Extracellular spermine triggers a rapid intracellular phosphatidic acid response in arabidopsis, involving PLDδ activation and stimulating ion flux

Polyamines, such as putrescine (Put), spermidine (Spd), and spermine (Spm), are low-molecular-weight polycationic molecules found in all living organisms. Despite the fact that they have been implicated in various important developmental and adaptative processes, their mode of action is still largely unclear. Here, we report that Put, Spd, and Spm trigger a rapid increase in the signaling lipid, phosphatidic acid (PA) in Arabidopsis seedlings but also mature leaves. Using time-course and dose-response experiments, Spm was found to be the most effective; promoting PA responses at physiological (low μM) concentrations. In seedlings, the increase of PA occurred mainly in the root and partly involved the plasma membrane polyamine-uptake transporter (PUT), RMV1. Using a differential 32Pi-labeling strategy combined with transphosphatidylation assays and T-DNA insertion mutants, we found that phospholipase D (PLD), and in particular PLDδ was the main contributor of the increase in PA. Measuring non-invasive ion fluxes (MIFE) across the root plasma membrane of wild type and pldδ-mutant seedlings, revealed that the formation of PA is linked to a gradual- and transient efflux of K+. Potential mechanisms of how PLDδ and the increase of PA are involved in polyamine function is discussed

Hydrogen peroxide-induced root Ca2+ and K+ fluxes correlate with salt tolerance in cereals: towards the cell-based phenotyping

Salinity stress-induced production of reactive oxygen species (ROS) and associated oxidative damage is one of the major factors limiting crop production in saline soils. However, the causal link between ROS production and stress tolerance is not as straightforward as one may expect, as ROS may also play an important signaling role in plant adaptive responses. In this study, the causal relationship between salinity and oxidative stress tolerance in two cereal crops—barley (Hordeum vulgare) and wheat (Triticum aestivum)—was investigated by measuring the magnitude of ROS-induced net K+ and Ca2+ fluxes from various root tissues and correlating them with overall whole-plant responses to salinity. We have found that the association between flux responses to oxidative stress and salinity stress tolerance was highly tissue specific, and was also dependent on the type of ROS applied. No correlation was found between root responses to hydroxyl radicals and the salinity tolerance. However, when oxidative stress was administered via H2O2 treatment, a significant positive correlation was found for the magnitude of ROS-induced K+ efflux and Ca2+ uptake in barley and the overall salinity stress tolerance, but only for mature zone and not the root apex. The same trends were found for wheat. These results indicate high tissue specificity of root ion fluxes response to ROS and suggest that measuring the magnitude of H2O2-induced net K+ and Ca2+ fluxes from mature root zone may be used as a tool for cell-based phenotyping in breeding programs aimed to improve salinity stress tolerance in cereals

Tissue-specific regulation of Na+ and K+ transporters explains genotypic differences in salinity stress tolerance in rice

Rice (Oryza sativa) is a staple food that feeds more than half the world population. As rice is highly sensitive to soil salinity, current trends in soil salinization threaten global food security. To better understand the mechanistic basis of salinity tolerance in rice, three contrasting rice cultivars - Reiziq (tolerant), Doongara (moderately tolerant), and Koshihikari (sensitive) - were examined and the differences in operation of key ion transporters mediating ionic homeostasis in these genotypes were evaluated. Tolerant varieties had reduced Na+ translocation from roots to shoots. Electrophysiological and quantitative reverse transcription PCR experiments showed that tolerant genotypes possessed 2-fold higher net Na+ efflux capacity in the root elongation zone. Interestingly, this efflux was only partially mediated by the plasma membrane Na+/H+ antiporter (OsSOS1), suggesting involvement of some other exclusion mechanisms. No significant difference in Na+ exclusion from the mature root zones was found between cultivars, and the transcriptional changes in the salt overly sensitive signaling pathway genes in the elongation zone were not correlated with the genetic variability in salinity tolerance amongst genotypes. The most important hallmark of differential salinity tolerance was in the ability of the plant to retain K+ in both root zones. This trait was conferred by at least three complementary mechanisms: (1) its superior ability to activate H+-ATPase pump operation, both at transcriptional and functional levels; (2) reduced sensitivity of K+ efflux channels to reactive oxygen species; and (3) smaller upregulation in OsGORK and higher upregulation of OsAKT1 in tolerant cultivars in response to salt stress. These traits should be targeted in breeding programs aimed to improve salinity tolerance in commercial rice cultivars

GABA operates upstream of H+-ATPase and improves salinity tolerance in Arabidopsis by enabling cytosolic K+ retention and Na+ exclusion

