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ABSCISIC ACID INSENSITIVE5 (ABI5), a key regulator of the abscisic acid (ABA) signalling pathway, plays a fundamental role in seed germination and post-germinative development. However, the molecular mechanism underlying the repression function of ABI5 remains to be elucidated. In this study, we demonstrate that the conserved eukaryotic WD40 repeat protein Receptor for Activated C Kinase 1 (RACK1) is a novel negative regulator of ABI5 in Arabidopsis. The RACK1 loss-of-function mutant is hypersensitive to ABA, while this phenotype is rescued by a mutation in ABI5. Moreover, overexpression of RACK1 suppresses ABI5 transcriptional activation activity for ABI5-targeted genes. RACK1 may also physically interact with ABI5 and facilitate its degradation. Furthermore, we found that RACK1 and the two substrate receptors CUL4-based E3 ligases (DWA1 and DWA2) function together to mediate the turnover of ABI5, thereby efficiently reducing ABA signalling in seed germination and post-germinative growth. In addition, molecular analyses demonstrated that ABI5 may bind to the promoter of RACK1 to repress its expression. Collectively, our findings suggest that RACK1 and ABI5 might form a feedback loop to regulate the homeostasis of ABA signalling in acute seed germination and early plant development., (© The Author(s) 2024. Published by Oxford University Press on behalf of the Society for Experimental Biology. All rights reserved. For commercial re-use, please contact reprints@oup.com for reprints and translation rights for reprints. All other permissions can be obtained through our RightsLink service via the Permissions link on the article page on our site—for further information please contact journals.permissions@oup.com.)

Plants have evolved a delicate molecular system to fine-tune their growth and development in response to dynamically changing light environments. In this study, we found that BBX28, a B-box domain protein, negatively regulates photomorphogenic development in a dose-dependent manner in Arabidopsis thaliana BBX28 interferes with the binding of transcription factor HY5 to the promoters of its target genes through physical interactions, thereby repressing its activity and negatively affecting HY5-regulated gene expression. In darkness, BBX28 associates with CONSTITUTIVELY PHOTOMORPHOGENIC1 (COP1) and undergoes COP1-mediated degradation via the 26S proteasome system. Collectively, these results demonstrate that BBX28 acts as a key factor in the COP1-HY5 regulatory hub by maintaining proper HY5 activity to ensure normal photomorphogenic development in plants., (© 2018 American Society of Plant Biologists. All rights reserved.)

Phosphorus deficiency in soil is one of the major limiting factors for plant growth. Plasma membrane H+-ATPase (PM H+-ATPase) plays an important role in the plant response to low-phosphorus stress (LP). However, few details are known regarding the action of PM H+-ATPase in mediating root proton (H+) flux and root growth under LP. In this study, we investigated the involvement and function of different Arabidopsis PM H+-ATPase genes in root H+ flux in response to LP. First, we examined the expressions of all Arabidopsis PM H+-ATPase gene family members (AHA1-AHA11) under LP. Expression of AHA2 and AHA7 in roots was enhanced under this condition. When the two genes were deficient in their respective Arabidopsis mutant plants, root growth and responses of the mutants to LP were highly inhibited compared with the wild-type plant. AHA2-deficient plants exhibited reduced primary root elongation and lower H+ efflux in the root elongation zone. AHA7-deficient plants exhibited reduced root hair density and lower H+ efflux in the root hair zone. The modulation of H+ efflux by AHA2 or AHA7 was affected by the action of 14-3-3 proteins and/or auxin regulatory pathways in the context of root growth and response to LP. Our results suggest that under LP conditions, AHA2 acts mainly to modulate primary root elongation by mediating H+ efflux in the root elongation zone, whereas AHA7 plays an important role in root hair formation by mediating H+ efflux in the root hair zone., (© The Author 2017. Published by Oxford University Press on behalf of the Society for Experimental Biology.)

