RESUMO
Plant nitrate (NO3(-)) acquisition depends on the combined activities of root high- and low-affinity NO3(-) transporters and the proton gradient generated by the plasma membrane H(+)-ATPase. These processes are coordinated with photosynthesis and the carbon status of the plant. Here, we present the characterization of romaine lettuce (Lactuca sativa 'Conquistador') plants engineered to overexpress an intragenic gain-of-function allele of the type I proton translocating pyrophosphatase (H(+)-PPase) of Arabidopsis (Arabidopsis thaliana). The proton-pumping and inorganic pyrophosphate hydrolytic activities of these plants are augmented compared with control plants. Immunohistochemical data show a conspicuous increase in H(+)-PPase protein abundance at the vasculature of the transgenic plants. Transgenic plants displayed an enhanced rhizosphere acidification capacity consistent with the augmented plasma membrane H(+)-ATPase proton transport values, and ATP hydrolytic capacities evaluated in vitro. These transgenic lines outperform control plants when challenged with NO3(-) limitations in laboratory, greenhouse, and field scenarios. Furthermore, we report the characterization of a lettuce LsNRT2.1 gene that is constitutive up-regulated in the transgenic plants. Of note, the expression of the LsNRT2.1 gene in control plants is regulated by NO3(-) and sugars. Enhanced accumulation of (15)N-labeled fertilizer by transgenic lettuce compared with control plants was observed in greenhouse experiments. A negative correlation between the level of root soluble sugars and biomass is consistent with the strong root growth that characterizes these transgenic plants.
Assuntos
Proteínas de Arabidopsis/metabolismo , Pirofosfatase Inorgânica/metabolismo , Lactuca/metabolismo , Nitrogênio/metabolismo , Ácidos/metabolismo , Proteínas de Transporte de Ânions/genética , Proteínas de Transporte de Ânions/metabolismo , Arabidopsis/enzimologia , Proteínas de Arabidopsis/genética , Biomassa , Carboidratos/análise , Carbono/metabolismo , Fertilizantes , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , Engenharia Genética , Imuno-Histoquímica , Pirofosfatase Inorgânica/genética , Lactuca/efeitos dos fármacos , Lactuca/genética , Lactuca/crescimento & desenvolvimento , Transportadores de Nitrato , Nitratos/farmacologia , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/metabolismo , Plantas Geneticamente Modificadas , SolubilidadeRESUMO
Coordinate regulation of transporters at both the plasma membrane and vacuole contribute to plant cell's ability to adapt to a changing environment and play a key role in the maintenance of the chemiosmotic circuits required for cellular growth. The plasma membrane (PM) Naâº/H⺠antiporter (SOS1) is involved in salt tolerance, presumably in sodium extrusion; the vacuolar type I Hâº-PPase AVP1 is involved in vacuolar sodium sequestration, but its overexpression has also been shown to alter the abundance and activity of the PM Hâº-ATPase. Here we investigate the relationship between these transporters utilizing loss-of-function mutants of SOS1 (sos1) and increased expression of AVP1 (AVP1OX). Heightened expression of AVP1 enhances pyrophosphate-dependent proton pump activity, salt tolerance, ion vacuolar sequestration, K⺠uptake capacity, root hair development, osmotic responses, and PM ATPase hydrolytic and proton pumping activities. In sos1 lines overexpressing AVP1, these phenotypes are negatively affected demonstrating that sos1 is epistatic to AVP1. Enhanced AVP1 protein levels require SOS1 and this regulation appears to be post-translational.
Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Epistasia Genética , Pirofosfatase Inorgânica/metabolismo , Tolerância ao Sal/fisiologia , Trocadores de Sódio-Hidrogênio/metabolismo , Arabidopsis/genética , Arabidopsis/crescimento & desenvolvimento , Proteínas de Arabidopsis/genética , Eletroforese em Gel de Poliacrilamida , Immunoblotting , Pirofosfatase Inorgânica/genética , Fenótipo , Raízes de Plantas/crescimento & desenvolvimento , Raízes de Plantas/metabolismo , Reação em Cadeia da Polimerase em Tempo Real , Tolerância ao Sal/genética , Trocadores de Sódio-Hidrogênio/genética , Regulação para CimaRESUMO
It is widely reported that some humic substances behave as exogenous auxins influencing root growth by mechanisms that are not yet completely understood. This study explores the hypothesis that the humic acids' effects on root development involve a nitric oxide signaling. Maize seedlings were treated with HA 20 mg C L(-1), IAA 0.1 nM, and NO donors (SNP or GSNO), in combination with either the auxin-signaling inhibitor PCIB, the auxin efflux inhibitor TIBA, or the NO scavenger PTIO. H(+)-transport-competent plasma membrane vesicles were isolated from roots to investigate a possible link between NO-induced H(+)-pump and HA bioactivity. Plants treated with either HA or SNP stimulated similarly the lateral roots emergence even in the presence of the auxin inhibitors, whereas NO scavenger diminished this effect. These treatments induced H(+)-ATPase stimulation by threefold, which was abolished by PTIO and decreased by auxin inhibitors. HA-induced NO synthesis was also detected in the sites of lateral roots emergence. These data depict a new scenario where the root development stimulation and the H(+)-ATPase activation elicited by either HA or exogenous IAA depend essentially on mechanisms that use NO as a messenger induced site-specifically in the early stages of lateral root development.
