A de novo genome assembly of Solanum bulbocastanum Dun., a Mexican diploid species reproductively isolated from the A-genome species, including cultivated potatoes.

Potato and its wild relatives are distributed mainly in the Mexican highlands and central Andes of South America. The South American A-genome species, including cultivated potatoes, are reproductively isolated from Mexican diploid species. Whole-genome sequencing has disclosed genome structure and similarity, mostly in cultivated potatoes and their closely related species. In this study, we generated a chromosome-scale assembly of the genome of a Mexican diploid species, Solanum bulbocastanum Dun., using PacBio long-read sequencing, optical mapping, and Hi-C scaffolding technologies. The final sequence assembly consisted of 737.9 Mb, among which 647.0 Mb were anchored to the 12 chromosomes. Compared with chromosome-scale assemblies of S. lycopersicum (tomato), S. etuberosum (non-tuber-bearing species with E-genome), S. verrucosum, S. chacoense, S. multidissectum, and S. phureja (all four are A-genome species), the S. bulbocastnum genome was the shortest. It contained fewer transposable elements (56.2%) than A-genome species. A cluster analysis was performed based on pairwise ratios of syntenic regions among the seven chromosome-scale assemblies, showing that the A-genome species were first clustered as a distinct group. Then, this group was clustered with S. bulbocastanum. Sequence similarity in 1,624 single-copy orthologous gene groups among 36 Solanum species and clones separated S. bulbocastanum as a specific group, including other Mexican diploid species, from the A-genome species. Therefore, the S. bulbocastanum genome differs in genome structure and gene sequences from the A-genome species. These findings provide important insights into understanding and utilizing the genetic diversity of S. bulbocastanum and the other Mexican diploid species in potato breeding.

Chromosomal Study on Selected Small Araneomorph Spiders from Brazil, Including the First Records in Palpimanidae and Theridiosomatidae (Araneae, Araneomorphae).

Among the 50,474 spider species, only 849 have chromosomal data available in the literature. Fifty spider families remain unknown from a cytogenetic perspective. The aim of this study was to analyze chromosomally selected araneomorph spiders from Brazil, to contribute to the cytotaxonomy and chromosome evolution of this group. The karyotypes of 12 species belonging to families Corinnidae, Linyphiidae, Oonopidae, Palpimanidae, Theridiidae, Theridiosomatidae, Trachelidae, and Zodariidae were analyzed, including the first chromosomal record for the first two families. Specimens (with the abdomen content partially exposed by perforation) were subjected to colchicine, hypotonization, and fixation. In most cases, the total content of the abdomen was used to prepare slides by spreading of cell suspension and subsequent Giemsa staining. The results were as follows: Cinetomorpha simplex Simon, 1892 (Oonopidae) 2n♂ = 9, X0; Neotrops sp. and Neoxyphinus termitophilus (Oonopidae), 2n♂ = 7, X0; Otiothops birabeni (Palpimanidae); Agyneta sp. (Linyphiidae), 2n♂ = 24, X1X20; Coleosoma floridanum, Thymoites sp.1 and Thymoites sp.2 (Theridiidae), 2n♂ = 22, X1X20; Naatlo sp. (Theridiosomatidae), 2n♂ = 30, X1X20; Orthobula sp. (Trachelidae) 2n♂ = 21, X0; Falconina sp. (Corinnidae), 2n♂ = 28, X1X20; Epicratinus sp. (Zodariidae) 2n♂ = 42, X1X20. The chromosomal morphology was determined for all the samples except for Oonopidae. Most species exhibited telocentric chromosomes, with the exception of Palpimanidae and Theridiosomatidae. The main findings: 1) support an hypothesis on ancestral karyotype of Zodariidae and Oonopidae; 2) reveal a relatively high chromosome number in Palpimanidae that supports an idea on relatively high ancestral chromosome number (2n♂ = 42) of entelegyne spiders (Palpimanoidea is sister group of entelegyne spiders); 3) show that the karyotype found in Theridiosomatidae is exceptional within the Araneoidea.

Insights into the Genetic Diversity of Leishmania (Viannia) panamensis in Panama, Inferred via Multilocus Sequence Typing (MLST).

