Short daily ultraviolet exposure enhances intrinsic water-use efficiency and delays senescence in Micro-Tom tomato plants.

Ultraviolet (UV) radiation, unless present at high doses, is recognised as a regulator of plant growth and some specific processes. The present study investigated the influence of short daily UV irradiation (15min/day, 11days) on leaf gas exchange and some biochemical and molecular markers of leaf senescence (such as stomata movements, chlorophyll breakdown, anthocyanin production, senescence-associated genes) in Micro-Tom tomato plants. The UV-induced reduction of g s (stomatal conductance) during the treatment was associated with the modified expression of some genes involved in the control of stomatal movements. We hypothesise a two-step regulation of stomatal closure involving salicylic and abscisic acid hormones. The temporal changes of g s and A net (net photosynthetic CO2 assimilation rate) along with the pigment behaviour, suggest a possible delay of leaf senescence in treated plants, confirmed by the expression levels of genes related to senescence such as SAG113 and DFR . The UV potential to induce a persistent partial inhibition of g s without severely affecting A net led to an increased iWUE (intrinsic water-use efficiency) during the 11-day treatment, suggesting a priming effect of short daily UV radiation towards drought conditions potentially useful in reducing the excess water use in agriculture.

Exogenous miRNAs induce post-transcriptional gene silencing in plants.

Plants seem to take up exogenous RNA that was artificially designed to target specific genes, followed by activation of the RNA interference (RNAi) machinery. It is, however, not known whether plants use RNAs themselves as signalling molecules in plant-to-plant communication, other than evidence that an exchange of small RNAs occurs between parasitic plants and their hosts. Exogenous RNAs from the environment, if taken up by some living organisms, can indeed induce RNAi. This phenomenon has been observed in nematodes and insects, and host Arabidopsis cells secrete exosome-like extracellular vesicles to deliver plant small RNAs into Botrytis cinerea. Here we show that micro-RNAs (miRNAs) produced by plants act as signalling molecules affecting gene expression in other, nearby plants. Exogenous miRNAs, such as miR156 and miR399, trigger RNAi via a mechanism requiring both AGO1 and RDR6. This emphasizes that the production of secondary small interfering RNAs is required. This evidence highlights the existence of a mechanism in which miRNAs represent signalling molecules that enable communication between plants.

Conservation of ethanol fermentation and its regulation in land plants.

Ethanol fermentation is considered as one of the main metabolic adaptations to ensure energy production in higher plants under anaerobic conditions. Following this pathway, pyruvate is decarboxylated and reduced to ethanol with the concomitant oxidation of NADH to NAD+. Despite its acknowledgement as an essential metabolic strategy, the conservation of this pathway and its regulation throughout plant evolution have not been assessed so far. To address this question, we compared ethanol fermentation in species representing subsequent steps in plant evolution and related it to the structural features and transcriptional regulation of the two enzymes involved: pyruvate decarboxylase (PDC) and alcohol dehydrogenase (ADH). We observed that, despite the conserved ability to produce ethanol upon hypoxia in distant phyla, transcriptional regulation of the enzymes involved is not conserved in ancient plant lineages, whose ADH homologues do not share structural features distinctive for acetaldehyde/ethanol-processing enzymes. Moreover, Arabidopsis mutants devoid of ADH expression exhibited enhanced PDC activity and retained substantial ethanol production under hypoxic conditions. Therefore, we concluded that, whereas ethanol production is a highly conserved adaptation to low oxygen, its catalysis and regulation in land plants probably involve components that will be identified in the future.

Ascophyllum nodosum Seaweed Extract Alleviates Drought Stress in Arabidopsis by Affecting Photosynthetic Performance and Related Gene Expression.

Drought represents one of the most relevant abiotic stress affecting growth and yield of crop plants. In order to improve the agricultural productivity within the limited water and land resources, it is mandatory to increase crop yields in presence of unfavorable environmental stresses. The use of biostimulants, often containing seaweed extracts, represents one of the options for farmers willing to alleviate abiotic stress consequences on crops. In this work, we investigated the responses of Arabidopsis plants treated with an extract from the brown alga Ascophyllum nodosum (ANE), under drought stress conditions, demonstrating that ANE positively influences Arabidopsis survival. Pre-treatment with ANE induced a partial stomatal closure, associated with changes in the expression levels of genes involved in ABA-responsive and antioxidant system pathways. The pre-activation of these pathways results in a stronger ability of ANE-treated plants to maintain a better photosynthetic performance compared to untreated plants throughout the dehydration period, combined with a higher capacity to dissipate the excess of energy as heat in the reaction centers of photosystem II. Our results suggest that drought stressed plants treated with ANE are able to maintain a strong stomatal control and relatively higher values of both water use efficiency (WUE) and mesophyll conductance during the last phase of dehydration. Simultaneously, the activation of a pre-induced antioxidant defense system, in combination with a more efficient energy dissipation mechanism, prevents irreversible damages to the photosynthetic apparatus. In conclusion, pre-treatment with ANE is effective to acclimate plants to the incoming stress, promoting an increased WUE and dehydration tolerance.

A reassessment of the role of sucrose synthase in the hypoxic sucrose-ethanol transition in Arabidopsis.

