Academic literature on the topic 'Molecular markers ISSR'

Fifty-seven accessions of torch ginger (Etlingera elatior) collected from seven states in Peninsular Malaysia were evaluated for their molecular characteristics using ISSR and SSR markers to assess the pattern of genetic diversity and association among the characteristics. Diversity study through molecular characterization showed that high variability existed among the 57 torch ginger accessions. ISSR and SSR molecular markers revealed the presence of high genetic variability among the torch ginger accessions. The combination of different molecular markers offered reliable and convincing information about the genetic diversity of torch ginger germplasm. This study found that SSR marker was more informative compared to ISSR marker in determination of gene diversity, polymorphic information content (PIC), and heterozygosity in this population. SSR also revealed high ability in evaluating diversity levels, genetic structure, and relationships of torch ginger due to their codominance and rich allelic diversity. High level of genetic diversity discovered by SSR markers showed the effectiveness of this marker to detect the polymorphism in this germplasm collection.

Inter-simple sequence repeat (ISSR) markers were used for cultivar identification and for determination of the phenetic relationships among 24 pear cultivars (Pyrus communis L.). The ability of several molecular marker systems including randomly amplified polymorphic DNA (RAPD), amplified fragment length polymorphisms (AFLP), inter-simple sequence repeats (ISSR), simple sequence repeats (SSR), and selective amplification of microsatellite polymorphic loci (SAMPL) to detect variation among clones of the most significant Portuguese cultivar, Rocha, was also investigated. Each of the eight ISSR primers tested was able to distinguish the 24 pear cultivars. The ISSR primers generated 337 markers, 79.5% of which were polymorphic. The cultivar dendrogram obtained with the ISSR marker data was very similar to that obtained with previous RAPD+AFLP analysis, confirming the genetic divergence of `Pérola', `Carvalhal' and `Lawson' from the other cultivars. Eight out of 15 apple [Malus sylvestris (L.) Mill. var domestica (Borkh.) Mansf.] SSR primers tested also amplified microsatellites in pear. None of the five molecular marker systems analyzed (with a total of 1082 markers) detected reproducible polymorphisms among the nine `Rocha' clones, in spite of the presence of clear phenotypic differences.

The identification and genetic relationships of 23 coffee species and one coffee-related species Canthium diccocum were studied using ISSR and SRAP markers. The average polymorphism information content of SRAP primers (0.81) was lower than ISSR primers (0.86), whereas the average resolving power of the SRAP primers (9.74) is higher than the ISSR primers (8.64). The genetic similarity among the species ranged from 0.30 to 0.89 using ISSR and 0.11 to 0.90 using SRAP marker systems. Based on marker analysis, all twenty three coffee species were clustered into two major groups. Both the markers amplified species-specific fragments and are useful in genetic diversity analysis of coffee.

Red maple (Acer rubrum) and silver maple (A. saccharinum) are sister species that readily hybridize in nature. No genetic or barcoding markers have been tested in these species. The main objective of the present study is to develop and characterize molecular markers for distinguishing A. rubrum and A. saccharinum and to validate the hybridity of A. freemanii derived from their crossings using the ISSR marker system. Thirteen A. rubrum and seven A. saccharinum populations were used. Four ISSR primers including ISSR 5, ISSR 8, ISSR 10, and ISSR UBC 825 were selected to amplify genomic DNA from the two species and their hybrids. Each primer generated at least one species-diagnostic ISSR marker for a total of six. Analysis of A. freemanii collected from North Dakota (USA) confirmed that the genotypes screened were true hybrids between A. rubrum and A. saccharinum. These markers were cloned and sequenced. Successful sequences were converted to SCAR markers using specifically designed primers. Overall, the developed diagnostic and specific ISSR and SCAR markers are useful in the certification of these two maple species and their hybrids. They can be used in tracking the introgression of A. rubrum and A. saccharinum DNA in other hybrid trees or populations.

Molecular genetic diversity and relationships among 86 Chrysopogon aciculatus (Retz.) Trin. accessions were assessed using intersimple sequence repeat (ISSR) and sequence-related amplified polymorphism (SRAP) markers. Twenty-five ISSR markers generated 283 amplification bands, of which 266 were polymorphic. In addition, 576 polymorphic bands were detected from 627 bands amplified using 30 SRAP primers. Both marker types revealed a high level of genetic diversity, with ISSR markers showing a higher proportion of polymorphic loci (PPL; 94%) than SRAP markers (91.87%). The ISSR and SRAP data were significantly correlated (r = 0.8023). Cluster analysis of the separate ISSR and SRAP data sets clustered the accessions into three groups, which generally were consistent with geographic provenance. Cluster analysis of the combined ISSR and SRAP data set revealed four major groups similar to those based solely on ISSR or SRAP markers. The findings demonstrate that ISSR and SRAP markers are reliable and effective tools for analysis of genetic diversity in C. aciculatus.

AbstractThe aim of this study is investigation the applicability of SSR and ISSR markers in evaluating the genetic relationships in twenty accessions ofAegilopsandTriticumspecies with D genome in different ploidy levels. Totally, 119 bands and 46 alleles were detected using ten primers for ISSR and SSR markers, respectively. Polymorphism Information Content values for all primers ranged from 0.345 to 0.375 with an average of 0.367 for SSR, and varied from 0.29 to 0.44 with the average 0.37 for ISSR marker. Analysis of molecular variance (AMOVA) revealed that 81% (ISSR) and 84% (SSR) of variability was partitioned among individuals within populations. Comparing the genetic diversity ofAegilopsandTriticumaccessions, based on genetic parameters, shows that genetic variation ofAe. crassaandAe. tauschiispecies are higher than other species, especially in terms of Nei’s gene diversity. Cluster analysis, based on both markers, separated total accessions in three groups. However, classification based on SSR marker data was not conformed to classification according to ISSR marker data. Principal co-ordinate analysis (PCoA) for SSR and ISSR data showed that, the first two components clarified 53.48% and 49.91% of the total variation, respectively. This analysis (PCoA), also, indicated consistent patterns of genetic relationships for ISSR data sets, however, the grouping of accessions was not completely accorded to their own geographical origins. Consequently, a high level of genetic diversity was revealed from the accessions sampled from different eco-geographical regions of Iran.

