Abstract NAC transcription factors are plant-specific proteins involved in many processes during the plant life cycle and responses to biotic and abiotic stresses. Previous studies have shown that stress-induced OsNAC5 from rice (Oryza sativa L.) is up-regulated by senescence and might be involved in control of iron (Fe) and zinc (Zn) concentrations in rice seeds. Aiming a better understanding of the role of OsNAC5 in rice plants, we investigated a mutant line carrying a T-DNA insertion in the promoter of OsNAC5, which resulted in enhanced expression of the transcription factor. Plants with OsNAC5 enhanced expression were shorter at the seedling stage and had reduced yield at maturity. In addition, we evaluated the expression level of OsNAC6, which is co-expressed with OsNAC5, and found that enhanced expression of OsNAC5 leads to increased expression of OsNAC6, suggesting that OsNAC5 might regulate OsNAC6 expression. Ionomic analysis of leaves and seeds from the OsNAC5 enhanced expression line revealed lower Fe and Zn concentrations in leaves and higher Fe concentrations in seeds than in WT plants, further suggesting that OsNAC5 may be involved in regulating the ionome in rice plants. Our work shows that fine-tuning of transcription factors is key when aiming at crop improvement. plantspecific specific stresses stressinduced stress induced OsNAC Oryza L. L upregulated up regulated (Fe (Zn plants TDNA T DNA factor maturity addition coexpressed co expressed finetuning fine tuning improvement

Abstract We investigated the immobilization of a tetrameric Kluyveromyces lactis β-galactosidase (EC: 3.2.1.23) (KL-Gal) on Immobead 150 using different support modification strategies. Immobead support was modified using an acid solution of H2SO4:HNO3 (3:1) (Immobead-Ac) or 5 % (v/v) glutaraldehyde (Immobead-Glu). Its unmodified form (Immobead) was also tested. Immobilization yields and efficiencies were evaluated by testing protein loads from 10 to 200 mg.g-1 support. The thermal, physico-chemical, textural and catalytic properties of the supports (modified and unmodified) and their derivatives (Immobead-KL-Gal, Immobead-Ac-KL-Gal and Immobead-Glu-KL-Gal) were analyzed. The highest immobilization yields and efficiencies were achieved with a protein load of 100 mg.g-1 support. Surface and pore areas of the Immobead support were greatly decreased after modification. Michaelis constant of the immobilized β-galactosidase increased in the derivatives. Maximum velocity decreased approximately 2.8 times for Immobead-KL-Gal and Immobead-Glu-KL-Gal, and approximately 1.4 times for Immobead-Ac-KL-Gal. In batch processes, the three derivatives could be reused successfully at least 15 times, maintaining high residual enzymatic activity during the lactose hydrolysis (in both cheese whey and milk). The tetrameric K. lactis β-galactosidase immobilized on Immobead supports via the tested treatments was stabilized and is an alternative tool for lactose hydrolysis in the dairy industry.

Abstract Oryza sativa, the common cultivated rice, is one of the most important crops for human consumption, but production is increasingly threatened by abiotic stresses. Although many efforts have resulted in breeding rice cultivars that are relatively tolerant to their local environments, climate changes and population increase are expected to soon call for new, fast generation of stress tolerant rice germplasm, and current within-species rice diversity might not be enough to overcome such needs. The Oryza genus contains other 23 wild species, with only Oryza glaberrima being also domesticated. Rice domestication was performed with a narrow genetic diversity, and the other Oryza species are a virtually untapped genetic resource for rice stress tolerance improvement. Here we review the origin of domesticated Oryza sativa from wild progenitors, the ecological and genomic diversity of the Oryza genus, and the stress tolerance variation observed for wild Oryza species, including the genetic basis underlying the tolerance mechanisms found. The summary provided here is important to indicate how we should move forward to unlock the full potential of these germplasms for rice improvement.

Resumo A contaminação de hortaliças por micro-organismos patogênicos está diretamente relacionada com a qualidade das águas de irrigação utilizadas na agricultura e com o solo onde se encontram. Com o objetivo de se verificar as condições empregadas na produção de alface (Lactuca sativa L.) em três pequenas propriedades do interior do Rio Grande do Sul (Brasil), foram avaliadas a condição físico-química das águas de irrigação, e a condição microbiológica das águas, solo e folhas de alface. As análises bacteriológicas das águas de irrigação, hortaliças e do solo foram realizadas usando a técnica dos tubos múltiplos ou por contagem em placa, e levou em conta os coliformes totais, coliformes termotolerantes, e presença ou ausência de Escherichia coli, Salmonella sp. e Listeria sp.. As análises físico-químicas realizadas nas águas de irrigação foram pH, turbidez, oxigênio dissolvido, demanda bioquímica de oxigênio, carbono orgânico total e nitrogênio total. As análises bacteriológicas nas águas de irrigação indicaram a ausência de Salmonella sp. e Listeria sp.. Entretanto, foi detectada a presença de E. coli e valores acima do permitido pela legislação para coliformes termotolerantes. Desta forma, estas águas se encontram fora dos padrões estabelecidos pela Resolução CONAMA n° 357. A pesquisa também mostrou a presença de E. coli, Salmonella sp. e Listeria sp. nas hortaliças irrigadas, sugerindo que a fonte de contaminação por Salmonella sp. e Listeria sp. deve ser a partir do solo, e não a partir da água de irrigação. Os dados físico-químicos mostraram que apenas uma das propriedades analisadas apresentou todos os parâmetros adequados, segundo a legislação vigente para águas doces de classe 1. Portanto, apenas uma das propriedades apresentou condições físico-químicas para ser utilizada na irrigação de alface, embora nenhuma tenha apresentado condições higiênico-sanitárias para tal uso.

