Genetic variation within and among species of five sections of the genus Arachis L. (Leguminosae) using RAPDs

Some Arachis species are widely used as commercial plants, e.g. the groundnut A. hypogaea, an important source of good quality protein and oil, and A. pintoi and A. glabrata, that are utilized as forage species. Germplasm of most Arachis species is available in germplasm banks. However, little it is known about the genetic attributes of this germplasm, and mainly about its genetic variability, which is very important for its maintenance. In the present study RAPDs were used to assay the genetic variation within and among 48 accessions of five sections of the genus Arachis and to establish the genetic relationships among these accessions. Ten of 34 primers tested were selected for DNA amplification reactions since they yielded the largest numbers of polymorphic loci. A dendrogram was constructed based on data from the 10 primers selected. Eighty RAPD polymorphic bands were analyzed among the accessions studied. The relationships among species based on RAPDs were similar to those previously reported based on morphological, cytological and crossability data; demonstrating that RAPDs can be used to determine the genetic relationships among species of the different sections of the genus Arachis. In general, wide variation was found among accessions and low variation was found within the accessions that had two or more plants analyzed. However, higher polymorphism was found in the section Trierectoides and in one accession of A. major, indicating that generalizations should be avoided and each species should be analyzed in order to establish collection and maintenance strategies.

Genetic relationships among Arachis hypogaea L. (AABB) and diploid Arachis species with AA and BB genomes

The cultivated peanut (Arachis hypogaea L.) is an allotetraploid, with two types of genomes, classified as AA and BB, according to cytogenetic characters. Similar genomes to those of A. hypogaea are found in the wild diploid species of section Arachis, which is one of the nine Arachis sections. The wild species have resistances to pests and diseases that affect the cultivated peanut and are a potential source of genes to increase the resistance levels in peanut. The aim of this study was to analyze the genetic variability within AA and BB genome species and to evaluate how they are related to each other and to A. hypogaea, using RAPD markers. Eighty-seven polymorphic bands amplified by ten 10-mer primers were analyzed. The species were divided into two major groups, and the AA and the BB genome species were, in general, separated from each other. The results showed that high variation is available within species that have genomes similar to the AA and the BB genomes of A. hypogaea.

Genetic variation within and among species of genus Arachis, section Rhizomatosae

The genus Arachis is endemic to South America and comprises 80 species, 69 of which have already been described and eleven not yet published. The genus includes the cultivated peanut ( A. hypogaea) and several forage species, the most important ones being A. glabrata and A. pintoi. Accessions of section Rhizomatosae, including three tetraploid species 2n = 4x = 40 (A. glabrata, A. pseudovillosa and A. nitida nom. nud.) and one diploid species 2n = 2x = 20 (A. burkartii), were evaluated using RAPD markers to assay genetic variability within and among species. The ten random primers used yielded a total of 113 polymorphic bands. The data were scored as the presence or absence of each band in each sample. A distance matrix and dendrogram were obtained using Link's coefficient and the neighbor-joining method. Most accessions analyzed grouped into two major clusters: the first comprised most accessions of A. glabrata and accessions of A. nitida, and the second cluster comprised accessions of A. burkartii. Arachis pseudovillosa and a few accessions of A. glabrata and A. nitida were placed between these major clusters. The diploid and tetraploid species were grouped quite separately, suggesting that the tetraploids did not originate from the diploid species analyzed.

