Cystoliths of Parietaria judaica can serve as an internal source of CO2 for photosynthetic assimilation when stomata are closed.

The recently reported 'alarm photosynthesis' acts as a biochemical process that assimilates CO2 derived from the decomposition of calcium oxalate crystals. This study examined whether CaCO3 cystoliths could also serve as CO2 pools, fulfilling a similar role. Shoots of Parietaria judaica were subjected to carbon starvation, abscisic acid (ABA), or bicarbonate treatments, and the volume of cystoliths and the photochemical parameters of photosystem II (PSII) were determined. The size of cystoliths was reduced under carbon starvation or ABA treatments, whereas it was restored by xylem-provided bicarbonate. Under carbon starvation, ABA, or bicarbonate treatments, the photochemical efficiency of PSII was higher, while non-photochemical quenching, representing the safe dissipation of excess PSII energy due to lack of electron sinks, was lower in treated samples compared with controls. This observation suggests the involvement of ABA or other carbon starvation cues in the release of subsidiary CO2 for photosynthesis, inevitably from an internal source, which could be the cystoliths. Carbon remobilized from cystoliths can be photosynthetically assimilated, thus acting as a safety valve under stress. Together with alarm photosynthesis, these results show a tight link between leaf carbon deposits and photosynthesis.

Changes in sugar metabolism associated to stem bark thickening partially assist young tissues of Eriobotrya japonica seedlings under boron stress.

Boron (B) toxicity frequently affects plant performances and productivity, especially in arid and semi-arid environments. In this experiment, loquat seedlings were subjected to 25 µM (control) or 400 µM B (B excess) to test the hypothesis that (i) B alters sugar/polyol metabolism in polyol-producing tree species as loquat and (ii) changes of leaf and stem anatomy assist young tissues against toxic effect of B. Gas exchange was monitored from the beginning of the experiment (FBE) till one week after the first visible symptoms of B toxicity appeared in the upper part of the stems (147 d FBE). At 147 FBE, plant biometric parameters and pattern of B accumulation, leaf and stem anatomy, chlorophyll a fluorescence kinetics as well as biochemical measurements were assessed in top (asymptomatic) leaves and upper stem bark. Boron accumulated principally (in the row) in top leaves > top bark > top wood in B-stressed plants, but no changes in allocation pattern were found between controls and B-stressed plants. Excess B promoted the increase in the spongy layer of top leaves and caused the development of cork and numerous collenchyma cells with increased cell wall thickness. This mechanism, which has never been described before, can be considered an attempt to store excessive B in tissues where B ions are less harmful. The accumulation of sorbitol (B-complexing polyol) in top leaves and stem bark can be considered as a further attempt to detoxify B excess. However, B toxicity drastically affects the photosynthetic rate of top leaves, mainly due to non-stomatal limitations, i.e., reduction of ambient CO2 use efficiency and of photosystem II (PSII) efficiency, modification of the partitioning excess energy dissipation in PSII, thus leading to an increased level of lipid peroxidation. Our results suggest that changes in sugar metabolism associated with leaf and stem bark thickening partially assist (but not totally preserve) young tissues of loquat plants under B stress.

High-quality permanent draft genome sequence of the extremely osmotolerant diphenol degrading bacterium Halotalea alkalilenta AW-7(T), and emended description of the genus Halotalea.

Members of the genus Halotalea (family Halomonadaceae) are of high significance since they can tolerate the greatest glucose and maltose concentrations ever reported for known bacteria and are involved in the degradation of industrial effluents. Here, the characteristics and the permanent-draft genome sequence and annotation of Halotalea alkalilenta AW-7(T) are described. The microorganism was sequenced as a part of the Genomic Encyclopedia of Type Strains, Phase I: the one thousand microbial genomes (KMG) project at the DOE Joint Genome Institute, and it is the only strain within the genus Halotalea having its genome sequenced. The genome is 4,467,826 bp long and consists of 40 scaffolds with 64.62 % average GC content. A total of 4,104 genes were predicted, comprising of 4,028 protein-coding and 76 RNA genes. Most protein-coding genes (87.79 %) were assigned to a putative function. Halotalea alkalilenta AW-7(T) encodes the catechol and protocatechuate degradation to ß-ketoadipate via the ß-ketoadipate and protocatechuate ortho-cleavage degradation pathway, and it possesses the genetic ability to detoxify fluoroacetate, cyanate and acrylonitrile. An emended description of the genus Halotalea Ntougias et al. 2007 is also provided in order to describe the delayed fermentation ability of the type strain.

