Genetic variability of garlic accessions as revealed by agro-morphological traits evaluated under different environments.

The cultivated garlic (Allium sativum L.) displays a wide phenotypic diversity, which is derived from natural mutations and phenotypic plasticity, due to dependence on soil type, moisture, latitude, altitude and cultural practices, leading to a large number of cultivars. This study aimed to evaluate the genetic variability shown by 63 garlic accessions belonging to Instituto Agronômico de Campinas and the Escola Superior de Agricultura "Luiz de Queiroz" germplasm collections. We evaluated ten quantitative characters in experimental trials conducted under two localities of the State of São Paulo: Monte Alegre do Sul and Piracicaba, during the agricultural year of 2007, in a randomized blocks design with five replications. The Mahalanobis distance was used to measure genetic dissimilarities. The UPGMA method and Tocher's method were used as clustering procedures. Results indicated significant variation among accessions (P < 0.01) for all evaluated characters, except for the percentage of secondary bulb growth in MAS, indicating the existence of genetic variation for bulb production, and germplasm evaluation considering different environments is more reliable for the characterization of the genotypic variability among garlic accessions, since it diminishes the environmental effects in the clustering of genotypes.

Chloroplast diversity of Casearia grandiflora in the Cerrado of Piauí State.

Casearia grandiflora (Salicaceae) is a typical Cerrado species adapted to disturbed environments, making it useful for restoration projects. Knowledge of genetic diversity is important for establishing conservation strategies for this species. This study aimed to compare chloroplast haplotype diversity and structure of C. grandiflora, under the assumption that protected areas hold greater genetic diversity than disturbed areas. The populations studied are from Parque Nacional de Sete Cidades Conservation Unit and from the surroundings of the city of Cocal de Telha, both located in the State of Piauí. Molecular analyses were performed with seven chloroplast microsatellite loci. The number of private haplotypes and haplotype diversity were higher in the conservation unit, which reinforces the importance of these areas in maintaining biodiversity. Analysis of molecular variance showed that most of the genetic variation is found within populations, with a moderate divergence between them (FST = 0.14). The Bayesian analysis and discriminant analysis of principal components suggested that populations are not structured, revealing that a set of individuals from Parque Nacional de Sete Cidades were more divergent within populations than between them. Since literature has little information on C. grandiflora, the results of this study provide important contribution to a better understanding of the specie's genetic diversity.

Genetic diversity of Casearia sylvestris populations in remnants of the Atlantic Forest.

Guaçatonga (Casearia sylvestris) is a native plant of the Atlantic Forest, with high medicinal potential and relevance for reforestation programs. The aim of this study was to characterize, with microsatellite markers, two populations of C. sylvestris from remaining areas of the Atlantic Forest in the State of São Paulo. High allelic variation was found in both populations (NA = 101 and 117; AR = 12.5 and 14.4), although with high endogamy coefficients (f = 0.640 and 0.363). Estimates of genetic structure suggested the presence of considerable genetic divergence between the populations (FST = 0.103); however, there was no spatial genetic structure within the populations. Genetic divergence may have occurred due to decreased gene flow between the fragmented populations as the result of deforestation. The results of this study demonstrate the importance of genetic diversity and its characterization in native plants within remaining forest areas for the management and restoration of such areas.

Genetic divergence in a soybean (Glycine max) diversity panel based on agro-morphological traits.

Owing to the narrow genetic basis of soybean (Glycine max), the incorporation of new sources of germplasm is indispensable when searching for alleles that contribute to a greater diversity of varieties. The alternative is plant introduction, which may increase genetic variability within breeding programs. Multivariate techniques are important tools to study genetic diversity and allow the precise elucidation of variability in a set of genotypes of interest. The agro-morphological traits of 93 soybean accessions from various continents were analyzed in order to assess the genetic diversity present, and to highlight important traits. The experimental design was incomplete blocks (Alpha lattice, 8 x 12) with three replicates. Nine agro-morphological traits were analyzed, and principal component analysis and cluster analysis were performed, the latter by Ward's method. The dendrogram obtained contained eight subgroups, confirming the genetic diversity among the accessions and revealing similarities between 11 national genotypes. The geographical origin of the accessions was not always related to the clusters. The traits evaluated, and the methods used, facilitated the distinction and characterization of genotypes between and within groups, and could be used in Brazilian soybean breeding programs.