. Molecular genetic analysis of three plants of thistle family has been carried out in the scope of this paper (Artemisia karatavica Krasch. & Abolin ex Poljakov, Artemisia cina Berg ex Poljakov, Artemisia porrecta Krasch. ex Poljakov). The plants have been collected in Turkestan region, Shardarinsky district, 15 km north-east of the village Komsomol, and along the road in Turkestan region, Baydibeksky district, 4.5 km south-east of the village Shakpak, and in the steppes in Turkestan region, Aryssky district, 1 km north-east of the village Darmino. In this paper, we used modern methods of molecular biology in order to determine genetic relatedness. ISSR analysis using universal primers has been conducted. ISSR-markers are the most common markers, and they are used for phylogenic analysis. This method is based on amplification of sequences limited by two microsatellite repeats using the primer that is complementary to the sequence of this microsatellite (4-12 repeat units). ISSR (region of the genome between two adjacent, oppositely oriented microsatellites) the sequence of microsatellite medullar part with some (1-3) nucleotides adjacent to the repeat tandem are used as primers. Tens of fragments of locus variety received in PCR are separated by electrophoresis and assessed for the presence or absence (due to marker dominance) of the fragments of a particular size. The main advantage of this type of markers - lack of necessity for knowledge of the sequences during primer designing.

Nineteen cashew accessions were analysed with 50 random primers, 12 ISSR primers and 6 AFLP primer pairs to compare the efficiency and utility of these techniques for detecting variation in cashew germplasm. Each marker system could discriminate between all of the accessions, albeit with varied efficiency of polymorphism detection. AFLP exhibited maximum discrimination efficiency with a genotype index of 1. The utility of each molecular marker technique, expressed as marker index, was estimated as a function of average band informativeness and effective multiplex ratio. Marker index was calculated to be more than 10 times higher in AFLP than in RAPD and ISSR. Similarity matrices were determined based on the data generated by molecular and morphometric analyses, and compared for congruency. AFLP displayed no correspondence with RAPD and ISSR. Correlation between ISSR and RAPD similarity matrices was low but significant (r = 0.63; p < 0.005). The similarity matrix based on morphometric markers exhibited no correlation with any of the molecular markers. AFLP, with its superior marker utility, was concluded to be the marker of choice for cashew genetic analysis.Key words: Anacardium occidentale, DNA fingerprinting, RAPD, ISSR, AFLP, morphometric.

Molecular markers are the most ideal approach to study genetic diversity. Consequently, we utilized both ISSR and RAPD markers to assess genetic diversity and relationships among three different populations of Farafra, Ossimi and Rahmani Egyptian sheep breeds. Both ISSR and RAPD gave moderate polymorphism 41.3% and 48.51%, respectively. Besides, this value was consistent with the moderate value of the mean of polymorphism information content (0.16 and 0.20, respectively). Farafra-F and Farafra-D populations had the highest similarity which was 0.92 for ISSR and 0.90 for the RAPD marker. Furthermore, ISSR and RAPD constructed dendrogram separated all the studied sheep into two main clusters. All the three populations of Farafra breed combined into one main cluster, while the second cluster contained both Rahmani and Ossimi breeds. The used molecular markers were able to discriminate among evaluated sheep and displayed that Farafra breed more closely related to Ossimi than Rahmani breed.

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Approximately 80-90% of grassland areas in Brazil consist of the forage Urochloa, genus and the apomictic species Urochloa brizantha [syn. Brachiaria brizantha (Hochst. ex A. Rich.) Stapf.] the most used. Some genotypes of Urochloa have being widely used with a wrong nomenclature, even for species as for cultivars. In this way, the Urochloa cultivar identification is primordial for breeding programs and seed production. Considering the importance of genetic purity in comercialized seed lots, the present study aimed to investigate the potential of ISSR markers for discrimination of U. brizantha (Xaraés; Piatã, Basilisk; MG4; MG5; Marandú) in order to determine the degree of contamination of seed lots. ISSR markers showed a low degree of polymorphism . However, results showed it is possible to identify cultivars in pure samples of seeds Urochloa, requiring only ten primers. The cultivar Basilisk was confirmed as U. brizantha cultivar. It was not possible to differentiate samples intentionally contaminated at levels stipulated in the work. Further studies with other primers and other contamination levels will be important to detect varietal mixtures in cultivars U. brizanhta.
Aproximadamente 80 a 90% das áreas de pastagens no Brasil são constituídas por forrageiras do gênero Urochloa, sendo a espécie apomítica Urochloa brizantha [syn. Brachiaria brizantha (Hochst. ex A. Rich.) Stapf.] a mais utilizada. Alguns genótipos de Urochloa têm sido amplamente distribuídos com a nomenclatura equivocada, tanto para espécies como para cultivares. A identificação dos cultivares de Urochloa é fundamental para os programas de melhoramento e produção de sementes. Considerando a importância da pureza varietal em lotes de sementes comercializadas, o presente trabalho teve o objetivo verificar o potencial dos marcadores ISSR para a discriminação de U. brizantha (Xaraés; Piatã, Basilisk; MG4; MG5; Marandú) com a finalidade de determinar o grau de contaminação de lotes de sementes. Os marcadores ISSR apresentaram um baixo grau de polimorfismo. Todavia, os resultados mostraram ser possível identificar os cultivares em amostras puras de sementes de Urochloa, sendo necessários somente dez primers. O cultivar Basilisk foi confirmado como U. brizantha. Não foi possível diferenciar amostras contaminadas propositalmente nos níveis estipulados no trabalho. Novos estudos com outros primers e outros níveis de contaminação serão importantes para detectar misturas varietais em cultivares de U.brizantha.

