Identification of snails within the Bulinus africanus group from East Africa by multiplex SNaPshotTManalysis of single nucleotide polymorphisms within the cytochrome oxidase subunit I

Identification of populations of Bulinus nasutus and B. globosus from East Africa is unreliable using characters of the shell. In this paper, a molecular method of identification is presented for each species based on DNA sequence variation within the mitochondrial cytochrome oxidase subunit I (COI) as detected by a novel multiplexed SNaPshotTM assay. In total, snails from 7 localities from coastal Kenya were typed using this assay and variation within shell morphology was compared to reference material from Zanzibar. Four locations were found to contain B. nasutus and 2 locations were found to contain B. globosus. A mixed population containing both B. nasutus and B. globosus was found at Kinango. Morphometric variation between samples was considerable and UPGMA cluster analysis failed to differentiate species. The multiplex SNaPshotTM assay is an important development for more precise methods of identification of B. africanus group snails. The assay could be further broadened for identification of other snail intermediate host species.

Classification of immature mosquito species according to characteristics of the larval habitat in the subtropical province of Chaco, Argentina

To classify mosquito species based on common features of their habitats, samples were obtained fortnightly between June 2001-October 2003 in the subtropical province of Chaco, Argentina. Data on the type of larval habitat, nature of the habitat (artificial or natural), size, depth, location related to sunlight, distance to the neighbouring houses, type of substrate, organic material, vegetation and algae type and their presence were collected. Data on the permanence, temperature, pH, turbidity, colour, odour and movement of the larval habitat's water were also collected. From the cluster analysis, three groups of species associated by their degree of habitat similarity were obtained and are listed below. Group 1 consisted of Aedes aegypti. Group 2 consisted of Culex imitator, Culex davisi, Wyeomyia muehlensi and Toxorhynchites haemorrhoidalis separatus. Within group 3, two subgroups are distinguished: A (Psorophora ferox, Psorophora cyanescens, Psorophora varinervis, Psorophora confinnis, Psorophora cingulata, Ochlerotatus hastatus-oligopistus, Ochlerotatus serratus, Ochlerotatus scapularis, Culex intrincatus, Culex quinquefasciatus, Culex pilosus, Ochlerotatus albifasciatus, Culex bidens) and B (Culex maxi, Culex eduardoi, Culex chidesteri, Uranotaenia lowii, Uranotaenia pulcherrima, Anopheles neomaculipalpus, Anopheles triannulatus, Anopheles albitarsis, Uranotaenia apicalis, Mansonia humeralis and Aedeomyia squamipennis). Principal component analysis indicates that the size of the larval habitats and the presence of aquatic vegetation are the main characteristics that explain the variation among different species. In contrast, water permanence is second in importance. Water temperature, pH and the type of larval habitat are less important in explaining the clustering of species.

Análise da riqueza da fauna de Braconidae (Hymenoptera, Ichneumonoidea) em remanescentes naturais da Área de Proteção Ambiental (APA) de Descalvado, SP

Analysis of the Braconidae (Hymenoptera: Ichneumonoidea) fauna richness in natural remnants of the Área de Proteção Ambiental (APA) of Descalvado, SP. A survey of the Braconidae fauna on fragmentary natural environments at northeast São Paulo State was conducted using Malaise traps installed in five sites of Área de Proteção Ambiental of Descalvado. A total of 2,262 specimens, representing 22 subfamilies and 94 genera, was sampled throughout a period of sixteen months (from May, 1999 to August, 2000). Biological data were obtained from current literature and behavior patterns of host utilization for the studied fauna, mainly Braconidae which have been successful in control of agricultural insect pests. Statistical analyses indicated that the asymptote of the genera richness has been approached using this sampling method. This is the most complete survey yet available from natural fragmentary areas of the Descalvado Braconidae fauna. Patterns of distribution and richness of the Braconidae genera were established. Cluster Analysis was adopted taking as attribute the number of Braconidae genera.