Genetic diversity of Lippia sidoides Cham. and L. gracilis Schauer germplasm.

The conservation of plants in germplasm banks ensures the characterization and availability of these resources for future generations. The present study used DNA markers to obtain genetic information about germplasm collections of Lippia sidoides and L. gracilis, which are maintained in an Active Germplasm Bank (AGB). Genetic variability of samples in the AGB was assessed using 12 combinations of amplified fragment length polymorphism (AFLP) primers (EcoRI/MseI). Twenty simple sequence repeat primers designed for L. alba were tested to determine their transferability in L. sidoides and L. gracilis. The AFLP markers generated 789 markers. The assessed loci exhibited a moderate Shannon diversity index (I = 0.42) in both species, suggesting that the conserved accessions possess an intermediate level of genetic diversity. Twelve microsatellite loci amplified satisfactorily, and nine loci were polymorphic in each species. A total of 23, 22, and 36 alleles, with an average of 2.5, 2.4, and 3.27 alleles per locus were identified for L. sidoides and L. gracilis accessions in the AGB, and Lippia sp sampled plants, respectively. Analyses of genetic structure permitted the identification of three different groups using both sets of markers, of which two were representative of L. sidoides. The information generated in this study may help to create, expand, and maintain collections of these species and may assist in genetic-breeding programs.

QTL mapping of soybean oil content for marker-assisted selection in plant breeding program.

The present study was undertaken to detect and map the quantitative trait loci (QTL) related to soybean oil content. We used 244 progenies derived from a bi-parental cross of the Lineage 69 (from Universidade Estadual Paulista "Júlio de Mesquita Filho"/Faculdade de Ciências Agrárias e Veterinárias - Breeding Program) and Tucunaré cultivar. A total of 358 simple sequence repeat (SSR; microsatellite) markers were used to investigate the polymorphism between the parental lines, and for the polymorphic lines all the F2 individuals were tested. Evaluation of the oil content and phenotype was performed with the aid of a Tango equipment by near infra-red reflectance spectroscopy, using single F2 seeds and F2:3 progenies, in triplicate. The data were analyzed by QTL Cartographer program for 56 SSR polymorphic markers. Two oil-content related QTLs were detected on K and H linkage groups. The total phenotypic variation explained by QTLs ranged from 7.8 to 46.75% for oil content. New QTLs were identified for the oil content in addition to those previously identified in other studies. The results reported in this study show that regions different from those already known could be involved in the genetic control of soybean oil content.

Molecular characterization of patchouli (Pogostemon spp) germplasm.

Patchouli [Pogostemon cablin (Blanco) Benth.] is an aromatic, herbaceous plant belonging to the Lamiaceae family native to Southeast Asia. Its leaves produce an essential oil regularly used by the perfume and cosmetics industries. However, since patchouli from the Philippines and India were described and named Pogostemon patchouli, there has been a divergence in the identity of these species. The objective of the current study was to study the genetic diversity of patchouli accessions in the Active Germplasm Bank of Universidade Federal de Sergipe using microsatellite and inter simple sequence repeat markers. The results of both types of molecular markers showed that there are two well-defined clusters of accessions that harbor exclusive alleles. It was observed that these two clusters are genetically distant, suggesting that they belong to two different species. Based on the results, two accessions were classified as Pogostemon heyneanus and the remaining accessions were classified as P. cablin.

Molecular and chemical characterization of vetiver, Chrysopogon zizanioides (L.) Roberty, germplasm.

Due to the economic interests in vetiver, Chrysopogon zizanioides (L.) Roberty, molecular and chemical studies are essential to generate information for its sustainable exploitation. The aim of this study was to undertake a molecular and chemical characterization of vetiver accessions of the active germplasm bank of the Universidade Federal de Sergipe. The molecular characteristics of the accessions were studied using amplified fragment length polymorphism markers, with a total of 14 primer combinations that generated 442 loci, allowing us to observe that these accessions have similar genomes. The vetiver accessions were divided into three distinct groups, where accession UFS-VET005 was the most differentiated and accession UFS-VET004 had the lowest essential oil content (0.70%). The content of the chemical constituents of the essential oils was observed to vary, with a predominance of khusimol, which ranged from 18.97 to 25.02%. It was possible to divide the vetiver accessions into two groups based on chemical composition, and these groups do not correlate with the molecular grouping. Therefore, it is necessary to perform molecular and chemical analyses to characterize vetiver accessions.