This study aims to investigate the antimicrobial susceptibility, molecular characteristics and virulence genes of community-acquired methicillin-resistant ITALIC! Staphylococcus aureus(CA-MRSA) isolates with skin and soft tissue infections (SSTIs). Outpatients with SSTIs visiting five medical and health institutions were enrolled from 2011 to 2013. Available ITALIC! S. aureus isolates were characterized by antimicrobial susceptibility testing, and detection of PVL genes. For CA-MRSA isolates, we performed typing of staphylococcal cassette chromosome ITALIC! mec(SCC ITALIC! mec), multi locus sequence typing (MLST) and carriage of 27 virulence genes. A total of 203 ITALIC! S. aureusstrains were isolated from 1400 outpatients with SSTIs, and 21 (10.3%) were CA-MRSA isolates. The positive rate of PVL genes among ITALIC! S. aureus, CA-MRSA and methicillin-susceptible ITALIC! S. aureus(MSSA) isolates were 39.4%, 71.4% and 35.7%, respectively. CA-MRSA strains had greater sensitivity to non-β-lactam antimicrobial agents. All CA-MRSA isolates belonged to SCC ITALIC! mecIV and V, accounting for 47.6% and 52.4%, respectively. ST59 was the most common lineage accounting for 76.2%; ST59-SCC ITALIC! mecIVa-PVL-positive clone was found to be the predominant clone, accounting for 38.1%. All CA-MRSA isolates were found to be positive for one or more virulence genes, 28.6% of isolates carried PVL, ITALIC! seb, ITALIC! sek, ITALIC! seq, ITALIC! hla, ITALIC! hlb, ITALIC! hldand ITALIC! hlg-2. CA-MRSA infections were relatively uncommon in outpatients with SSTIs, but they carried many virulence genes, ST59-SCC ITALIC! mecIV a-PVL-positive clone was the predominant clone in Wuhan, China., (© FEMS 2016. All rights reserved. For permissions, please e-mail: journals.permissions@oup.com.)

AT14A has a small domain that has sequence similarities to integrins from animals. Integrins serve as a transmembrane linker between the extracellular matrix and the cytoskeleton, which play critical roles in a variety of biological processes. Because the function of AT14A is unknown, Arabidopsis thaliana AT14A, which is a transmembrane receptor for cell adhesion molecules and a middle member of the cell wall-plasma membrane-cytoskeleton continuum in plants, has been described. AT14A, co-expressed with green fluorescent protein (GFP), was found to localize mainly to the plasma membrane. The mutant Arabidopsis at14a-1 cells exhibit various phenotypes with cell shape, cell cluster size, thickness, and cellulose content of cell wall, the adhesion between cells, and the adhesion of plasma membrane to cell wall varied by plasmolysis. Using direct staining of filamentous actin and indirect immunofluorescence staining of microtubules, cortical actin filaments and microtubules arrays were significantly altered in cells, either where AT14A was absent or over-expressed. It is concluded that AT14A may be a substantial middle member of the cell wall-plasma membrane-cytoskeleton continuum and play an important role in the continuum by regulating cell wall and cortical cytoskeleton organization.

Rice (Oryza sativa L.) inflorescence (panicle) architecture is an important agronomic trait for rice breeding. A number of high-yielding japonica rice strains, characterized by an erect panicle (EP) of their architecture, have been released as commercial varieties in China. But no EP-type indica varieties are released so far. Here, we identified two allelic erect-panicle mutants in indica rice, erect panicle2-1 (ep2-1) and erect panicle2-2 (ep2-2), exhibiting the characteristic erect panicle phenotype. Both mutants were derived from spontaneous mutation. We cloned the EP2 gene by way of a map-based cloning strategy, and a transgenic complementation test rescued the phenotype of ep2-1. Anatomical investigations revealed that the ep2 mutants have more vascular bundles and a thicker stem than that of wild-type plants, explaining the panicle erectness phenotype in ep2 mutants. It was shown that EP2 was specifically expressed in the vascular bundles of internodes by GUS staining and RT-PCR. EP2 encodes a novel plant-specific protein, which localizes to the endoplasmic reticulum with unknown biochemical function. In addition, EP2 also regulates other panicle characteristics, such as panicle length and grain size, but grain number per panicle shows little change, indicating that the mutation of the ep2 gene could be applied in EP-type indica rice breeding.

