57 research outputs found

    kNACking on heaven’s door: how important are NAC transcription factors for leaf senescence and Fe/Zn remobilization to seeds?

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    Senescence is a coordinated process where a plant, or a part of it, engages in programmed cell death to salvage nutrients by remobilizing them to younger tissues or to developing seeds. As Fe and Zn deficiency are the two major nutritional disorders in humans, increased concentration of these nutrients through biofortification in cereal grains is a long-sought goal. Recent evidences point to a link between the onset of leaf senescence and increased Fe and Zn remobilization. In wheat, one member of the NAC (NAM, ATAF, and CUC) transcription factor (TF) family (NAM-B1) has a major role in the process, probably regulating key genes for the early onset of senescence, which results in higher Fe and Zn concentrations in grains. In rice, the most important staple food for nearly half of the world population, the NAM-B1 ortholog does not have the same function. However, other NAC proteins are related to senescence, and could be playing roles on the same remobilization pathway. Thus, these genes are potential tools for biofortification strategies in rice. Here we review the current knowledge on the relationship between senescence, Fe and Zn remobilization and the role of NAC TFs, with special attention to rice. We also propose a working model for OsNAC5, which would act on the regulation of nicotianamine (NA) synthesis and metal–NA remobilization

    Transcriptional plasticity bufers genetic variation in zinc homeostasis

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    In roots of Arabidopsis thaliana, Zn can be either loaded into the xylem for translocation to the shoot or stored in vacuoles. Vacuolar storage is achieved through the action of the Zn/Cd transporter HMA3 (Heavy Metal Atpase 3). The Col-0 accession has an HMA3 loss-of-function allele resulting in high shoot Cd, when compared to accession CSHL-5 which has a functional allele and low shoot Cd. Interestingly, both Col-0 and CSHL-5 have similar shoot Zn concentrations. We hypothesize that plants sense changes in cytosolic Zn that are due to variation in HMA3 function, and respond by altering expression of genes related to Zn uptake, transport and compartmentalisation, in order to maintain Zn homeostasis. The expression level of genes known to be involved in Zn homeostasis were quantified in both wildtype Col-0 and Col-0::HMA3CSHL-5 plants transformed with the functional CSHL-5 allele of HMA3. We observed significant positive correlations between expression of HMA3 and of genes known to be involved in Zn homeostasis, including ZIP3, ZIP4, MTP1, and bZIP19. The results support our hypothesis that alteration in the level of function of HMA3 is counterbalanced by the fine regulation of the Zn homeostasis gene network in roots of A. thaliana

    Reference gene selection for quantitative reverse transcription-polymerase chais reaction normalization during in vitro adventitious rooting in Eucaliptus globulus Labill

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    Background: Eucalyptus globulus and its hybrids are very important for the cellulose and paper industry mainly due to their low lignin content and frost resistance. However, rooting of cuttings of this species is recalcitrant and exogenous auxin application is often necessary for good root development. To date one of the most accurate methods available for gene expression analysis is quantitative reverse transcription-polymerase chain reaction (qPCR); however, reliable use of this technique requires reference genes for normalization. There is no single reference gene that can be regarded as universal for all experiments and biological materials. Thus, the identification of reliable reference genes must be done for every species and experimental approach. The present study aimed at identifying suitable control genes for normalization of gene expression associated with adventitious rooting in E. globulus microcuttings. Results: By the use of two distinct algorithms, geNorm and NormFinder, we have assessed gene expression stability of eleven candidate reference genes in E. globulus: 18S, ACT2, EF2, EUC12, H2B, IDH, SAND, TIP41, TUA, UBI and 33380. The candidate reference genes were evaluated in microccuttings rooted in vitro, in presence or absence of auxin, along six time-points spanning the process of adventitious rooting. Overall, the stability profiles of these genes determined with each one of the algorithms were very similar. Slight differences were observed in the most stable pair of genes indicated by each program: IDH and SAND for geNorm, and H2B and TUA for NormFinder. Both programs indentified UBI and 18S as the most variable genes. To validate these results and select the most suitable reference genes, the expression profile of the ARGONAUTE1 gene was evaluated in relation to the most stable candidate genes indicated by each algorithm. Conclusion: Our study showed that expression stability varied between putative reference genes tested in E. globulus. Based on the AGO1 relative expression profile obtained using the genes suggested by the algorithms, H2B and TUA were considered as the most suitable reference genes for expression studies in E. globulus adventitious rooting. UBI and 18S were unsuitable for use as controls in qPCR related to this process. These findings will enable more accurate and reliable normalization of qPCR results for gene expression studies in this economically important woody plant, particularly related to rooting and clonal propagation