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Conselho Nacional de Desenvolvimento Científico e Tecnológico
The Zabrotes subfasciatus Boheman (Coleoptera: Bruchidae) is a species probably originated from Central America. It became an agricultural pest since it was established and continually began to reproduce in stored seeds and later diffused throughout tropical and subtropical regions. In stores, those insects cause damages to the grains, by boring them and giving them an unpleasant flavor, therefore depreciating their commercial value. In seeds, the pest consumes the reserves of the cotyledons, therefore endangering germination. This study was conducted to estimate the genetic diversity in Z. subfasciatus populations, by using the molecular markers ISSR (Inter Simple Sequence Repeat). Twelve populations of Z. subfasciatus amostradas were sampled in eight Brazilian states as totaling 269 individuals, then they were evaluate. Five primers ISSR (UBC 807, UBC 808, UBC 809, UBC 811, UBC 891) were used, whereas a total of 51 polymorphic bands averaging 10 bands by each primer were amplified. The percent polymorphism within each population ranged from 74.51 to 92.16, with an average percentage of 83.82. The expected heterozygozite corrected by Nei in 1978 ranged from 0.2253 to 0.;3281 with some 0.2885 average, whereas the genetic diversity index by Shannon and Weaver (HE) ranged from 0.2908 to 0.4805, as averaging 0.4167. At species level, those two indexes showed the values 0.3636 and 0.5393 respectively. The FST values between the population pairs obtained by molecular variance analysis (AMOVA) ranged from 0.06804 to 0.6165. The genetic distance by Nei used in estimation of the genetic divergence among populations ranged from 0.0563 to 0.3250. The AMOVA allowed for the partition of the genetic variation into two levels: either inside and among populations. Higher genetic variation was observed inside population, with 66% of the total variation. Only 34% genetic variation were observed among populations. The Mantel` test showed low correlation between geographical distance and FST, genetic identity and FST, and between the Nei´genetic distance and FST. According to the results, the following conclusions were drawn: the genetic variability of the species is still considered as low in Brazil, probably due to the recent introduction of the pest; and those Zabrotes subfasciatus populations showed to be not geographically structured in Brazil.
Zabrotes subfasciatus Boheman (Coleoptera: Bruchidae), espécie originária provavelmente na América Central, se tornou uma praga agrícola quando se estabeleceu e passou a se reproduzir continuamente em sementes armazenadas, difundindo-se posteriormente pelas regiões tropicais e subtropicais. Em armazéns, esses insetos causam danos aos grãos, perfurando-os e conferindo-lhes sabor desagradável, depreciando o seu valor comercial. Nas sementes, a praga consome as reservas dos cotilédones, comprometendo a germinação. O objetivo deste trabalho foi estimar a diversidade genética de populações de Z. subfasciatus por meio de marcadores moleculares ISSR (Inter Simple Sequence Repeat). Foram avaliadas 12 populações de Z. subfasciatus, amostradas em oito estados brasileiros, totalizando 269 indivíduos. Cinco primers ISSR foram utilizados (UBC 807, UBC 808, UBC 809, UBC 811, UBC 891), sendo amplificadas um total de 51 bandas polimórficas com média de 10 bandas por primer. A porcentagem de polimorfismo dentro de cada população variou de 74,51 a 92,16, com porcentagem média de 83,82. A heterozigosidade esperada corrigida de Nei (HE), variou de 0,2253 a 0,3281, com média de 0,2885 e o índice de diversidade genética de de Shannon e Weaver (I) variou de 0,2908 a 0,4805, com média de 0,4167. Em nível de espécie, estes dois índices apresentaram valores de 0,3636 e 0,5393 respectivamente. Os valores de FST entre pares de populações obtidos pela análise de variância molecular (AMOVA) variaram de 0,06804 a 0,6165. A distância genética de Nei (1978), usada para estimar a divergência genética entre populações, variou de 0,0563 a 0,3250. A AMOVA permitiu uma partição da variação genética em dois níveis: dentro de populações e entre populações. Foi observada maior variação genética dentro da população, com 66% da variação total. Apenas 34% da variação genética foi observada entre populações. O teste de Mantel revelou baixa correlação entre distância geográfica e FST, identidade genética e FST e entre distância genética de Nei e FST. Pode-se concluir que a variabilidade genética da espécie ainda é considerada baixa no Brasil, provavelmente devido à introdução recente da praga. As populações de Zabrotes subfasciatus avaliadas não se apresentam geograficamente estruturadas no Brasil.

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Fundação de Amparo a Pesquisa do Estado de Minas Gerais
The Bank of Germplasm of Vegetables (BGH) of the Federal University of Viçosa, it maintains about 870 accesses of the gender Lycopersicon, many of them still no characterized. That culture is attacked by countless curses, where the flywhite (Bemisia tabaci) one of the most important is considered. Starting from 1994, it happened the introduction of a new biotype of B. tabaci in Brazil (biotype B), responsible for the spread of new begomovírus species in tomato. One of those new species, Tomato yellow spot virus (ToYSV), it causes severe symptoms and with high precocity in the tomato. Inside of this context, our work had as objectives: (1) to evaluate 96 tomato accesses (Lycopersicon esculentum Mill.) as the resistance to the gemivirus Tomato yellow spot virus and (2) to characterize the genetic variability starting from molecular data. The resistance to ToYSV was evaluated through a selection being inoculated the accesses through agroinoculação and he saw biobalística, using the isolated of ToYSV Bi2. As a result of that selection it was selected the accesses that manifested the largest resistance pattern and again inoculated. The accesses that showed as resistant they were selected again and inoculated the same conditions low, being planted 20 plants by accesses. It was evaluated her witnesses from the virus in a visual way to the 10, 20 and 30 days after inoculation. The visual evaluation was confirmed through PCR and hybridization. The accesses were fenotipados tends as base the percentage of infected plants confirmed by PCR and hybridization, of the total of inoculated plants. The molecular variability was characterized using molecular markers ISSR. The accesses were sowed in vegetation house and leaves of three plants by access were collected for extraction in bulk of DNA. Ten molecular markers anchored ISSR were used. The bands analyzed for each employed primer allowed the construction of the head office of binary data. The head office was used in I calculate it of the dissimilar using the complement of the coefficient of similarity of Jaccard. The head office was used in the accomplishment of the groupings by the method of optimization of Tocher and hierarchical UPGMA. Of the 96 accesses inoculated through agroinoculação, 31 accesses were selected classified as highly resistant (AR), being inoculated through biobalistica and selected among of them 4 accesses. The 4 selected accesses were inoculated again and as result he stood out the access BGH-224 that was classified as AR with only 10% of infected plants, and suitable as promising access for obtaining of you cultivate with resistance to ToYSV. The markers ISSR generated, together, 53 bands polymorphic of a total of 144 amplified. The primer 840 generated the largest number of bands polimórficas, with 18% (13 bands) of the 144 bands obtained by the 10 used primers. The size of the amplified fragments varied from 250 to 2000 pb. By the evaluation of the dendrogram obtained by the grouping method UPGMA and the grouping for the method Tocher, it was possible to differentiate the accesses. The access BGH- 980 was classified separately, being the most divergent of the tested accesses. They were classified in groups of 2 accesses the equal of accesses BGH- 674 and BGH-991, BGH-616 and BGH-970, that constitute duplicated accesses. The markers ISSR were useful in the characterization of the variability of the accesses of Lycopersicon esculentum, amplifying relatively high number of locos for primer, being enough to discriminate the appraised accesses.
O Banco de Germoplasma de Hortaliças (BGH) da Universidade Federal de Viçosa, mantém cerca de 870 acessos do gênero Lycopersicon, muitos deles ainda não caracterizados. Essa cultura é atacada por inúmeras pragas, onde a mosca-branca (Bemisia tabaci) é considerada uma das mais importantes. A partir de 1994, ocorreu a introdução de um novo biótipo de B. tabaci no Brasil (biótipo B), responsável pela disseminação de novas espécies de begomovírus em tomateiro. Uma dessas novas espécies, o Tomato yellow spot virus (ToYSV), causa sintomas severos e com alta precocidade no tomateiro. Dentro deste contexto, nosso trabalho teve como objetivos: (1) avaliar 96 acessos de tomateiro (Lycopersicon esculentum Mill.) quanto a resistência ao gemivirus Tomato yellow spot virus (ToYSV) e (2) caracterizar a variabilidade genética a partir de dados moleculares. A resistência ao ToYSV foi avaliada através de uma triagem, sendo inoculados os acessos via agroinoculação e via biobalística, empregando o isolado de ToYSV Bi2. Como resultado dessa triagem foi selecionado os acessos que manifestaram o maior padrão de resistência e foram novamente inoculados. Os acessos que se manifestaram como resistentes foram selecionados novamente e inoculados baixo as mesmas condições, sendo plantadas 20 plantas por acessos. Foi avaliada a presencia do vírus de modo visual aos 10, 20 e 30 dias após inoculação. Foi confirmada a avaliação visual via PCR e hibridização. Os acessos foram fenotipados tendo como base a porcentagem de plantas infectadas confirmadas por PCR e hibridização, do total de plantas inoculadas. A variabilidade molecular foi caracterizada empregando marcadores moleculares ISSR. Os acessos foram semeados em casa de vegetação e folhas de três plantas por acesso foram coletadas para extração em bulk do DNA. Dez marcadores moleculares ISSR ancorados foram empregados. As bandas analisadas para cada primer empregado permitiu a construção da matriz de dados binários. A matriz foi empregada no calculo da dissimilaridade utilizando o complemento do coeficiente de similaridade de Jaccard. A matriz foi empregada na realização dos agrupamentos pelo método de otimização de Tocher e hierárquico UPGMA. Dos 96 acessos inoculados via agroinoculação, foram seleccionados 31 acessos classificados como Altamente Resistente (AR), sendo inoculados via biobalística e selecionados dentre deles 4 acessos. Os 4 acessos selecionados foram inoculados novamente e como resultado destacou-se o acesso BGH-224 que foi classificado como AR, com apenas 10% de plantas infectadas, e indicado como acesso promissor para obtenção de cultivares com resistência ao ToYSV. Os marcadores ISSR geraram, em conjunto, 53 bandas polimórficas de um total de 144 amplificadas. O primer 840 gerou o maior número de bandas polimórficas, com 18 % (13 bandas) das 144 bandas obtidas pelos 10 primers empregados. O tamanho dos fragmentos amplificados variou de 250 a 2000 pb. Mediante a avaliação do dendrograma obtido pelo método de agrupamento UPGMA e o agrupamento pelo método Tocher, foi possível diferenciar os acessos. O acesso BGH-980 foi classificado isoladamente, sendo o mais divergente dos acessos testados. Foram classificados em grupos de 2 acessos os pares de acessos BGH-674 e BGH-991, BGH-616 e BGH-970, ainda que semelhantes não constituem acessos duplicados. Os marcadores ISSR foram úteis na caracterização da variabilidade dos acessos de Lycopersicon esculentum, amplificando número relativamente elevado de locos por primer, sendo suficiente para discriminar os acessos valiados.