High quality draft genome sequence of Olivibacter sitiensis type strain (AW-6(T)), a diphenol degrader with genes involved in the catechol pathway.

Olivibacter sitiensis Ntougias et al. 2007 is a member of the family Sphingobacteriaceae, phylum Bacteroidetes. Members of the genus Olivibacter are phylogenetically diverse and of significant interest. They occur in diverse habitats, such as rhizosphere and contaminated soils, viscous wastes, composts, biofilter clean-up facilities on contaminated sites and cave environments, and they are involved in the degradation of complex and toxic compounds. Here we describe the features of O. sitiensis AW-6(T), together with the permanent-draft genome sequence and annotation. The organism was sequenced under the Genomic Encyclopedia for Bacteria and Archaea (GEBA) project at the DOE Joint Genome Institute and is the first genome sequence of a species within the genus Olivibacter. The genome is 5,053,571 bp long and is comprised of 110 scaffolds with an average GC content of 44.61%. Of the 4,565 genes predicted, 4,501 were protein-coding genes and 64 were RNA genes. Most protein-coding genes (68.52%) were assigned to a putative function. The identification of 2-keto-4-pentenoate hydratase/2-oxohepta-3-ene-1,7-dioic acid hydratase-coding genes indicates involvement of this organism in the catechol catabolic pathway. In addition, genes encoding for ß-1,4-xylanases and ß-1,4-xylosidases reveal the xylanolytic action of O. sitiensis.

The thanatos mutation in Arabidopsis thaliana cellulose synthase 3 (AtCesA3) has a dominant-negative effect on cellulose synthesis and plant growth.

Genetic functional analyses of mutants in plant genes encoding cellulose synthases (CesAs) have suggested that cellulose deposition requires the activity of multiple CesA proteins. Here, a genetic screen has led to the identification of thanatos (than), a semi-dominant mutant of Arabidopsis thaliana with impaired growth of seedlings. Homozygous seedlings of than germinate and grow but do not survive. In contrast to other CesA mutants, heterozygous plants are dwarfed and display a radially swollen root phenotype. Cellulose content is reduced by approximately one-fifth in heterozygous and by two-fifths in homozygous plants, showing gene-dosage dependence. Map-based cloning revealed an amino acid substitution (P578S) in the catalytic domain of the AtCesA3 gene, indicating a critical role for this residue in the structure and function of the cellulose synthase complex. Ab initio analysis of the AtCesA3 subdomain flanking the conserved proline residue predicted that the amino acid substitution to serine alters protein secondary structure in the catalytic domain. Gene dosage-dependent expression of the AtCesA3 mutant gene in wild-type A. thaliana plants resulted in a than dominant-negative phenotype. We propose that the incorporation of a mis-folded CesA3 subunit into the cellulose synthase complex may stall or prevent the formation of functional rosette complexes.

Role of Lon1 protease in post-germinative growth and maintenance of mitochondrial function in Arabidopsis thaliana.

Maintenance of protein quality control and turnover is essential for cellular homeostasis. In plant organelles this biological process is predominantly performed by ATP-dependent proteases. Here, a genetic screen was performed that led to the identification of Arabidopsis thaliana Lon1 protease mutants that exhibit a post-embryonic growth retardation phenotype. Translational fusion to yellow fluorescent protein revealed AtLon1 subcellular localization in plant mitochondria, and the AtLon1 gene could complement the respiratory-deficient phenotype of the yeast PIM1 gene homolog. AtLon1 is highly expressed in rapidly growing plant organs of embryonic origin, including cotyledons and primary roots, and in inflorescences, which have increased mitochondria numbers per cell to fulfill their high energy requirements. In lon1 mutants, the expression of both mitochondrial and nuclear genes encoding respiratory proteins was normal. However, mitochondria isolated from lon1 mutants had a lower capacity for respiration of succinate and cytochrome c via complexes II and IV, respectively. Furthermore, the activity of key enzymes of the tricarboxylic acid (TCA) cycle was significantly reduced. Additionally, mitochondria in lon1 mutants had an aberrant morphology. These results shed light on the developmental mechanisms of selective proteolysis in plant mitochondria and suggest a critical role for AtLon1 protease in organelle biogenesis and seedling establishment.