Leishmaniasis is a disease caused by parasites of the genus Leishmania and transmitted by sand fly vectors. Tegumentary leishmaniasis is the most prevalent clinical outcome in Latin America, afflicting people from 18 countries. In Panama, the annual incidence rate of leishmaniasis is as high as 3000 cases, representing a major public health problem. In endemic regions, L. panamensis is responsible for almost eighty percent of human cases that present different clinical outcomes. These differences in disease outcomes could be the result of the local interplay between L. panamensis variants and human hosts with different genetic backgrounds. The genetic diversity of L. panamensis in Panama has only been partially explored, and the variability reported for this species is based on few studies restricted to small populations and/or with poor resolutive markers at low taxonomic levels. Accordingly, in this study, we explored the genetic diversity of sixty-nine L. panamensis isolates from different endemic regions of Panama, using an MLST approach based on four housekeeping genes (Aconitase, ALAT, GPI and HSP70). Two to seven haplotypes per locus were identified, and regional differences in the genetic diversity of L. panamensis were observed. A genotype analysis evidenced the circulation of thirteen L. panamensis genotypes, a fact that might have important implications for the local control of the disease.

Chromosomal study on selected small araneomorph spiders from Brazil, including the first records in Palpimanidae and Theridiosomatidae (Araneae, Araneomorphae)

Among the 50,474 spider species, only 849 have chromosomal data available in the literature. Fifty spider families remain unknown from a cytogenetic perspective. The aim of this study was to analyze chromosomally selected araneomorph spiders from Brazil, to contribute to the cytotaxonomy and chromosome evolution of this group. The karyotypes of 12 species belonging to families Corinnidae, Linyphiidae, Oonopidae, Palpimanidae, Theridiidae, Theridiosomatidae, Trachelidae, and Zodariidae were analyzed, including the first chromosomal record for the first two families. Specimens (with the abdomen content partially exposed by perforation) were subjected to colchicine, hypotonization, and fixation. In most cases, the total content of the abdomen was used to prepare slides by spreading of cell suspension and subsequent Giemsa staining. The results were as follows: Cinetomorpha simplex Simon, 1892 (Oonopidae) 2n♂ = 9, X0; Neotrops sp. and Neoxyphinus termitophilus (Oonopidae), 2n♂ = 7, X0; Otiothops birabeni (Palpimanidae); Agyneta sp. (Linyphiidae), 2n♂ = 24, X1X20; Coleosoma floridanum, Thymoites sp.1 and Thymoites sp.2 (Theridiidae), 2n♂ = 22, X1X20; Naatlo sp. (Theridiosomatidae), 2n♂ = 30, X1X20; Orthobula sp. (Trachelidae) 2n♂ = 21, X0; Falconina sp. (Corinnidae), 2n♂ = 28, X1X20; Epicratinus sp. (Zodariidae) 2n♂ = 42, X1X20. The chromosomal morphology was determined for all the samples except for Oonopidae. Most species exhibited telocentric chromosomes, with the exception of Palpimanidae and Theridiosomatidae. The main findings: 1) support an hypothesis on ancestral karyotype of Zodariidae and Oonopidae; 2) reveal a relatively high chromosome number in Palpimanidae that supports an idea on relatively high ancestral chromosome number (2n♂ = 42) of entelegyne spiders (Palpimanoidea is sister group of entelegyne spiders); 3) show that the karyotype found in Theridiosomatidae is exceptional within the Araneoidea.

Phenotypic and molecular analyses in diploid and tetraploid genotypes of Solanum tuberosum L. reveal promising genotypes and candidate genes associated with phenolic compounds, ascorbic acid contents, and antioxidant activity.

Potato tubers contain biochemical compounds with antioxidant properties that benefit human health. However, the genomic basis of the production of antioxidant compounds in potatoes has largely remained unexplored. Therefore, we report the first genome-wide association study (GWAS) based on 4488 single nucleotide polymorphism (SNP) markers and the phenotypic evaluation of Total Phenols Content (TPC), Ascorbic Acid Content (AAC), and Antioxidant Activity (AA) traits in 404 diverse potato genotypes (84 diploids and 320 tetraploids) conserved at the Colombian germplasm bank that administers AGROSAVIA. The concentration of antioxidant compounds correlated to the skin tuber color and ploidy level. Especially, purple-blackish tetraploid tubers had the highest TPC (2062.41 ± 547.37 mg GAE), while diploid pink-red tubers presented the highest AA (DDPH: 14967.1 ± 4687.79 µmol TE; FRAP: 2208.63 ± 797.35 mg AAE) and AAC (4.52 mg ± 0.68 AA). The index selection allowed us to choose 20 promising genotypes with the highest values for the antioxidant compounds. Genome Association mapping identified 58 SNP-Trait Associations (STAs) with single-locus models and 28 Quantitative Trait Nucleotide (QTNs) with multi-locus models associated with the evaluated traits. Among models, eight STAs/QTNs related to TPC, AAC, and AA were detected in common, flanking seven candidate genes, from which four were pleiotropic. The combination in one population of diploid and tetraploid genotypes enabled the identification of more genetic associations. However, the GWAS analysis implemented independently in populations detected some regions in common between diploids and tetraploids not detected in the mixed population. Candidate genes have molecular functions involved in phenolic compounds, ascorbic acid biosynthesis, and antioxidant responses concerning plant abiotic stress. All candidate genes identified in this study can be used for further expression analysis validation and future implementation in marker-assisted selection pre-breeding platforms targeting fortified materials. Our study further revealed the importance of potato germplasm conserved in national genebanks, such as AGROSAVIA's, as a valuable genetic resource to improve existing potato varieties.

