Resumo O objetivo deste trabalho foi analisar a resposta androgênica de genótipos brasileiros de trigo a diferentes pré-tratamentos das espigas, antes da cultura de micrósporos isolados, e ao efeito de um agente gelificante no meio de cultura de indução. Cinco genótipos foram avaliados quanto à formação de embriões, regeneração de plantas verdes e à duplicação espontânea dos cromossomos. Espigas de trigo foram submetidas a dois pré-tratamentos: frio, a 4ºC por 21 dias; e ácido 2-hidroxinicotínico, a 32ºC por dois dias. Os meios de cultura foram avaliados com ou sem Ficoll como agente gelificante. O frio produziu mais embriões e plantas verdes do que o pré-tratamento químico, em quatro dos cinco genótipos testados. Apenas dois genótipos tratados com ácido 2-hidroxinicotínico foram capazes de produzir plantas, e um deles produziu uma única planta albina. O meio com Ficoll produziu mais embriões do que o meio líquido e gerou maior número de plantas. A duplicação espontânea dos cromossomos varia entre os genótipos e os pré-tratamentos e apresenta alta variabilidade.

Abstract The objective of this work was to analyze the androgenic response of Brazilian wheat genotypes to different pretreatments of the spikes, prior to the culture of isolated microspores, and to the effect of a gelling agent in the induction culture medium. Five genotypes were evaluated for embryo formation, green plant regeneration, and spontaneous chromosome duplication. Wheat spikes were subjected to two pretreatments: cold, at 4ºC for 21 days; and 2-hydroxynicotinic acid, at 32ºC for two days. Culture media were evaluated with or without Ficoll as a gelling agent. Cold produced more embryos and green plants than the chemical pretreatment in four out of five genotypes. Only two genotypes treated with 2-hydroxynicotinic acid were able to produce plants, and one of them produced a single albino plant. Medium containing Ficoll produced more embryos than liquid medium and promoted a higher number of plants. Spontaneous chromosome duplication varies between genotypes and pretreatments, and shows high variability.

Abstract Low transformation efficiency is one of the main limiting factors in the establishment of genetic transformation of wheat via Agrobacterium tumefaciens. To determine more favorable conditions for T-DNA delivery and explant regeneration after infection, this study investigated combinations of acetosyringone concentration and pH variation in the inoculation and co-cultivation media and co-culture temperatures using immature embryos from two Brazilian genotypes (BR 18 Terena and PF 020037). Based on transient expression of uidA, the most favorable conditions for T-DNA delivery were culture media with pH 5.0 and 5.4 combined with co-culture temperatures of 22 °C and 25 °C, and a 400 μM acetosyringone supplement. These conditions resulted in blue foci in 81% of the embryos. Media with more acidic pH also presented reduced A. tumefaciens overgrowth during co-culture, and improved regeneration frequency of the inoculated explants. BR 18 Terena was more susceptible to infection by A. tumefaciens than PF 020037. We found that it is possible to improve T-DNA delivery and explant regeneration by adjusting factors involved in the early stages of A. tumefaciens infection. This can contribute to establishing a stable transformation procedure in the future.

Yellow dwarf disease, one of the most important diseases of cereal crops worldwide, is caused by virus species belonging to the Luteoviridae family. Forty-two virus isolates obtained from oat (Avena sativa L.), wheat (Triticum aestivum L.), barley (Hordeum vulgare L.), corn (Zea mays L.), and ryegrass (Lolium multiflorum Lam.) collected between 2007 and 2008 from winter cereal crop regions in southern Brazil were screened by polymerase chain reaction (PCR) with primers designed on ORF 3 (coat protein - CP) for the presence of Barley yellow dwarf virus and Cereal yellow dwarf virus (B/CYDV). PCR products of expected size (~357 bp) for subgroup II and (~831 bp) for subgroup I were obtained for three and 39 samples, respectively. These products were cloned and sequenced. The subgroup II 3' partial CP amino acid deduced sequences were identified as BYDV-RMV (92 - 93 % of identity with "Illinois" Z14123 isolate). The complete CP amino acid deduced sequences of subgroup I isolates were confirmed as BYDV-PAV (94 - 99 % of identity) and established a very homogeneous group (identity higher than 99 %). These results support the prevalence of BYDV-PAV in southern Brazil as previously diagnosed by Enzyme-Linked Immunosorbent Assay (ELISA) and suggest that this population is very homogeneous. To our knowledge, this is the first report of BYDV-RMV in Brazil and the first genetic diversity study on B/CYDV in South America.