Populations of Erythrina velutina Willd. at risk of extinction.

The goal of this study was to characterize the structure of two natural populations of the coral tree using RAPD and ISSR markers. The study evaluated all individuals in two different areas in the northeastern region of Brazil: the first was in the riparian area, 10 km x 100 m along the edge of the lower São Francisco River, and the second was in the municipality of Pinhão, in a semiarid region between the municipalities of Neópolis and Santana do São Francisco. We used all the coral trees present in those two areas (37 individuals). The results of the RAPD and ISSR markers were highly congruent, supporting the reliability of the techniques used. Similarity was estimated using the Jaccard arithmetic complement index. A dendrogram was constructed using the unweighted pair group method with arithmetic mean cluster algorithm, and the robustness of the data was bootstrapped with 5000 replicates. A principal coordinate analysis was performed on the basis of Jaccard coefficients. The total genetic variation observed was 21%, corresponding to the variation between the populations, and 79% of the variation was observed within the populations.

Development of a novel set of microsatellite markers for Lippia alba (Verbenaceae).

Microsatellite primers were developed and optimized for Lippia alba to characterize the L. alba germplasm bank of Universidade de São Paulo. A genomic library enabled the design of 9 microsatellite primers. Six of the 9 primers yielded polymorphic products, which defined 2 groups in the bank. The data provide support to characterize germplasm banks, genetic breeding programs for L. alba, and other genetic diversity studies and classifications of species in the genus Lippia.

New loci of Lychnophora ericoides and transferability to Lychnophora pinaster, endangered medicinal species from Brazil.

Lychnophora ericoides and Lychnophora pinaster are species used in popular medicine as analgesic or anti-inflammatory agents to treat contusions, rheumatism, and insect bites. In this study, 21 simple sequence repeat loci of L. ericoides were developed and transferred to L. pinaster. Three populations of L. ericoides and 2 populations of L. pinaster were evaluated; they were collected in the State of Minas Gerais. Population parameters were estimated, and the mean values of observed and expected heterozygosity were 0.297 and 0.408 (L. ericoides) and 0.228 and 0.310 (L. pinaster), respectively. Greater genetic variability was observed within populations than between populations of L. ericoides (62 and 37%) and L. pinaster (97 and 2.8%). These results provide information for genetic conservation and taxonomic studies of these endangered species.

Isolation and characterization of microsatellite loci from the tick Amblyomma aureolatum (Acari: Ixodidae).

Amblyomma aureolatum (Pallas) is the main vector of the bacterium Rickettsia rickettsii, the etiological agent of Brazilian spotted fever. This disease is the most lethal human spotted fever rickettsiosis in the world. Microsatellite loci were isolated from a dinucleotide-enriched library produced from A. aureolatum sampled in Southeastern Brazil. Eight polymorphic microsatellites were further characterized among 38 individuals sampled from São Paulo metropolitan region. The number of observed alleles ranged from 2 to 9, observed heterozygosity was 0.184-0.647, and expected heterozygosity was 0.251-0.747. Cross-species amplifications suggested that these loci will be useful for other Amblyomma species.

Cross-species transferability of microsatellite markers in the genus Lippia.

The cross-species transferability of 20 microsatellite markers was tested in the genus Lippia. Eleven markers were polymorphic after screening 19 accessions of Lippia sidoides and Lippia gracilis maintained in the Active Germplasm Bank (AGB) from Universidade Federal de Sergipe. Additionally, 40 accessions of Lippia spp were collected in Sergipe to increase the germplasm bank. A total of 23, 22, and 36 alleles were identified, with an average of 2.3, 2.2, and 3.27 alleles per locus, respectively, for each group. The markers that were used were efficient tools to access genetic diversity in the germplasm bank and will be useful for further research aiming at the conservation and management of these important aromatic species.

Development and characterization of novel microsatellite markers in Hyptis pectinata (Lamiaceae).

A microsatellite-enriched library was constructed and a set of 19 SSR markers were developed to characterize a germplasm collection of Hyptis pectinata (L.) Poit., maintained at the Universidade Federal de Sergipe (UFS). Fifteen markers of 19 ranged from moderately to highly polymorphic. A total of 113 alleles were identified, with a mean of 7.52 alleles per locus. The mean HO and HE were 0.582 and 0.657, respectively. The primers developed were efficient tools for accessing the genetic diversity of the germplasm collection analyzed and may also be useful for other studies involving this species and other species in the genus Hyptis.