Environmental and Genetic Factors Affecting Apospory Expressivity in Diploid Paspalum rufum.

In angiosperms, gametophytic apomixis (clonal reproduction through seeds) is strongly associated with polyploidy and hybridization. The trait is facultative and its expressivity is highly variable between genotypes. Here, we used an F1 progeny derived from diploid apomictic (aposporic) genotypes of Paspalum rufum and two F2 families, derived from F1 hybrids with different apospory expressivity (%AES), to analyze the influence of the environment and the transgenerational transmission of the trait. In addition, AFLP markers were developed in the F1 population to identify genomic regions associated with the %AES. Cytoembryological analyses showed that the %AES was significantly influenced by different environments, but remained stable across the years. F1 and F2 progenies showed a wide range of %AES variation, but most hybrids were not significantly different from the parental genotypes. Maternal and paternal genetic linkage maps were built covering the ten expected linkage groups (LG). A single-marker analysis detected at least one region of 5.7 cM on LG3 that was significantly associated with apospory expressivity. Our results underline the importance of environmental influence in modulating apospory expressivity and identified a genomic region associated with apospory expressivity at the diploid level.

Differential Epigenetic Marks Are Associated with Apospory Expressivity in Diploid Hybrids of Paspalum rufum.

Apomixis seems to emerge from the deregulation of preexisting genes involved in sexuality by genetic and/or epigenetic mechanisms. The trait is associated with polyploidy, but diploid individuals of Paspalum rufum can form aposporous embryo sacs and develop clonal seeds. Moreover, diploid hybrid families presented a wide apospory expressivity variation. To locate methylation changes associated with apomixis expressivity, we compare relative DNA methylation levels, at CG, CHG, and CHH contexts, between full-sib P. rufum diploid genotypes presenting differential apospory expressivity. The survey was performed using a methylation content-sensitive enzyme ddRAD (MCSeEd) strategy on samples at premeiosis/meiosis and postmeiosis stages. Based on the relative methylation level, principal component analysis and heatmaps, clearly discriminate samples with contrasting apospory expressivity. Differential methylated contigs (DMCs) showed 14% of homology to known transcripts of Paspalum notatum reproductive transcriptome, and almost half of them were also differentially expressed between apomictic and sexual samples. DMCs showed homologies to genes involved in flower growth, development, and apomixis. Moreover, a high proportion of DMCs aligned on genomic regions associated with apomixis in Setaria italica. Several stage-specific differential methylated sequences were identified as associated with apospory expressivity, which could guide future functional gene characterization in relation to apomixis success at diploid and tetraploid levels.

Physiological, Biochemical and Yield-Component Responses of Solanum tuberosum L. Group Phureja Genotypes to a Water Deficit.

Water deficits are the major constraint in some potato-growing areas of the world. The effect is most severe at the tuberization stage, resulting in lower yield. Therefore, an assessment of genetic and phenotypic variations resulting from water deficits in Colombia germplasm is required to accelerate breeding efforts. Phenotypic variations in response to a water deficit were studied in a collection of Solanum tuberosum Group Phureja. A progressive water deficit experiment on the tuberization stage was undertaken using 104 genotypes belonging to the Working Collection of the Potato Breeding Program at the Universidad Nacional de Colombia. The response to water deficit conditions was assessed with the relative chlorophyll content (CC), maximum quantum efficiency of PSII (Fv/Fm), relative water content (RWC), leaf sugar content, tuber number per plant (TN) and tuber fresh weight per plant (TW). Principal Component Analysis (PCA) and cluster analysis were used, and the Drought Tolerance Index (DTI) was calculated for the variables and genotypes. The soluble sugar contents increased significantly under the deficit conditions in the leaves, with a weak correlation with yield under both water treatments. The PCA results revealed that the physiological, biochemical and yield-component variables had broad variation, while the yield-component variables more powerfully distinguished between the tolerant and susceptible genotypes than the physiological and biochemical variables. The PCA and cluster analysis based on the DTI revealed different levels of water deficit tolerance for the 104 genotypes. These results provide a foundation for future research directed at understanding the molecular mechanisms underlying potato tolerance to water deficits.

