Avaliação da qualidade fisiológica de sementes de açaí

Quarenta lotes de sementes de açaizeiro (Euterpe oleracea Mart.) colhidos na coleção de germoplasma de palmeiras do Instituto Agronômico de Campinas (Ubatuba, Estado de São Paulo) foram submetidos aos seguintes testes para a avaliação de qualidade fisiológica das sementes: grau de umidade, germinação, primeira contagem de germinação (emissão do botão germinativo aos quatro dias da semeadura), índice de velocidade de germinação (IVG) e condutividade elétrica. O grau de umidade das sementes apresentou valores de 32,2 a 45,5%. A avaliação da qualidade das sementes de açaizeiro pode fundamentar-se nos testes de germinação, de primeira contagem de germinação, de IVG e de condutividade elétrica quando interpretados em conjunto.

Total and individual carotenoid profiles in Solanum phureja of cultivated potatoes: I. Concentrations and relationships as determined by spectrophotometry and HPLC

Total and individual carotenoid concentrations were determined by spectro photometry and HPLC, in raw tubers of a sample of 23 accessions of Solanum phureja potatoes taken at random from the world germplasm collection following its stratification on tuber flesh color. Lutein, zeaxanthin, violaxanthin, antheraxanthin and beta-carotene were detected in all accessions and three distinct patterns of carotenoid accumulation were evidenced by cluster analysis. Accessions in group 1 showed the highest concentrations of total carotenoids (1258-1840 mu g 100 g(-1) FW) comprised largely of zeaxanthin (658-1290 mu g 100 g(-1) FW) with very low or no presence of beta-carotene (below 5.4 mu g 100 g(-1) FW). Accessions in group 2 presented moderate total carotenoid concentrations with violaxanthin, antheraxanthin, lutein and zeaxanthin as the major carotenoids. Accessions in group 3 showed low concentrations of total carotenoids (97-262 mu g 100 g(-1) FW) and very low or no zeaxanthin, with lutein and violaxanthin as the predominant carotenoids and relatively high concentrations of beta-carotene(up to 27 mu g 100 g(-1) FW). Five accessions with significant concentrations of zeaxanthin were identified with the accession 703566 showing the highest concentration (1290 p g 100 g(-1) FW). This value is to our knowledge higher than any value previously reported for potatoes, including those achieved through genetic modification. For the 23 S. phureja accessions, total carotenoid concentration was positively and significantly correlated with antheraxanthin and zeaxanthin concentrations, and negatively and significantly correlated with beta-carotene concentration. (C) 2008 Elsevier B.V. All rights reserved.

Similaridade genética entre acessos de goiabeiras e araçazeiros baseada em marcadores moleculares AFLP

Espécies do gênero Psidium, como goiabeira e araçazeiros, são economicamente importantes e têm como área de diversidade genética primária o Brasil. Foram determinadas as relações genéticas, com base no marcador AFLP, para acessos do Banco Ativo de Germoplasma (BAG) de Psidium da Empresa Brasileira de Pesquisa Agropecuária (EMBRAPA) Semiárido, para orientar trabalhos de melhoramento e de manejo de recursos genéticos do gênero. Foram analisados 88 acessos, sendo 64 de goiabeira e 24 de araçazeiros, coletados em dez Estados brasileiros, adotando-se para agrupamento o dendrograma UPGMA, considerando a matriz de similaridade do coeficiente de Jaccard de 149 bandas polimórficas de AFLP de 16 combinações dos iniciadores EcoRI e MseI. A análise da variância de dados moleculares foi realizada considerando a variação entre e dentro das populações de goiabeira dos dez Estados. O dendrograma apresentou boa definição, com coeficiente de correlação cofenética de 0,94. Foram observados dois grandes grupos: um formado por acessos de goiabeira e outro com acessos de araçazeiros, com inclusão de alguns acessos de goiabeira. Foram observados agrupamentos específicos no dendograma apenas para os indivíduos de goiabeira coletados em Goiás e Roraima. Os acessos estudados apresentaram similaridade variando de 28 a 98%, evidenciando a alta variabilidade genética dos mesmos. A variação entre acessos foi estimada em 0,16 (ΦST), indicando diferenciação genética moderada entre as populações de goiabeira dos dez Estados. Para aumentar a variabilidade genética do BAG estudado, sugere-se a coleta de um número maior de acessos nos Estados de Goiás e Roraima, dado o alto índice de similaridade entre os acessos provenientes destes Estados, bem como coletas amplas em outros Estados brasileiros. Sugerem-se ainda cruzamentos entre os poucos acessos de goiabeira posicionados no grupo dos araçazeiros para o desenvolvimento de híbridos interespecificos no gênero Psidium.

