PEROXIDASE ACTIVITY IN MAIZE INBRED LINES RESISTANT OR SUSCEPTIBLE TO MAIZE DWARF MOSAIC VIRUS

Peroxidase (EC 1.11.1.7) activity levels were analyzed in maize seedling inbred lines contrasting with their resistance to maize dwarf mosaic virus (MDMV). Groups of noninoculated resistant and susceptible inbred lines showed different levels of guaiacol peroxidase activity at the seedling stage, with 78 % of susceptible inbred lines presenting enzyme activity values below 91.46 A470 min–1g FW –1. However, the zymogram patterns of peroxidase did not allow the differentiation of non-inoculated resistant or susceptible inbred lines. The virus complex induced a general enhancement of enzyme activity, without qualitative changes in the isoenzymes, when inoculated into resistant or susceptible inbred lines. However, a quantitative change, with 19% OD average increase in a moderately anionic isoform was observed in some resistant inbred lines in response to virus inoculation. Our results could suggest that peroxidase activity prior to infection, and the increase in activity of an specific anionic isoform in some resistant inbred lines, due to virus inoculation, could be related to a defense mechanism against MDMV

Optimization of particle bombardment parameters for the genetic transformation of Brazilian maize inbred lines

O objetivo deste trabalho foi desenvolver um sistema de transformação genética para genótipos tropicais de milho pelo bombardeamento de embriões imaturos com micropartículas. O bombardeamento compartículas foi realizado utilizando-se uma construção gênica contendo os genes bar e uidA sob o controle do promotor CaMV35S. As melhores condições para transformação das linhagens de milho tropical L3 e L1345foram obtidas após o cultivo de embriões imaturos durante 4 horas, antes do bombardeamento, em alta osmolaridade, e bombardeados com a pressão de 1.100 psi de gás hélio, dois tiros por placa e uma distância de vôo da micropartícula de 6,6 cm. As freqüências de transformação obtidas variaram entre 0,9 e 2,31%. A integração dos genes heterólogos no genoma das plantas transgênicas de milho foi confirmada por análises de Southern blot e expressão dos genes bar e uidA. O protocolo de transformação genética de milho desenvolvido neste estudo irá aumentar, provavelmente, a eficiência de produção de novas linhagens tropicais de milho expressando características agronômicas desejáveis.The objective of this work was to develop a genetic transformation system for tropical maize genotypes via particle bombardment of immature zygotic embryos. Particle bombardment was carried out using a genetic construct with bar and uidA genes under control of CaMV35S promoter. The best conditions to transform maize tropical inbred lines L3 and L1345 were obtained when immature embryos were cultivated, prior to the bombardment, in higher osmolarity during 4 hours and bombarded at an acceleration helium gas pressure of 1,100 psi, two shots per plate, and a microcarrier flying distance of 6.6 cm. Transformation frequencies obtained using these conditionsranged from 0.9 to 2.31%. Integration of foreign genes into the genome of maize plants was confirmed by Southern blot analysis as well as bar and uidA gene expressions. The maize genetic transformation protocol developed in this work will possibly improve the efficiency to produce new transgenic tropical maize lines expressing desirable agronomic characteristics

Optimization of particle bombardment parameters for the genetic transformation of Brazilian maize inbred lines Otimização dos parâmetros de bombardeamento de partículas para a transformação genética de linhagens brasileiras de milho

The objective of this work was to develop a genetic transformation system for tropical maize genotypes via particle bombardment of immature zygotic embryos. Particle bombardment was carried out using a genetic construct with bar and uidA genes under control of CaMV35S promoter. The best conditions to transform maize tropical inbred lines L3 and L1345 were obtained when immature embryos were cultivated, prior to the bombardment, in higher osmolarity during 4 hours and bombarded at an acceleration helium gas pressure of 1,100 psi, two shots per plate, and a microcarrier flying distance of 6.6 cm. Transformation frequencies obtained using these conditions ranged from 0.9 to 2.31%. Integration of foreign genes into the genome of maize plants was confirmed by Southern blot analysis as well as bar and uidA gene expressions. The maize genetic transformation protocol developed in this work will possibly improve the efficiency to produce new transgenic tropical maize lines expressing desirable agronomic characteristics.O objetivo deste trabalho foi desenvolver um sistema de transformação genética para genótipos tropicais de milho pelo bombardeamento de embriões imaturos com micropartículas. O bombardeamento com partículas foi realizado utilizando-se uma construção gênica contendo os genes bar e uidA sob o controle do promotor CaMV35S. As melhores condições para transformação das linhagens de milho tropical L3 e L1345 foram obtidas após o cultivo de embriões imaturos durante 4 horas, antes do bombardeamento, em alta osmolaridade, e bombardeados com a pressão de 1.100 psi de gás hélio, dois tiros por placa e uma distância de vôo da micropartícula de 6,6 cm. As freqüências de transformação obtidas variaram entre 0,9 e 2,31%. A integração dos genes heterólogos no genoma das plantas transgênicas de milho foi confirmada por análises de Southern blot e expressão dos genes bar e uidA. O protocolo de transformação genética de milho desenvolvido neste estudo irá aumentar, provavelmente, a eficiência de produção de novas linhagens tropicais de milho expressando características agronômicas desejáveis

Optimization of particle bombardment parameters for the genetic transformation of Brazilian maize inbred lines

The objective of this work was to develop a genetic transformation system for tropical maize genotypes via particle bombardment of immature zygotic embryos. Particle bombardment was carried out using a genetic construct with bar and uidA genes under control of CaMV35S promoter. The best conditions to transform maize tropical inbred lines L3 and L1345 were obtained when immature embryos were cultivated, prior to the bombardment, in higher osmolarity during 4 hours and bombarded at an acceleration helium gas pressure of 1,100 psi, two shots per plate, and a microcarrier flying distance of 6.6 cm. Transformation frequencies obtained using these conditions ranged from 0.9 to 2.31%. Integration of foreign genes into the genome of maize plants was confirmed by Southern blot analysis as well as bar and uidA gene expressions. The maize genetic transformation protocol developed in this work will possibly improve the efficiency to produce new transgenic tropical maize lines expressing desirable agronomic characteristics