Dry matter productivity and bromatological quality of ryegrass genotypes cultivated in southern Brazil / [Produtividade de matéria seca e qualidade bromatológica de genótipos de azevém cultivados no Sul do Brasil]

The goal of this work was to assess the biomass production and bromatological quality of ryegrass genotypes in ten municipalities of the Western and North Plateau regions of the State of Santa Catarina, Brazil. The cultivars La Estanzuela 284 (diploid), Bar HQ, Barjumbo, INIA Escorpio, Potro, and Winter Star (tetraploids) were compared, distributed in a randomized block design, in which the municipalities constituted the blocks, with three replications. The cuts were performed when the plants reached 30cm, leaving a residue of 10cm. In three cuts, the cultivars Barjumbo and Bar HQ were the most productive, exceeding 4.6 t ha-1 of dry matter. In the places in which five cuts were performed, the production of these cultivars exceeded 7.3 t ha-1, placing them again ahead of the others. The average crude protein content in three cuts was greater than 25% in all cultivars. There was no difference between the genotypes in the content of neutral detergent fiber and total digestible nutrients. There was a significant correlation between quantitative and qualitative productive variables. The assessed cultivars represented good options for composing short-term or long-term winter-feeding systems, adjusted to the integration with annual crops or warm-season pastures.(AU)

O objetivo deste trabalho foi avaliar a produção de biomassa e qualidade bromatológica de genótipos de azevém, em dez municípios das regiões Oeste e Planalto Norte Catarinense, Estado de Santa Catarina, Brasil. Foram comparados os cultivares La Estanzuela 284 (diploide), Bar HQ, Barjumbo, INIA Escorpio, Potro e Winter Star (tetraploides), distribuídos em um delineamento blocos casualizados, em que os municípios constituíram os blocos, com três repetições. Os cortes foram realizados quando as plantas atingiram 30cm, deixando um resíduo de 10cm. Sob três cortes, os cultivares Barjumbo e Bar HQ foram os mais produtivos, ultrapassando 4,6 t ha-1 de matéria seca. Nos locais em que ocorreram cinco cortes, a produção destes cultivares superou 7,3 t ha-1, posicionando-os novamente à frente dos demais. O teor médio de proteína bruta em três cortes foi superior a 25% em todos os cultivares. Não houve diferença entre os genótipos no teor de fibra detergente neutro e de nutrientes digestíveis totais. Verificou-se correlação significativa entre variáveis produtivas quantitativas e qualitativas. Os cultivares testados representam boas opções para compor sistemas forrageiros hiberno-primaveris de curta ou longa duração, ajustando-se à integração com lavouras ou pastagens anuais de estação quente.(AU)

Unrevealing the Karyotypic Evolution and Cytotaxonomy of Armored Catfishes (Loricariinae) with Emphasis in Sturisoma, Loricariichthys, Loricaria, Proloricaria, Pyxiloricaria, and Rineloricaria.

This study provides new insight into the chromosomal diversification in Loricariinae. We analyzed nine species from different Brazilian hydrographic basins, using conventional and molecular cytogenetic methods, aiming to understand the karyotypic diversification, and contribute with cytotaxonomic markers in this group considered one of the most diverse of Loricariidae. Our results evidenced a high karyotypic variability in diploid number (2n) ranging from 2n = 54 (Loricariichthys platymetopon and Loricariichthys anus), 2n = 60 (Rineloricaria reisi and Rineloricaria parva), 2n = 62 (Proloricaria prolixa), 2n = 64 (Loricaria cataphracta complex species), 2n = 66 (Sturisoma barbatum), and 2n = 68 (Pyxiloricaria menezesi). Different patterns of 18S and 5S ribosomal DNA (rDNA) were also identified, while slight divergences in heterochromatin distribution were observed. This high variability is probably related with independent events of Robertsonian translocations, pericentric inversions, and different mechanisms of rDNA sites dispersion (nonreciprocal translocation and transposable element [TEs] co-localization). In addition, our study provides a set of efficient chromosomal markers for the characterization of all analyzed species, and certainly, in future analyzes, will contribute as a useful cytotaxonomic tool in groups where the traditional taxonomy based on morphological data are not sufficient to clarify their relationship.

Molecular Cytogenetic Analysis in Freshwater Prawns of the Genus Macrobrachium (Crustacea: Decapoda: Palaemonidae).

