Genetic diversity among air yam (Dioscorea bulbifera) varieties based on single sequence repeat markers.

Dioscorea is the largest genus in the Dioscoreaceae family, and includes a number of economically important species including the air yam, D. bulbifera L. This study aimed to develop new single sequence repeat primers and characterize the genetic diversity of local varieties that originated in several municipalities of Brazil. We developed an enriched genomic library for D. bulbifera resulting in seven primers, six of which were polymorphic, and added four polymorphic loci developed for other Dioscorea species. This resulted in 10 polymorphic primers to evaluate 42 air yam accessions. Thirty-three alleles (bands) were found, with an average of 3.3 alleles per locus. The discrimination power ranged from 0.113 to 0.834, with an average of 0.595. Both principal coordinate and cluster analyses (using the Jaccard Index) failed to clearly separate the accessions according to their origins. However, the 13 accessions from Conceição dos Ouros, Minas Gerais State were clustered above zero on the principal coordinate 2 axis, and were also clustered into one subgroup in the cluster analysis. Accessions from Ubatuba, São Paulo State were clustered below zero on the same principal coordinate 2 axis, except for one accession, although they were scattered in several subgroups in the cluster analysis. Therefore, we found little spatial structure in the accessions, although those from Conceição dos Ouros and Ubatuba exhibited some spatial structure, and that there is a considerable level of genetic diversity in D. bulbifera maintained by traditional farmers in Brazil.

Distribution, management and diversity of the endangered Amerindian yam (Dioscorea trifida L.).

The objective of this study was to verify the occurrence of Dioscorea trifida in Brazil and to obtain information concerning its distribution, management and diversity. Farmers from 21 communities were interviewed in the states of São Paulo, Santa Catarina and Mato Grosso. During the visits, semi-structured interviews were conducted to collect socio-economic, management and diversity data for this crop. Fifty-one collected accessions, plus two accessions obtained at local markets of Amazonas, were characterized using 12 morphological traits. Most the interviewed farmers were men (75%) with a mean age of 59.5 years. Just a few young people and labor force were available for agricultural activities, with an average of only three individuals per farm. Most farmers (56%) grew only one variety of D. trifida, although 44% had more than one variety in their fields, which aims to provide greater assurance at harvest. Many popular names were observed for D. trifida, and cará roxo (purple yam) was the name most used by farmers (43.4%). Characters referring to the tuber, such as skin and flesh color, were most relevant for the distinction of the accessions. The results of this study may collaborate to develop strategies for conservation, both ex situ and in situ, within the view of on farm conservation.

Distribution, management and diversity of the endangered Amerindian yam (Dioscorea trifida L) / Distribuição, manejo e diversidade da espécie ameaçada de inhame ameríndio (Dioscorea trifida L)

The objective of this study was to verify the occurrence of Dioscorea trifida in Brazil and to obtain information concerning its distribution, management and diversity. Farmers from 21 communities were interviewed in the states of São Paulo, Santa Catarina and Mato Grosso. During the visits, semi-structured interviews were conducted to collect socio-economic, management and diversity data for this crop. Fifty-one collected accessions, plus two accessions obtained at local markets of Amazonas, were characterized using 12 morphological traits. Most the interviewed farmers were men (75%) with a mean age of 59.5 years. Just a few young people and labor force were available for agricultural activities, with an average of only three individuals per farm. Most farmers (56%) grew only one variety of D. trifida, although 44% had more than one variety in their fields, which aims to provide greater assurance at harvest. Many popular names were observed for D. trifida, and cará roxo (purple yam) was the name most used by farmers (43.4%). Characters referring to the tuber, such as skin and flesh color, were most relevant for the distinction of the accessions. The results of this study may collaborate to develop strategies for conservation, both ex situ and in situ, within the view of on farm conservation.(AU)

