Increased firmness and modified cell wall composition by ethylene were reversed by the ethylene inhibitor 1-methylcyclopropene (1-MCP) in the non-climacteric olives harvested at dark green stage - Possible implementation of ethylene for olive quality.

This study aimed to investigate the firmness retention by ethylene treatment in olive fruit, as observed earlier. Ethylene concentrations up to 1000 µL L-1 were applied to dark green 'Konservolia' olives harvested shortly before the green maturation and exposed to 20 °C for up to 9 d. Surprisingly, the results indicated a tendency to fruit firmness increases in concentration-dependent manner in a non-climacteric fruit. The highest concentration increased the firmness within 12 h by approximately 1.35-fold, but transiently for approximately up to 5 d; all ethylene inhibitors tested, either of synthesis (ethoxyvinyl glycine or AVG), or perception (1 -methyl-cyclopropene or 1-MCP, and silver nitrate) prevented the firmness increase. Texture was evaluated by firmness and changes in lignin, cellulose (CL), total pectins (TPC), water soluble pectins (WSP) and total non-cellulosic sugars (total sugars) concentrations, and in pectin esterification degree (DE) in the alcohol insoluble residue (AIR) of 'Konservolia' fruit pericarp during 1.5-d, 5-d and 10-d treatments with 1000 µL L-1 ethylene at 20 °C. Pectins in AIR were also extracted sequentially with cyclohexane-trans-1,2-diaminetetra-acetate (CDTA), Na2CO3, 1 M and 4 M KOH. The results showed that on day 1.5, the increased firmness was consistent with increased CL (crystalline formation, as observed by microscopy), total sugars and DE levels, but reduced WSP, whereas softening reversed the changes and lowered TPC and CDTA-soluble pectins in all fruit on day 10. However, on day 5 ethylene-treated olives exhibited a transitional phase during softening, characterized by retention of high TPC concentration and energy demand, as indicated by elevated respiration rates. The inhibitor 1-MCP, applied before ethylene, did inhibit the responses to ethylene treatment. Ethylene firming effect and the respective cell wall changes in olives are demonstrated for first time. The experiments could be used for research on perception and transcription responses to ethylene in olive, a non-climacteric fruit. In practice, high ethylene concentrations could also be beneficial for firmness increase and/or short storage of dark green olives.

Changes in the properties of calcium-carbon inclusions during leaf development and their possible relationship with leaf functional maturation in three inclusion-bearing species.

In many plant species, carbon-calcium inclusion (calcium oxalate crystals or cystoliths containing calcium carbonate) formation is a fundamental part of their physiology even necessary for normal growth and development. Despite the long-standing studies on carbon-calcium inclusions, the alterations in their properties during leaf development and their possible association with the maturation of the photosynthetic machinery have not been previously examined. In order to acquire more insights into this subject, we examined three of the most common species bearing abundant inclusions of different types, i.e., Amaranthus hybridus, Vitis vinifera, and Parietaria judaica. Results of our study showed that, irrespective of species and type of inclusion, similar patterns in the alterations of their properties are observed during leaf maturation, except for some differences in cell differentiation and distribution between raphides and druses in Vitis vinifera. As expected, inclusion formation has taken place at very early developmental stages and maximum density was observed in very young leaves. Inclusion properties are changing in a coordinated way with leaf area and these modifications are compatible with the concept that each idioblast or lithocyst "services" a finite number/area of adjacent cells. This tight coordination is also evident at the whole leaf level. Moreover, we observed an association of the properties of carbon-calcium inclusions and gas exchange, suggesting a possible implication of these structures in photosynthesis.

Functional analysis of the cyclophilin PpiB role in bacterial cell division.

Escherichia coli PpiB is a peptidyl-prolyl cis/trans isomerase (PPIase, EC: 5.2.1.8) with chaperone activity. Here, we show that the ΔppiB deletion strain and the PpiB over-expression wild-type strain are both characterized by defects in cell division involving milder or severe cell filamentation, respectively. Using various PpiB mutants, we show that the PPIase activity of PpiB is necessary for the observed cell filamentation, whereas other structural features apart from the active site are also important for this phenotype. Early divisome components zipA and ftsZ showed decreased expression in ΔppiB cells, whereas the corresponding proteins partially suppressed the division phenotype of ΔppiB cells as well. Although PpiB itself has no obvious specific affinity for the septal ring as a GFP translational fusion showed a diffuse cytoplasmic localization, it interacts with FtsZ employing the C-terminal FtsZ domain, decreases its GTPase activity and when over-expressed shows an inhibitory effect on the proper FtsZ localization at future division sites. Furthermore, additional putative PpiB prey proteins are able to partially restore the ΔppiB phenotype indicating that PpiB is able to control bacterial cell division by probably modulating the function of various other proteins which are indirectly associated with the process.

