Abstract: Authentication of cell lines is of paramount importance to validate the results from their use in biomedical research. Although isoenzyme polymorphism is the standard method, molecular methods based on mitochondrial DNA (mtDNA) have been developed to replace it. The aim of this study was the improvement of our isoenzyme electrophoretic analysis and the validation of one molecular technique targeted at mtDNA for the authentication of our animal cell lines. The combined method of cellular lysing through osmotic shock, followed by freezing-thawing in N2 to obtain isoenzyme extracts, and with 42 × 106 cells maintained the best efficiency. The superior electrophoretic conditions were PAGE run at 200 V. All cell lines had isoenzymatic mobility corresponding to their species to lactate dehydrogenase, malate-dehydrogenase, and glucose-6-phosphate dehydrogenase isoenzymes, and could be distinguished from each other. Two molecular techniques based on mtDNA were tested, one on the cytochrome b gene and other on cytochrome c oxidase I subunit gene. Due to difficulties in distinguishing all cell lines using only one these techniques, we merged the primers of two methods in such a way that there was a sufficient differentiation of all DNA fragments. The sequencing of these PCR products was also performed to validate these data.

No Brasil, Anthurium andraeanum tem se destacado como espécie importante para produção de flores e folhas de corte, assim como para cultivo em vaso. Devido ao importante papel das plantas ornamentais na diversificação da agricultura tropical, há grande interesse em conhecer as causas dos problemas que afetam o cultivo de antúrio para melhorar seu potencial produtivo. Este trabalho teve como objetivo identificar o agente causal do mosaico intenso e deformação foliar observados em plantas de antúrios em plantios comerciais em quatro municípios do estado de São Paulo; avaliar a extensão da ocorrência do virus nesses municípios e os métodos para a sua detecção, assim como analisar a reação de genótipos de antúrios em relação a este vírus. Foram amostrados antúrios cultivados em Mogi das Cruzes, Pariquera-Açu, Iguape e Registro. Folhas e espatas de plantas sintomáticas foram utilizadas em ensaios de transmissão mecânica. O vírus induziu sintomas semelhantes aos observados por Cucumber mosaic virus (CMV), em plantas indicadoras, e só foi transmitido quando espatas foram utilizadas como inóculo. Técnicas de PTA-ELISA e RT-PCR foram realizadas para a identificação do CMV em antúrios de todas as propriedades amostradas. Antúrios das variedades IAC, 'Astral', 'Eidibel', 'Garoa' e 'Juréia', e das cultivares 'Caipira' e 'Branco' foram inoculados com CMV e mostraram-se suscetíveis. Alguns dos sintomas observados em antúrios infectados com CMV, embora se assemelhem aos de Dasheen mosaic virus, são mais drásticos por induzir deformação foliar e mosaico intenso. Ambos os vírus são cosmopolitas e transmitidos de modo não persistente por afídeos. Embora o círculo de hospedeiras do CMV seja amplo, o antúrio não consta dessa lista. A eliminação de plantas invasoras, principalmente Commelina sp. e Tradescantia sp., hospedeiras naturais do CMV, a remoção de antúrios sintomáticos e o controle da população de afídeos são recomendações para a cultura de antúrio, uma vez que não há genótipos resistentes disponíveis.

In Brazil, Anthurium andraeanum (flamingo flower) stands out as an important species for cut leaves and flowers productions, as well as potted cultivation. Due to the fact that ornamental plants play an important part in the tropical agriculture diversification, there is great interest to know the problems that affect anthurium cultivation in order to improve its productive potential. The aims of this study were identify the causal agent of intense mosaic and leaf deformation observed in anthurium commercial crops from four cities of the São Paulo state, Brazil; to assess the extent of the virus occurrence in these cities and the methods for its detection, as well as to analyse the anthurium genotypes reactions in relation to the virus. Anthuriums were sampled from the following cities: Mogi das Cruzes, Pariquera-Açu, Iguape and Registro. Leaves and spathes from symptomatic plants were used in mechanical transmission assays. The virus was transmitted only when spathes were used to prepare the inocula, and it induced Cucumber mosaic virus (CMV)-like symptoms in the indicator hosts. PTA-ELISA and RT-PCR were performed to identify the virus species, since CMV was detected in all agricultural areas sampled. Anthuriums from IAC varieties, 'Jureia', 'Garoa', 'Eidibel' and 'Astral' and from 'Caipira' and 'Branco' cultivars were inoculated with CMV and all of them were susceptible to the virus. Some of the symptoms observed in CMV infected anthuriums although similar to those induced by Dasheen mosaic virus, are more severe for inducing leaf deformation and intensive mosaic. Both viruses are worldwide spread and transmitted by aphid in a non-persistent manner. Although CMV has a wide host range, it was not found in the literature the presence of anthurium as a host to the virus. The eradication of weed plants, mainly Commelina and Tradescantia that are CMV natural hosts, the elimination of the symptomatic anthuriums and control of the aphid population should also be done in the anthurium crop, since anthurium genotypes with resistance for CMV are not available.

El presente trabajo se efectuó con tres tipos de cacao, Theobroma cacao L., Criollo (Porcelana, Guasare y Criollo Merideño San Juan-CMSJ) colectados en el estado Mérida; Trinitario (ICS-1) y Foras tero (IMC-67xOC-61), cosechados en el estado Aragua, con la finalidad de evaluar la calidad del proceso de secado y el comportamiento físico del grano. Se efectuaron registros diarios de peso, pH y acidez titulable (AT) en las almendras , así como porcentaje de testa y acidez volátil (AV) al final del secado. El protocolo utilizado fue un secado gradual, en gavetas de madera durante 6 d. Los resultados muestran que la pérdida de peso en los tipos evaluados, se ajusta a una función polinomial Y = a Xm - b X + c con un coeficiente de correlación entre 0,96 y 0,99. Con relación a la AT en cotiledones de los diferentes tipos evaluados (P>0,001) y porcentaje de cascarilla, el Criollo Porcelana presentó valores superiores a las demás variedades. La humedad se mantuvo constante en todos los cacaos .

Three kinds of cocoa were used in this investigation: Criollos (Porcelana, Guasare y Criollo Merideño San Juan) collected in Mérida State; Trinitario: ICS-1 and the Forastero IMC-67xOC-61, harvested in Aragua State. Registrations of Daily weigh, pH and tritable acidity (TA) in the beans percentage of testa (test/head) and volatile acidity (VA) when the dried stage is over were done. The process used was a gradual dried into wooden drawers during six days. The results have shown that the weigh lost in the samples evaluated are set to a polimonial function Y = a Xm - b X + c, with a correlation rate between 0.96 and 0.99. About the TA to the cotyledons of the evaluated cocoas (P>0,001), the Criollo Porcelana had higher values than the rest of the cocoas, humidity remained constant in all cocoas and the highest percentage of hulls was cocoa presented Porcelana.