Freshwater prawns of the genus Macrobrachium are one of the important components of circumtropical marine, estuarine, and freshwater environments. They have been extensively exploited for human consumption for many years. More than 250 species reflect the evolutionary success of this highly diversified group, with a complex and challenging taxonomy due to morphological variations and vast geographical distribution. Although genetic approaches have been used to clarify phylogenetic and taxonomic aspects of Macrobrachium species, cytogenetic information is still very scarce and mostly focused on chromosome number and morphology. Here, we present chromosome data for three species from the Neotropical region, M. carcinus, M. acanthurus, and M. amazonicum, and one species from the Oriental region, M. rosenbergii. Using conventional cytogenetic approaches and chromosome mapping of repetitive DNAs by fluorescence in situ hybridization (FISH), we identified numerical diversification of the diploid set, within and between both zoogeographic regions. These included M. acanthurus and M. amazonicum sharing diploid chromosomes of 98, while M. carcinus has 94, and M. rosenbergii has 118 chromosomes. Argentophilic sites are also variable in number, but they occur in a much higher number than 18S rDNA, representing two to 10 sites within the study species. Microsatellites repeat motifs are also abundant in the chromosomes, with a co-localization and uniform distribution along the chromosome arms, but completely absent in the AT-rich centromeric regions. As a whole, our study suggests that the 2n divergence was followed by a considerable rDNA diversification. The abundance of the exceptional amount of microsatellite sequences in the chromosomes also suggests that they are essential components of the Macrobrachium genome and, therefore, maintained as a shared feature by the species, the reason for which is yet unknown.

Queen Execution, Diploid Males, and Selection For and Against Polyandry in the Brazilian Stingless Bee Scaptotrigona depilis.

Female mating frequency varies. Determining the causes of this variation is an active research area. We tested the hypothesis that in stingless bees, Meliponini, single mating is due to the execution of queens that make a matched mating at the complementary sex determination locus and have diploid male offspring. We studied the Brazilian species Scaptotrigona depilis. We made up 70 test colonies so that 50% (single matched mating), 25% (double mating), 12.5% (quadruple mating), or 0% (single nonmatched mating) of the emerging brood were diploid males. Queen execution following diploid male emergence was equal and high in colonies producing 50% (77% executed) and 25% (75%) diploid males versus equal and low in colonies producing 12.5% (7%) and 0% (0%) diploid males. These results show that queens that mate with two males with similar paternity suffer an increased chance of being executed, which selects against double mating. However, double mating with unequal paternity (e.g., 25∶75), which occasionally occurs in S. depilis, is selectively neutral. Single mating and double mating with unequal paternity form one adaptive peak. The results show a second adaptive peak at quadruple mating. However, this is inaccessible via gradual evolutionary change in a selective landscape with reduced fitness at double mating.

Screening and analysis on the differentially expression genes between diploid and autotetraploid watermelon by using of digital gene expression profile / Triagem e análise da expressão diferencial dos genes entre melancia diplóide e autotetraplóide através do perfil de expressão gênica digital

Abstract Synthetic polyploids are key breeding materials for watermelon. Compared with diploid watermelon, the tetraploid watermelon often exhibit wide phenotypic differences and differential gene expression. Digital gene expression (DGE) profile technique was performed in this study to present gene expression patterns in an autotetraploid and its progenitor diploid watermelon, and deferentially expressed genes (DEGs) related to the abiotic and biotic stress were also addressed. Altogether, 4,985 DEGs were obtained in the autotetraploid against its progenitor diploid, and 66.02% DEGs is up-regulated. GO analysis shows that these DEGs mainly distributed in 'metabolic process', 'cell' and 'catalytic activity'. KEGG analysis revealed that these DEGs mainly cover 'metabolic pathways', 'secondary metabolites' and 'ribosome'. Moreover, 134 tolerance related DEGs were identified which cover osmotic adjustment substance, protective enzymes/protein, signaling proteins and pathogenesis-related proteins. This study present the differential expression of stress related genes and global gene expression patterns at background level in autotetraploid watermelons. These new evidences could supplement the molecular theoretical basis for the better resistance after the genome doubling in the gourd family.

Resumo Poliploides sintéticos são materias fundamentais para melhoramento genético da melancia. Comparativamente ao seu homólogo diploide, a melancia tetraploide apresenta amplas diferenças genotípica e fenotípica e diferença de expressão gênica. A expressão gênica digital ou DGE (digital gene expression) foi utilizada neste estudo para representar o perfil de expressão gênica da melancia autotetraploide e seu progenitor diploide e a expressão diferencial de genes relacionados ao estresse biótico e abiótico. Os resultados mostraram que 4.985 DEGs foram observados no organismo autotetraploide, sendo que, deste total, 66.02%foram supra-regulados. A análise de ontologia gênica (GO) mostrou que estes DEGs estão relacionados principalmente com processos metabólicas, célula e atividade catalítica, abrangendo de acordo com a análise de genes e genoma (KEGG) rotas metabólicas, metabolismo secundário e ribossomos. Além disso, 134 genes de defesa foram identificados, abrangendo substâncias de ajuste osmótico, enzimas/proteínas de proteção, proteínas sinalizadoras e proteínas relacionadas à patogênese. Este estudo mostrou a expressão diferencial de genes relacionados ao estresse e o perfil global de expressão gênica de melancia autotetraploide, estes resultados podem complementar, a nível molecular, o entendimento do fator resistência após a duplicação do genoma em cucurbitáceas.