Reevaluation of the plant "gemstones": Calcium oxalate crystals sustain photosynthesis under drought conditions.

Land plants face the perpetual dilemma of using atmospheric carbon dioxide for photosynthesis and losing water vapors, or saving water and reducing photosynthesis and thus growth. The reason behind this dilemma is that this simultaneous exchange of gases is accomplished through the same minute pores on leaf surfaces, called stomata. In a recent study we provided evidence that pigweed, an aggressive weed, attenuates this problem exploiting large crystals of calcium oxalate as dynamic carbon pools. This plant is able to photosynthesize even under drought conditions, when stomata are closed and water losses are limited, using carbon dioxide from crystal decomposition instead from the atmosphere. Abscisic acid, an alarm signal that causes stomatal closure seems to be implicated in this function and for this reason we named this path "alarm photosynthesis." The so-far "enigmatic," but highly conserved and widespread among plant species calcium oxalate crystals seem to play a crucial role in the survival of plants.

Alarm Photosynthesis: Calcium Oxalate Crystals as an Internal CO2 Source in Plants.

Calcium oxalate crystals are widespread among animals and plants. In land plants, crystals often reach high amounts, up to 80% of dry biomass. They are formed within specific cells, and their accumulation constitutes a normal activity rather than a pathological symptom, as occurs in animals. Despite their ubiquity, our knowledge on the formation and the possible role(s) of these crystals remains limited. We show that the mesophyll crystals of pigweed (Amaranthus hybridus) exhibit diurnal volume changes with a gradual decrease during daytime and a total recovery during the night. Moreover, stable carbon isotope composition indicated that crystals are of nonatmospheric origin. Stomatal closure (under drought conditions or exogenous application of abscisic acid) was accompanied by crystal decomposition and by increased activity of oxalate oxidase that converts oxalate into CO2 Similar results were also observed under drought stress in Dianthus chinensis, Pelargonium peltatum, and Portulacaria afra Moreover, in A. hybridus, despite closed stomata, the leaf metabolic profiles combined with chlorophyll fluorescence measurements indicated active photosynthetic metabolism. In combination, calcium oxalate crystals in leaves can act as a biochemical reservoir that collects nonatmospheric carbon, mainly during the night. During the day, crystal degradation provides subsidiary carbon for photosynthetic assimilation, especially under drought conditions. This new photosynthetic path, with the suggested name "alarm photosynthesis," seems to provide a number of adaptive advantages, such as water economy, limitation of carbon losses to the atmosphere, and a lower risk of photoinhibition, roles that justify its vast presence in plants.

A splicing mutation in the novel mitochondrial protein DNAJC11 causes motor neuron pathology associated with cristae disorganization, and lymphoid abnormalities in mice.

Mitochondrial structure and function is emerging as a major contributor to neuromuscular disease, highlighting the need for the complete elucidation of the underlying molecular and pathophysiological mechanisms. Following a forward genetics approach with N-ethyl-N-nitrosourea (ENU)-mediated random mutagenesis, we identified a novel mouse model of autosomal recessive neuromuscular disease caused by a splice-site hypomorphic mutation in a novel gene of unknown function, DnaJC11. Recent findings have demonstrated that DNAJC11 protein co-immunoprecipitates with proteins of the mitochondrial contact site (MICOS) complex involved in the formation of mitochondrial cristae and cristae junctions. Homozygous mutant mice developed locomotion defects, muscle weakness, spasticity, limb tremor, leucopenia, thymic and splenic hypoplasia, general wasting and early lethality. Neuropathological analysis showed severe vacuolation of the motor neurons in the spinal cord, originating from dilatations of the endoplasmic reticulum and notably from mitochondria that had lost their proper inner membrane organization. The causal role of the identified mutation in DnaJC11 was verified in rescue experiments by overexpressing the human ortholog. The full length 63 kDa isoform of human DNAJC11 was shown to localize in the periphery of the mitochondrial outer membrane whereas putative additional isoforms displayed differential submitochondrial localization. Moreover, we showed that DNAJC11 is assembled in a high molecular weight complex, similarly to mitofilin and that downregulation of mitofilin or SAM50 affected the levels of DNAJC11 in HeLa cells. Our findings provide the first mouse mutant for a putative MICOS protein and establish a link between DNAJC11 and neuromuscular diseases.