Isolation and characterization of microsatellite loci for Bixa orellana, an important source of natural dyes.

Annatto (Bixa orellana) is a plant native from the American continental tropical zone. The seeds are used to produce a carotenoid-based yellow to orange food coloring. Microsatellite markers were developed for the Brazilian native species Bixa orellana to describe its genetic diversity and structure as well as to support conservation studies. Twenty-five microsatellite loci were isolated and characterized using an enriched genomic library. Ten loci were polymorphic in the 50 accessions sampled in this study, while 15 were considered monomorphic. The mean number of alleles per locus was 3.8, ranging from 2 to 6 alleles per locus. Mean values for the observed and expected heterozygosities were 0.541 (ranging from 0 to 0.658) and 0.639 (ranging from 0.422 to 0.787), respectively. All markers described in this study will be useful in further studies evaluating the genetic diversity, population dynamics, and conservation genetics of Bixa orellana.

A modified acidic approach for DNA extraction from plant species containing high levels of secondary metabolites.

Purified genomic DNA can be difficult to obtain from some plant species because of the presence of impurities such as polysaccharides, which are often co-extracted with DNA. In this study, we developed a fast, simple, and low-cost protocol for extracting DNA from plants containing high levels of secondary metabolites. This protocol does not require the use of volatile toxic reagents such as mercaptoethanol, chloroform, or phenol and allows the extraction of high-quality DNA from wild and cultivated tropical species.

Isolation and characterization of microsatellites for the yam Dioscorea cayenensis (Dioscoreaceae) and cross-amplification in D. rotundata.

Dioscorea cayenensis and Dioscorea rotundata are among the most important yam species for the humid and sub-humid tropics. We isolated nine polymorphic microsatellite markers using a microsatellite-enriched genomic library technique. The nine primer pairs were validated in 22 D. cayenensis accessions, and were tested for transferability in 26 D. rotundata accessions. The number of bands ranged from 2 to 4, with a mean of 3.11. D. cayenensis gave primer polymorphism information content values ranging from 0.37 to 0.62, while for D. rotundata the values ranged from 0.15 to 0.66. The D parameter in D. cayenensis ranged from 0.14 to 0.40, while in D. rotundata it ranged from 0.05 to 0.34. These SSR markers will be useful to characterize genetic diversity in D. cayenensis and D. rotundata accessions.

Microsatellite markers for the Cabreúva tree, Myroxylon peruiferum (Fabaceae), an endangered medicinal species from the Brazilian Atlantic Forest.

The Cabreúva tree, Myroxylon peruiferum, is an endangered tropical species from Brazil used in forest restoration projects. It is known for its medicinal properties. Eleven microsatellite markers were developed for this species, from a microsatellite-enriched library. Nine of these markers, characterized in 30 individuals from a semideciduous forest remnant population in southeast Brazil, were polymorphic, with allele numbers ranging from 2 to 8 per locus; expected and observed heterozygosities ranged from 0.103 to 0.757 and 0.107 to 0.704, respectively. One locus (Mpe-C04) showed significant deviation from Hardy-Weinberg equilibrium, probably due to null alleles. Two other loci (Mpe-E09 and Mpe-H07) were monomorphic in this population. These microsatellite loci should be useful for future population genetic studies of this species.

Produção de açafrão em função de acessos e do peso de rizomas-semente / Influence of the weight of seed-rhizomes in the production of turmeric