Soybean (Glycine max) is an important crop in the world in terms of total production and usage. It is also among the least diverse species. The main objectives of the present study were 1) to determine differences between ISSR and RAPD marker systems in detecting genetic variation in soybeans and 2) to identify and characterize accession- diagnostic molecular markers in G. max accessions. Genomic DNAs from 108 G. max accessions from 11 different gene pools were analyzed using several ISSR and RAPD primers. The levels of polymorphic loci detected with the two marker systems were in general moderate and similar.. Overall, 82% of genetic distance values were above 0.40 based on ISSR analysis. However, RAPD data revealed that the accessions from different countries are closely related with 64% genetic distance values below 0.40. The dendrograms constructed with ISSR data revealed that the South Korean accessions formed an out-group while the RAPD analysis showed that accessions from Sweden were separate from the other 10 gene pools. One variety-diagnostic marker generated with ISSR 5 primer was identified in the accession Kao Chien Tao from China. This marker was cloned, and sequenced. Although RAPD and ISSR marker systems detected similar levels of genetic variability, they target different regions of the soybean genome, resulting in different clustering of the 11 gene pools indicating different genetic relatedness among them. This finding demonstrates the usefulness of both marker systems in assessing diversity and relatedness among Glycine max gene pools.

O pinhão-manso (Jatropha curcas) é uma espécie arbórea de ampla distribuição geográfica e com qualidades que a tornam importante do ponto de vista natural, ecológico e principalmente sócio-econômico, pois seus frutos são uma valiosa fonte de óleo vegetal com potencial para produção de biodiesel, proporcionando vantagens ambientais, econômicas e sociais. Este trabalho teve como objetivo avaliar a diversidade genética no germoplasma de pinhão-manso e para isto foram utilizados marcadores moleculares microssatélites e ISSR. A partir de uma biblioteca enriquecida com locos microssatélites foram desenvolvidos 18 pares de primers para a espécie, sendo estes utilizados, juntamente com 30 pares de primers SSR desenvolvidos no CBMEG, visando à caracterização e estudo da estrutura genética populacional. Os acessos dos bancos de germoplasma do CPQBA (Centro Pluridisciplinar de Pesquisas Químicas, Biológicas e Agrícolas) da UNICAMP e da UFS (Universidade Federal de Sergipe) foram avaliados. O germoplasma pertencente ao CPQBA está organizado em 12 populações e o da UFS representado por 17 acessos únicos. Não foi observado polimorfismo entre as populações inviabilizando o estudo populacional. A caracterização dos grupos formados pelos acessos dos bancos de germoplasma foi realizada utilizando 14 marcadores ISSR, revelando que 86,64% da variação genética encontram-se dentro dos grupos e 13,36% entre eles. O número médio total de alelos (na) foi de 1,99 alelos por loco e o número efetivo de alelos (ne) foi de 1,42 alelos por loco. A diversidade genética de Nei (1973) indicou uma baixa diversidade genética dentro dos grupos (0,26), assim como o Índice de Shannon (I) para os acessos (0,41), considerado um baixo valor de diversidade genética. A análise bayesiana alocou todos os acessos avaliados em quatro grupos, todos os acessos apresentaram Q > 0,8. Os grupos formados não apresentaram nenhuma relação com a origem dos acessos. O índice médio de similaridade de Jaccard indicou que existem 30% de similaridade entre os grupos e a amplitude de similaridade variou de 0,23 a 0,94. O dendrograma formou os mesmos quatro grupos de acessos que o formado pela análise bayesiana, tornando ainda mais consistente os resultados obtidos na presente análise. O estudo revela a necessidade e importância de reunir o maior número possível de acessos de diferentes regiões e países para formar o banco de germoplasma da espécie viabilizando a conservação e programas de melhoramento da espécie, haja vista seu promissor potencial para produção de bicombustível.
Physic nut (Jatropha curcas) is a geographically widespread perennial plant species. It is ecologically important in natural communities and economically due to the oil extracted from its fruits that exhibit high potential for biodiesel production, thus, providing environmental, economical and social advantages. The current work aimed to evaluate the genetic diversity in physic nut germplasm using microsatellites and ISSR molecular markers. From a microsatelliteenriched library, 18 primer pairs were developed for the species and were used along with 30 SSR primer pairs developed at CBMEG to characterize and study the population genetic structure. Acessions from the germplasm banks at CPQBA (Centro Pluridisciplinar de Pesquisas Químicas, Biológicas e Agrícolas) from UNICAMP and from UFS (Universidade Federal de Sergipe) were evaluated. The germplasm from CPQBA is organized in 12 populations whereas the accessions from UFS represent 17 soloist accessions. The polymorphism observed between the populations does not impair population genetic studies. The clusters of accessions from the germplasm Banks were characterized using 14 ISSR markers, revealing 86.64% of the genetic diversity are found within the clusters whereas between them, it corresponds to 13.36%. The total average number of alleles per locus (na) corresponded to 1.99 and the effective number of alleles (ne) was of 1.42 alleles per locus. The genetic diversity, investigated as in Nei (1973), indicated a low genetic diversity within the groups (0.26). Shannon index (I) for the accessions evidenced a low value of genetic diversity (0.41). Bayesian analyses of all investigated accessions in four groups demonstrated that all the accessions exhibit Q > 0.8. The clustering patterns did not indicated origin relationships among the accessions. Jaccard average index indicated 30% of similarity between the groups and the amplitude of similarity ranged from 0.23 to 0.94. The dendrogram analysis grouped the four clusters generated by the Bayesian analysis, confirming the consistency of the results. The current study reveals the necessity and importance of gathering as many germplasm accession as possible for the species in order to allow the establishment of conservation and breeding program strategies, considering the potential of the species for biofuel production.