Abstract The contamination of greenery by pathogenic microorganisms is directly related to the quality of irrigation water used in agriculture and to the soil where the greenery is located. In order to verify the conditions surrounding lettuce (Lactuca sativa L.) production at three small properties of Rio Grande do Sul State, Brazil, we evaluated the physico-chemical conditions of irrigation water as well as the microbiological condition of water, soil and lettuce leaves. Bacteriological analyses of the irrigation water, greenery and soil were performed using the multiple tubes or plate-counting techniques, and the total coliforms, thermo-tolerant coliforms, and presence or absence of Escherichia coli, Salmonella sp. and Listeria sp. was evaluated. The irrigation water was analyzed to determine pH, turbidity, dissolved oxygen, biochemical oxygen demand, total organic carbon and total nitrogen. Bacteriological analysis on the irrigation water indicated the absence of Salmonella sp. and Listeria sp.. However, we detected the presence of E. coli and higher thermo-tolerant coliform levels than that established by current legislation. Thus, these water samples do not conform to the standards established by the CONAMA n° 357 Resolution. Our analyses also showed the presence of E. coli, Salmonella sp. e Listeria sp. on the irrigated greenery, suggesting that the source of the Salmonella sp. and Listeria sp. contamination must be the soil, rather than the irrigation water. Physico-chemical analyses showed that only one of the analyzed properties conforms to the standards established by current legislation for Class 1 fresh water. Therefore, only one of the properties presented the physico-chemical conditions appropriate for lettuce irrigation, although no property presented hygienic sanitary conditions for such use.

Resumo O Vale do Taquari apresenta uma grande quantidade de indústrias alimentícias e curtumes, que lançam seus efluentes no Rio Taquari, além de um cemitério em Área de Preservação Permanente, localizado no município de Muçum. Este trabalho teve como objetivo realizar análises físico-químicas e microbiológicas da água do Rio Taquari e comparar pontos à montante e à jusante de três locais considerados potencialmente poluidores. Os locais escolhidos foram um cemitério e um curtume no município de Muçum, e um curtume seguido de um frigorífico de frangos no município de Roca Sales. Foram realizadas análises de pH, turbidez, oxigênio dissolvido (O2), nitrogênio total (N) e coliformes termotolerantes. Realizou-se também o isolamento de Escherichia coli seguido de testes de resistência aos antibióticos neomicina, rifampicina, clindamicina e penicilina. As análises físico-químicas realizadas neste estudo mostraram que somente a turbidez, em quatro dos seis pontos analisados, apresentou valores acima do permitido pela legislação para águas de classe I, sendo que estes valores elevados provavelmente têm origem inorgânica. As bactérias isoladas no ponto à jusante do cemitério e à montante e à jusante das indústrias de Roca Sales foram sensíveis à neomicina, rifampicina e clindamicina, e resistentes à penicilina. Pode-se concluir, portanto, que mesmo com a liberação dos poluentes, não existe uma interferência significativa na qualidade da água nos três locais analisados do Rio Taquari.

Abstract The Taquari Valley has several food industries and tanneries which release their effluents into the Taquari River, as well as a cemetery in a Permanent Preservation Area in Muçum town. This study performed physicochemical and microbiological analyses of water from before and after three Taquari River sites considered potentially polluting and compared the results of those samples. The chosen sites were a cemetery and a tannery in Muçum and a tannery next to a poultry processing plant located in Roca Sales. We analyzed pH, turbidity, dissolved oxygen (O2), total nitrogen (N) and thermotolerant coliforms. We also isolated Escherichia coli and performed resistance tests to neomycin, rifampicin, clindamycin and penicillin antibiotics. The physicochemical analyses showed that only turbidity, in four out of the six analyzed spots, had values above the established threshold of class I water. These elevated values are probably of inorganic origin. Bacteria isolated from the samples collected after the cemetery and both before and after the Roca Sales industries were sensitive to neomycin, rifampicin and clindamycin, and resistant to penicillin. It can be concluded, therefore, that even the release of the pollutant effluents does not substantially interfere with the water quality of the three analyzed sites.

In the state of Rio Grande do Sul, Brazil, yerba mate (Ilex paraguariensis A. St.-Hil.) is an economically and culturally important species. Because of the difficulty in obtaining plantlets from seeds, micropropagation has emerged as an alternative that could increase production. This work aimed to evaluate the effect that treatment with 6-benzyladenine (BA), at concentrations of 2.2, 8.9, and 20 µM and associated with naphthaleneacetic acid (NAA, 0.5 µM), has on the in vitro growth and leaf emission of I. paraguariensis nodal segments cultivated in 0.5 X Murashige and Skoog (MS) medium or 0.5 X Woody Plant Medium (WPM). The most efficient treatment was MS medium with 8.9 µM BA+NAA, in which I. paraguariensis presented greater shoot elongation and higher leaf counts than in the other treatments. Therefore, use of the MS medium with 8.9 µM BA+NAA is recommended in order to increase efficiency in the initial development of I. paraguariensis explants.