Halotalea alkalilenta gen. nov., sp. nov., a novel osmotolerant and alkalitolerant bacterium from alkaline olive mill wastes, and emended description of the family Halomonadaceae Franzmann et al. 1989, emend. Dobson and Franzmann 1996.

A novel Gram-negative, motile, nonsporulating, rod-shaped bacterium isolated from alkaline sludge-like wastes ('alpeorujo' or 'alperujo') of two-phase olive oil extraction is described. The strain, designated AW-7(T), is an obligate aerobe that is halotolerant (tolerating up to 15 % w/v NaCl), sugar-tolerant (tolerating up to 45 % and 60 % w/v (+)-d-glucose and maltose respectively; these are the highest concentrations tolerated by any known members of the Bacteria domain) and alkalitolerant (growing at a broad pH range of 5-11). Strain AW-7(T) is chemo-organotrophic. Ubiquinone-9 was detected in the respiratory chain of strain AW-7(T). The major fatty acids present are C(18 : 1)omega7c, C(16 : 0), C(19 : 0) cyclo omega8c, C(12 : 0) 3-OH and C(16 : 1)omega7c/iso-C(15 : 0) 2-OH. Based on 16S rRNA gene sequence analysis, strain AW-7(T) showed almost equal phylogenetic distances from Zymobacter palmae (95.6 % similarity) and Carnimonas nigrificans (95.4 % similarity). In addition, low DNA-DNA relatedness values were found for strain AW-7(T) against Carnimonas nigrificans CECT 4437(T) (22.5-25.4 %) and Z. palmae DSM 10491(T) (11.9-14.4 %). The DNA G+C content of strain AW-7(T) is 64.4 mol%. Physiological and chemotaxonomic data further confirmed the differentiation of strain AW-7(T) from the genera Zymobacter and Carnimonas. Thus, strain AW-7(T) represents a novel bacterial genus within the family Halomonadaceae, for which the name Halotalea gen. nov. is proposed. Halotalea alkalilenta sp. nov. (type strain AW-7(T)=DSM 17697(T)=CECT 7134(T)) is proposed as the type species of the genus Halotalea gen. nov. A reassignment of the descriptive 16S rRNA signature characteristics of the family Halomonadaceae permitted the placement of the novel genus Halotalea into the family; in contrast, the genus Halovibrio possessed only 12 out of the 18 signature characteristics proposed, and hence it was excluded from the family Halomonadaceae.

Olivibacter sitiensis gen. nov., sp. nov., isolated from alkaline olive-oil mill wastes in the region of Sitia, Crete.

A novel, Gram-negative, non-motile, non-sporulating, rod-shaped bacterium isolated from a viscous two-phase olive-oil mill waste ('alpeorujo') is described. The strain, designated AW-6T, is an obligate aerobe, forming irregular, pigmented creamy white colonies. The pH and temperature ranges for growth were pH 5-8 and 5-45 degrees C, with optimal pH and temperature for growth of pH 6-7 and 28-32 degrees C, respectively. Strain AW-6T was chemo-organotrophic and utilized mostly D+ -glucose, protocatechuate and D+ -xylose, followed by L-cysteine, D- -fructose, D+ -galactose, L-histidine, lactose, sorbitol and sucrose. Menaquinone-7 was detected in the respiratory chain of strain AW-6T. The major fatty acids of strain AW-6T were C(16 : 1)omega7c and/or iso-C(15 : 0) 2-OH, iso-C(15 : 0), iso-C(17 : 0) 3-OH and C(16 : 0). The closest phylogenetic relative of strain AW-6T was clone BIti35 (89.7 % 16S rRNA gene sequence similarity), while Sphingobacterium thalpophilum DSM 11723T was the closest recognized relative within the Sphingobacteriaceae (88.2 % similarity). Strain AW-6T showed a low level of DNA-DNA relatedness to S. thalpophilum DSM 11723T (33.8-37.0 %). The DNA G+C content of strain AW-6T was 45.6 mol%. Physiological and chemotaxonomic data further confirmed the distinctiveness of strain AW-6T from members of the genera Sphingobacterium and Pedobacter. Thus, strain AW-6T is considered to represent a novel species of a new genus within the family Sphingobacteriaceae, for which the name Olivibacter sitiensis gen. nov., sp. nov. is proposed. The type strain is AW-6T =DSM 17696T=CECT 7133T).