Dioscorea trifida L. é uma espécie de inhame comestível originária da América do Sul, que em conjunto com outras espécies importantes economicamente do gênero Dioscorea é mantida por pequenos agricultores tradicionais. Assim, observa-se que as comunidades tradicionais exercem um papel fundamental na manutenção e geração da diversidade genética de inhame.O objetivo deste trabalho foi obter informações a respeito da distribuição, manejo e diversidade genética de D. trifida no Brasil. Para tanto, foram visitados e entrevistados agricultores dos Estados de São Paulo, Santa Catarina e Mato Grosso. Durante as visitas, foram coletados 51 acessos, os quais, juntamente com dois acessos adquiridos em feiras livres no Estado do Amazonas, foram caracterizados por meio de 16 descritores morfológicos, 16 ISSR e oito SSR.Observou-se que o cultivo de D. trifida ocorre na maioria das vezes em roças com menos de dois hectares (92%) mantidas por agricultores tradicionais, com a produção ocorrendo em baixa escala, visando principalmente a subsistência das pessoas envolvidas com o cultivo e manutenção da espécie. Dentre as denominações encontradas para a espécie, as mais citadas foram \"cará roxo\", em 43,4% das unidades amostrais, \"cará\" e \"cará branco\", ambos observados em 9,4%, e \"cará mimoso\", com 7,6%. Observou-se também uma regionalização dessas denominações, onde \"cará roxo\" e \"cará branco\" foram atribuídos à espécie pelos agricultores dos Estados de São Paulo e Mato Grosso, sendo que \"cará\" e \"cará mimoso\" foram atribuídos pelos agricultores de Santa Catarina. Os nomes \"cará roxo\" e \"cará\" foram também atribuídos aos dois acessos da Amazônia. Além dessas, várias outras denominações para a espécie foram encontradas, porém com baixa frequência. Na caracterização morfológica observou-se que as cores da casca e da polpa foram os caracteres morfológicos mais relevantes para a distinção dos acessos. O nível de polimorfismo entre os acessos foi elevado, 95% para SSR e 76% para ISSR. O coeficiente de similaridade de Jaccard, bem como os resultados obtidos com as análises de agrupamento, coordenadas principais e bayesiana para os marcadores SSR e ISSR, separaram os acessos em três grupos principais: I - acessos de Ubatuba-SP; II - acessos de Iguape-SP e SantaCatarina; III - acessos de Mato Grosso. Os dois acessos do Amazonas variaram de posição de acordo com a região genômica analisada. A maior parte da diversidade genética foi observada dentro dos grupos formados (66,5% e 60,6% para ISSR and SSR, respectivamente), embora a diversidade entre grupos tenha sido de considerável magnitude, mostrando a estruturação dos acessos de acordo com sua origem, o que foi comprovado pela correlação baixa, mas significativa entre as distâncias genéticas e geográficas dos acessos. Portanto, os resultados obtidos para os marcadores SSR e ISSR demonstraram que a diversidade genética dos acessos de D. trifida mantidos por pequenos agricultores tradicionais do Brasil está levemente estruturada no espaço geográfico amostrado. Estes resultados poderão auxiliar na elaboração de estratégias de conservação para a espécie, tanto ex situ como in situ, dentro da visão de conservação on farm.
Dioscorea trifida L. is a species of edible yams originated in South America, which together with other economically important species of the genus Dioscorea is cultivated by small traditional farmers. Thus, it is observed that traditional communities play a key role in the generation and maintenance of yams genetic diversity. The aim of this study was to obtain information about the distribution, management and genetic diversity of D. trifida in Brazil. So, were visited and interviewed farmers in the states of São Paulo, Santa Catarina and Mato Grosso. During the visits, we collected 51 accessions, which, together with two accessions purchased at fairs in the state of Amazonas, were characterized using 16 morphological descriptors, 16 ISSR and eight SSR markers.We observed that D. trifida occurs most often in swidden fields with less than two hectares (92%) maintained by traditional farmers, with production occurring on a small scale, mainly targeting the livelihood of the people involved with the cultivation and maintenance of the species. Among the names found for the species, the most cited were \"cará roxo\" in 43.4% of the sample units, \"cará\" and \"cará branco\", both observed in 9.4% and \"cará mimoso\" with 7.6%. There is also a regionalization of these denominations, where \"inhame roxo\" and \"inhame branco\" were assigned by farmers to the species in the states of São Paulo and Mato Grosso, where \"cará\" and \"cará mimoso\" were allocated to farmers of Santa Catarina. The names \"cará roxo\" and \"cará\" were also awarded to two accessions from the Amazon. Besides these, several other names for the species were found, but with low frequency. In the morphological characterization, the skin and flesh color were the most relevant traits for the distinction of accessions. The polymorphism level between the accessions was high, 95% for SSR and 76% for ISSR. The Jaccard similarity coefficient and the results obtained in the cluster analysis, principal coordinates and Bayesian analyses for ISSR and SSR markers, separated the accessions into three main groups: I - accessions from Ubatuba-SP; II - accessions from Iguape-SP and Santa Catarina; III - accessions from Mato Grosso. The accessions from Amazonas ranged their position according to the genomic region analyzed. The majority of genetic diversity was observed within groups (66.5% and 60.6% for ISSR and SSR, respectively), although differences between groups was of considerable magnitude, showing the structure of the accessions according to their origin, which was confirmed by correlation between genetic and geographic distances of accessions. Therefore, the results obtained for the SSR and ISSR markers showed that the genetic diversity of accessions of D. trifida maintained by small traditional farmers in Brazil is slightly structured in the geographic area sampled. These results may help in developing conservation strategies for the species, both ex situ and in situ, within the vision of on farm conservation.