O objetivo deste trabalho foi verificar a ocorrência de Dioscorea trifida no Brasil e obter informações a respeito da sua distribuição, manejo e diversidade. Para tanto, foram visitados e entrevistados agricultores de 21 comunidades, nos Estados de São Paulo, Santa Catarina, Mato Grosso. Durante as visitas, foram realizadas entrevistas semi-estruturadas visando obter dados sócioeconômicos, de manejo e diversidade da cultura. Cinquenta e um acessos, além de dois acessos adquiridos em feiras no Estado do Amazonas, foram caracterizados por meio de 12 descritores morfológicos. A maioria dos entrevistados era homem (75%) com média de idade de 59,5 anos. Observou-se um número reduzido de jovens e mão de obra disponível para as atividades agrícolas, cerca de três indivíduos por roça. A maior parte dos agricultores (56%) cultiva apenas uma variedade de D. trifida, embora 44% tenham declarado o cultivo de mais de uma variedade, procedimento que visa dar maior garantia de colheita. Vários nomes populares foram observados para D. trifida, sendo cará roxo, a denominação mais utilizada pelos agricultores (43,4%). Os caracteres referentes às túberas, como cor da casca e da polpa, foram os mais relevantes para a distinção dos acessos. Os resultados obtidos poderão colaborar para elaborar estratégias de conservação, tanto ex situ como in situ, dentro da visão de conservação on farm.(AU)

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.

Distribution, management and diversity of yam local varieties in Brazil: a study on Dioscorea alata L.

Widely spread in the tropics, yams were introduced into Brazil during the colonial period and are currently grown throughout the country. Despite its importance as a pharmacological and food source, there is a lack of studies describing how and where this tuber is grown in Brazil. The aim of this study was to provide an overview of the cultivation and distribution of Dioscorea alata in different Brazilian regions. A total of 63 farmers were visited in different municipalities and communities in four regions in the country: South, Southeast, Northeast and Midwest. Semi-structured interviews were conducted to collect socio-economic, use, management and diversity data for this crop. The majority of interviewers were men, married, with children, using retirement benefits and agriculture as income and family labour as the main support in the yam cultivation. A wide distribution of this species was found, with the occurrence of D. alata in the four sampled regions. A variety of vernacular names for this species was collected, differing according to the region where it is cultivated. Most farmers cultivate yams in fields, however an increased usage of home gardens for the cultivation of this tuber was found. Also, most farmers cultivate yams in association with other crops in areas of different sizes and slash and burn practices, although mostly disappearing, are still being used by many farmers. The results of this study provide more concrete data on the distribution and diversity of this important crop.

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.

Genetic structure and diversity in the Dioscorea cayenensis/D. rotundata complex revealed by morphological and isozyme markers.

Of the 600 known yam species, only 10 are utilized as food, and the Dioscorea cayenensis/D. rotundata species complex is among the most cultivated. In Brazil, these species are commercially cultivated in the northeast region and are cultivated in the south and southeast regions as subsistence crops by traditional agriculturists. This study aimed to evaluate the genetic diversity of 21 local varieties of D. cayenensis and 2 D. rotundata accessions using 7 isozymic loci and 24 morphological markers, and to investigate the diversity distribution in different levels of organization, such as swidden fields and communities of Vale do Ribeira. Cluster analyses for both the isozymic and morphological data separated the 2 D. rotundata accessions from the D. cayenensis accessions from Vale do Ribeira. The analysis with morphological characteristics showed the presence of 2 subgroups (Iguape and Cananéia) within group I, which included all of the local varieties from Vale do Ribeira; this result may indicate the influence of the cultural units on the morphological variation. Molecular analysis of variance indicated that most of the isozymic variability was concentrated among swiddens within communities (42.5%) and within communities (40.3%). Most of the morphological variability was also concentrated among swidden fields within communities (44.8%). The correlation between geographic and genetic distances indicated that neither morphological (r = 0.17) nor isozymic diversity (r = -0.15) is structured in space. Thus, the traditional agriculturists of Vale do Ribeira maintain and manage a great diversity of D. cayenensis varieties in their communities.

