Mitochondrial carrier family (MCF) proteins catalyze the specific transport of various substrates, such as nucleotides, amino acids and cofactors. Although some of the mitochondrial transporters have been identified, many of these proteins have not yet been completely characterized. Likewise, the proteic machinery and mechanisms involved in the mitochondrial alternative respiration is still not well known. In this context, this work first presents a study of a previously identified but uncharacterized mitochondrial transporter AtSFC1, a potential succinate/fumarate carrier. Hence, to obtain the biochemical role of AtSFC1, we carried out substrate specificity and investigated its physiological function using 35S antisense transgenic lines in Arabidopsis thaliana. Briefly, the functional integration of AtSFC1 in the cytoplasmic membrane of intact Escherichia coli cells reveals a high specificity for a citrate/isocitrate in a counter exchange mode. Additionally, we discussed the potential role for AtSFC1 in the provision of intermediates of tricarboxylic acid cycle to provide carbon and energy to support growth in heterotrophic tissues. In the second part of this thesis, we investigated the function of alternative electron donors to the mitochondrial electron transport chain (mETC) during carbon deprivation as well as after the supply of amino acids. The breakdown products of branched chain amino acids can provide electrons to the mETC via the ETF/ETFQO (electron transfer flavoprotein: flavoprotein ubiquinone oxidoreductase) complex. This system is located in the mitochondria and induced at the level of transcription during stress situations. Thus, in order to obtain a comprehensive picture of how alternative respiration pathway interacts with other pathways and adjust to different cellular and metabolic requirements, we performed metabolic and physiological approaches using Arabidopsis cell culture ETFQO T-DNA insertion mutants. The results discussed here support that the ETF/ETFQO system is an essential pathway able to donate electrons to the ubiquinone pool. In addition, the behavior of the respiratory complexes suggest new electrons entry points, which must be elucidated. | A família de transportadores mitocondriais (MCF) catalisam transportes específicos de vários substratos, tais como nucleotídeos, aminoácidos e cofatores. Embora alguns dos transportadores tenham sido identificados, muitas destas proteínas ainda não foram completamente caracterizadas. Do mesmo modo, o metabolismo mitocondrial sob estresse ainda não é completamente conhecido. Neste contexto, este trabalho apresenta inicialmente um estudo de um transportador mitocondrial previamente identificado e ainda não caracterizado, designado como AtSFC1, um potencial transportador de succinato/fumarato. Assim, para identificar a especificidade de substrato, a proteína AtSFC1 foi integrada em liposomas e ensaios bioquímicos foram realizados e para investigar a função fisiológica deste transportador, foram utilizadas plantas transgênicas de Arabidospis thaliana para o gene AtSFC1, cuja expressão foi reduzida pela técnica antisenso. Brevemente, a integração funcional do AtSFC1 na membrana citoplasmática de células de Escherichia coli revelou uma especificidade para citrato/isocitrato do tipo antiporte. Além disso, discutimos o potencial papel para AtSFC1 no fornecimento de intermediários para o ciclo ácido dos tricarboxílicos para suportar o crescimento nos tecidos heterotróficos. Na segunda parte desta tese, investigou-se a função de doadores de elétrons alternativos para a cadeia mitocondrial transportadora de elétrons (mCTE) sob deficiência de carbono, bem como após o fornecimento de aminoácidos. Os produtos de degradação de aminoácidos de cadeia ramificada podem doar elétrons para o mCTE através do complexo ETF/ETFQO (electron transfer flavoprotein: flavoprotein ubiquinone oxidoreductase). Este sistema está localizado na mitocôndria e induzido ao nível de transcrição em situações de estresse. Assim, a fim de obter um design detalhado de como essa via interage com outras e como ela se ajusta a diferentes requisitos celulares e metabólicas, foram realizadas abordagens metabólicas e fisiológicas utilizando cultura de células de Arabidopsis mutantes com inserção de T-DNA na região codificante do gene ETFQO. Os resultados são aqui discutidos, confirmam que o sistema ETF/ETFQO é uma via essencial capaz de doar elétrons para o pool de ubiquinona. Além disso, o comportamento dos complexos respiratórios sugere novos pontos de entrada de elétrons, os quais devem ser elucidados. | Conselho Nacional de Desenvolvimento Científico e Tecnológico