Plants under low-oxygen availability adapt their metabolism to compensate for the lower ATP production that arises from the limited respiratory activity in mitochondria. Anaerobic glycolysis requires continuous fuelling of carbon units, also provided from sucrose. The anaerobic catabolism of sucrose is thought to require the activity of sucrose synthase, being this enzymatic reaction more energetically favourable than that of invertase. The role of sucrose synthases (SUS) for aerobic sucrose catabolism in Arabidopsis has been recently questioned since SUS mutants fail to show altered phenotype or metabolic profile. In the present paper, we analysed the role of SUS1 and SUS4, both induced by low oxygen, in plant survival and ethanol production. The results showed that mutants lacking both SUS were as tolerant to low oxygen as the wild type in most of the experimental conditions tested. Only under conditions of limiting sugar availability the requirement of SUS1 and SUS4 for ethanol production was evident, although partly compensated by invertase activities, as revealed by the use of a double mutant lacking the two major cytosolic invertases. We conclude that, contrary to general belief, the sucrose synthase pathway is not the preferential route for sucrose metabolism under hypoxia.

Low oxygen response mechanisms in green organisms.

Low oxygen stress often occurs during the life of green organisms, mostly due to the environmental conditions affecting oxygen availability. Both plants and algae respond to low oxygen by resetting their metabolism. The shift from mitochondrial respiration to fermentation is the hallmark of anaerobic metabolism in most organisms. This involves a modified carbohydrate metabolism coupled with glycolysis and fermentation. For a coordinated response to low oxygen, plants exploit various molecular mechanisms to sense when oxygen is either absent or in limited amounts. In Arabidopsis thaliana, a direct oxygen sensing system has recently been discovered, where a conserved N-terminal motif on some ethylene responsive factors (ERFs), targets the fate of the protein under normoxia/hypoxia. In Oryza sativa, this same group of ERFs drives physiological and anatomical modifications that vary in relation to the genotype studied. The microalga Chlamydomonas reinhardtii responses to low oxygen seem to have evolved independently of higher plants, posing questions on how the fermentative metabolism is modulated. In this review, we summarize the most recent findings related to these topics, highlighting promising developments for the future.

Transcript profiling of chitosan-treated Arabidopsis seedlings.

In nature, plants can recognize potential pathogens, thus activating intricate networks of defense signals and reactions. Inducible defense is often mediated by the detection of microbe or pathogen associated molecular pattern elicitors, such as flagellin and chitin. Chitosan, the deacetylated form of chitin, plays a role in inducing protection against pathogens in many plant species. We evaluated the ability of chitosan to confer resistance to Botrytis cinerea in Arabidopsis leaves. We subsequently treated Arabidopsis seedlings with chitosan and carried out a transcript profiling analysis using both ATH1 GeneChip microarrays and quantitative RT-PCR. The results showed that defense response genes, including camalexin biosynthesis genes, were up-regulated by chitosan, both in wild-type and in the chitin-insensitive cerk1 mutant, indicating that chitosan is perceived through a CERK1-independent pathway.

HRE1 and HRE2, two hypoxia-inducible ethylene response factors, affect anaerobic responses in Arabidopsis thaliana.

Plants often experience challenging hypoxic conditions imposed by soil waterlogging or complete flooding. In rice, Sub1A, a flooding-induced ethylene responsive factor (ERF) plays a crucial role in submergence tolerance. In this study, we examined two Arabidopsis Hypoxia Responsive ERF genes (HRE1 and HRE2), belonging to the same ERF group as Sub1A. Transgenic Arabidopsis plants, which over-expressed HRE1, showed an improved tolerance of anoxia, whereas a double-knockout mutant hre1hre2 was more susceptible than the wild type. HRE1 over-expressing plants showed an increased activity in the fermentative enzymes pyruvate decarboxylase and alcohol dehydrogenase together with increased ethanol production under hypoxia, but not in normoxia. Whole-genome microarray analyses suggested that an over-expression of HRE1, but not HRE2, increased the induction of most anaerobic genes under hypoxia. Real-time quantitative (q)PCR analyses confirmed a positive effect of HRE1 over-expression on several anaerobic genes, whereas the double-knockout mutant hre1hre2 showed a decreased expression in the same genes after 4 h of hypoxia. Single-knockout mutants did not show significant differences from the wild type. We found that the regulation of HRE1 and HRE2 by low oxygen relies on different mechanisms, since HRE1 requires protein synthesis to be induced while HRE2 does not. HRE2 is likely to be regulated post-transcriptionally by mRNA stabilization. We propose that HRE1 and HRE2 play a partially redundant role in low oxygen signalling in Arabidopsis thaliana, thus improving the tolerance of the plant to the stress by enhancing anaerobic gene expression and ethanolic fermentation.

Heat acclimation and cross-tolerance against anoxia in Arabidopsis.

Arabidopsis seedlings are highly sensitive to low oxygen and they die rapidly when exposed to anoxia. Tolerance to anoxia depends on the ability to efficiently use carbohydrates through the fermentative pathway, as highlighted by the lower tolerance displayed by a mutant devoid of alcohol dehydrogenase. Other mechanisms of tolerance are also possible and may include a role for heat-induced genes. In fact, heat shock proteins (HSPs) are induced by anoxia. This suggests that there may be a cross-adaptation mechanism between heat and anoxic stress, and in this work, we studied the acclimation of Arabidopsis seedlings both to low oxygen and heat. The results show that seedlings subjected to hypoxia or heat pretreatment survive anoxia much better. Interestingly, we also observed an increased anoxia tolerance in heat-treated alcohol dehydrogenase (adh) mutant plants. On the other hand, anoxic pretreatment does not confer tolerance to heat stress. The success of the induction of HSPs by anoxia is in direct relation to the amount of sucrose available, and this in turn relates to how well seedlings will survive under anoxia. HSP transcripts were also detected during seed development and germination, two hypoxia-prone processes, suggesting that hypoxia-induced HSP expression is physiologically relevant.