Genetic variation between several species of sections Extranervosae, Caulorrhizae, Heteranthae, and Triseminatae (genus Arachis) estimated by DNA polymorphism

Genetic variation within and among accessions of the genus Arachis representing sections Extranervosae, Caulorrhizae, Heteranthae, and Triseminatae was evaluated using RFLP and RAPD markers. RAPD markers revealed a higher level of genetic diversity than did RFLP markers, both within and among the species evaluated. Phenograms based on various band-matching algorithms revealed three major clusters of similarity among the sections evaluated. The first group included the species from section Extranervosae, the second group consisted of sections Triseminatae, Caulorrhizae, and Heteranthae, and the third group consisted of one accession of Arachis hypogaea, which had been included as a representative of section Arachis. The phenograms obtained from the RAPD and RFLP data were similar but not identical. Arachis pietrarellii, assayed only by RAPD, showed a high degree of genetic similarity with Arachis villosulicarpa. This observation supported the hypothesis that these two species are closely related. It was also shown that accession V 7786, previously considered to be Arachis sp. aff. pietrarellii, and assayed using both RFLPs and RAPDs, was possibly a new species from section Extranervosae, but very distinct from A. pietrarellii.

Resistência de genótipos de amendoim (Arachis hypogaea L.) de hábitos de crescimento ereto e rasteiro a Anticarsia gemmatalis Hübner, 1818 (Lepidoptera: Noctuidae)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Resistência de germoplasma silvestre de amendoim (arachis spp.) a Enneothrips flavens Moulton, 1941 (Thysanoptera: Thripidae) e Stegasta bosquella (Chambers, 1875) (Lepidoptera: Gelechiidae)

Coordenação de Aperfeiçoamento de Pessoal de Nível Superior (CAPES)

Seletividade do imazapic para dois cultivares de amendoim (Arachis hypogaea) cultivados na ausência e na presença de palha de cana-de-açúcar

Neste trabalho, objetivou-se estudar a seletividade de imazapic a dois cultivares de amendoim (Tatu Vermelho e IAC-5), em condições de ausência e de presença de palha de cana-de-açúcar. O experimento foi instalado em vasos, em delineamento inteiramente casualizado, com os tratamentos dispostos em esquema fatorial 3x2, com quatro repetições. Constituíram os fatores principais três doses de imazapic (0, 98 e 140 g ha-1), aplicadas em pré-emergência sobre duas condições de palha de cana-de-açúcar (0 e 15 t ha-1). Constatou-se que a camada de palha possui potencial em reter o herbicida, especialmente sob períodos de pouca precipitação ou irrigação; as plantas de amendoim sob esta condição apresentaram menos sintomas de intoxicação. Entretanto, na ausência da palha, as plantas, independentemente do cultivar, apresentaram maiores injúrias visuais e redução na altura, massa seca da parte aérea e raiz.

EFFECTS of MINERAL NUTRITION on INTER- and INTRASPECIFIC INTERFERENCE of PEANUT (Arachis hypogaea L.) and HAIRY BEGGARTICKS (Bidens pilosa L.)

The present work was carried out to study the effects of mineral nutrition on peanut (Arachis hypogaea L) cv. IAC Runner-886 and hairy beggarticks (Bidens pilosa L) growth, when submitted to inter- and intraspecific competition. The treatments consisted of two peanut plants per pot, two hairy beggarticks per pot and one plant of each species per pot. The plants were nourished with Hoagland and Arnon (1950) complete solution, or without potassium, or without phosphorus or without nitrogen. Sixty days after planting, no inter- or intraspecific competition effect on growth characteristics of peanut was verified and nutrition was not a limiting factor to the culture. No interaction between competition and nutrition effects was observed for both species. The weed suffered more negative effects from intraspecific competition and nutrition. The absence of N had a pronounced effect compared to the other elements, resulting in a reduction in all the evaluated characteristics. The deficiency of nutrients and competition affected the weed more than the crop, showing that peanut was more competitive than hairy beggarticks.