Seasonal and spatial species richness variation of dung beetle (Coleoptera, Scarabaeidae s. str.) in the Atlantic Forest of southeastern Brazil

The knowledge on Atlantic Forest scarab beetle fauna is quite limited. This biome is strongly degraded and these insects can be used as bioindicators since they are sensitive to forest destruction and show distinct organizational patterns in forest fragments or in areas that have been deteriorated by human activity. Thus, a study of the Scarabaeidae (sensu stricto) dung beetles fauna that inhabit Serra do Japi, São Paulo, Brazil (23º12'-23º22' S and 46º53'-47º03'W) was carried out; the monthly species richness was analyzed in six areas during one year and the vegetation's structural physiognomy was described. The areas included a conserved and a degraded valley, a northward and a southward hillside, a hilltop, and an area of secondary forest growing under eucalyptus trees. The specimens were collected using four pitfall traps baited with human feces, which remained at each spot during 48 hours. Between September, 1997 and August, 1998, 3524 individuals of 39 species were collected; the most abundant were: Canthidium trinodosum, Eurysternus cyanescens, Uroxys kratochvili, Scybalocanthon nigriceps, Uroxys lata, Canthonella sp., Dichotomius assifer, Deltochilum furcatum, Canthidium sp.2, Canthon latipes, Deltochilum rubripenne, Eurysternus sp., and Dichotomius sp.1. The number of individuals and species was greater in the hot, rainy season, when there was a correlation between the number of species and the mean annual temperature [r²= 0.69; p<0.01]. The lower winter richness was most pronounced in the conserved valley, while richness remained relatively constant in the degraded valley; abundance was much higher in the degraded valley. The cluster analysis showed that the valleys and hillsides are the most similar in relation to species composition and abundance, yet different from the secondary forest with eucalypts and the hilltop.

Spatial estimation of foliar phosphorus in different species of the genus Coffea based on soil properties

Information underlying analyses of coffee fertilization systems should consider both the soil and the nutritional status of plants. This study investigated the spatial relationship between phosphorus (P) levels in coffee plant tissues and soil chemical and physical properties. The study was performed using two arabica and one canephora coffee variety. Sampling grids were established in the areas, and the points georeferenced. The assessed properties of the soil were levels of available phosphorus (P-Mehlich), remaining phosphorus (P-rem) and particle size, and of the plant tissue, phosphorus levels (foliar P). The data were subjected to descriptive statistical analysis, correlation analysis, cluster analysis, and probability tests. Geostatistical and trend analyses were only performed for pairs of variables with significant linear correlation. The spatial variability for foliar P content was high for the variety Catuai and medium for the other evaluated plants. Unlike P-Mehlich, the variability in P-rem of the soil indicated the nutritional status of this nutrient in the plant.

Isoenzymatic variability in wild potatoes

Two species of wild potato Solanum commersonii, subspecies commersonii and malmeanum, and S. chacoense, subspecies muelleri occur in southern Brazil. Their rusticity and easy adaptation to extreme climatic conditions make them valuable for breeding programs. The objective of this work was to analyze the isoenzymatic variability of 113 clones of wild potato subspecies. They were collected and maintained at Embrapa-Centro de Pesquisa Agropecuária de Clima Temperado, at Pelotas, RS, Brazil. Enzymes involved in energetic (group I) or in peripherical (group II) metabolism constituted the material used. Polyacrylamide horizontal gel electrophoresis was used to analyze peroxidase, aspartate transaminase, phosphoglucomutase and isocitrate dehydrogenase isoenzymes. Solanum spp. has considerable genetic variability for isoenzymatic patterns. Cluster analysis classified the clones into 51 subgroups, based on electrophoretic variants of group I enzymes, and into 89, when group II enzyme variants were added. Genotypic differentiation of S. chacoense muelleri in relation to S. commersonii commersonii and S. commersonii malmeanum is evident when expressed through similarity and cluster analysis.

Genetic diversity and relationship in American and African oil palm as revealed by RFLP and AFLP molecular markers

The objective of this work was to evaluate the genetic diversity, its organization and the genetic relationships within oil palm (Elaeis oleifera (Kunth) Cortés, from America, and E. guineensis (Jacq.), from Africa) germplasm using Restriction Fragment Length Polymorphism (RFLP) and Amplified Fragment Length Polymorphism (AFLP). In complement to a previous RFLP study on 241 E. oleifera accessions, 38 E. guineensis accessions were analyzed using the same 37 cDNA probes. These accessions covered a large part of the geographical distribution areas of these species in America and Africa. In addition, AFLP analysis was performed on a sub-set of 40 accessions of E. oleifera and 22 of E. guineensis using three pairs of enzyme/primer combinations. Data were subjected to Factorial Analysis of Correspondence (FAC) and cluster analysis, with parameters of genetic diversity being also studied. Results appeared congruent between RFLP and AFLP. In the E. oleifera, AFLP confirmed the strong structure of genetic diversity revealed by RFLP, according to geographical origin of the studied material, with the identification of the same four distinct genetic groups: Brazil, French Guyana/Surinam, Peru, north of Colombia/Central America. Both markers revealed that genetic divergence between the two species is of the same magnitude as that among provenances of E. oleifera. This finding is in discrepancy with the supposed early tertiary separation of the two species.