Genetic diversity between and within populations of Handroanthus heptaphyllus (Vell.) Mattos using microsatellite markers

Handroanthus heptaphyllus (Vell.) Mattos, popularmente conhecida por ipê-roxo, é uma espécie pertencente à família Bignoneaceae, muito apreciada por sua beleza, madeira de excelente qualidade e utilizada em produtos medicinais e programas de reflorestamento de áreas degradadas, paisagismo e restauração. Neste trabalho, objetivou-se estudar a diversidade genética entre e dentro das populações de H. heptaphyllus por meio de marcadores microssatélites. Foram estudadas 192 plântulas, formadas a partir de sementes colhidas de duas populações, em um total de 30 árvores, de fragmentos florestais naturais na região de Botucatu - SP. Foram analisados oito locos microssatélites, com polimorfismo alélico, variando de seis alelos para o loco TAU22 a 14 alelos para os locos TAU12, TAU30 e TAU31, com número efetivo médio de alelos por loco (Âe) igual a 4,9. As médias para a heterozigosidade esperada (Ĥe), para as duas populações foi de 0,785, a heterozigosidade observada (Ĥo)foi de 0,609 e o índice de fixação (^F) variou pouco entre as populações, com média de 0,222. O valor médio da divergência genética entre as duas populações (Ĝst') foi de 0,100. Conclui-se que a maior diversidade genética ocorre dentro das populações; portanto, em programa de coleta de germoplasma, para a região de Botucatu, é recomendado realizar uma maior amostragem de indivíduos dentro das populações, o que possibilitaria uma boa representatividade genética. As populações estudadas possuem diversidade genética para subsidiar programas de melhoramento genético e conservação de germoplasma.

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.

Development of an analytical method for the quantification of pfaffic acid in Brazilian ginseng (Hebanthe eriantha)

Hebanthe eriantha (Poir.) Pedersen (Amaranthaceae), which is known as Brazilian ginseng is widely used in folk medicine as an aphrodisiac and antidiabetic tonic. The anti-tumor activity, attributed to the pfaffic acid present in roots of H. eriantha, is responsible for the great interest in the commercialization of this species. In Brazil, the species H. eriantha is mainly used in commercial preparations, although other plants of the genus Pfaffia and Hebanthe have been marketed as Pfaffia paniculata or Brazilian ginseng. The pfaffic acid present in the roots is mainly conjugated with sugars (pfaffosides) and can be used as an active marker of H. eriantha, which helps to differentiate this species from others marketed as Brazilian ginseng. The main objective of this study was to develop and validate a liquid chromatographic method to quantify pfaffic acid in the roots of H. eriantha. The extraction and hydrolysis conditions were optimized using an univariate and experimental design, respectively, and the quantification of pfaffic acid by high performance liquid chromatography with diode-array detection (HPLC-DAD) was validated. This method was used to evaluate the pfaffic acid content in 30 different genotypes of the species from a germplasm collection. The content of pfaffic acid ranged from 0.97 to 4.29% (w/w) on a dry weight basis. © 2013 Elsevier B.V.

Genetic erosion risk by environmental and antropic factor applied on strategies for Lychnophora ericoides conservation

This study evaluated the genetic erosion risk factors and the strategic points for the conservation of Lychnophora ericoides population in “Paraíso Perdido” farm, Serra da Canastra (20° 37’ 54” S; 46° 19’ 37” W; 833 m height) in São João Batista do Glória City, Minas Gerais State, Brazil. The number of young and adult plants, the soil and the phenology were evaluated in two sample areas of 125 m2. Information about the species utilization was obtained with local informants. Data on the region were obtained through literature review, in loco evaluation, GPS and geo-referenced map. In addition, local use of the plant for mixtures of drug was evaluated. According to the results obtained, the soil of the population is lithic with a weathered portion of frank-sandy texture, very acidic and dystrophic. The population density is 0.16 individuals/m2, 0.078 young/adult plant. The predominant phenophase was fruiting (100% plants) followed by flowering (21.62% plants). The local community uses the leaves of the plant in the form of hydroalcoholic extracts, as anti-inflammatory. Based on the evaluated parameters, the population is at 73% risk of genetic erosion. The detected key points were the development of activities including the participation of the local community for habitat protection as well as germplasm collection, seedlings production and reintroduction, together with environmental education, supervision, and reduction in the propensity for fire.