Evaluation in situ of genotoxic and cytotoxic response in the diploid/polyploid complex Odontophrynus (Anura: Odontophrynidae) inhabiting agroecosystems.

Polyploidization has been documented across a wide range of vertebrates. Gene duplication could promote better adaptation to environmental changes and to chronic injury or stress. We investigated if genotoxic and cytotoxic responses to agricultural impact are affected by ploidy. We evaluate syntopic populations of the cryptic diploid/polyploid complex Odontophrynus cordobae/O. americanus breeding in an agroecosystem from Central Argentina. The blood of 72 adult anurans was analysed. We used erythrometry to distinguish Odontophrynus individuals with different ploidy levels. We calculated micronucleus frequencies (Mn) and erythrocytic nuclear abnormalities (ENAs) as genotoxic effects and enucleated, mitotic, pyknotic and immature erythrocytes as cytotoxic endpoints (CYT). Mn, ENAs and CYT frequencies were significantly different between diploid and polyploid organisms. The higher frequencies of Mn and CYT were recorded in polyploid organisms, and the higher frequency of ENAs was recorded in diploids. These results indicate that stress response, as indicated by most genotoxic and cytotoxic endpoints, was higher in polyploids respect to diploids. Polyploidy could provide greater genetic flexibility increasing buffering against exogenous DNA-damaging agents and thus confer an advantage over diploids under certain environmental conditions.

Screening and analysis on the differentially expression genes between diploid and autotetraploid watermelon by using of digital gene expression profile / Triagem e análise da expressão diferencial dos genes entre melancia diplóide e autotetraplóide através do perfil de expressão gênica digital

Synthetic polyploids are key breeding materials for watermelon. Compared with diploid watermelon, the tetraploid watermelon often exhibit wide phenotypic differences and differential gene expression. Digital gene expression (DGE) profile technique was performed in this study to present gene expression patterns in an autotetraploid and its progenitor diploid watermelon, and deferentially expressed genes (DEGs) related to the abiotic and biotic stress were also addressed. Altogether, 4,985 DEGs were obtained in the autotetraploid against its progenitor diploid, and 66.02% DEGs is up-regulated. GO analysis shows that these DEGs mainly distributed in metabolic process, cell and catalytic activity. KEGG analysis revealed that these DEGs mainly cover metabolic pathways, secondary metabolites and ribosome. Moreover, 134 tolerance related DEGs were identified which cover osmotic adjustment substance, protective enzymes/protein, signaling proteins and pathogenesis-related proteins. This study present the differential expression of stress related genes and global gene expression patterns at background level in autotetraploid watermelons. These new evidences could supplement the molecular theoretical basis for the better resistance after the genome doubling in the gourd family.(AU)

Poliploides sintéticos são materias fundamentais para melhoramento genético da melancia. Comparativamente ao seu homólogo diploide, a melancia tetraploide apresenta amplas diferenças genotípica e fenotípica e diferença de expressão gênica. A expressão gênica digital ou DGE (digital gene expression) foi utilizada neste estudo para representar o perfil de expressão gênica da melancia autotetraploide e seu progenitor diploide e a expressão diferencial de genes relacionados ao estresse biótico e abiótico. Os resultados mostraram que 4.985 DEGs foram observados no organismo autotetraploide, sendo que, deste total, 66.02%foram supra-regulados. A análise de ontologia gênica (GO) mostrou que estes DEGs estão relacionados principalmente com processos metabólicas, célula e atividade catalítica, abrangendo de acordo com a análise de genes e genoma (KEGG) rotas metabólicas, metabolismo secundário e ribossomos. Além disso, 134 genes de defesa foram identificados, abrangendo substâncias de ajuste osmótico, enzimas/proteínas de proteção, proteínas sinalizadoras e proteínas relacionadas à patogênese. Este estudo mostrou a expressão diferencial de genes relacionados ao estresse e o perfil global de expressão gênica de melancia autotetraploide, estes resultados podem complementar, a nível molecular, o entendimento do fator resistência após a duplicação do genoma em cucurbitáceas.(AU)

Micropropagación y determinación del número cromosómico de Puya trianae con fines de conservación y uso ornamental / Micropropagation and determination of the chromosome number of Puya trianae for conservation and ornamental use