O açafrão-da-terra (Curcuma longa L.) é originário do sudeste asiático e subcontinente indiano. É uma herbácea de caule subterrâneo, alaranjado, com vários rizomas secundários aproveitados na indústria alimentícia e farmacêutica devido às características de cor, sabor, odor, produção de óleos essenciais, e corantes. Na escolha do melhor material propagativo deve-se levar em consideração o material genético, o peso, tamanho, idade, capacidade de reserva acumulada, sanidade, dentre outros fatores. O objetivo neste experimento foi avaliar a influencia de diferentes acessos e pesos de rizomas-sementes na produção de açafrão. Empregou-se o delineamento em blocos casualizados em parcelas subdivididas com quatro repetições, tendo como tratamento principal os acessos (C-06, C-36 e C-38) e como tratamento secundário as classes de rizomas-sementes (peso): pequeno, médio e grande, ±5, ±10 e ±15 g/rizoma, respectivamente. Após a colheita, os rizomas foram distribuídos sobre tela suspensa para secagem à sombra com ventilação natural, por 20 dias. Posteriormente, para avaliar a produção, os rizomas foram classificados em 4 classes: A (> 15g ), B (±10 g), C (±5 g) e D(<5g). A interação entre os acessos e o tamanho do rizoma-semente foi significativa para todas as variáveis, com exceção da Classe D. Com o uso de rizoma-semente grande os acessos C-38 e o C-06 apresentaram maior produtividade total, 0,834 e 0,812kg/planta, respectivamente. O descarte gerado foi menor no acesso C-38 do que no C-06, representando 7,8 e 12,8% da produção total, respectivamente. O uso de rizomas-semente maiores aumentou significativamente a produção total. No acesso C-06 a produção passou de 0,481 para 0,812 kg/planta, ou seja, um aumento de 70%. O ganho relativo na produção de rizoma (kg/planta) no acesso C-06 para o plantio de rizomas com ±15 g, foi de 28%.

Turmeric (Curcuma longa L) originated in Southeast Asia and the Indian subcontinent. It is an herbaceous plant with underground, orange stem with several secondary rhizomes used in the food and pharmaceutical industries, because of its characteristics of color, taste, smell, production of essential oils and dyes. In cultivation, the best choice of propagation material must take account the genetic material, weight, age, accumulated reserve capacity, sanity, among other factors. In this study, seed-rhizomes of three weight categories - small, medium and large -, ± 05, ± 10 and ± 15 g / rhizome, respectively, of three genetic materials - C-06, C-36 and C-38 - from the Germplasm Bank of the ESALQ / USP were cultured from December 2009 to August 2010, at a spacing of 0.5 mx 0.2 m. After harvest, they were distributed on canvas suspended for drying in the shade and natural ventilation for 20 days. Later, to evaluate the production, they were classified into four categories: A (> 15g), B (± 10 g), C (± 5 g) and D (<5g) .The interaction between accessions and size of seed-rhizomes was significant for all variables, except for category D. With the use of large seed-rhizomes, C-38 and C-06 had a higher total yield, with 0.834 and 0.812 kg/plant, respectively. The use of large seed-rhizomes increased significantly the total production. In C-06, the production increased from 0.481 to 0.812 kg/plant, i.e. an increase of 70%. Also in C-06, the relative gain in the production of rhizome (kg / plant) for the planting of seed-rhizomes with ± 15 g was 28%.

Population variability of Bemisia tabaci (Genn.) in different hosts.

The silverleaf whitefly, Bemisia tabaci (Genn.) (Hemiptera: Aleyrodidae), is a cryptic species complex that contains some of the most damaging pests in tropical and subtropical regions. Recent studies have indicated that this complex is composed of at least 24 distinct and morphologically indistinguishable species that mainly differ in their ability to transmit phytoviruses, adapt to hosts, and induce physiological changes in certain hosts. The importance of this species has been increasing worldwide, because it serves as a phytovirus vector, particularly for geminiviruses, in economically important crops. Here, we aimed to examine the population variability of B. tabaci populations inhabiting 6 agricultural crops grown in 5 regions of Brazil and 1 region of the USA; BRrep [Brasília (DF, Brazil) - cabbage], ILsoj [Urbana (IL/USA) - soybean], BJabo [Bom Jesus da Lapa (BA, Brazil) - pumpkin], CPsoj [Campinas (SP, Brazil) - soybean], UBman [Ubatuba (SP, Brazil) - cassava], and PEmel [Petrolina (PE, Brazil) - melon]. Thirteen polymorphic loci with 50 alleles were observed, with an average of 2.37 (range: 2.00-2.91) alleles per population. The UBman and PEmel B. tabaci populations were the most differentiated, which was probably caused by insect adaptation to the host plant and the use of insecticides. A 33.87% inter-population variation was observed, indicating that microsatellites may be used to measure differentiation among these B. tabaci populations. Based on the comparison of microsatellites in the current study, only the Middle East-Asia Minor 1 population of B. tabaci was found in the six populations.