Genetic variability of banana with ornamental potential

The Musa germplasm collection at Embrapa Cassava and Fruits detains accessions from different sections of the Musa genus. The objective of the present study was to identify and morphologically characterize banana accessions from the banana germplasm with ornamental potential, as well as to quantify their genetic variability; and identify possible progenitors to be used in breeding aiming to achieve ornamental crossbreeds. The accessions were evaluated with the use of 32 morphological descriptors. Then, they were the following grouped into categories: landscape plants, cut flower, potted plants, and male inflorescence minifruits. The pre-selected accessions presented great genetic variability and ornamental potential for different uses. The accessions of the Rhodochlamys and Callimusa sections were selected to be used as landscape plants, cut flowers, potted plants, male inflorescence and minifruits. Most of the diploids from the Eumusa section evaluated in this study are indicated for the production of ornamental minifruits, except for 'Lidi' and Cici, which can also be indicated as landscape plants. The BB diploids have great potential for the use of the male inflorescence in floral arrangements, and did not offer any other indication.

Molecular strategies for genetic diversity analysis and development of markers linked to resistance traits in apple

The goal of many plant scientists’ research is to explain natural phenotypic variation in term of simple changes in DNA sequence. DNA-based molecular markers are extensively used for the construction of genome-wide molecular maps and to perform genetic analysis for simple and complex traits. The PhD thesis was divided into two main research lines according to the different approaches adopted. The first research line is to analyze the genetic diversity in an Italian apple germplasm collection for the identification of markers tightly linked to targeted genes by an association genetic method. This made it possible to identify synomym and homonym accessions and triploids. The fruit red skin color trait has been used to test the reliability of the genetic approaches in this species. The second line is related to the development of molecular markers closely linked to the Rvi13 and Rvi5 scab resistance genes, previously mapped on apple’s chromosome 10 and 17 respectively by using the traditional linkage mapping method. Both region have been fine-mapped with various type of markers that could be used for marker-assisted selection in future breeding programs and to isolate the two resistance genes.

The isolation, characterization, and application of WRKY genes as useful molecular markers in tropical trees

The improvement of tropical tree crops using conventional breeding methods faces challenges due to the length of time involved. Thus, like most crops, there is an effort to utilize molecular genetic markers in breeding programs to select for desirable agronomic traits. Known as marker assisted breeding or marker assisted selection, genetic markers associated with a phenotype of interest are used to screen and select material reducing the time necessary to evaluate candidates. As the focus of this research was improving disease resistance in tropical trees, the usefulness of the WRKY gene superfamily was investigated as candidates for generating useful molecular genetic markers. WRKY genes encode plant-specific transcriptional factors associated with regulating plants' responses to both biotic and abiotic stress. ^ One pair of degenerate primers amplified 48 WRKY gene fragments from three taxonomically distinct, economically important, tropical tree crop species: 18 from Theobroma cacao L., 21 from Cocos nucifera L. and 9 from Persea americana Mill. Several loci from each species were polymorphic because of single nucleotide substitutions present within a putative non-coding region of the loci. Capillary array electrophoresis-single strand conformational polymorphism (CAE-SSCP) mapped four WRKY loci onto a genetic linkage map of a T. cacao F2 population segregating for resistance to witches' broom disease. Additionally, PCR primers specific for four T. cacao loci successfully amplified WRKY loci from 15 members of the Byttneriae tribe. A method was devised to allow the reliable discrimination of alleles by CAE-SSCP using only the mobility assigned to the sample peaks. Once this method was validated, the diversity of three WRKY loci was evaluated in a germplasm collection of T. cacao . One locus displayed high diversity in the collection, with at least 18 alleles detected from mobility differences of the product peaks. The number of WRKY loci available within the genome, ease of isolation by degenerate PCR, codominant segregation demonstrated in the F2 population, and usefulness for screening germplasm collections and closely related wild species demonstrates that the WRKY superfamily of genes are excellent candidates for developing a number of genetic molecular markers for breeding purposes in tropical trees. ^

Molecular analysis of evolutionary relationships of Phaseolus dumosus with the gene pool of common bean

