Removal of the N-linked glycan structure from the peanut peroxidase prxPNC2: Influence on protein stability and activity

Lines of transgenic tobacco have been generated that are transformed with either the wild-type peanut peroxidase prxPNC2 cDNA, driven by the CaMV3 5S promoter (designated 35S::prxPNC2-WT) or a mutated PNC2 cDNA in which the asparagine residue (Asn(189)) associated with the point of glycan attachment (Asn(189)) has been replaced with alanine (designated 35S::prxPNC2-M). PCR, using genomic DNA as template, has confirmed the integration of the 35S::prxPNC2-WT and 35::prxPNC2-M constructs into the tobacco genome, and western analysis using anti-PNC2 antibodies has revealed that the prxPNC2-WT protein product (PNC2-WT) accumulates with a molecular mass of 34,670 Da, while the prxPNC2-M protein product (PNC2-M) accumulates with a molecular mass of 32,600 Da. Activity assays have shown that both PNC2-WT and PNC2-M proteins accumulate preferentially in the ionically-bound cell wall fraction, with a significantly higher relative accumulation of the PNC2-WT isoenzyme in the ionically-bound fraction when compared with the PNC2-M isoform. Kinetic analysis of the partially purified PNC2-WT isozyme revealed an affinity constant (apparent K-m) of 11.2 mM for the reductor substrate guaiacol and 1.29 mM for H2O2, while values of 11.9 mM and 1.12 mM were determined for the PNC2-M isozyme. A higher Arrenhius activation energy (E,,) was determined for the PNC2-M isozyme (22.9 kJ mol(-1)), when compared with the PNC2-WT isozyme (17.6 kJ mol(-1)), and enzyme assays have determined that the absence of the glycan influences the thermostability of the PNC2-M isozyme. These results are discussed with respect to the proposed roles of N-linked glycans attached to plant peroxidases. (c) 2005 Elsevier Ltd. All rights reserved.

HERITABILITY FOR RESISTANCE TO ROSETTE DISEASE IN EXOTIC VALENCIA GROUNDNUTS

Groundnut rosette disease (GRD) is the most destructive virus disease of Valencia groundnuts ( Arachis hypogaea L.) in sub-Saharan Africa. Cultural, biological and chemical control measures have received limited success due to small scale farmers’ inability to use them. Use of host plant resistance provides the most effective and economically viable management option for the resource poor farmers. This study was conducted to determine heritability for resistance to GRD in Valencia groundnuts. Six crosses; Valencia C (P1) × ICGV-SM 90704 (P2), Valencia C (P1) × ICGV-SM 96801(P2), Valencia C (P1) × ICGV-SM 99566 (P2), NuMex-M3 (P1) × ICGV-SM 90704 (P2), NuMex-M3 × ICGV-SM 96801 (P2), and NuMex-M3 (P1) × ICGV-SM 99566 (P2), were made to generate F1, F2, BC1P1 and BC1P2 populations. Data on GRD severity were collected on a 1-9 score scale. Genetic Advance as a percentage of the mean (GAM) and heritability were estimated using variance components. Phenotypic Coefficient of Variation (PCV) and Genotypic Coefficient of Variation (GCV) estimates were high (20.04-70.1%) in the six crosses, except for Valencia C × ICGV-SM 96801(18.1%) and NuMex-M3 × ICGV-SM 96801(17.1%), which exhibited moderate GCV values. Broad and narrow sense heritability estimates for GRD disease score ranged from 64.1 to 73.7% and 31 to 41.9%, respectively, in all the crosses. GAM was high in all the crosses (21-50.7%), except for Valencia C x ICGV-SM 96801 (14.67), M3 x ICGV-SM 99566 (18%) and NuMex-M3 x ICGV-SM 96801 (13.5%) crosses that exhibited moderate GAM. The study revealed the presence of variability of GRD resistance, implying that genetic improvement of these exotic materials is possible.