Gelasine Herb. (Tigridieae: Iridaceae) é composto por sete espécies nativas da América do Sul, sendo três delas encontradas no Rio Grande do Sul (Brasil): G. coerulea (Vell.) Ravenna, G. elongata (Graham) Ravenna e G. uruguaiensis Ravenna. São plantas bulbosas de folhas plicadas, flores perfeitas azuis ou roxas e compostas por dois conjuntos de tépalas desiguais. Gelasine elongata e G. coerulea encontram-se na lista de espécies ameaçadas para o RS, sendo a primeira delas ameaçada e a segunda criticamente ameaçada. Apesar de seu atual estado de vulnerabilidade, Gelasine é um gênero ainda pouco estudado, não havendo nenhuma informação quanto à variabilidade e diversidade genética. Os dados citogenéticos são também ainda escassos. Assim, a presente dissertação tem por objetivo caracterizar as três espécies de Gelasine ocorrentes no sul do Brasil quanto a aspectos moleculares, citogenéticos, além de compreender suas relações. Para a caracterização da diversidade genética foram usadas duas populações de G. coerulea e duas de G. elongata; não foi possível a utilização de G. uruguaiensis em função do número restrito de indivíduos. As amostras de DNA foram obtidas a partir de folhas das espécies mencionadas e empregada a técnica de ISSR (Inter Simple Sequence Repeat). Foram testados 44 primers para ISSR, destes, 12 apresentaram um bom padrão de amplificação que em conjunto geraram 91 loci. A quantidade de bandas por primer variou em média de 7,5. Este trabalho resultou em dados inéditos para o gênero Gelasine quanto à variabilidade genética inter e intra-populacional. Os resultados indicam que a variação genética intrapopulacional é muito baixa e que a maior diversidade encontrada para estas espécies ocorreu entre as populações. Não foi verificada correlação significativa com a distância geográfica entre as populações. Tais resultados indicam que o sistema reprodutivo, o método de dispersão de sementes e a presença de descendência clonal oriunda da divisão dos bulbos subterrâneos são fatores de grande influência na diversidade. Para caracterização citogenética das espécies foi empregada coloração convencional e bandeamento CMA/DAPI, e realizadas medidas cromossômicas. Foi também estimado o tamanho do genoma por citometria de fluxo a partir de folhas frescas das três espécies. As análises citogenéticas se mostraram bastante eficientes para a diferenciação das três espécies de Gelasine investigadas. Gelasine coerulea e G. uruguaiensis apresentam o mesmo número cromossômico básico e somático (2n = 2x = 14), não sendo encontrados citótipos poliploides. Ambas têm cariótipos relativamente simétricos, porém se mostram bastante distintas quanto ao tamanho dos cromossomos e seu padrão de bandeamento, com uma grande variação na ocorrência e distribuição de sequências de DNA repetitivo (bandas CMA/DAPI). O conteúdo de DNA também permite a clara diferenciação dessas espécies, tendo G. coerulea 2C = 11,30 pg e G. uruguaiensis 2C = 16,88 pg. Gelasine elongata possui número cromossômico básico diferente das anteriores (2n = 2x = 12) e cariótipo claramente bimodal. Os cromossomos têm menor tamanho, o que, consequentemente reflete no menor tamanho de genoma (2C = 3,45 pg). Além disso, o padrão de bandas CMA/DAPI é notadamente mais simples que das outras duas espécies, onde o maior par de cromossomos (par I) exibe as únicas bandas CMA+ presentes na região da constrição secundária. Os dados obtidos para G. elongata apontam para uma maior semelhança dessa espécie com outras duas do gênero Eleuthenine (2n = 2x = 12), o que reforça os dados filogenéticos existentes, onde G. elongata está separada de G. coerulea e agrupada no mesmo ramo de Eleutherine. Não foi observado heteromorfismo cromossômico para Gelasine elongata e nem para as outras duas espécies investigadas, embora tal situação tenha sido reportada para aquela espécie. Os dados obtidos para Gelasine com o uso dos fluorocromos CMA e DAPI, bem como os demais parâmetros citogenéticos investigados permitiram a clara diferenciação entre as espécies. Associados a uma abordagem filogenética, tais resultados auxiliam a compreensão das relações entre essas espécies e sua evolução.
Gelasine Herb. (Tigridieae: Iridaceae) comprises seven native species from South America, three of them are found in Rio Grande do Sul (Brasil): G. coerulea (Vell.) Ravenna, G. elongata (Graham) Ravenna and G. uruguaiensis Ravenna. These species are bulbous plants with plicate leaves and blue or violet perfect flowers which are composed of two unequal groups of tepals. Gelasine elongata and G. coerulea are included in the list of endangered species from RS, the first one is considered endangered and the latter, critically endangered. Notwithstanding its current vulnerability status, Gelasine is still a poorly studied genus and genetic variability and diversity information concerning its species are lacking. Cytogenetic data are also scarce. Thus the present dissertation aims to characterize the three Gelasine species occurring in Southern Brazil regarding its molecular and cytogenetic aspects in addition to understand their relationships. To characterize their genetic diversity, two populations of G. coerulea and two of G. elongata were used; it was not possible to investigate G. uruguaiensis due to its restricted number of individuals. DNA samples were obtained from leaves of the aforementioned species and ISSR (Inter Simple Sequence Repeat) technique was employed. Fourty-four ISSR primers were tested, 12 of these presented good amplification pattern which generated a total of 97 loci. The number of bands per primer had an average of 7.5. The present study resulted in novelty data for Gelasine concerning its inter and intrapopulation genetic variability. The results indicate a very low intrapopulation genetic variation and most of the diversity found in these species occurred among their populations. No significant correlation was verified between geographical distances of populations. Such results indicate that reproductive system, seed dispersal mechanisms and presence of clonal descendants generated from divisions of subterranean bulbs are factors that greatly influence in diversity. For cytogenetic characterization of the species, conventional staining and CMA/DAPI banding were employed and chromosome measurements were made. Also, genome size was estimated through flow cytometry using fresh leaves from the three species. Cytogenetic analyses were very efficient to differentiate all investigated species of Gelasine. Gelasine coerulea and G. uruguaiensis have the same basic and somatic chromosome number (2n = 2x = 14); polyploid cytotypes were not found. Both species display fairly symmetric karyotypes, however they are very distinct with respect to chromosome sizes and banding patterns, with a great variation in the occurrence and distribution of repetitive DNA sequences (CMA/DAPI bands). DNA content also allows clear differentiation of these species; G. coerulea has 2C = 11,30 pg and G. uruguaiensis has 2C = 16,88 pg. Gelasine elongata has a different base chromosome number than both former species (2n = 2x = 12) and a clearly bimodal karyotype. Its chromosomes are smaller which, consequently, reflects on the smaller genome size (2C = 3,45 pg). Furthermore, its CMA/DAPI band pattern is markedly simpler than the ones from the other two species, where the largest chromosome pair (pair I) contains the only CMA+ bands present in the secondary constriction region. Data obtained from G. elongata points out a larger resemblance between this species and two others belonging to Eleuthenine (2n = 2x = 12), which supports the phylogenetic data where G. elongata is separate from G. coerulea and groups with Eleutherine. Chromosome heteromorphism was not observed in Gelasine elongata nor in the two other investigated species, even though it had been reported for the first one. Data obtained from Gelasine with the use of CMA and DAPI fluorochromes, along with the other cytogenetic parameters investigated, allowed clear differentiation between species. Allied to a phylogenetic approach, these results can bring better understanding to the relations between these species and their evolution.