Puya trianae se caracteriza por desarrollar una vistosa inflorescencia que la caracteriza como planta de uso ornamental. Aunque actualmente no se considera amenazada, debido al rápido deterioro de los ecosistemas donde habita, la supervivencia de sus poblaciones sí está amenazada; por dichas razones se desarrolló un protocolo de micropropagación, aplicable para su conservación y para producción masiva de plántulas con fines ornamentales, así como a la determinación de su número cromosómico. Para la multiplicación se utilizaron plántulas provenientes de semillas germinadas in vitro realizando cuatro ensayos para determinar tipo, concentración y tiempo de exposición de algunos reguladores, además de la escisión parcial o total de explantes. Durante la fase de enraizamiento se cultivaron brotes en MS/2 solo y/o suplementado con AIB y ANA; la aclimatización de las plántulas obtenidas se realizó en diferentes mezclas de tierra:turba:capote, en cuarto de crecimiento e invernadero. Los resultados mostraron que en MS+2.0 mgL-1 de TDZ durante 45 días se produjo la mayor cantidad de yemas y brotes que terminaron su desarrollo después de 90 días. Los brotes enraizaron en MS/2 cuantificándose 70% de enraizamiento. Se registró una viabilidad del 90% y una longitud de plántulas de 3.6 cm en tierra:turba después de 60 días de aclimatización. El conteo cromosómico se realizó en células meristemáticas radicales tratadas con colchicina al 0.5%, fijadas en carnoy, hidrolizadas con HCL y con enzimas y teñidas con orceína acética, obteniéndose un número cromosómico de 50, con cromosomas muy pequeños (aprox. 1.2μm).

Puya trianae is characterized for developing a colorful inflorescence, which turned it into an ornamental plant. Is not currently considered threatened, due to the fast habitat deterioration, populations survival is threatened; due to the last, we developed a micropropagation protocol, focused on its conservation and mass production for ornamental purpose, as well as the chromosome counting. For multiplication, seedlings from germinated seeds were used in vitro and we made four tests to determine the type, concentration and time of exposure of some regulators, as well as the partial or total explants excised. Throughout the rooting phase, shoots were grown in MS/2 alone and/or supplemented with IBA and NAA; the acclimatization of the obtained seedlings was carried out in different mixtures land:peat:capote, in a growth room and a greenhouse. Buds and shoots that finished their development during 45 days in MS+2.0 mgL-1 of TDZ were the majority, after three months of development. The shoots rooted in MS/2 quantifying a 70% of rooting. A 90% viability and a plantlets length of 3.6 cm in soil: peat after 60 days of transfer. We made a radical meristematic cells chromosomal counting treated with a 0.5% of colchicine fixed in carnoy, hydrolyzed with HCL and enzymes and stained with acetic orcein, obtaining a chromosome number of 2n=50, with very small chromosomes (aprox. 1.2μm).

Divergent Chromosome Evolution in Hypostominae Tribes (Siluriformes: Loricariidae): Correlation of Chromosomal Data with Morphological and Molecular Phylogenies.

Hypostominae is the largest subfamily of Loricariidae, and is widely distributed throughout the Neotropic. In the present article, we analyze three Loricariidae species that were considered part of Hypostominae, from three different tribes, to discuss chromosome evolution in this fish group and to review the existent data for the subfamily. Rhinelepis aspera had 54 chromosomes (20m + 26sm+8st), whereas Pterygoplichthys ambrosettii and Megalancistrus parananus had 52 chromosomes, with 16m+24sm+8st+4a and 18m+24sm+10st, respectively. The karyological data were compared with existent phylogenetic hypotheses, indicating a common ancestor with 2n = 52 chromosomes for the Acanthicus, Hemiancistrus, and Peckoltia clades, as well as for Hypostomini. Shared recurrent characteristics of the tribes are discussed, as well as peculiarities of genera Ancistrus and Hypostomus. We propose that the occurrence of fragile sites demonstrated for Ancistrus facilitated chromosomal rearrangements that decreased the proportion of metacentric/submetacentric chromosomes and the diploid number in many species from this genus. Although Hypostominae is usually considered a subfamily with derived chromosome features, our revision shows that this is valid only for Hypostomini and Ancistrini, which have a divergent chromosome evolution from other tribes that seems to conserve plesiomorphic features.

Synergy from reproductive division of labor and genetic complexity drive the evolution of sex.

Computer experiments that mirror the evolutionary dynamics of sexual and asexual organisms as they occur in nature were used to test features proposed to explain the evolution of sexual recombination. Results show that this evolution is better described as a network of interactions between possible sexual forms, including diploidy, thelytoky, facultative sex, assortation, bisexuality, and division of labor between the sexes, rather than a simple transition from parthenogenesis to sexual recombination. Diploidy was shown to be fundamental for the evolution of sex; bisexual reproduction emerged only among anisogamic diploids with a synergistic division of reproductive labor; and facultative sex was more likely to evolve among haploids practicing assortative mating. Looking at the evolution of sex as a complex system through individual-based simulations explains better the diversity of sexual strategies known to exist in nature, compared to classical analytical models.