Efeito sacietogênico de um novo inibidor de tripsina da paçoca do amendoim com aumento plasmático de colecistocinina (cck)

Obesity is increasing, reaching epidemic levels in many regions of the world. Studies have shown that consumption of peanuts influences on weight control and this influence may be due to the action of trypsin inhibitors sacietogênica that condition increased plasma colescistocinina (CCK). Moreover, the peanut has other health benefits, and these assignments are guaranteed to increase their production and consumption of several of its products, including the paçoca peanut. The aim of this study was to identify the presence of a trypsin inhibitor in paçoca peanut and evaluate its effect on food intake, weight gain and histomorphological changes in swiss mice (n = 8) and Wistar rats (n = 6). Experimental diets were prepared based on the AIN-93G and supplemented with tack or peanut trypsin inhibitor partially purified paçoca peanut (AHTI). After each treatment, the animals were anesthetized and euthanized, their bloods were collected by cardiac puncture for the determination of CCK and other biochemical parameters (glucose, triglycerides, total cholesterol, high density lipoprotein, low density lipoprotein, glutamic-pyruvic transaminase, glutamic oxaloacetic transaminase and albumin) and their pancreas removed for histologic and morphometric analysis. The supplementation with paçoca peanut and the AHTI showed a decrease of body weight gain and food intake in both mice and rats, due to the satiety, since the animals showed no evidence of impairment of nutritional status conditioned by consumption the AHTI. There were also observed biochemical or morphological important when compared with controls. However, AHTI led to increased secretion of CCK, a peptide sacietogênico. Thus, these results indicate that AHTI present in paçoca peanut, is able to enhance the secretion of plasma CCK and thereby reduce the weight gain associated with lower food intake of experimenta animals

Características agronômicas e produtividade de amemdoim em diferentes formas de cultivo no Recôncavo Baiano.

O presente trabalho foi desenvolvido com o objetivo de avaliar as características agronômicas e a produtividade do amendoim, cultivar Havana, em cultivo solteiro e consorciado com pinhão manso e girassol, na região do Recôncavo Baiano.

Annual crop rotation of tropical pastures with no-till soil as affectedby lime surface application.

Soil acidity and low natural fertility are the main limiting factors for grain production in tropical regionssuch as the Brazilian Cerrado. The application of lime to the surface of no-till soil can improve plant nutrition, dry matter production, crop yields and revenue. The present study, conducted at the Lageado Experimental Farm in Botucatu, State of São Paulo, Brazil, is part of an ongoing research project initi-ated in 2002 to evaluate the long-term effects of the surface application of lime on the soil?s chemical attributes, nutrition and kernel/grain yield of peanut (Arachis hypogaea), white oat (Avena sativa L.) and maize (Zea mays L.) inter cropped with palisade grass (Urochloa brizantha cv. Marandu), as well as the forage dry matter yield of palisade grass in winter/spring, its crude protein concentration, estimated meat production, and revenue in a tropical region with a dry winter during four growing seasons. The experiment was designed in randomized blocks with four replications. The treatments consisted of four rates of lime application (0, 1000, 2000 and 4000 kg ha−1), performed in November 2004. The surface application of limestone to the studied tropical no-till soil was efficient in reducing soil acidity from the surface down to a depth of 0.60 m and resulted in greater availability of P and K at the soil surface. Ca and Mg availability in the soil also increased with the lime application rate, up to a depth of 0.60 m. Nutrient absorption was enhanced with liming, especially regarding the nutrient uptake of K, Ca and Mg by plants.Significant increases in the yield components and kernel/grain yields of peanut, white oat and maize were obtained through the surface application of limestone. The lime rates estimated to achieve the maximum grain yield, especially in white oat and maize, were very close to the rates necessary to increase the base saturation of a soil sample collected at a depth of 0?0.20 m to 70%, indicating that the surface liming of 2000 kg ha−1is effective for the studied tropical no-till soil. This lime rate also increases the forage dry matter yield, crude protein concentration and estimated meat production during winter/spring in the maize-palisade grass inter cropping, provides the highest total and mean net profit during the four growing seasons, and can improve the long-term sustainability of tropical agriculture in the Brazilian Cerrado.

Reação de etnovariedades de amendoim à mancha preta (Passalora personata) em Rio Branco, Acre.