Análise de crescimento em amendoinzeiro (Arachis hypogaea L.) submetido à matocompetição

Foram verificados os efeitos de diferentes períodos de controle e convivência de plantas daninhas (0; 14; 28; 42; 56; 70; 84 e 98 dias) na cultura do amendoinzeiro (Arachis hypogaea L.) em Moji-Guaçu (SP). As principais espécies invasoras da área experimental foram: Acanthospermum australe (Loef.) O. Kuntz, Waltheria indica L., Borreria alata DC., Richardia scabra L., Rhynchelytrum repens (Nees) Stapf. et Hubb. e Diodia teres Walt. de maneira geral, durante quase todo o ciclo, a espécie de maior importância relativa foi A. australe, sendo suplantada por outras espécies apenas na colheita, quando já estava encerrando seu ciclo de desenvolvimento. O período inicial de controle das plantas daninhas, além de reduzir a densidade da comunidade infestante, diminuiu sua riqueza em espécies e alterou a importância relativa das populações. A matocompetição resultou na redução do crescimento do amendoinzeiro, principalmente a partir de 42 dias de convivência, afetando a altura das plantas, a área foliar, o índice de área foliar e o acúmulo epígeo de matéria seca. Não foram afetados os números de ramos e de folíolos por planta, nem a qualidade dos grãos e o vigor das sementes do amendoim. A produção de frutos foi reduzida ao redor de 31,5% em decorrência da interferência das plantas daninhas. O período total de prevenção da interferência na produção foi de 14 dias e o período anterior à interferência foi de 70 dias.

Inhibition of the myotoxic activity of Bothrops jararacussu venom and its two major myotoxins, BthTX-I and BthTX-II, by the aqueous extract of Tabernaemontana catharinensis A. DC. (Apocynaceae)

Partial neutralization of the myotoxic effect of Bothrops jararacussu venom (BV) and two of its myotoxins [bothropstoxin-I (BthTX-I), catalytically inactive, and II (BthTX-II), showing low PLA(2) activity], by the lyophilized aqueous extract of Tabernaemontana catharinensis (AE), was studied in rat isolated soleus muscle preparations (in vitro) and through i.m. injection in the gastrocnemius muscle (in vivo) by determination of creatine kinase (CK) activity and histopathological analysis. Incubation of soleus muscle for 1 h with BV or toxins (20 mug/ml) plus AE (400 mug/ml) added immediately after BV, BthTX-I or BthTX-II reduced CK levels by 53%, 37% and 56%, respectively. The myonecrotic effects of BV (20 mug/ml) upon soleus muscle was reduced 24%, 35% and 36% when AE (400 mug/ml) was added 1 h after BV and CK was evaluated 30 min, 1 and 2 h later, respectively. For BthTX-I these values were 46%, 48% and 47%, while for BthTX-II no inhibitory effect was detected. Histological analysis of soleus muscle after incubation with AE (400 mug/ml, I h) did not reveal any change in muscle fibers, but severe necrosis induced by -BV or toxins (20 mug/ml) was clearly in evidence, and decreased significantly when soleus muscle was protected by AE. This protection was also observed when AE was administered 1 h after BV or BthTX-I, but not after BthTX-II. AE did not inhibit the catalytic PLA(2), activity of BthTX-II or BV and did not change the PAGE pattern of BV, BthTX-I or BthTX-II. In vivo assays were performed in 100-g rats and maximal CK release was attained at a dose of 100 mug of BV, 3 h after injection. AE was not effective when injected 20 s after BV or toxins. However, injecting BV or toxins (100 mug), which were pre-incubated with AE (2 mg) caused an inhibition of 57%, 59% and 51%, respectively, with zero time pre-incubation, but was less effective with I h pre-incubation. This plant represents a potential source of promising myotoxin inhibitors. (C) 2004 Elsevier GmbH. All rights reserved.