A batata-da-serra, Ipomoea serrana Sim.-Bianch. & L.V. Vasconcelos, é uma liana endêmica da Chapada Diamantina, Bahia, cuja raiz tuberosa e consumida por populações humanas ha muitos anos. Apesar da espécie, classificada como vulnerável pela IUCN (União Internacional para a Conservação da Natureza), estar submetida à pressão antrópica devido à exploração de raízes tuberosas, para uso na alimentação, são raros os estudos com a espécie, razão pela qual e de grande importância conhecer a diversidade e estrutura genética da espécie. Estudos sobre diversidade e estrutura genética a partir de marcadores moleculares são importantes por fornecerem dados sobre impactos da exploração antrópica sobre as populações, podendo oferecer subsídio para planos de manejo e conservação de espécie. Cinco populações da Chapada Diamantina, constituindo um total de 142 indivíduos, foram investigados com quatro iniciadores Inter Simple Sequence Repeats (ISSR), resultando em 34 bandas, das quais 25 foram polimórficas. A analise dos parâmetros genéticos mostrou que as populações apresentam variabilidade moderada, com 73,8% de bandas polimórficas, 0,264 de índice de diversidade de Nei e 0,389 de índice de Shannon (valores médios). A maior parte da variação ocorreu dentro das populações (77%), estimado pela analise de variância molecular (AMOVA), enquanto que a variação entre populações foi de 23%, o que corroborou os resultados de estruturação obtidos pelo programa Structure, analise de coordenadas principais (PCoA) e agrupamento estimado pelo método Neighbor-Joining, a partir do coeficiente de dissimilaridade de Jaccard. A analise Bayesiana separou os indivíduos em quatro grupos, sendo que as populações Andaraí e Capão foram alocadas em grupos distintos, enquanto as outras três populações compartilharam indivíduos distribuídos em outros dois grupos. Este estudo, por seu caráter pioneiro com relação aos marcadores moleculares, constituiu o primeiro passo para o conhecimento da diversidade genética da espécie. Estudos futuros poderão ampliar o conhecimento sobre a espécie podendo oferecer subsidio para a elaboração de um plano de manejo para esta espécie que tem sido explorada na região.
batata-da-serra, Ipomoea serrana Sim.-Bianchi. & L.V. Vasconcelos, is an endemic liana from the Chapada Diamantina, Bahia, whose tuberous roots have been consumed by human populations for many years. Although the species, classified as vulnerable by the IUCN (International Union for Conservation of Nature), is subject to anthropic pressure due to the exploration of tuberous roots for food consumption, few studies have been conducted on the species, which is why it is of great importance to know the diversity and genetic structure of the species. Studies on genetic diversity and structure with molecular markers are important for providing data on the impacts of anthropogenic exploitation and can be useful for the species management and conservation. Five populations of Chapada Diamantina, consisting a total of 142 individuals were studied with four ISSR primers, resulting in 34 bands, 25 of which were polymorphic. The genetic diversity analysis showed that populations have a moderate variability, with 73.8% of polymorphic bands, Nei\'s unbiased gene diversity (He) was 0.264; Shannon diversity index (I) was 0.389, average values. Most of the variation was within populations (77%), as estimated by the analysis of molecular variance (AMOVA), whereas the variation between populations was 23% of the total, which corroborated the results of program Structure, principal co-ordinate analysis (PcoA) and cluster analyses, using the Neighbor-Joining method, and the dissimilarity coefficient of Jaccard. The Bayesian analysis separated the individuals into four groups, with populations Andarai and Capao allocated into different groups, while the other three populations shared individuals in two other groups. Considering the pioneering characteristic of this study at the molecular level, it represents the first step towards the knowledge on the genetic diversity of the species. Future studies will increase the knowledge on the genetics of the species and may provide subsidy for the development of a management plan for a species that has been explored in the region.