    The combined strategy for iron uptake is not exclusive to domesticated rice

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    Iron (Fe) is an essential micronutrient that is frequently inaccessible to plants. Rice (Oryza sativa L.) plants employ the Combined Strategy for Fe uptake, which is composed by all features of Strategy II, common to all Poaceae species, and some features of Strategy I, common to non-Poaceae species. To understand the evolution of Fe uptake mechanisms, we analyzed the root transcriptomic response to Fe defciency in O. sativa and its wild progenitor O. rufpogon. We identifed 622 and 2,017 diferentially expressed genes in O. sativa and O. rufpogon, respectively. Among the genes up-regulated in both species, we found Fe transporters associated with Strategy I, such as IRT1, IRT2 and NRAMP1; and genes associated with Strategy II, such as YSL15 and IRO2. In order to evaluate the conservation of these Strategies among other Poaceae, we identifed the orthologs of these genes in nine species from the Oryza genus, maize and sorghum, and evaluated their expression profle in response to low Fe condition. Our results indicate that the Combined Strategy is not specifc to O. sativa as previously proposed, but also present in species of the Oryza genus closely related to domesticated rice, and originated around the same time the AA genome lineage within Oryza diversifed. Therefore, adaptation to Fe2+ acquisition via IRT1 in fooded soils precedes O. sativa domestication

    Calagem como forma de redução da toxidez por cobre em aveia preta

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    Soils which are cultivated with grapevines have high available copper (Cu) content, which can be toxic to cover crops cohabiting vineyards, such as black oats. This study aimed to assess the effect of liming in reducing Cu toxicity in black oats grown in sandy soils. Samples of a Typic Hapludalf were collected at 0-20cm, dried and subjected to the addition of Cu (0 to 50Mg kg-1) and limestone (0, 1.5, and 3.0Mg ha-1). The soil was placed in a rhizobox and black oats were grown for 30 days. We assessed root and shoot dry matter production, copper (Cu), calcium (Ca) and magnesium (Mg) contents in the tissues; Cu content in the root symplast and apoplast, as well as Cu, carbon and pH values in the rhizosphere and bulk soil. Liming reduced Cu toxicity in black oats. Cu was preferentially accumulated in the roots, mostly in the apoplast, which may be the result of a plant tolerance mechanism to prevent the transport of Cu to the shoots. Key words: heavy metal, phytotoxicity, limestone, rhizosphere.Solos cultivados com videiras possuem alto teor de cobre (Cu) disponível, que pode ser tóxico às plantas de cobertura do solo que coabitam vinhedos, como a aveia preta. O estudo objetivou avaliar o efeito da calagem na redução da toxidez por Cu em plantas de aveia preta cultivadas em solo arenoso. Amostras de um Argissolo Vermelho foram coletadas na camada de 0-20cm, secas e submetidas à adição de duas doses de Cu (0 e 50Mg kg-1) e três de calcário (0, 1,5 e 3,0Mg ha-1). O solo foi acondicionado em rhizobox e submetido ao cultivo de aveia preta durante 30 dias. Avaliaram-se a produção de matéria seca das raízes e da parte aérea, o teor de cobre (Cu), cálcio (Ca) e magnésio (Mg) nos tecidos; o teor de Cu no simplasto e apoplasto das raízes, e os teores de Cu, de carbono e valores de pH no solo rizosférico e não rizosférico. A aplicação de calcário reduziu a toxidez por Cu na aveia preta. O Cu foi preferencialmente acumulado nas raízes, especialmente no apoplasto, o que pode ser resultado de mecanismo de tolerância das plantas para evitar o transporte de parte do elemento para a parte aérea

    AtGRP3 is implicated in root size and aluminum response pathways in Arabidopsis

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    AtGRP3 is a glycine-rich protein (GRP) from Arabidopsis thaliana shown to interact with the receptor-like kinase AtWAK1 in yeast, in vitro and in planta. In this work, phenotypic analyses using transgenic plants were performed in order to better characterize this GRP. Plants of two independent knockout alleles of AtGRP3 develop longer roots suggesting its involvement in root size determination. Confocal microscopy analysis showed an abnormal cell division and elongation in grp3-1 knockout mutants. Moreover, we also show that grp3-1 exhibits an enhanced Aluminum (Al) tolerance, a feature also described in AtWAK1 overexpressing plants. Together, these results implicate AtGRP3 function root size determination during development and in Al stress
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