Mammalian receptor for activated C kinase 1 (RACK1) is a versatile scaffold protein, playing regulatory roles in multiple signal transduction pathways. Moreover, RACK1 interacts with the heterotrimeric G-proteins (G-proteins) and regulates some specific functions of Gbetagamma. Although the protein sequences of both RACK1 and G-proteins are highly conserved in Arabidopsis, their relationship remains elusive. Here we provide genetic and biochemical evidence that Arabidopsis RACK1 and G-proteins may act through a mechanism that is distinct from their counterparts in mammals. Loss-of-function alleles of RACK1A (the most abundantly expressed RACK1 gene in Arabidopsis) do not appear to share morphological and developmental phenotypes with loss-of-function alleles of GPA1 (encoding the sole Galpha in Arabidopsis) or AGB1 (encoding the sole Gbeta in Arabidopsis). The analysis of gpa1 rack1a and agb1 rack1a double mutants suggested that the effect of RACK1A on morphological and developmental traits may occur independently of the presence or absence of the G-protein subunits. Although both RACK1A and G-protein subunits are negative regulators of ABA responses in the ABA inhibition of early seedling development, an additive ABA hypersensitivity was observed in gpa1 rack1a and agb1 rack1a double mutants. Biochemical analysis suggested that unlike their counterparts in mammals, RACK1 may not physically interact with AGB1. Taken together, these findings revealed some fundamental differences in the relationship of RACK1 and G-proteins between Arabidopsis and mammals.

Receptor for Activated C Kinase 1 (RACK1) is viewed as a versatile scaffold protein in mammals. The protein sequence of RACK1 is highly conserved in eukaryotes. However, the function of RACK1 in plants remains poorly understood. Accumulating evidence suggested that RACK1 may be involved in hormone responses, but the precise role of RACK1 in any hormone signalling pathway remains elusive. Molecular and genetic evidence that Arabidopsis RACK1 is a negative regulator of ABA responses is provided here. It is shown that three RACK1 genes act redundantly to regulate ABA responses in seed germination, cotyledon greening and root growth, because rack1a single and double mutants are hypersensitive to ABA in each of these processes. On the other hand, plants overexpressing RACK1A displayed ABA insensitivity. Consistent with their proposed roles in seed germination and early seedling development, all three RACK1 genes were expressed in imbibed, germinating and germinated seeds. It was found that the ABA-responsive marker genes, RD29B and RAB18, were up-regulated in rack1a mutants. Furthermore, the expression of all three RACK1 genes themselves was down-regulated by ABA. Consistent with the view that RACK1 negatively regulates ABA responses, rack1a mutants lose water significantly more slowly from the rosettes and are hypersensitive to high concentrations of NaCl during seed germination. In addition, the expression of some putative RACK1-interacting, ABA-, or abiotic stress-regulated genes was mis-regulated in rack1a rack1b double mutants in response to ABA. Taken together, these findings provide compelling evidence that RACK1 is a critical, negative regulator of ABA responses.

Rice grain filling is a process of conversion of sucrose into starch catalysed by a series of enzymes. Sucrose synthase (SUS) is considered as a key enzyme regulating this process. This study investigated the possible roles of sucrose and abscisic acid (ABA) in mediating the activity and expression of SUS protein of grains during grain filling in rice (Oryza sativa). Field-grown rice plants and detached cultured panicles were used as experimental materials. Several treatments, including spikelet thinning, leaf cutting, and applications of different concentrations of exogenous sucrose and ABA, were imposed during grain filling. A higher SUS activity was found in superior grains than in inferior grains in the earlier stage of grain filling, which was significantly and closely related to a higher grain filling rate and starch accumulation. An increase in sucrose concentration in grains as a result of different treatments increased both SUS activity and SUS protein expression in grains. An increase in ABA concentration gave similar results. Furthermore, effects of interactions between sucrose and ABA on the activity and expression of SUS protein in grains were also found. It was suggested that sucrose- and ABA-mediated rice grain filling is largely due to an increase in SUS activity and SUS protein expression.