Wettability, polarity, and water absorption of holm oak leaves: effect of leaf side and age.

Plant trichomes play important protective functions and may have a major influence on leaf surface wettability. With the aim of gaining insight into trichome structure, composition, and function in relation to water-plant surface interactions, we analyzed the adaxial and abaxial leaf surface of holm oak (Quercus ilex) as a model. By measuring the leaf water potential 24 h after the deposition of water drops onto abaxial and adaxial surfaces, evidence for water penetration through the upper leaf side was gained in young and mature leaves. The structure and chemical composition of the abaxial (always present) and adaxial (occurring only in young leaves) trichomes were analyzed by various microscopic and analytical procedures. The adaxial surfaces were wettable and had a high degree of water drop adhesion in contrast to the highly unwettable and water-repellent abaxial holm oak leaf sides. The surface free energy and solubility parameter decreased with leaf age, with higher values determined for the adaxial sides. All holm oak leaf trichomes were covered with a cuticle. The abaxial trichomes were composed of 8% soluble waxes, 49% cutin, and 43% polysaccharides. For the adaxial side, it is concluded that trichomes and the scars after trichome shedding contribute to water uptake, while the abaxial leaf side is highly hydrophobic due to its high degree of pubescence and different trichome structure, composition, and density. Results are interpreted in terms of water-plant surface interactions, plant surface physical chemistry, and plant ecophysiology.

Effects of Lactobacillus salivarius, Lactobacillus reuteri, and Pediococcus acidilactici on the nematode Caenorhabditis elegans include possible antitumor activity.

This study examined the effects of three lactic acid bacteria (LAB) strains on the nematode Caenorhabditis elegans. Lactobacillus salivarius, Lactobacillus reuteri, and Pediococcus acidilactici were found to inhibit the development and growth of the worm. Compared to Escherichia coli used as the control, L. reuteri and P. acidilactici reduced the lifespan of wild-type and short-lived daf-16 worms. On the contrary, L. salivarius extended the lifespan of daf-16 worms when used live, but reduced it as UV-killed bacteria. The three LAB induced the expression of genes involved in pathogen response and inhibited the growth of tumor-like germ cells, without affecting DAF16 localization or increasing corpse cells. Our results suggest the possible use of C. elegans as a model for studying the antitumor attributes of LAB. The negative effects of these LAB strains on the nematode also indicate their potential use against parasitic nematodes.

Characterization of a Mesorhizobium loti alpha-type carbonic anhydrase and its role in symbiotic nitrogen fixation.

Carbonic anhydrase (CA) (EC 4.2.1.1) is a widespread enzyme catalyzing the reversible hydration of CO(2) to bicarbonate, a reaction that participates in many biochemical and physiological processes. Mesorhizobium loti, the microsymbiont of the model legume Lotus japonicus, possesses on the symbiosis island a gene (msi040) encoding an alpha-type CA homologue, annotated as CAA1. In the present work, the CAA1 open reading frame from M. loti strain R7A was cloned, expressed, and biochemically characterized, and it was proven to be an active alpha-CA. The biochemical and physiological roles of the CAA1 gene in free-living and symbiotic rhizobia were examined by using an M. loti R7A disruption mutant strain. Our analysis revealed that CAA1 is expressed in both nitrogen-fixing bacteroids and free-living bacteria during growth in batch cultures, where gene expression was induced by increased medium pH. L. japonicus plants inoculated with the CAA1 mutant strain showed no differences in top-plant traits and nutritional status but consistently formed a higher number of nodules exhibiting higher fresh weight, N content, nitrogenase activity, and delta(13)C abundance. Based on these results, we propose that although CAA1 is not essential for nodule development and symbiotic nitrogen fixation, it may participate in an auxiliary mechanism that buffers the bacteroid periplasm, creating an environment favorable for NH(3) protonation, thus facilitating its diffusion and transport to the plant. In addition, changes in the nodule delta(13)C abundance suggest the recycling of at least part of the HCO(3)(-) produced by CAA1.

Photosynthetic characteristics of olive tree (Olea europaea) bark.