Background Strawberry is a pseudofruit mainly cultivated in temperate climate regions. Considering its high levels of ascorbic acid and phenolic compounds, the consumption of strawberry fruit can be beneficial to health. The Brazilian strawberry production revolves around 3000 tons per year, significantly influencing the food market and generating income to farmers. However, this production can be partially impaired by two-spotted spider mite (TSSM) Tetranychus urticae Koch infestations, due to decreases in the quality and quantity of fruit. Since there are no data in the literature about alterations caused by TSSM infestation in strawberry plants, our work aimed towards evaluating nutritional and physicochemical parameters of TSSM-infested strawberry plants, along with the related chemical treatment (CT) (acaricide) or biological treatment (predatory mite Phytoseiulus macropilis Banks). Results Strawberry fruit from TSSM-infested plants present the highest levels of acidity and exhibit low levels of anthocyanin and phenolic compounds, while fruit from TSSM-infested plants + biological control using predatory mite shows high levels of soluble solids, phenolic compounds and ascorbic acid, along with a high soluble solid content/titratable (SSC/TA) acidity ratio, which indicates high quality fruit. Conclusions Our results suggest that TSSM infestation decreases fruit quality and that the biological control of TSSM using a predatory mite is a suitable alternative to organic production, since the presence of predatory mite does not affect fruit quality and development.

Deficiência de ferro (Fe) ocorre freqüentemente em plantas, uma vez que este mineral é pouco disponível em condições aeróbicas. Plantas de arroz cultivadas sob alagamento, no entanto, estão sujeitas ao excesso de Fe, que pode ser extremamente tóxico. Alguns cultivares de arroz são resistentes a altas concentrações de ferro, mas os mecanismos fisiológicos responsáveis por essa resistência são pouco conhecidos. A ferritina é uma proteína de ampla distribuição e capaz de armazenar ferro, sendo considerada importante para a homeostase deste metal. Acúmulo de ferritina em condições de alta disponibilidade de ferro já foi descrito em algumas espécies vegetais. Entretanto, o papel da ferritina no mecanismo de tolerância de plantas de arroz ao excesso de ferro não é conhecido. Neste trabalho, expressamos ferritina de arroz em E. coli, produzimos um anticorpo policlonal anti-ferritina de arroz e este foi utilizado para avaliar o acúmulo de ferritina em dois cultivares de arroz (Oryza sativa) considerados suscetível (BR-IRGA 409) e tolerante (EPAGRI 108) ao excesso de ferro. O anticorpo foi capaz de reconhecer ferritina purificada de sementes de ervilha, assim como ferritina de folhas de arroz. Aumentos nos níveis de mRNA e proteína foram observados nos dois cultivares sob excesso de ferro, com maior acúmulo da proteína no cultivar EPAGRI 108. Quando submetidas a excesso do elemento, plantas deste mesmo cultivar atingiram concentrações de Fe mais baixas do que plantas do cultivar BR-IRGA409, principalmente nas partes aéreas. Sugere-se que o mecanismo de tolerância ao excesso de ferro no cultivar EPAGRI 108 inclui limitação da translocação de Fe e aumento do acúmulo de ferritina. Este é o primeiro trabalho que mostra maior acúmulo da proteína ferritina em um cultivar de Oryza sativa tolerante ao excesso de Fe, fornecendo evidência de um possível papel desta proteína nos mecanismos de tolerância a este metal.

Plants ordinarily face iron (Fe) deficiency, since this mineral is poorly available in soils under aerobic conditions. Nonetheless, wetland and irrigated rice plants can be exposed to excess, highly toxic Fe. Ferritin is a ubiquitous Fe-storage protein, important for iron homeostasis. Increased ferritin accumulation resulting from higher Fe availability was shown in some plant species. However, the role of ferritin in tolerance mechanisms to Fe overload in rice is yet to be established. In this study, recombinant rice ferritin was expressed in Escherichia coli, producing an anti-rice ferritin polyclonal antibody which was used to evaluate ferritin accumulation in two rice (Oryza sativa L.) cultivars, either susceptible (BR-IRGA 409) or tolerant (EPAGRI 108) to Fe toxicity. Increased ferritin mRNA and protein levels resulting from excess Fe treatment were detected in both cultivars, with higher ferritin protein accumulation in EPAGRI 108 plants, which also reached lower shoot Fe concentrations when submitted to iron overload. The tolerance mechanism to excess Fe in EPAGRI 108 seems to include both restricted Fe translocation and increased ferritin accumulation. This is the first work that shows higher accumulation of the ferritin protein in an iron-excess tolerant Oryza sativa cultivar, providing evidence of a possible role of this protein in iron tolerance mechanisms.