This study describes the use of erythrocytes (RBCs) of loggerhead turtles as in vitro models for evaluating their toxicity to methylmercury. Blood samples of loggerhead turtles that were born in the Colombian Caribbean were used. The LC50 of RBCs to methylmercury was determined at 96 h using methylmercury concentrations of 0.5–100 mg L−1. Next, the viability of the RBCs and the activity of the enzymes superoxide dismutase (SOD), glutathione S-transferase (GST), and lipid peroxidation by malondialdehyde (MDA) at 6 and 12 h of exposure to acute concentrations of 0, 1, and 5 mg L−1 were evaluated. The LC50 for loggerhead turtle RBCs was 8.32 mg L−1. The cell viability bioassay of RBCs exposed for 12 h only showed 100% cell viability. Increasing in vitro MeHg concentrations caused a corresponding increase in MDA concentration as well as decreases in the activities of SOD and GST. The RBCs represent an excellent model for ecotoxicological studies and SOD, GST, and MDA are biomarkers of environmental pollution and oxidative stress in loggerhead turtles. This was the first study conducted on loggerhead turtle where the response of RBCs to MeHg-induced oxidative stress is evaluated.
Management of the South American tomato leafminer, Tuta absoluta Meyrick, with insecticides has led to the widespread development of insect resistance. Mass trapping using traps baited with the female-produced sex pheromone is an attractive alternative for the management of this pest. The current study evaluated several commercial trap designs for capture of T. absoluta. Based on its small size and ease of handling, the most effective trap is a small plastic container with entry windows cut on the sides filled with motor oil over water. These traps are most effective when placed near ground level. Tests of septa containing 0.1 or 0.2 mg of the pheromone (95:5) E4, Z8-14Ac/E4,Z8,Z11-14Ac were slightly more attractive than septa loaded with 0.5, 1.0, or 2 mg during the first week of use, but the latter three loadings were slightly more attractive than the first two loadings after 9 weeks. Ideal trap baits were loaded with 0.5 mg of pheromone. Higher numbers of T. absoluta were captured near upwind borders of tomato fields suggesting that treatments against T. absoluta should be concentrated near upwind parts of fields. Comparisons of conventional insecticide treatment versus mass trapping to manage T. absoluta damage in three different test sites showed that even when initial captures in monitoring traps were high (>35 males trap(-1) day(-1)), mass trapping at 48 traps/ha reduced leaf damage more efficiently than conventional insecticide treatment. Based on the typical insecticide recommendations against T. absoluta, mass trapping is an economically viable alternative.
Colombia is one of the leading producers of yellow passion fruit but the genetic studies based on molecular markers from commercial plantations have not been considered to select interesting market material. The goal of this study was to assess the genetic variability and the population structure of 51 Colombian commercial yellow passion fruit accessions (102 individuals), and to provide the necessary information for prospective selection and breeding programs. Thus, a total of six microsatellites were amplified with 58 alleles identified and an average of 9.66 alleles per locus, including nine private and 31 rare. Diversity indexes showed polymorphic information content values of 0.74 (PIC), an observed (Ho) and expected (He) heterozygosity average of 0.52 and 0.78, respectively. Spatial distribution showed the greatest allelic richness (11 to 14) in most of the Valle del Cauca accessions. The average genetic distance among accessions was 0.68, and the cluster analysis showed three main groups poorly supported (bootstrap <50%), with slight geographical structure and high differentiation between individuals of the same accession. Structure analysis indicated K=4 as the genetic structure's uppermost hierarchical level, while Bayesian clustering showed a division of individuals into four genetically distinct groups. The low geographic structure and high variability of the accessions could be explained by allogamy and seed exchange frequency among farmers. Results issued suggest a complementary agro-morphological assessment to establish total genetic variability and implement a breeding program through assisted selection of superior genotypes in search of more productive and resistant cultivars to phytosanitary problems.
Loggerhead sea turtle Caretta caretta is widely distributed in the oceans of tropical and subtropical latitude. This turtle is an endangered species due to anthropic and natural factors that have decreased their population levels. In this study, RNA sequencing and de-novo assembly of genes expressed in blood were performed. The raw FASTQ files have been deposited on NCBI's SRA database with accession number SRX2629512. A total of 5.4 Gb raw sequence data were obtained, corresponding to 48,257,019 raw reads. Trinity pipeline was used to perform a de-novo assembly, we were able to identify 64,930 transcripts for female loggerhead turtle transcriptome with an N50 of 1131 bp. The obtained transcriptome data will be useful for further studies of the physiology, biochemistry and evolution in this species.
Lymphocyte culture and partial karyotype of the marine turtle Caretta caretta (Testudines: Cheloniidae) in Santa Marta, Colombian Caribbean. Over the past few years an important reduction in the number of nesting marine turtle Caretta caretta individuals has been registered in the Colombian Caribbean, raising the question of a possible extinction in the medium term. A conservation plan is needed. We studied the culture requirements for C. caretta lymphocytes and preliminary karyotype analysis for cytogenetic identification, immunological study and toxicology without the need to kill individuals. Peripheral blood samples were obtained from 47 individuals in Santa Marta, Colombia and tests were made until optimal conditions were established for lymphocyte culture. The karyotype had 56 chromosomes, 32 macrochromosomes and 24 microchromosomes. An ideogram showed that C. caretta has four groups of chromosomes. Sexual chromosomes were not observed. These results do not coincide with the karyotype described from the Pacific (Japan). The present study is the first to include a complete description of the chromosome morphology of turtles from the Atlantic Ocean. it is possible that one of the adaptive strategies of this species is genetic interchange with other species of the family, producing viable hybrids. individuals in this study might be viable hybrids of C. caretta and further molecular studies are needed.
