Abstract Detection of pathogens associated with seed requires to count with representative and homogeneous samples to obtain reliable results. This research objective was to evaluate whether: 1) the level of infection (5, 10 and 15%); 2) the increase in the number of single sample (10, 15, 20 and 25) used to form composite samples; and 3) increasing and reducing the usual sample size from 10% (established at CIMMYT for diagnosis of pests and diseases) to 15 and 5%, affect the predictive capability of DAS-ELISA test to barley stripe mosaic virus (BSMV). From seed lots of bread wheat (Triticum aestivum L.), one naturally infected and another healthy, 210 single samples (MS), 20 g each, divided into equal groups of 70 MS samples with 5, 10 and 15% level of infection (NI) to form sample sizes (TM) of 15, 10 and 5%, and composite samples (MC) with 10, 15, 20 and 25 MS samples by NI and TM were formed. The results indicated that reducing the level of NI in MS samples from 15% to 10 and 5% decreased significantly (α= 0.05) in 21 and 31.5% respectively, the sensitivity of the test to BSMV virus. Similarly, reducing the number of MS samples to form MC samples significantly affected (α= 0.05) the sensitivity of the test to the virus and increased the likelihood of false negative record. Although the increase and reduction of TM from 10% to 15 and 5% significantly affected (α= 0.05) the sensitivity of the test to the virus, not finding a linear relationship between TM from the MC sample with the absorbance value. This study demonstrates the importance of the level of infection from the BSMV virus in the seed, of TM and MS number in MS in MC samples to obtain reliable results by DAS-ELISA.

Resumen La detección de patógenos asociados a semilla requiere contar con muestras representativas y homogéneas para obtener resultados confiables. Esta investigación tuvo como objetivos evaluar si: 1) el nivel de infección ( 5, 10 y 15 %); 2) el incremento del número de muestra simples (10, 15, 20 y 25), utilizadas para formar muestras compuestas; y 3) el incremento y la reducción del tamaño de muestra usual de 10% (establecido en el CIMMYT para el diagnóstico de plagas y enfermedades) a 15 y 5%, afectan la capacidad predictiva de la prueba DAS-ELISA al virus del mosaico estriado de la cebada (BSMV, por sus siglas en ingles). A partir de lotes de semilla de trigo harinero (Triticum aestivum L.), uno naturalmente infectado y otro sano, se formaron 210 muestras simples (MS), de 20 g cada una, divididas en grupos iguales de 70 muestras MS con 5, 10 y 15% de nivel de infección (NI) para formar tamaños de muestra (TM) de 15, 10 y 5%, y muestras compuestas (MC) con 10, 15, 20 y 25 muestras MS por NI y TM. Los resultados indicaron que al reducir el nivel NI en muestras MS de 15% a 10 y 5% decreció significativamente (α= 0.05) en 21 y 31.5%, respectivamente, la sensibilidad de la prueba al virus BSMV. Similarmente, la reducción del número de muestras MS para formar muestras MC afectó significativamente (α= 0.05) la sensibilidad de la prueba al virus e incrementó la probabilidad de registrar falsos negativos. Aunque el incremento y la reducción del TM de 10% a 15 y 5% afectó significativamente (α= 0.05) la sensibilidad de la prueba al virus no se encontró una relación lineal entre el TM de la muestra MC con el valor de absorbancia. Este estudio evidencia la importancia del nivel de infección del virus BSMV en la semilla, del TM y del número de MS en muestras MC para la obtención de resultados confiables por la prueba DAS-ELISA.

The use of transgenic crops is steadily increasing around the world, led by soybean (based on total area) and maize (in terms of total number of countries). Transgenic maize is grown in at least 17 countries across four continents: Africa, America, Asia and Europe. The comprehensive global spread of transgenic maize has significant implications for organizations involved in germplasm conservation and genetic enhancement; particularly as some countries require a GMO-free declaration when receiving shipments of maize germplasm. This article describes the protocol used by the International Maize and Wheat Improvement Center (CIMMYT) for monitoring unintentional transgene flow in maize genebank and breeding plots. The protocol is based on polymerase chain reaction (PCR) markers for detecting specific recombinant DNA sequences in bulked samples collected from sentinel plots. To date, no unintentional transgene flow has been detected in CIMMYT fields of maize genebank accessions or breeding materials.

Antes do uso em larga escala de linhagens não-patogênicas de Fusarium oxysporum como agentes de biocontrole da murcha-de-Fusarium, o seu comportamento e seus impactos potenciais no solo devem ser estudados como parte da avaliação de riscos. O objetivo do presente trabalho foi avaliar os efeitos de linhagens antagonistas de F. oxysporum, geneticamente manipuladas (T26/6) ou não (233/1), na biomassa e atividade microbiana do solo. Os efeitos foram avaliados no noroeste da Itália em dois solos de Albenga, sendo um natural e outro previamente solarizado, e em um terceiro solo obtido numa plantação de 10 anos de Popolus sp., em Carignano. Não foram observados efeitos significativos na quantidade de ATP, hidrólise de diacetato de fluoresceina e biomassa P, após a introdução dos antagonistas. Um aumento transitório foi observado na evolução de dióxido de carbono e no carbono da biomassa em resposta à adição dos antagonistas. Embora os resultados tenham demonstrado apenas efeitos transitórios, outros estudos são necessários para a avaliação dos efeitos em populações específicas de microrganismos.

Before planning the large-scale use of nonpathogenic strains of Fusarium oxysporum as biocontrol agents of Fusarium wilt, their behaviour and potential impact on soil ecosystems should be carefully studied as part of risk assessment. The aim of this work was to evaluate the effects of antagonistic F. oxysporum strains, genetically manipulated (T26/6) or not (233/1), on soil microbial biomass and activity. The effects were evaluated, in North-western Italy, in two soils from different sites at Albenga, one natural and the other previously solarized, and in a third soil obtained from a 10-year-old poplar stand (Popolus sp.), near Carignano. There were no detectable effects on ATP, fluorescein diacetate hydrolysis, and biomass P that could be attributed to the introduction of the antagonists. A transient increase of carbon dioxide evolution and biomass C was observed in response to the added inoculum. Although the results showed only some transient alterations, further studies are required to evaluate effects on specific microorganism populations.