Assuntos
Membrana Celular/enzimologia , Substâncias Húmicas , Óxido Nítrico/farmacologia , Raízes de Plantas/efeitos dos fármacos , Raízes de Plantas/crescimento & desenvolvimento , ATPases Translocadoras de Prótons/metabolismo , Zea mays/efeitos dos fármacos , Membrana Celular/efeitos dos fármacos , Ativação Enzimática/efeitos dos fármacos , Sequestradores de Radicais Livres/farmacologia , Concentração de Íons de Hidrogênio/efeitos dos fármacos , Hidrólise/efeitos dos fármacos , Ácidos Indolacéticos/farmacologia , Óxido Nítrico/biossíntese , Nitroprussiato/farmacologia , Raízes de Plantas/citologia , Bombas de Próton/metabolismo , Zea mays/citologia , Zea mays/enzimologia , Zea mays/crescimento & desenvolvimentoRESUMO
Banana, papaya and pineapple are the most consumed tropical fruits in the world, being Brazil one of the main producers. Fungi Colletotrichum musae, Colletotrichum gloeosporioides and Fusarium subglutinans f.sp. ananas cause severe post harvest diseases and losses in fruits quality. The aim of this work was to evaluate the effectiveness of five monoterpenes to inhibit the mycelial growth and conidia germination of these three phytopathogens. The monoterpenes citral, citronellal, L-carvone, isopullegol and a-pinene were diluted in ethanol to final concentrations from 0.2 to 1 percent. All monoterpenes were found to inhibit the growth of the three studies fungi in a dose-dependent manner. Citral was the most effective of the oils tested and showed potent fungicidal activity at concentrations above 0.5 percent. Also, in vivo evaluation with these tropical fruits demonstrated the efficiency of citral to inhibit fungal growth. These results indicate the potential use of citral as a natural pesticide control of post-harvest fruit diseases.
Banana, mamão e abacaxi são as frutas tropicais mais consumidas no mundo, sendo o Brasil um dos principais produtores. Os fungos Colletotrichum musae, Colletotrichum gloeosporioides e Fusarium subglutinans f.sp ananas são os principais causadores de doenças e perdas em pós-colheita de frutas. A proposta deste estudo foi avaliar a eficácia de cinco monoterpenos em inibir o crescimento micelial e a germinação dos conídios destes três fitopatógenos. Os monoterpenos citral, citronelal, L-carvona, isopulegol e a-pineno foram diluídos em etanol à concentração final de 0,2 a 1 por cento. Todos os monoterpenos testados inibiram os três fungos estudados de maneira dose-dependente. Citral foi o mais efetivo dos óleos testados e apresentou uma potente atividade fungicida em concentrações acima de 0,5 por cento. Mais ainda, avaliação in vivo com estas frutas tropicais demonstrou a eficácia de citral como inibidor do crescimento fúngico. Estes resultados indicam o uso em potencial de citral como um pesticida natural no controle das doenças em pós-colheita de frutas tropicais.
Assuntos
Colletotrichum/genética , Colletotrichum/isolamento & purificação , Fungicidas Industriais/análise , Técnicas In Vitro , Monoterpenos/análise , Óleos de Plantas/análise , Controle Biológico de Vetores , Doenças das Plantas , Métodos , Micelas , VirulênciaRESUMO
Banana, papaya and pineapple are the most consumed tropical fruits in the world, being Brazil one of the main producers. Fungi Colletotrichum musae, Colletotrichum gloeosporioides and Fusarium subglutinans f.sp. ananas cause severe post harvest diseases and losses in fruits quality. The aim of this work was to evaluate the effectiveness of five monoterpenes to inhibit the mycelial growth and conidia germination of these three phytopathogens. The monoterpenes citral, citronellal, L-carvone, isopullegol and α-pinene were diluted in ethanol to final concentrations from 0.2 to 1%. All monoterpenes were found to inhibit the growth of the three studies fungi in a dose-dependent manner. Citral was the most effective of the oils tested and showed potent fungicidal activity at concentrations above 0.5%. Also, in vivo evaluation with these tropical fruits demonstrated the efficiency of citral to inhibit fungal growth. These results indicate the potential use of citral as a natural pesticide control of post-harvest fruit diseases.