Genetic diversity and virulence pattern in field populations of Pyricularia grisea from rice cultivar Metica-1

Rice blast is a major yield constraint of the irrigated rice in the State of Tocantins, Brazil. The objective of this investigation was to study the phenotypic and genetic diversity within the pathogen population of Pyricularia grisea in samples collected from four individual farms of rice cultivar Metica-1, under epidemic conditions of leaf blast. A set of 87 isolates was tested on 32 rice genotypes including eight international differentials. Considering 80% similarity in virulence, two groups comprising a total of 81 isolates were recognized, independently of the farms from which they were collected. Eighty percent of the isolates pertained to pathotype ID-14, indicating high cultivar specificity and narrow diversity of virulence in the sample population. The virulence in pathogen population on rice cultivars BR-IRGA 409 and Rio Formoso was low. Analysis of P. grisea isolates using rep-PCR with two primer sequences from Pot2 generated fingerprint profiles of one to nine bands. Cluster analysis revealed the occurrence of six fingerprint groups with similarities ranging from 0.09 to 1. There was no straight relationship between virulence of the isolates based on reaction pattern on 32 genotypes and grouping based on Pot2 rep-PCR analysis of P. grisea isolates collected from 'Metica-1'.

Análise de crescimento de girassol em Latossolo com diferentes níveis de saturação por bases no Cerrado

Com o objetivo de avaliar o comportamento de cinco cultivares de girassol em um Latossolo Vermelho argiloso, foram realizados quatro ensaios sob quatro níveis de saturação por bases (18%, 35%, 56% e 77%) obtidos com aplicação de calcário dolomítico. Os dados semanais de crescimento foram ajustados pelas funções logística, Weibull de três parâmetros e a logística generalizada de quatro parâmetros empregando-se o procedimento NLIN do SAS, bem como os processos interativos do método DUD. A função logística forneceu os melhores ajustes aos dados de crescimento. As cultivares foram categorizadas de acordo com o porte das plantas e tendências de crescimento a partir da análise de agrupamentos (cluster analysis), usando o método das ligações completas e a métrica da distância euclidiana. Todas as cultivares tiveram o crescimento reduzido quando o nível de saturação de bases foi de 18%. A cultivar M734 apresentou bom desempenho no nível de 35% de saturação por bases. Nos demais casos, os melhores resultados foram obtidos nos níveis de saturação por bases entre 56% e 77%. As cultivares EMBRAPA 122 e M738 tiveram redução de crescimento quando a saturação foi elevada para 77%. 'EMBRAPA 122' apresentou maior crescimento inicial e menor altura ao final do ciclo vegetativo.

Characterization of potato genotypes using molecular markers

The objective of this work was to characterize 27 potato genotypes, using molecular markers. Polyacrylamide gel electrophoresis, RAPD techniques and isozymes of esterase, phosphoglucomutase and soluble proteins were analyzed in tubers, and isocitrate dehydrogenase, aspartate transaminase, phosphoglucomutase and peroxidase, in leaves. Eighteen primers were tested and four were chosen, kits OPX (01, 04 and 09) and OPY (07), to analyze RAPD markers in leaf extracts. Similarity and cluster analysis were conducted using Jaccard coefficient and the unweighted pair-group method using arithmetic average. Despite the differences detected in the analysis of proteins and isozymes in the tubers, as well as of isozymes in the leaves, the characterization of all genotypes through gel electrophoresis was not possible, while RAPD markers were efficient to characterize all the 27 genotypes.

Reaction of arracacha genotypes to the root soft rot caused by Pectobacterium chrysanthemi

The purpose of this paper was to screen thirty-two arracacha genotypes for their reaction to root soft rot. Twenty roots of each genotype were inoculated with two Pectobacterium chrysanthemi isolates in a randomized experiment (10 roots/isolate). After inoculation, roots were individually wrapped with PVC film and kept at 26ºC in closed plastic bags. Soft rot lesions were recorded after 36 hours and genotypes were grouped in four classes of susceptibility by cluster analysis: 10 were less susceptible, 16 intermediate, 3 susceptible and 3 very susceptible. All the tested arracacha genotypes showed only variation in the degree of susceptibility.