The non-protein amino acid γ-aminobutyric acid (GABA) rapidly accumulates in plant tissues in response to salinity. However, the physiological rationale for this elevation remains elusive. This study compared electrophysiological and whole-plant responses of salt-treated Arabidopsis mutants pop2-5 and gad1,2, which have different abilities to accumulate GABA. The pop2-5 mutant, which was able to overaccumulate GABA in its roots, showed a salt-tolerant phenotype. On the contrary, the gad1,2 mutant, lacking the ability to convert glutamate to GABA, showed oversensitivity to salinity. The greater salinity tolerance of the pop2-5 line was explained by: (i) the role of GABA in stress-induced activation of H+-ATPase, thus leading to better membrane potential maintenance and reduced stress-induced K+ leak from roots; (ii) reduced rates of net Na+ uptake; (iii) higher expression of SOS1 and NHX1 genes in the leaves, which contributed to reducing Na+ concentration in the cytoplasm by excluding Na+ to apoplast and sequestering Na+ in the vacuoles; (iv) a lower rate of H2O2 production and reduced reactive oxygen species-inducible K+ efflux from root epidermis; and (v) better K+ retention in the shoot associated with the lower expression level of GORK channels in plant leaves

Radio Galaxy Zoo: Knowledge Transfer Using Rotationally Invariant Self-Organising Maps

With the advent of large scale surveys the manual analysis and classification of individual radio source morphologies is rendered impossible as existing approaches do not scale. The analysis of complex morphological features in the spatial domain is a particularly important task. Here we discuss the challenges of transferring crowdsourced labels obtained from the Radio Galaxy Zoo project and introduce a proper transfer mechanism via quantile random forest regression. By using parallelized rotation and flipping invariant Kohonen-maps, image cubes of Radio Galaxy Zoo selected galaxies formed from the FIRST radio continuum and WISE infrared all sky surveys are first projected down to a two-dimensional embedding in an unsupervised way. This embedding can be seen as a discretised space of shapes with the coordinates reflecting morphological features as expressed by the automatically derived prototypes. We find that these prototypes have reconstructed physically meaningful processes across two channel images at radio and infrared wavelengths in an unsupervised manner. In the second step, images are compared with those prototypes to create a heat-map, which is the morphological fingerprint of each object and the basis for transferring the user generated labels. These heat-maps have reduced the feature space by a factor of 248 and are able to be used as the basis for subsequent ML methods. Using an ensemble of decision trees we achieve upwards of 85.7% and 80.7% accuracy when predicting the number of components and peaks in an image, respectively, using these heat-maps. We also question the currently used discrete classification schema and introduce a continuous scale that better reflects the uncertainty in transition between two classes, caused by sensitivity and resolution limits

Radio Galaxy Zoo: Cosmological Alignment of Radio Sources

We study the mutual alignment of radio sources within two surveys, FIRST and TGSS. This is done by producing two position angle catalogues containing the preferential directions of respectively $30\,059$ and $11\,674$ extended sources distributed over more than $7\,000$ and $17\,000$ square degrees. The identification of the sources in the FIRST sample was performed in advance by volunteers of the Radio Galaxy Zoo project, while for the TGSS sample it is the result of an automated process presented here. After taking into account systematic effects, marginal evidence of a local alignment on scales smaller than $2.5\deg$ is found in the FIRST sample. The probability of this happening by chance is found to be less than $2$ per cent. Further study suggests that on scales up to $1.5\deg$ the alignment is maximal. For one third of the sources, the Radio Galaxy Zoo volunteers identified an optical counterpart. Assuming a flat $\Lambda$CDM cosmology with $\Omega_m = 0.31, \Omega_\Lambda = 0.69$, we convert the maximum angular scale on which alignment is seen into a physical scale in the range $[19, 38]$ Mpc $h_{70}^{-1}$. This result supports recent evidence reported by Taylor and Jagannathan of radio jet alignment in the $1.4$ deg$^2$ ELAIS N1 field observed with the Giant Metrewave Radio Telescope. The TGSS sample is found to be too sparsely populated to manifest a similar signal

Distinct evolutionary origins of intron retention splicing events in NHX1 antiporter transcripts relate to sequence specific distinctions in Oryza species

The genome of Asian cultivated rice (Oryza sativa L.) shows the presence of six organelle-specific and one plasma membrane (OsNHX1-7) NHX-type cation proton antiporters. Of these, vacuolar-localized OsNHX1 is extensively characterized. The genus Oryza consists of 27 species and 11 genome-types, with cultivated rice, diploid O. sativa, having an AA-type genome. Oryza NHX1 orthologous regions (gene organization, 5′ upstream cis elements, amino acid residues/motifs) from closely related Oryza AA genomes cluster distinctly from NHX1 regions from more ancestral Oryza BB, FF and KKLL genomes. These sequence-specific distinctions also extend to two separate intron retention (IR) events involving Oryza NHX1 transcripts that occur at the 5′ and 3′ ends of the NHX1 transcripts. We demonstrate that the IR event involving the 5′ UTR is present only in more recently evolved Oryza AA genomes while the IR event governing retention of the 13th intron of Oryza NHX1 (terminal intron) is more ancient in origin, also occurring in halophytic wild rice, Oryza coarctata (KKLL). We also report presence of a retro-copy of the OcNHX1 cDNA in the genome of O. coarctata (rOcNHX1). Preferential species and tissue specific up- or down-regulation of the correctly spliced NHX1 transcript/5′ UTR/13th intron-retaining splice variants under salinity was observed. The implications of IR on NHX1 mRNA stability and ORF diversity in Oryza spp. is discussed