O amendoim (Arachis hypogaea L.) é importante fonte de proteína para várias aldeias indígenas na Amazônia. Esta pesquisa teve como objetivo avaliar em campo a reação de cinco variedades de amendoim coloridos cultivados na Terra Indígena Kaxinawa de Nova Olinda, situada no rio Envira, Jordão, AC em relação à mancha preta causada por Passalora personata.

Nodulation, gas exchanges and production of peanut cultivated with Bradyrhizobium in soils with different textures.

Nitrogen fertilization from biological source is an uncommon practice for peanut growers due to the limited results, mainly in environments with water restriction. In this study, the response of a commercial Bradyrhizobium was evaluated on the nodulation and production of peanuts grown in sandy and medium textured soils. Two experiments using different soils were carried out in the field during the dry season, in Campina Grande, Paraíba State, Brazil. Three peanut genotypes were submitted to the following treatments: 1-no nitrogen fertilization (control), 2- chemical fertilization (ammonium sulfate) and 3- inoculation with Bradyrhizobium [commercial strain BR 1405 (SEMIA 6144)]. A completely randomized 3x3 factorial design was adopted with five repetitions for both experiments. The evaluates variables were: height of the main stem, number of nodes/plant, root length, root dry weight, weight of pods/plant and number of pods/plant. In addition, gas exchanges were estimated using IRGA apparatus. Both genotypes (BRS Havana and L7 Bege) were benefited in relation to production due to an inoculation with SEMIA 6144. No physiological response was verified in genotypes or N-treatments to gas exchange, excepting for the Ci/Ca ratio in the medium textured soil experiment. BRS Havana showed low Ci/Ca ratio in Bradyrhizobium treatment, indicating that SEMIA 6144 improved the plants photosynthetic efficiency.

Study of the genetic variability and similarity among and within Arachis villosulicarpa, A-pietrarellii and A-hypogaea through isoenzyme analysis

Arachis villosulicarpa is a perennial species cultivated for its soft and tasty seeds by indigenous inhabitants of the Mate Grosso State, Brazil. Besides A. hypogaea, this species is considered as the only species of Arachis which represents a valuable food source for human consumption. Due to the lack of knowledge concerning the genetic diversity of A. villosulicarpa, this study was conducted to evaluate the genetic variability of the accessions from the Germplasm Collection of CENARGEN/EMBRAPA (Brasilia, DF, Brazil) and Institute Agronomico (IAC, Campinas, SP, Brazil). In addition, the genetic similarity between A. villosulicarpa, the related wild species A. pietrarellii, and the cultivated peanut A. hypogaea cv. Tatu was evaluated. From the entire sample analyzed of A. villosulicarpa, the accession from Institute Agronomico showed the highest indices of diversity for both enzymatic systems analyzed, pointing this accession as a promising source of genetic variability that must be preserved in the Germplasm Bank. A high level of genetic similarity was observed between A. pietrarellii and A. villosulicarpa, supporting previous suggestions that A. pietrarellii could be the ancestral progenitor species of A. villosulicarpa or that both species originated from a common ancestor.

Genetic relationships among Arachis species based on AFLP

Amplified Fragment Length Polymorphism (AFLP) was used to establish the genetic relationships among 20 species from seven of the nine sections of genus Arachis. The level of polymorphism among nine accessions of the cultivated peanut, A. hypogaea L., was also evaluated. Three combinations of primers were used to amplify the AFLPs. The fragments were separated in 6% denaturing acrylamide gels. A total of 408 fragments were analyzed. An average of 135.3 fragments per primer combination were scored, and the largest number of fragments was 169 using primer combination Eco RI - ACC / Mse I - CTG, while the lowest was 108, with Eco RI - ACT / Mse I - CTT. In general, the genetic relationships established using AFLPs agreed with the classification established using morphology and crossability data. The results indicated that AFLPs are good markers for establishing the relationships among Arachis species. The polymorphism detected in A. hypogaea by this method was higher than the one found with other markers, like RAPDs and RFLPs. However, our data suggest that the polymorphism detected be using AFLP with only three primer combinations is still too low to be used for any kind of genetic study in this species.