Simple-sequence repeat markers of Cattleya coccinea (Orchidaceae), an endangered species of the Brazilian Atlantic Forest.

Microsatellite markers were developed for the endangered Brazilian orchid species Cattleya coccinea to describe its genetic diversity and structure and to support conservation studies. Nine microsatellite loci were isolated and characterized using an enriched genomic library. All loci are polymorphic at least in the 2 populations sampled, except for loci Cac05 and Cac09 for the Petrópolis population. The mean number of alleles per locus was 8.8 between populations. The mean values of the observed and expected heterozygosities were 0.541 (ranging from 0 to 1) and 0.639 (ranging from 0 to 0.9), respectively. Cross-amplifications were performed in 7 additional Epidendroideae species, and at least 2 loci were successful in 3 additional Cattleya species, Epidendrum secundum, and Brasiliorchis gracilis. All markers described herein will be useful in further studies evaluating the genetic diversity, population dynamics, and conservation genetics of C. coccinea and related species.

Microsatellite polymorphisms in cassava landraces from the Cerrado biome, Mato Grosso do sul, Brazil.

Using nine microsatellite loci, we investigated genetic structure and diversity in 83 Brazilian cassava accessions, including several landraces, in the Cerrado biome in Mato Grosso do Sul, Brazil. All nine loci were polymorphic, averaging 6.00 alleles per locus. Treating each of seven municipalities as a cassava group or population, they averaged 3.5 alleles per locus, with 97% polymorphic loci, high values for observed heterozygosity (0.32) and gene diversity (0.56). Total genetic variability was high (0.668), and most of this genetic variability was concentrated within municipalities (0.577). Cluster and structure analyses divided accessions into two major clusters or populations (K = 2). Also, a significant genetic versus geographic correlation was found (r = 0.4567; P < 0.0260). Migratory routes in the Cerrado are considered main contributors to the region's high cassava diversity and spatial genetic structure, amplifying interactions between traditional farmers and the evolutionary dynamics of this crop.

Variation in the loss of seed dormancy during after-ripening of wild and cultivated rice species.

BACKGROUND AND AIMS: The aim of this paper was to verify the variation in the loss of seed dormancy during after-ripening and the interspecific and interpopulation variability in the degree of dormancy of seven wild and two cultivated rice species comprising 21 populations and two cultivars. METHODS: Four wild rice species from South America, Oryza glumaepatula, O. latifolia, O. grandiglumis and O. alta, and two O. sativa cultivars were tested in one experiment. In a second experiment, five wild species, O. punctata, O. eichingeri, O.rufipogon, O. latifolia and O. glumaepatula, and one cultivated species (O. glaberrima) were evaluated. Initial germination tests were performed soon after the seeds were harvested and subsequently at 2-month intervals, for a total of six storage periods in the first experiment and three in the second. All tests were conducted in the dark at a temperature of 27 degrees C. KEY RESULTS: Different patterns of after-ripening among populations within and between species were observed. CONCLUSIONS: The cultivated species (O. sativa and O. glaberrima) and, amongst the wild species, the tetraploids O. latifolia, O. grandiglumis and the diploids O. eichingeri and O. punctata, had weak dormancy, losing it completely 2 months after harvest, while O. rufipogon and O. glumaepatula exhibited pronounced dormancy. The latter showed different patterns of after-ripening between populations indigenous to the Amazon region and those originating in the Paraguay River system. Seeds of Solimoes (Amazon) and Japura origin showed weak dormancy whereas those of Paraguay origin showed deep dormancy. Ecological differences among natural habitats may be involved in such differentiation.