Zinc and iron are two essential micronutrients for plants. The homeostasis networks of the two metals are intertwined. Arabidopsis halleri is a zinc- and cadmium-tolerant and zinc-hyperaccumulating species, which also present adaptation of its iron homeostasis(1,4). Transcriptomic studies identified genes which are constitu-tively over-expressed in Arabidopsis halleri compared to Arabidopsis thaliana and which may have a role in metal tolerance or accumulation(2-4). Among them, a candidate gene encodes the FRD3 (FERRIC REDUCTASE DEFECTIVE 3) protein, a member of the MATE family of membrane transporters. FRD3 is a citrate transporter involved in iron homeostasis(5-7) and plays a role in zinc tolerance in A. thaliana(8). The FRD3 gene displays a complex regulation. In A. thaliana, alternative transcript initiation for FRD3 determines two transcripts, which dif-fer in their 5'UTRs and have differential translation efficiency. The two transcripts are selectively regulated under stress conditions: iron and zinc depletion, zinc excess or cadmium presence(9). In A. halleri, a single highly ex-pressed FRD3 transcript with high translation efficiency is present(9).

Zinc and iron are two essential micronutrients for plants. The homeostasis networks of the two metals are intertwined. Arabidopsis halleri is a zinc- and cadmium-tolerant and zinc-hyperaccumulating species, which also present adaptation of its iron homeostasis(1,4). Transcriptomic studies identified genes which are constitu-tively over-expressed in Arabidopsis halleri compared to Arabidopsis thaliana and which may have a role in metal tolerance or accumulation(2-4). Among them, a candidate gene encodes the FRD3 (FERRIC REDUCTASE DEFECTIVE 3) protein, a member of the MATE family of membrane transporters. FRD3 is a citrate transporter involved in iron homeostasis(5-7) and plays a role in zinc tolerance in A. thaliana(8). The FRD3 gene displays a complex regulation. In A. thaliana, alternative transcript initiation for FRD3 determines two transcripts, which dif-fer in their 5'UTRs and have differential translation efficiency. The two transcripts are selectively regulated under stress conditions: iron and zinc depletion, zinc excess or cadmium presence(9). In A. halleri, a single highly ex-pressed FRD3 transcript with high translation efficiency is present(9). We are further examining the FRD3 function in zinc and iron homeostasis in A. thaliana and A. halleri. We will present data (i) on the high expression of FRD3 in A. halleri, (ii) on the functional characterization of the two alternative FRD3 transcripts and their role in metal homeostasis in A. thaliana in comparison with the A. halleri FRD3 transcript and (iii) on the zinc phenotypes of the frd3 A. thaliana mutant.

This year marks the 20th anniversary of the discovery and characterization of the two Arabidopsis PHT1 genes encoding the phosphate transporter in Arabidopsis thaliana. So far, multiple inorganic phosphate (Pi) transporters have been described, and the molecular basis of Pi acquisition by plants has been well characterized. These genes are involved in Pi acquisition, allocation and/or signal transduction. This review summarizes how Pi is taken up by the roots and further distributed within two plants: Arabidopsis thaliana and Oryza sativa L. by plasma membrane phosphate transporters PHT1 and PHO1 as well as by intracellular transporters: PHO1, PHT2, PHT3, PHT4, PHT5 (VPT1), SPX-MFS and phosphate translocators family. We also describe the role of the PHT1 transporters in mycorrhizal roots of rice as an adaptive strategy to cope with limited phosphate availability in soil.