Reação de plantas de amendoim (Arachis hypogaea L. ) ao trifluralin e mudanças na morfologia da parte subterrânea

Com o objetivo de reproduzir sintomas de malformação de raízes, observados em campo, foram estuda dos os efeitos do trifluralin sobre Arachis hypogaea L., cultivar Tatu, em condições de vaso, em Jaboticabal, SP. Quatro formulações de trifluralin foram aplicadas em solo Latossol Roxo - série Jaboticabal, barro argiloso, com 2,7% de matéria orgânica, em dosagem correspondentes a 1,5; 3,0 e 6,0 litros da formulação comercial por ha. Cada vaso contendo 2,5 1 -de solo, recebeu duas sementes pré -germinada s e foi mantido sob condições de casa-de-vegetação durante 29 dias. Foi adotado o delineamento inteiramente casualizado com treze tratamentos e quatro repetições, perfazendo total de 52 parcelas. Aos 29 dias as plantas, foram retiradas e determinadas as seguintes características: peso da matéria seca das raízes, dos caules, dos pecíolos e dos folíolos; número de folíolos; área foliar e comprimento da haste principal. Alterações morfológicas externas e internas foram observadas. Os resultados mostraram que as partes subter rân eas das plantas foram afetadas, com uma correlação linear negativa e significativa entre o aumento da dose dos herbicidas e peso da matéria. As folhas se mostraram pouco sensíveis, sendo que nenhuma das formulações diminuiu o número de foliolos da planta. A área foliar e o peso da matéria seca dos folíolos somente foram afetados pelo aumento da dose de T. Fecotrigo com va lores de F iguais a 9,62* e 7,61*, respectivamente. de modo geral,po de-se concluir que o maior efeito ocorre devido ao incremento nas doses e não devido a possiveis diferenças entre os tipos de formula ções testadas. Quanto a alterações morfológicas observou-se que nas doses mais elevadas houve grande diminuição na quantidade de raízes, com raízes secundárias curtas e grossas. O hipocótilo emitiu raízes até a altura do nó cotiledonar. Na dose de 3,0 1/ha já foram notadas raízes secundárias um pouco mais espessas que na dose de 1,5 1/ha. As lâminas histológicas de amostras da re gião do hipocótilo mostraram parên quima cortical bastante espesso, de vido ao maior numero de camadas de células, porém não se notou anormalidade na forma das células, nem nos demais tecidos da região estudada.

Behavior of Leishmania major metacyclic promastigotes during the course of infection and immune response development in resistant versus susceptible hosts

Pouco se conhece sobre os epítopos derivados de promastigotas metacíclicos de Leishmania que são importantes para a regulação ou destruição do parasita, como alvos de ação imunológica no hospedeiro vertebrado. Neste estudo, nós investigamos um método alternativo para obter promastigotas metacíclicos de Leishmania major, pela avaliação do curso da infecção e reação de hipersensibilidade do tipo retardado (HTR) em hospedeiros resistentes e susceptíveis. Promastigotas não-infectantes (procíclicos) de L. major, recentemente isolados de amastigotas, foram selecionados pela adesão a colunas de lã de vidro negativamente carregadas, enquanto que promastigotas metacíclicos não se aderem à coluna e podem ser recuperados com facilidade. Condições ótimas de cromatografia foram validadas por análise estatística. O rendimento médio de parasitas obtidos após separação em colunas de lã de vidro e a viabilidade dos promastigotas foram estimados por microscopia óptica. Os promastigotas metacíclicos tiveram um rendimento médio de 43,5% a 57,5%. Camundongos BALB/c (susceptíveis) e camundongos C57BL/6 (resistentes) apresentaram padrões distintos de lesões cutâneas, os primeiros com lesões mais agressivas, induzidas por promastigotas metacíclicos. As respostas à reação de HTR foram maiores nos grupos de camundongos C57BL/6, submetidos à infecção com promastigotas metacíclicos. Estes resultados indicam que o novo método poderia ser integrado aos protocolos existentes para estudar a metaciclogênese de parasitas do gênero Leishmania in vivo.