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Conselho Nacional de Pesquisa e Desenvolvimento Científico e Tecnológico - CNPq
Habitats fragmentation is the greatest cause of biodiversity erosion in tropical forests among the anthropic action. Due to the high degree of fragmentation and the isolation of the remaining Atlantic forest, many populations are being extinguished locally while others are suffering losses of its genetic variability. The Bromeliaceae family has a great variety of tropical ornamental species. Eleven species of the Aechmea genus occur in the State of Pernambuco, and although it is of great economic and ecological importance, there has been few studies on its genetic diversity. Aechmea fulgens is a native Atlantic forest species, which natural populations may be suffering losses on the genetic variability. Plants genetic diversity may be estimated in many ways. Modern techniques on molecular biology allow observing the polymorphism directly on the organism genic sequence. Molecular markers broaden the horizons on researches for the conservation of species and have been widely used to monitor the genetic variability. Among many molecular markers available, SSR and ISSR are relevant. With the of aim evaluating the degree of genic diversity on Aechmea fulgens populations, providing from three different fragments of Atlantic forest in Pernambuco, SSR and ISSR markers were used. Twelve pairs of microsatellites oligonucleotides developed for Tillandsia, Guzmania and Pictairnia bromeliad genera were tested. From these, only five pairs had polymorphism, showing transference of these primers to the Aechmea gender. Twenty ISSR oligonucleotides were used to amplify Aechmea fulgens sample, from which eight primers that had the most consistent polymorphism. The analysis on variance results revealed greater differences inside populations than among them. The results suggest that both markers may be used for genetic variability evaluation, contributing for the success of future studies and species conservation programs.
A fragmentação de habitats é a maior causa da erosão da biodiversidade nas florestas tropicais junto com a ação antrópica. Devido ao alto grau de fragmentação e ao isolamento dos remanescentes atuais da floresta atlântica, várias populações estão se extinguindo localmente e outras sofrendo perda de sua variabilidade genética. A família Bromeliaceae inclui uma grande variedade de espécies ornamentais tropicais. O gênero Aechmea tem significativa representatividade no Estado de Pernambuco, onde ocorrem onze espécies e, apesar de sua grande importância ecológica e econômica, existem poucos estudos sobre sua diversidade genética. Aechmea fulgens é uma especie nativa da Mata Atlântica, cujas populações naturais podem estar sofrendo perda de sua variabilidade genética. A diversidade genética em plantas pode ser estimada de várias maneiras. As modernas técnicas de biologia molecular permitem a observação de polimorfismo diretamente na seqüência gênica de organismos. Os marcadores moleculares abriram novas perspectivas para pesquisas em conservação de espécies e têm sido amplamente utilizados no monitoramento da variabilidade genética. Dentre os diversos marcadores moleculares disponíveis atualmente, destacam-se as seqüências simples repetidas (SSR) e repetições entre seqüências simples (ISSR). Com o objetivo de avaliar o nível de diversidade genética de populações de Aechmea fulgens (Brongn.) provenientes de três fragmentos distintos da mata atlântica de Pernambuco, foram utilizados marcadores SSR e ISSR. Um total de 12 pares de oligonucleotídeos de microssatélites, desenvolvidos para bromeliáceas dos gêneros Tillandsia, Guzmania e Pitcairnia foram testados. Desse total apenas cinco pares apresentaram polimorfismo demonstrando a transferência desses primers para o gênero Aechmea. Adicionalmente, 20 oligonucleotídeos ISSR foram utilizados para amplificar as amostras de A. fulgens, tendo sido selecionados oito primers por apresentarem polimorfismo mais consistente. Os resultados das análises de variância revelaram que a maior divergência esteve presente dentro das populações do que entre elas. Os resultados sugerem que os dois tipos de marcadores são adequados para a avaliação da variabilidade genética, colaborando para o sucesso de futuros estudos e programas de conservação dessa espécie.

O gênero Paspalum L. compreende aproximadamente 400 espécies no mundo e cerca de 220 no Brasil. Paspalum é ecologicamente e economicamente importante e tem sido utilizado como pastagem. Paspalum notatum Flügge (grama-forquilha) é uma valorosa gramínea forrageira nos subtrópicos. Esta espécie consiste de vários biótipos sexuais (diplóides) e apomíticos (tetraplóides, ocasionalmente tri e pentaplóides). Neste trabalho, os Inter Simple Sequence repeat (ISSR) foram utilizados para acessar a diversidade genética da grama-forquilha (Paspalum notatum). Os tecidos vegetativos de 95 acessos de grama-forquilha foram obtidos de vários locais da América do Sul (Brasil, Argentina e Uruguai). Um total de 91 de fragmentos reproduzível ISSR foi observado. Oitenta e nove fragmentos (97,5% do total observado) foram polimórficos. A análise de agrupamento (UPGMA) foi realizada para o conjunto de dados ISSR. Os resultados ilustram as relações genéticas entre 95 acessos de Paspalum notatum. A comparação entre dados moleculares, morfológicos e nível de ploidia foi realizada. Em resumo, os marcadores moleculares ISSR mostraram-se eficientes para distinção dos genótipos analisados e observou-se uma variabilidade ampla para a espécie. Estes resultados adicionam novas informações sobre a diversidade genética em Paspalum notatum, conseqüentemente contribuindo para o conhecimento biológico desta espécie e fornecendo subsídios para futuros programas de melhoramento genético e para programas de conservação.
The genus Paspalum L. comprises approximately 400 species worldwide and about 220 in Brazil. Paspalum is ecologically and economically important, and has been very useful as pasture and Paspalum notatum Flügge (bahiagrass) is a valuable forage grass in the subtropics. This species consists of several sexual (diploid) and apomictic (tetraploid, ocasionally tri and pentaploids) biotypes. In this work, inter Simple Sequence Repeats (ISSR) markers were used to assess the genetic variability of a bahiagrass (Paspalum notatum) collection. Vegetative tissues of 95 bahiagrass accessions were obtained from various locations in South America (Brazil, Argentina and Uruguay). A total of 91 reproducible ISSR fragments were observed and eighty nine fragments (97.5% of the total observed) were polymorphic. Cluster analyses (UPGMA) were performed from the ISSR data set and the results illustrate the genetic relationships among the 95 accessions of Paspalum notatum. A comparison among molecular, morphological and ploidy levels data were done. ISSR markers were effective in distinguishing the genotypes analyzed, and a wide variability was observed for this species. These results add new information regarding the genetic diversity in Paspalum notatum, thus contributing towards the biological knowledge of this species, and providing with subsides for future plant breeding and conservation programs.

Medicinal plants are major sources of secondary metabolites for which they have been paid more attention by pharmaceutical industries. In order to produce these secondary metabolites, medicinal plants are cultivated and for that plant tissue or organ, culture can be a suitable alternative. However, these plants are treated with plant hormones and elicitors to enhance the secondary metabolites and such elicitation may lead to genetic or epigenetic changes which are known as somaclonal variations. Thus, a stringent method of monitoring is required to observe the true-to-types of these medicinal plants when multiplied through tissue culture. Molecular markers like Randomly Amplified Polymorphic DNA (RAPD), Inter-Simple Sequence Repeat (ISSR), and Simple Sequence Repeats (SSR) are highly suitable markers to assess clonal fidelity in micropropagated medicinal plants. In the present chapter, the execution of such markers to check somaclonal variations in tissue culture raised medicinal plants is discussed in detail.

Soil pollution with industrial leftovers is of real danger to living organisms since harmful effects can arise after exposure to the contaminants in the soil. In our study, we applied a plant bioassay battery to monitor soil genotoxicity after short-term exposure to the soil. The soil was collected in 3 rounds: at the central part of the brownfield before (S-I) and after (S-III) topsoil removal, and at the brownfield periphery (S-II). The permissible value of the total contamination index is &amp;amp;lt;16 and the corresponding values were 780 in S-I, 69 in S-II and 133 in S-III soil showing that whole brownfield territory is extremely polluted with heavy metals. Cytogenetic markers were recorded in Allium and Tradescantia test-systems and two types of molecular markers, RAPD and ISSR, were analysed in Allium. Our results revealed that the most polluted soil sample has induced an alarming increase of apoptotic cells in onion roots. Chromosome aberration and micronuclei frequency in Allium decreased inconsistently along with the pollution reduction in the soil. Increased frequencies of all cytogenetic markers were revealed in Tradescantia cuttings after exposure to the S-I soil extracts. Cluster analysis of Allium RAPD and ISSR markers showed that the most polluted soil samples induced genetic changes in onions different from those induced by the least polluted soil. Both plant test-systems in this study confirm that soil from the brownfield is harmful to plants and is potentially hazardous to humans.