The AthaMap database generates a map of predicted transcription factor binding sites (TFBS) and small RNA target sites for the whole Arabidopsis thaliana genome. With the advent of protein binding microarrays (PBM), the number of known TFBS for A. thaliana transcription factors (TFs) has increased dramatically. Using 113 positional weight matrices (PWMs) generated from a single PBM study and representing a total number of 68 different TFs, the number of predicted TFBS in AthaMap was now increased by about 3.8 × 107 to 4.9 × 107. The number of TFs with PWM-predicted TFBS annotated in AthaMap has increased to 126, representing a total of 29 TF families and newly including ARF, AT-Hook, YABBY, LOB/AS2 and SRS. Furthermore, links from all Arabidopsis TFs and genes to the newly established Arabidopsis Information Portal (AIP) have been implemented. With this qualitative and quantitative update, the improved AthaMap increases its value as a resource for the analysis of A. thaliana gene expression regulation at www.athamap.de.

Stadermann KB, Holtgräwe D, Weisshaar B. Chloroplast genome sequence of Arabidopsis thaliana accession Landsberg erecta assembled from Single-Molecule, Real-Time sequencing data. <em>Genome Announcements</em>. 2016;4(5): e00975-16. | A publicly available data-set from Pacific Biosciences was used to create an assembly of the chloroplast genome sequence of the Arabidopsis thaliana Landsberg erecta genotype. The assembly is solely based on SMRT sequencing data and hence provides high resolution of the two inverted repeat regions typically contained in chloroplast genomes.

MAIN CONCLUSION : Two TFL1 -like genes, CorfloTFL1 and CorcanTFL1 cloned from Cornus florida and C. canadensis, function in regulating the transition to reproductive development in Arabidopsis. TERMINAL FLOWER 1 (TFL1) is known to regulate inflorescence development in Arabidopsis thaliana and to inhibit the transition from a vegetative to reproductive phase within the shoot apical meristem. Despite the importance, TFL1 homologs have been functionally characterized in only a handful eudicots. Here we report the role of TFL1 homologs of Cornus L. in asterid clade of eudicots. Two TFL1-like genes, CorfloTFL1 and CorcanTFL1, were cloned from Cornus florida (a tree) and C. canadensis (a subshrub), respectively. Both are deduced to encode proteins of 175 amino acids. The amino acid sequences of these two Cornus TFL1 homologs share a high similarity to Arabidopsis TFL1 and phylogenetically more close to TFL1 paralogous copy ATC (Arabidopsis thaliana CENTRORADIALIS homologue). Two genes are overexpressed in wild-type and tfl1 mutant plants of A. thaliana. The over-expression of each gene in wild-type Arabidopsis plants results in delaying flowering time, increase of plant height and cauline and rosette leaf numbers, excessive shoot buds, and secondary inflorescence branches. The over-expression of each gene in the tfl1 mutant rescued developmental defects, such as the early determinate inflorescence development, early flowering time, and other vegetative growth defects, to normal phenotypes of wild-type plants. These transgenic phenotypes are inherited in progenies. All data indicate that CorfloTFL1 and CorcanTFL1 have conserved the ancestral function of TFL1 and CEN regulating flowering time and inflorescence determinacy.

In the present work, high quantities of arabidopsides were extracted and purified from Arabidopsis thaliana L.

The phenotypic changes in the root system of Arabidopsis thaliana seedlings in transgenic lines with overexpression and suppressed gene expression of serine-threonine protein kinase KIN10, under conditions of energy shortage and under normal conditions, were shown. The normal growth and development of KIN10 overexpressing plants under energy deficiency conditions were detected. The significant inhibition of the development of these plant lines was observed under normal conditions. The levels of KIN10 gene expression under normal conditions in different organs of Arabidopsis thaliana, particularly in the roots, stems, leaves and flowers were analyzed. The highest-level expression of the gene was found in the leaves.