Functional and structural characteristics of corticular photosynthesis of sun-exposed bark of olive tree (Olea europaea L.) were examined. Stomata are only sporadically present during stem primary growth. Light transmission through the phellem was age dependent, decreasing rapidly in stems older than five years of age. Light transmission was also low in pubescent 1-year-old stems. Light transmission was about 50% higher in wet phellem than in dry phellem. Photosynthetic capacity on a unit area basis (measured with an oxygen disc electrode at 27 degrees C and about 5% CO(2) on chlorophyllous tissue discs isolated from the stem) was higher in 1-, 20- and 30-year-old stems compared with 2-10-year-old stems. Low chlorophyll a/b ratio and light compensation points were recorded in olive stems with low phellem light transmission, in accordance with the shade acclimation hypothesis. The intrinsic photochemical efficiency of photosystem II of all stems, especially young stems, was less than that of the leaves. Our results show that olive tree bark possesses an efficient photosynthetic mechanism that may significantly contribute not only to the reduction in concentrations of CO(2) in the inner bark, but also to whole-tree carbon balance.

Boron remobilization at low boron supply in olive (Olea europaea) in relation to leaf and phloem mannitol concentrations.

For plant species in which a considerable portion of the photoassimilates are translocated in the phloem as sugar alcohols, boron is freely translocated from mature organs to growing tissues. However, the effects of decreased plant boron status on boron remobilization are poorly understood. We conducted a growth chamber experiment (CE) and a field experiment (FE) to study the effects of low boron supply on boron remobilization in olive (Olea europaea L.), a species that transports considerable amounts of mannitol in the phloem. For the CE, several physiological parameters were compared between control (B+) and boron-deficient olive plants (B-) during the expansion of new leaves. Boron remobilization was assessed by measuring boron content of selected leaves at the beginning and at the end of the CE. As expected, boron was remobilized from mature leaves to young leaves of B+ plants; however, considerable boron remobilization was also observed in B- plants, suggesting a mechanism whereby olive can sustain a minimum boron supply for growth of new tissues despite an insufficient external boron supply. Boron deficiency caused inhibition of new growth but had no effect on photosynthetic capacity per unit leaf surface area of young and mature leaves, thereby altering the carbon utilization pattern and resulting in carbon allocation to structures within the source leaves and accumulation of soluble carbohydrates. Specifically, in mature B- leaves in the CE and in B- leaves in the FE, mannitol concentration on a leaf water content basis increased by 48 and 27% respectively, compared with controls. Carbon export ability (assessed by both phloem anatomy and phloem exudate composition of FE leaves) was enhanced at low boron supply. We conclude that, at low boron supply, increased mannitol concentrations maintain boron remobilization from source leaves to boron-demanding sink leaves.

Photoacclimation in spathiphyllum.

We studied photoacclimation in Spathiphyllum grown at an irradiance of 40 or 420 micromol/m2 s (LL or HL, respectively). All parameters studied responded to acclimation. Leaves at LL, in contrast to HL, were thinner and oriented perpendicular to the incident light, had more chlorophyll per g f w, fewer stomata on the upper leaf surface and a reduced layer of mesophyll cells. Their chloroplasts at HL had wider grana with less thylakoids per granum, and better organized photosystems than at LL. PSI and PSII activities per mg chlorophyll ( Vmax ), and PSI and PSII content (total activity per g f w), were lower at LL than at HL and so was the light requirement for saturation of the PSI or PSII partial photoreactions, suggesting that fewer photosystems with larger antenna size prevail at LL, but many more with smaller antenna size at HL. Analysis of chlorophyll distribution among the thylakoid pigment-protein complexes showed less antenna chlorophyll serving PSII (CPa+LHCP1+LHCP3) than that serving PSI (CPIa+CPI+LHCP2) at LL as compared to HL, and thus a lower PSII/PSI ratio at LL, in agreement with the general finding that LL plants, with larger PSII antenna size, have lower PSII/PSI ratio. The increase in PSI antenna size at LL was correlated with the increase in the distribution of chlorophyll in pigment-protein complexes serving PSI, and a very large chlorophyll/protein molar ratio in the isolated CPI complex. On the other hand, the PSII antenna chlorophyll (CPa+LHCP1+LHCP3) on a g f w basis, and the chlorophyll a/b ratio remained more or less constant at LL or HL. This may reflect our finding that Spathiphyllum contains mainly the 27 kDa inner LHCII antenna protein, the size of which remains unaffected by photoacclimation. The increase in the distribution of chlorophyll in pigment-protein complexes serving PSII at HL, therefore, reflects the higher population of PSII at HL. Very high PSI activity was found at HL, which we attribute to the highly organized small in size PSI.