 68 a 76%. Análise filogenética realizada a partir das seqüências de aminoácidos agrupou os isolados em estudo junto a isolados de CABMV, distante de isolados de PWV. Em conjunto, os resultados indicam que os isolados de maracujá analisados constituem na verdade uma estirpe do CABMV.

Detection and partial characterization of a carlavirus causing stem necrosis of soybean in Brazil

Stunting and stem necrosis were noticed in soybeans (Glycine max) grown in 2000/2001 in West Central Brazil the same condition was also observed in the following year in plantations as far as 2,000 km from the initial area. Based on transmission (mechanical, graft, insect vector), purification and serology, electron microscopy and molecular studies the causal agent was determined to be a whitefly-borne carlavirus which is possibly related to Cowpea mild mottle virus (CpMMV).

Crotalaria paulinea, novo hospedeiro natural do vírus do mosaico severo do caupi

Amostras foliares de Crotalaria paulinea apresentando mosaico foram coletadas em São Luiz, MA, e enviadas ao Laboratório de Virologia Vegetal da UFC. As amostras foram testadas por Elisa indireto, contra anti-soros para Cowpea aphid-borne mosaic virus (CABMV) e Cucumber mosaic virus (CMV) e por dupla difusão em àgar contra anti-soro para Cowpea severe mosaic virus (CPSMV). As amostras reagiram somente com o anti-soro para CPSMV, indicando ser C. paulinea mais um hospedeiro natural do vírus. Extratos das folhas de C. paulinea foram inoculados em plantas de caupi (Vigna unguiculata subsp. unguiculata) mantidas em casa de vegetação. Dez dias após a inoculação, as plantas passaram a exibir sintomas de mosaico e a presença do CPSMV foi confirmada por sorologia. Nos estudos de gama de hospedeiros, envolvendo oito espécies botânicas, o isolado de CPSMV obtido de C. paulinea (CPSMV-Cp) infetou sistemicamente somente cultivares de caupi. Estudos de reações de RT-PCR revelaram a presença de uma banda no gel de agarose de 594 pb para o CPSNV-Cp semelhante às de outros isolados de CPSMV. O CPSMV-Cp foi multiplicado em caupi cv. Pitiúba e purificado por clarificação com n-butanol, precipitação viral com PEG e ultracentrifugação. A preparação purificada apresentou um espectro de absorção ultravioleta típico de núcleoproteína com uma razão A260/A280 de 1,7. Coelho da raça Nova Zelândia Branca imunizado com a preparação viral purificada, produziu anti-soro policlonal reativo com CPSMV em dupla difusão em àgar. Este é o primeiro relato sobre a infecção natural de CPSMV em C. paulinea.

Phytosociology in agricultural areas submitted to distinct wintercropping management

Studies related to weed dynamics are essential for agricultural sustainability in tropical soils, as their interference can cause significant yield losses, especially in crops with lower competitive ability. This study aimed to assess the composition of weed communities in the third cropping season in areas submitted to distinct wintercroppings for two consecutive years. Evaluations were made in terms of species composition, level of infestation, and severity of occurrence of each weed species in relation to the others within the same area. The wintercropping management systems were: (1) Brachiaria ruziziensis; (2) corn intercropped with B. ruziziensis; (3) corn and (4) cowpea. Plant communities under these wintercrop systems were evaluated in the third year in terms of relative frequency, relative dominance, relative abundance and importance index for each species. Areas were also compared in terms of species composition by the Sorensen's similarity coefficient. Two years of distinct wintercrop managements were not enough to cause a significant shift in terms of weed composition and severity in a given area. There is also evidence that some weed species are more favored by specific crops, although the causes of this selective behavior are not fully explored in this article. The presence of B. ruziziensis, either alone or intercropped with corn, resulted in lower similarity coefficients when compared to areas where this species was not present. It is suggested that several mechanisms briefly discussed may be involved in the impact of the presence of B. ruziziensis on the weed community.

Occurrence of weed species in Jatropha curcas intercropping systems

This study aimed to evaluate the level of infestation by weed species in a consolidated Jatropha plantation, as a function of the plant species grown in interrows. The experiment was installed in 2006 at the district of Itahum, city of Dourados, state of Mato Grosso do Sul, Brazil, made possible through a partnership between Embrapa Western Agriculture and Paraiso Farm. Treatments consisted of (1) Jatropha in monocrop, with no plant at the interrrows; or the following plants cultivaded at the interrows of Jatropha: (2) Stylosanthes spp.; (3) Brachiaria ruziziensis; (4) Brachiaria ruziziensis + Stylosanthess pp.; (5) Brachiaria humidicola; (6) Panicum maximum cv. massai; (7) Cajanus cajan cv. anão; (8) Crotalaria spectabilis; (9) Crop rotation system 1 - (maize second crop -Crambe abyssinica - soybean - peanut); and (10) Crop rotation system 2 - (cowpea - radish - maize - cowpea) conducted for two years. Phytosociological characterization of weed species was accomplished in 2011 based on the Ecological Approach. Estimations of relative abundance, frequency, dominance and Importance Value Index were obtained. Areas were also characterized by the diversity coefficients of Simpson and modified Shannon-Weiner, and then grouped by cluster analysis. Areas with low soil coverage resulted in higher infestation levels; crop rotation in the interrows of Jatropha produced a significant reduction in weed infestation, but the lowest infestation levels were observed when grasses were grown intercropped with Jatropha. Lower diversity coefficients were associated with occurrence of the most troublesome weed species. In the first years after planting Jatropha, species of Brachiaria or a crop rotation involving species with high mulching ability and whose biomass exhibit a high C:N ratio, should be established in the interrows to avoid problems with weeds infestation.