Biochemical and isoenzyme analyses of elephant grass, Pennisetum purpureum (Schum) varieties

This study characterized seven Pennisetum purpureum varieties, namely v. Anão, Bajra, Cameroon, Guaçu, Roxo, Taiwan A-144 and Uruckwami, through biochemical analyses, including protein, glucose and fructose contents, and polyacrylamide gel electrophoresis using the esterase system, by sampling 30, 60, 90, 120 and 150 day-old leaves. The number of nodes per stem and the percentage of bud emergence were also recorded. Variety Taiwan A-144 presented the highest number of nodes per stem and percentage of emerging buds. Protein concentration decreased gradually after 60 days for all varieties, except for Anão. Variety Guaçu presented the highest level of glucose in 90 day-old plants, whereas Cameroon presented the highest levels at 120 and 150 days. The esterase band patterns changed with plant age for all varieties, showing a tendency to increase the number of bands with time. The best age for discriminating between esterase bands of P. purpureum varieties was at 120 days, when most variation could be observed.

Foram analisadas sete variedades de P. purpureum, tais como: Anão, Bajra, Cameroon, Guaçu, Roxo, Taiwan A-144 e Uruckwami, através de testes bioquímicos determinando a concentração de proteínas, glicose e frutose, além de observar, através de gel de eletroforese, as bandas de esterase. Os extratos protéicos foram preparados amostrando-se folhas com 30, 60, 90, 120 e 150 dias. Os números de nós por colmo e a porcentagem de brotamento das gemas foram também avaliados. A variedade Taiwan A-144 apresentou o maior número de nós por colmo e porcentagem de brotamento das gemas. Com relação à concentração de proteínas, esta decresceu gradualmente após 60 dias para todas as variedades, com exceção da Anão. A maior concentração de glicose foi apresentada pela variedade Guaçu aos 90 dias e pela Cameroon aos 120 e 150 dias. Os padrões eletroforéticos da esterase se alteraram com a idade da planta para todas as variedades, mostrando tendência a aumentar o número de bandas com o tempo. A melhor idade para discriminar as variedades de capim elefante foi aos 120 dias, onde foi encontrada maior variação.

Biochemical and isoenzyme analyses of elephant grass, Pennisetum purpureum (Schum) varieties

This study characterized seven Pennisetum purpureum varieties, namely v. Anão, Bajra, Cameroon, Guaçu, Roxo, Taiwan A-144 and Uruckwami, through biochemical analyses, including protein, glucose and fructose contents, and polyacrylamide gel electrophoresis using the esterase system, by sampling 30, 60, 90, 120 and 150 day-old leaves. The number of nodes per stem and the percentage of bud emergence were also recorded. Variety Taiwan A-144 presented the highest number of nodes per stem and percentage of emerging buds. Protein concentration decreased gradually after 60 days for all varieties, except for Anão. Variety Guaçu presented the highest level of glucose in 90 day-old plants, whereas Cameroon presented the highest levels at 120 and 150 days. The esterase band patterns changed with plant age for all varieties, showing a tendency to increase the number of bands with time. The best age for discriminating between esterase bands of P. purpureum varieties was at 120 days, when most variation could be observed.

Foram analisadas sete variedades de P. purpureum, tais como: Anão, Bajra, Cameroon, Guaçu, Roxo, Taiwan A-144 e Uruckwami, através de testes bioquímicos determinando a concentração de proteínas, glicose e frutose, além de observar, através de gel de eletroforese, as bandas de esterase. Os extratos protéicos foram preparados amostrando-se folhas com 30, 60, 90, 120 e 150 dias. Os números de nós por colmo e a porcentagem de brotamento das gemas foram também avaliados. A variedade Taiwan A-144 apresentou o maior número de nós por colmo e porcentagem de brotamento das gemas. Com relação à concentração de proteínas, esta decresceu gradualmente após 60 dias para todas as variedades, com exceção da Anão. A maior concentração de glicose foi apresentada pela variedade Guaçu aos 90 dias e pela Cameroon aos 120 e 150 dias. Os padrões eletroforéticos da esterase se alteraram com a idade da planta para todas as variedades, mostrando tendência a aumentar o número de bandas com o tempo. A melhor idade para discriminar as variedades de capim elefante foi aos 120 dias, onde foi encontrada maior variação.