Valuable genetic variation for bean breeding programs is held within the common bean secondary gene pool which consists of Phaseolus albescens, P. coccineus, P. costaricensis, and P. dumosus. However, the use of close relatives for bean improvement is limited due to the lack of knowledge about genetic variation and genetic plasticity of many of these species. Characterisation and analysis of the genetic diversity is necessary among beans' wild relatives; in addition, conflicting phylogenies and relationships need to be understood and a hypothesis of a hybrid origin of P. dumosus needs to be tested. This thesis research was orientated to generate information about the patterns of relationships among the common bean secondary gene pool, with particular focus on the species Phaseolus dumosus. This species displays a set of characteristics of agronomic interest, not only for the direct improvement of common bean but also as a source of valuable genes for adaptation to climate change. Here I undertake the first comprehensive study of the genetic diversity of P. dumosus as ascertained from both nuclear and chloroplast genome markers. A germplasm collection of the ancestral forms of P. dumosus together with wild, landrace and cultivar representatives of all other species of the common bean secondary gene pool, were used to analyse genetic diversity, phylogenetic relationships and structure of P. dumosus. Data on molecular variation was generated from sequences of cpDNA loci accD-psaI spacer, trnT-trnL spacer, trnL intron and rps14-psaB spacer and from the nrDNA the ITS region. A whole genome DArT array was developed and used for the genotyping of P. dumosus and its closes relatives. 4208 polymorphic markers were generated in the DArT array and from those, 742 markers presented a call rate >95% and zero discordance. DArT markers revealed a moderate genetic polymorphism among P. dumosus samples (13% of polymorphic loci), while P. coccineus presented the highest level of polymorphism (88% of polymorphic loci). At the cpDNA one ancestral haplotype was detected among all samples of all species in the secondary genepool. The ITS region of P. dumosus revealed high homogeneity and polymorphism bias to P. coccineus genome. Phylogenetic reconstructions made with Maximum likelihood and Bayesian methods confirmed previously reported discrepancies among the nuclear and chloroplast genomes of P. dumosus. The outline of relationships by hybridization networks displayed a considerable number of interactions within and between species. This research provides compelling evidence that P. dumosus arose from hybridisation between P. vulgaris and P. coccineus and confirms that P. costaricensis has likely been involved in the genesis or backcrossing events (or both) in the history of P. dumosus. The classification of the specie P. persistentus was analysed based on cpDNA and ITS sequences, the results found this species to be highly related to P. vulgaris but not too similar to P. leptostachyus as previously proposed. This research demonstrates that wild types of the secondary genepool carry a significant genetic variation which makes this a valuable genetic resource for common bean improvement. The DArT array generated in this research is a valuable resource for breeding programs since it has the potential to be used in several approaches including genotyping, discovery of novel traits, mapping and marker-trait associations. Efforts should be made to search for potential populations of P. persistentus and to increase the collection of new populations of P. dumosus, P. albescens and P. costaricensis that may provide valuable traits for introgression into common bean and other Phaseolus crops.

Seleção e divergência genética de pessegueiros e nectarineiras e susceptibilidade a danos causados pelo frio na Estación Experimental de Aula Dei (CSIC)

The peaches and nectarines are highly appreciated by consumer, but it is climacteric fruits, with availability in the market in small time. It is necessary to invest to obtain genotypes with fruit quality and small perishability or that it presente less physiological disorders after storage. The aims of this work were i) to evaluate the genetic divergence among 40 peach and nectarine trees genotypes based on postharvest quality and select posible parents; ii) to evaluate the susceptibility to chilling injury in peaches and nectarines after cold storage; iii) to evaluate divergence of peaches and nectarines on the basis in the susceptibility for chiling injury and select superior genotypes; iv) evaluate the correlations between quality and susceptibility to chilling injury of peaches and nectarines v) select parents with the combination of lower susceptibility to chilling injury and higher quality fruit. The study was carried out in EEAD-CSIC, Zaragoza - Spain, during the production cycle 2013/2014. A total of 40 peaches and nectarines genotypes from germplasm collection were evaluated. The quality characteristics as flesh firmness, total soluble solids, titratable acidity, pH, rippining index and flesh color parameters were evaluated. The fruits were submitted to cold storage at 0 °C and 5 °C, with 95% average relative humidity. The evaluations were after 14 and 28 days, it being observed the presence of symptoms, such as wooliness through mealiness, flesh grainy, leatheriness and flesh color changes, through browning, bleeding and off flavor. As a selection parameter was adopted 20% of genotypes that had a higher frequency of superiority for quality characteristics, susceptibility to chilling injury and the combining of both. For quality characteristic presented greater divergence the ‘Queen Giant’, ‘Sudanel Blanco’ and ‘Borracho de Jarque’. Based on the quality the eight genotypes were selected, ‘Andross’, ‘San Jaime’, ‘San Lorenzo’, ‘Borracho de Jarque’, ‘Sudanell 1’, ‘Carson’, ‘Baby Gold 6’ and ‘Stanford’. All genotypes studied exhibited susceptibility to one or more symptoms caused by cold storage during 28 days, independent of temperature. For 14 days, the ‘Baby Gold 6’, ‘Flavortop’ and ‘Queen Giant’ genotypes did not show any physiological disorder caused by cold. In general, the temperature of 0 °C favored fruit postharvest conservation, it have a lower incidence and severity of symptoms caused by cold storage. The storage for 14 days contributed for the lower incidence of damage in the genotypes fruits studied. For 14 days, with both temperatures, it was observed divergence for ‘Queen Giant’, ‘Sudanell Blanco’, ‘Baby Gold 6’ ‘GF3’, ‘Baby Gold 8’, ‘Campiel’ and ‘Campiel Rojo’ genotypes. For 28 days, in the 5 °C condition, ‘Queen Giant’, ‘Big Top’, ‘Flavortop’ and ‘Redhaven’ genotypes were divergents. Based on susceptibility to chilling injury at 0 °C, the eight genotypes were selected, it being these, ‘Queen Giant’, ‘Keimoes’, ‘Flavortop’, ‘Big Top’, 'Redhaven', 'Sudanell 3', 'Bonet I' and ‘Carson’. The quality parameters as rippining index, soluble solids, firmness and titratable acidity presented correlation among them. These, also it had correlation with woolines and bowning, what it indicate that fruits with more ripening can have this symptoms more easily. The browning, mealiness, flesh grainy and off flavor variables were correlationed with the time period and temperartures, what it confirm that these symptoms are the main disorders caused by cold storage. The quality characteristics together susceptibility to chilling injury allowed selected ‘Baby Gold 6’, ‘Sarell’, ‘Keimoes’, ‘GF3’ ‘San Jaime’, ‘Big Top’, ‘Sudanell 1’, ‘Carson’, ‘Baby Gold 8’, and ‘San Lorenzo’ genotypes.