Composition and functional groups of epiedaphic ants (Hymenoptera: Formicidae) in irrigated agroecosystem and in nonagricultural areas

The objective of this work was to evaluate the species composition and functional groups of ants in nonagricultural (NA) and in irrigated areas (S, seasonal irrigation; P, irrigation with well water; W, irrigation with wastewater) in an arid agricultural region in central Mexico, throughout 2005 and 2006. A total of 52,358 ants belonging to 6 subfamilies, 21 genera and 39 species was collected using pitfall traps. The species best represented in all plots were: Forelius pruinosus, Pheidole obtusospinosa, Monomorium minimum and Dorymyrmex spp. NA plots recorded the highest density of ants. The highest values for diversity (H') and equitativity (J') were recorded in NA and P plots, while the lowest were recorded in W plots. Cluster analysis showed two different groups regarding species composition: NA-S and W-P. Functional groups recorded were: dominant Dolichoderinae, three species; subordinate Camponotini, five species; hot climate specialists, three species; tropical climate specialists, seven species; cold climate specialists, five species; cryptic species, one species; opportunists, six species; generalized Myrmicinae, nine species. Agricultural activity affects the structure of the ant community with epiedaphic forage, and the constant use of irrigation wastewater in conjunction with intense agricultural practices has negative effect upon species richness of epiedaphic ants.

Genetic diversity for gliadin patterns of durum wheat landraces in the Northwest of Iran and Azerbaijan

The objective of this study was to identify gliadin band patterns and the extent of genetic diversity in durum wheat genotypes from Northwestern Iran and the Republic of Azerbaijan. Gliadins from 46 landraces and four cultivars were evaluated through acid PAGE analyses. Sixty-six polymorphic bands and 81 patterns were identified. Twenty-four different motility bands and 22 patterns were found in the ω gliadin region with 14 polymorph bands and 20 patterns for α and γ gliadins, and 14 bands and 19 different patterns for β gliadins. The combination of these patterns generated 38 and 39 combinations for Gli-1 and Gli-2 loci, respectively. The genetic diversity index (H) was higher for α gliadins (0.924), followed by ω and γ gliadins (0.899 and 0.878, respectively), and for β gliadin patterns (0.866). Extensive polymorphism (H = 0.875) was observed in four gliadin pattern regions, with higher genetic diversity in the Iranian landraces than in the Azerbaijani ones. Each genotype had special identifying patterns in the gliadin acid PAGE analysis, and cluster analysis based on Jaccard's similarity coefficients formed six groups. Gliadin has a simple, repeatable and economic analysis, and can be used in genetic studies

Genetic diversity in table grapes based on RAPD and microsatellite markers

The objective of this work was to analyze the genetic diversity of 47 table grape accessions, from the grapevine germplasm bank of Embrapa Semiárido, using 20 RAPD and seven microsatellite markers. Genetic distances between pairs of accessions were obtained based on Jaccard's similarity index for RAPD data and on the arithmetic complement of the weighted index for microsatellite data. The groups were formed according to the Tocher's cluster analysis and to the unweighted pair‑group method with arithmetic mean (UPGMA). The microsatellite markers were more efficient than the RAPD ones in the identification of genetic relationships. Information on the genetic distance, based on molecular characteristics and coupled with the cultivar agronomic performance, allowed for the recommendation of parents for crossings, in order to obtain superior hybrids in segregating populations for the table grape breeding program of Embrapa Semiárido.

Morphological and molecular diversity among cassava genotypes

The objective of this work was to characterize morphologically and molecularly the genetic diversity of cassava accessions, collected from different regions in Brazil. A descriptive analysis was made for 12 morphological traits in 419 accessions. Data was transformed into binary data for cluster analysis and analysis of molecular variance. A higher proportion of white or cream (71%) root cortex color was found, while flesh colors were predominantly white (49%) and cream (42%). Four accession groups were classified by the cluster analysis, but they were not grouped according to their origin, which indicates that diversity is not structured in space. The variation was greater within regions (95.6%). Sixty genotypes were also evaluated using 14 polymorphic microsatellite markers. Molecular results corroborated the morphological ones, showing the same random distribution of genotypes, with no grouping according to origin. Diversity indices were high for each region, and a greater diversity was found within regions, with: a mean number of alleles per locus of 3.530; observed and expected heterozygosity of 0.499 and 0.642, respectively; and Shannon index of 1.03. The absence of spatial structure among cassava genotypes according to their origins shows the anthropic influence in the distribution and movement of germplasm, both within and among regions.