Differential antibody isotype expression to the major Paracoccidioides brasiliensis antigen in juvenile and adult form paracoccidioidomycosis

We investigated the relationship between antibody response to the major Paracoccidioides brasiliensis antigen, a 43-kDa glycoprotein, and the two paracoccidioidomycosis (PCM) clinical presentations, the juvenile and the adult forms. Total immunoglobulin G (IgG), IgG isotypes, and IgA anti-gp43 antibodies were determined by enzyme-linked immunosorbent assay in patients' sera. Juvenile PCM patients had higher (P =.003) IgG anti-gp43 levels than adult form patients. IgG1 subclass levels, however, were comparable between the two clinical forms. Patients with the juvenile form had higher (P <.001) IgG4, but lower(P =.03) IgG2 levels than patients with the adult form. The IgG4 isotype, regulated by interleukin 4, was found in all juvenile form patients but in only 12% of the adult form patients. In contrast, high levels of the IgG2 isotype, regulated by interferon-gamma, were found in 41% of the adult PCM patients, mainly those with a more benign disease, but in only 12% of the juvenile patients. IgG3 was either absent or detected at low levels. These results demonstrate, for the first time, specific IgG4 antibodies in the humoral immune response of patients with an endemic deep mycosis and suggest that the switch to the IgG subclasses in PCM is regulated by the patients' T-helper subset (Th-l or Th-2) dominant cytokine profile. A possible role for IgG4 in the immunopathogenesis of the juvenile, more severe form of the disease is discussed. Finally, IgA was found mainly in adult form patients, probably as a result of the chronic mucosal antigenic stimulation characteristic of this form. (C) Elsevier, Paris.

Major carotenoid composition of Brazilian Valencia orange juice: Identification and quantification by HPLC

The carotenoid composition of Brazilian Valencia orange juice was determined by open column chromatography (OCC) and high-performance liquid chromatography. Carotenoid pigments were extracted using acetone and saponified using 10% methanolic potassium hydroxide. Sixteen pigments were isolated by OCC and identified as alpha-carotene, zeta-carotene, beta-carotene, alpha-cryptoxanthin, beta-cryptoxanthin, lutein-5,6-epoxide, violaxanthin, lutein, antheraxanthin, zeaxanthin, luteoxanthin A, luteoxanthin B, mutatoxanthin A, mutatoxanthin B, auroxanthin B and trollichrome B. Thirteen carotenoid pigments were separated using a ternary gradient (acetonitrile-methanol-ethyl acetate) elution on a C-18 reversed-phase column. Among these, violaxanthin, lutein, zeaxanthin, beta-cryptoxanthin, zeta-carotene, alpha-carotene, and beta-carotene were quantified. The total carotenoid content was 12 +/- 6.7 mg/1, and the major carotenoids were lutein (23%), beta-cryptoxanthin (21%), and zeaxanthin (20%). 2005 Elsevier Ltd. All rights reserved.

Effect of chickpea (Cicer arietinum L.) germination on the major globulin content and in vitro digestibility

A germinação das sementes de grão-de-bico foi acompanhada por um período de 6 dias, no qual pequenas variações nos teores de nitrogênio e globulina total foram registradas. A globulina majoritária (tipo 11 S) apresentou maiores variações após o quarto dia de germinação. A natureza e distribuição da fração globulina majoritária isolada na cromatografia em Sepharose CL-6B mostrou pequenas modificações ao final do período de germinação. A eletroforese em gel de poliacrilamida com dodecilssulfato de sódio do pico eluído na cromatografia em Sepharose CL-6B demonstra modificações nas bandas de proteínas entre os pesos moleculares de 20 e 30 kDa e acima de 60 kDa, indicando degradação protéica durante o período. Atividade proteolítica foi detectada na fração albumina da semente que aumentou até o quarto dia, seguido de queda até o sexto dia de germinação, quando da utilização de globulina total isolada da semente e caseína como substratos. Farinha de grão-de-bico, frações albumina e globulina total isoladas não apresentaram aumento na digestibilidade in vitro; entretanto, a fração globulina majoritária isolada foi mais suscetível à hidrólise após germinação.