RFLP analysis of genetic variation in species of section Arachis, genus Arachis (Leguminosae)

Four A-genome species of the genus Arachis ( A. cardenasii, A. correntina, A. duranensis, A. kempff-mercadoi), three B genomes species ( A. batizocoi, A. ipaensis and A. magna), the AABB allotetraploid A. hypogaea (cultivated peanut) and introgression lines resulting from a cross between A. hypogaea and A. cardenasii were analyzed by RFLP. The A genome species (cytologically characterized by the presence of a small chromosome pair 'A') were closely similar to each other and shared a large number of restriction fragments. In contrast, the B genome species differed more from one another and shared few fragments. The results of this study indicate that the absence of the small chromosome pair is not a good criterion for grouping species of section Arachis as B genome species, since their genome might be quite distinct from the B genome of A. hypogaea. The lowest genetic variation was detected within accessions of A. duranensis (17 accessions), followed by A. batizocoi (4 accessions) and A. cardenasii (9 plants of accession GKP 10017). The high level of genetic variation found in A. cardenasii might indicate that not all accessions of wild species of Arachis are autogamous, as reported for 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.

Occurrence of Enneothrips flavens moulton and Stegasta bosquella (chambers) and its effects on agronomic traits of wild Arachis accessions

Some wild species of the genus Arachis have demonstrated potential for improvement of peanuts. This work was performed to evaluate the occurrence and symptoms of Enneothrips flavens and Stegasta bosquella and its effects on agronomic traits of wild Arachis accessions. Nine accessions of wild Arachis species and a commercial A. hypogaea variety were studied in a split plot statistical scheme with a completely randomized block design and four replications. The main plots consisted of plants sprayed or not sprayed for insect control, while the subplots comprised the peanut accessions. Accessions GKP10017 (A. cardenasii) and V7639 (A. kuhlmannii) showed the lowest percentages of leaflets with E. flavens and S. bosquella. Accessions V9912, V7639 and V8979 (all three A. kuhlmannii) and V13250 (A. kempff-mercadoi) showed the lowest responses to insecticide application among the various plant traits evaluated. These accessions are of interest for further studies to identify mechanisms of resistance, to be used in breeding programs for resistance to these insects.

Caracteres estruturais e dimensão fractal da folha de espécies de Arachis e sua relação com a cercosporiose

Conselho Nacional de Desenvolvimento Científico e Tecnológico (CNPq)

Acumulação de macronutrientes pelo amendoim(Arachis hypogea L.) cultivado no outono em época da seca

Plantas de amendoim da variedade Tatu (tipo vegetativo Valencia), cultivadas em um latossol-vermelho amarelo,foram colhidas em varios estágios de desenvolvimento. Sobre estas, foram determinadas concentrações e quantidades dos macronutrientes acumulados. Verificou-se que o nitrogênio, o potássio e o cálcio foram os nutrientes que mais se acumularam na planta. Quanto ao potássio, observou-se que este nutriente foi absorvido acen tuadamente apos o florescimento. Constatou-se um decréscimo na quantidade de enxofre na parte aerea apos o florescimento. As quantidades de nutrientes extraidos, em kg/ha, em base a uma população de 160.000 plantas/ha, foram, através das vagens (sementes + cascas): 142 kg N, 15 kg P, 30 kg K, 5 kg Ca, 10 kg Mg e 7,5 kg S e através da parte aerea ou vegetativa: 201 kg N, 16 kg P, 140 kg K, 113 kg Ca, 20 kg Mg e 16 kg S.

Peanut (Araohis hypogaea cv. Tatu-53) productivity under nutrient foliar sprays containing growth regulators

Two nutrient foliar sprays, namely Ferti-Foliage (21-21 -21) and Wuxal (9-9-7), were applied to peanut plants under field conditions. Both were applied 23 days after germination of seeds, at the beginning of flowering, and during flowering. Other treatments were application of NPK fertilizer (9-30-16, 250 kg/ha) into the soil and check (no fertilizer). The experiment was carried out on a latosolic B "Terra Roxa" soil, sowing being made on March 6th and harvest on July 10th. Statistical analysis showed no significant differences amongst treatments. However, certain treatments had better yields. For instance, application of Ferti-Foliage showed a tendency to increasing number of pods per plant and number of seeds per pod. Same product when applied at the beginning of flowering had a tendency to increase production of seeds and of forage. Application of NPK (9-30-16) into the soil showed similar results.