Bananas adrift in time – a case study in the Solomons

Diseases, pests and environmental constraints pose a major threat to the sustainability of banana production globally. To address these challenges, the discovery and study of new sources of genetic resistance and adaptability are required, along with the general conservation of diversity. The Solomon Islands, located in the south-western Pacific region near Papua New Guinea, are a major center of banana diversity. Some collections had been made by nationals of some of the diversity present but little was known internationally of the rich genetic resource present. Two separate visits to the Solomon Islands characterized banana collections, documented and collected germplasm, recommended conservation strategies and provided training in cultivar characterization. A remarkable range of genetic diversity was found, including: many AA and AAA cooking types somewhat like those present in Papua New Guinea; nine Fei cultivars in relatively common usage, and two undescribed wild species as well as five AAB Pacific Plantains and four ABB cooking bananas belonging to the Kalapua subgroup. About six of the unique cultivars were successfully collected and established in the regional in vitro germplasm collection of SPC in Suva, Fiji. Nine Solomon Islanders were trained in the finer points of characterizing banana cultivars. Further collecting and study/evaluation of this rich diversity will promote its appreciation and potential utilization for meeting the challenges and opportunities ahead. Future studies could also determine the spread of the Awawe species and variability of morphological traits in the population. Community-based conservation could promote awareness of dietary diversity for better nutrition, via using the Fei bananas described in this paper. Establishing a virus-free regional field collection could help in comprehensively characterizing and evaluating regional Musa genetic resources. Existing sites could embrace the broader unique diversity of the Solomon Islands, and facilitate sharing this diversity in conjunction with a regional virus-tested in vitro collection.

Assessment of genetic diversity and identification of core collection in sandalwood germplasm using RAPDS

Sandalwood is an economically important aromatic tree belonging to the family Santalaceae. The trees are used mainly for their fragrant heartwood and oil that have immense potential for foreign exchange. Very little information is available on the genetic diversity in this species. Hence studies were initiated and genetic diversity estimated using RAPD markers in 51 genotypes of Santalum album procured from different geographcial regions of India and three exotic lines of S. spicatum from Australia. Eleven selected Operon primers (10mer) generated a total of 156 consistent and unambiguous amplification products ranging from 200bp to 4kb. Rare and genotype specific bands were identified which could be effectively used to distinguish the genotypes. Genetic relationships within the genotypes were evaluated by generating a dissimilarity matrix based on Ward's method (Squared Euclidean distance). The phenetic dendrogram and the Principal Component Analysis generated, separated the 51 Indian genotypes from the three Australian lines. The cluster analysis indicated that sandalwood germplasm within India constitutes a broad genetic base with values of genetic dissimilarity ranging from 15 to 91 %. A core collection of 21 selected individuals revealed the same diversity of the entire population. The results show that RAPD analysis is an efficient marker technology for estimating genetic diversity and relatedness, thereby enabling the formulation of appropriate strategies for conservation, germplasm management, and selection of diverse parents for sandalwood improvement programmes.