Conserved number of U2 snDNA sites in Piabina argentea, Piabarchus stramineus and two Bryconamericus species (Characidae, Stevardiinae)

The chromosomal location of 5S rRNA and U2 snRNA genes of Piabina argentea, Piabarchus stramineus and two Bryconamericus species from two different Brazilian river basins were investigated, in order to contribute to the understanding of evolutionary characteristics of these repetitive DNAs in the subfamily Stevardiinae. The diploid chromosome number was 2n = 52 for Bryconamericus cf. iheringii, Bryconamericus turiuba, Piabarchus stramineus and Piabina argentea. The 5S rDNA clusters were located on one chromosome pair in P. stramineus and B. cf. iheringii, and on two pairs in B. turiuba and P. argentea. The U2 snDNA clusters were located on the one pair in all species. Two-color FISH experiments showed that the co-localization between 5S rDNA and U2 snDNA in P. stramineus can represent a marker for this species. Thus, the present study demonstrated that the number of U2 snDNA clusters observed for the four species was conserved, but particular characteristics can be found in the genome of each species.(AU)

A localização cromossômica dos genes de RNAr 5S e RNAsn U2 de Piabina argentea, Piabarchus stramineus e duas espécies de Bryconamericus provenientes de duas bacias hidrográficas foi investigada, com a intenção de contribuir com o entendimento de características evolutivas destes DNAs repetitivos na subfamília Stevardiinae. O número cromossômico diploide foi 2n = 52 para Bryconamericus cf. iheringii, Bryconamericus turiuba, Piabarchus stramineus e Piabina argentea. Os sítios de DNAr 5S foram localizados em um par cromossômico em P. stramineus e B. cf. iheringii, e em dois pares em B. turiuba e P. argentea. Os sítios de DNAsn U2 foram localizados em um par em todas as espécies. Experimentos de FISH com duas sondas mostraram que a co-localização entre os DNAr 5S e DNAsn U2 em P. stramineus pode representar um marcador para esta espécie. Portanto, o presente estudo demonstrou que o número de sítios de DNAsn U2 observado para as quatro espécies foi conservado, porém características particulares podem ser encontradas no genoma de cada espécie.(AU)

Screening and analysis on the differentially expression genes between diploid and autotetraploid watermelon by using of digital gene expression profile

Abstract Synthetic polyploids are key breeding materials for watermelon. Compared with diploid watermelon, the tetraploid watermelon often exhibit wide phenotypic differences and differential gene expression. Digital gene expression (DGE) profile technique was performed in this study to present gene expression patterns in an autotetraploid and its progenitor diploid watermelon, and deferentially expressed genes (DEGs) related to the abiotic and biotic stress were also addressed. Altogether, 4,985 DEGs were obtained in the autotetraploid against its progenitor diploid, and 66.02% DEGs is up-regulated. GO analysis shows that these DEGs mainly distributed in metabolic process, cell and catalytic activity. KEGG analysis revealed that these DEGs mainly cover metabolic pathways, secondary metabolites and ribosome. Moreover, 134 tolerance related DEGs were identified which cover osmotic adjustment substance, protective enzymes/protein, signaling proteins and pathogenesis-related proteins. This study present the differential expression of stress related genes and global gene expression patterns at background level in autotetraploid watermelons. These new evidences could supplement the molecular theoretical basis for the better resistance after the genome doubling in the gourd family.

Resumo Poliploides sintéticos são materias fundamentais para melhoramento genético da melancia. Comparativamente ao seu homólogo diploide, a melancia tetraploide apresenta amplas diferenças genotípica e fenotípica e diferença de expressão gênica. A expressão gênica digital ou DGE (digital gene expression) foi utilizada neste estudo para representar o perfil de expressão gênica da melancia autotetraploide e seu progenitor diploide e a expressão diferencial de genes relacionados ao estresse biótico e abiótico. Os resultados mostraram que 4.985 DEGs foram observados no organismo autotetraploide, sendo que, deste total, 66.02%foram supra-regulados. A análise de ontologia gênica (GO) mostrou que estes DEGs estão relacionados principalmente com processos metabólicas, célula e atividade catalítica, abrangendo de acordo com a análise de genes e genoma (KEGG) rotas metabólicas, metabolismo secundário e ribossomos. Além disso, 134 genes de defesa foram identificados, abrangendo substâncias de ajuste osmótico, enzimas/proteínas de proteção, proteínas sinalizadoras e proteínas relacionadas à patogênese. Este estudo mostrou a expressão diferencial de genes relacionados ao estresse e o perfil global de expressão gênica de melancia autotetraploide, estes resultados podem complementar, a nível molecular, o entendimento do fator resistência após a duplicação do genoma em cucurbitáceas.