RESUMEN-La granadilla es la segunda especie en importancia económica del género Passiflora y Colombia es el principal productor del mundo con 53.000 t/año. Son pocos los estudios sobre la diversidad intraespecifica en la especie que permitan establecer las relaciones genéticas entre individuos. El objetivo de esta investigación fue explorar la variabilidad genética de la granadilla cultivada en Colombia por medio de marcadores microsatélites. Diez marcadores microsatélites fueron evaluados en 41 accesiones (82 individuos) provenientes de los principales departamentos productores. Un total de cinco microsatélites fueron amplificados con 66 alelos identificados y un promedio de 12,2, entre ellos 7 únicos y 13 raros. Los índices de diversidad mostraron un contenido de información polimórfica de 0,74 (PIC), y una heterocigocidad promedio observada (Ho) y esperada (He) de 0,98 y 0,96 bajo condiciones de equilibrio de Hardy-Weinberg. La distancia genética promedio dentro y entre poblaciones fue de 0,65 y 0,80, siendo Boyacá, Valle del Cauca y Putumayo los más distantes (>0,87). Los análisis de clasificación arbórea (nj) y factorial de correspondencia múltiple (AFCM) revelaron poca estructuración geográfica de las accesiones y dispersión de los individuos de un mismo origen. La carencia de estructuración y la alta variabilidad intraespecífica podría explicarse por el fenómeno de alogamia presente en la especie y el intercambio de semillas entre productores. En conclusión, estos resultados sugieren una evaluación agromorfológica complementaria que permita establecer la variabilidad genética total e implementar un programa de mejoramiento genético por medio de la selección asistida de genotipos superiores en búsqueda de cultivares más productivos y resistentes a problemas fitosanitarios que afectan los cultivos. Palabras clave: Passiflora, fruta, marcadores moleculares, variabilidad intraespecífica, fitomejoramiento. CHARACTERIZATION AND ANALYSIS OF THE GENETIC VARIABILITY OF SWEET PASSION FRUIT (Passiflora ligularis Juss.) IN COLOMBIA USING MICROSATELLITE MARKERSABSTRACT-Sweet passion fruit is the second economically important species of the genus Passiflora and Colombia is the largest producer in the world with 53,000 t/year. There are however, few studies on intraspecific diversity within the species that establish genetic relationships between individuals. The objective of this research was to explore the genetic variability of cultivated sweet passion fruit in Colombia using microsatellite markers. Ten microsatellite markers were evaluated in 41 accessions (82 individuals) from the main producing departments in the country. A total of five microsatellites were amplified with 66 alleles identified and an average of 12.2, including 7 unique and 13 rare. Diversity indexes showed a polymorphic information content of 0.74 (PIC), and an observed (Ho) and expected (He) heterozygosity average of 0.98 and 0.96 under Hardy-Weinberg equilibrium conditions. The average genetic distance within and among populations was 0.65 and ...
Twenty-eight soil samples were obtained from open fields and greenhouses used for tomato cultivation in various regions of Colombia. For functional characterization, 99 Bacillus thuringiensis (Bt) strains were isolated and characterized by abundance and morphology of microscopic crystals, SDS-PAGE of protein extracts and M-PCR analyses of genes of the cry1 family, as well as for their insecticidal activity against Tuta absoluta second instar larvae. Native Bt strains had amorphous (5%), bi-pyramidal (27%), square (8%), spherical (38%) and triangular (22%) crystal forms. Based on the presence of 1-4 different crystal forms, 18 different profiles were established. The SDS-PAGE analyses of protein extracts established ten different strain groups based on their protein band weight and potential biological activity. The M-PCR technique identified 35 native Bt strains based on the presence of the 6 genes cry1Aa, cry1Ab, cry1Ac, cry1B, cry1C and cry1D, whose frequency of occurrence was 76, 26, 21, 35, 32 and 8.8%, respectively. Thirteen different PCR profiles were found in native Bt strains. Several gene combinations tended to cooccur with elevated frequency, such as the pairs cry1Ac/ cry1C, cry1Ab/cry1Ac and cry1Ab/cry1B, for which Pearson correlation coefficients were 0.69, 0.52 and 0.54, respectively. Native strains ZBUJTL39 and ZCUJTL11 had up to three times higher biological activity against T. absoluta second instar larvae than the reference strain Bt var. kurstaki HD1, with an LD 50 of 2.4 lg/ml (P \ 0.05) for native Bt strain ZCUJTL11. This study suggests a high biodiversity of native Bt strains from tomato growing regions in Colombia, which has important implications for designing biological control strategies for T. absoluta.
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