Microtubules play important roles in many physiological processes such as plant responses to drought stress. Abscisic acid (ABA) accumulates significantly in plants in response to drought conditions, which has been considered as a major response for plants to enhance drought tolerance. In this work, the focus was on the possible roles of microtubules in the induction of ABA biosynthesis in the roots of Zea mays when subjected to osmotic stress. The dynamic changes of microtubules in response to the stress were investigated by immunofluorescence staining, enzyme-linked immunosorbent assay, and a pharmacological approach. Disruption and stabilization of microtubules both significantly stimulated ABA accumulation in maize root cells, although this stimulation was markedly lower than that caused by osmotic stress. Cells in which the microtubule stability had been changed did not respond further to osmotic stress in terms of ABA biosynthesis. However, treatment with both a microtubule de-stabilizer and a stabilizer enhanced the sensitivity of cells to osmotic stress in terms of ABA accumulation. It is suggested that both osmotic stress and changes in microtubule dynamics would trigger maize root cells to biosynthesize ABA, and interactions between osmotic stress and microtubule dynamics would have an effect on ABA accumulation in root cells, although the exact mechanism is not clear at present.

Regulator of G-protein signalling (RGS) proteins identified recently in Arabidopsis have been involved in the regulation of several physiological processes, but largely nothing is known about their roles at both the physiological and the molecular level. In the experiments reported here, the overexpression approach was used to present evidence that RGS1 protein plays critical roles in plant development and in modulating abscisic acid (ABA) and drought stress signal transduction. RGS1 affected the shapes of leaves, the development of floral buds, the elongation of stems, siliques, and hypocotyls, and the time of flowering. Post-germination growth was inhibited by 1 microM ABA, and root growth was hypersensitive to ABA for 35S-RGS1 transgenic plants. RGS1 overexpression conferred more drought tolerance to transgenic plants, as compared with the wild type (Columbia). Reverse transcription-PCR (RT-PCR) results indicated that RGS1 overexpresssion significantly stimulated the expression of NCED and ABA2, that encode two key enzymes catalysing ABA biosynthesis. Furthermore, the expression of several stress-regulated genes was either up- or down-regulated in RGS1-overexpressing transgenic plants. Combining the results above with previous results, it is suggested that RGS1 exerted its effects on plant responsiveness to ABA and drought tolerance largely through changing the expression either of genes responsible for ABA biosynthesis, which leads to changes in endogenous ABA levels, or of stress-responsive genes.

The scaffold protein RACK1 (Receptor for Activated C Kinase 1) serves as an integrative point for diverse signal transduction pathways. The Arabidopsis genome contains three RACK1 orthologues, however, little is known about their functions. It is reported here that one member of this gene family, RACK1A, previously identified as the Arabidopsis homologue of the tobacco arcA gene, mediates hormone responses and plays a regulatory role in multiple developmental processes. RACK1A expresses ubiquitously in Arabidopsis. Loss-of-function mutations in RACK1A confer defects in multiple developmental processes including seed germination, leaf production, and flowering. rack1a mutants displayed reduced sensitivity to gibberellin and brassinosteroid in seed germination, hypersensitivity to abscisic acid in seed germination and early seedling development, and hyposensitivity to auxin in adventitious and lateral root formation. These results provide the first genetic evidence that RACK1A is involved in multiple signal transduction pathways.

Two tropical woody species, Acacia confusa Merrill and Litsea glutinosa (Lour.) C.B. Robinson, were grown under controlled conditions in PVC pipes filled with John Innes No. 2 soil. To investigate root distribution, physiological characteristics and hydraulic conductivity, four soil treatments were imposed-well-watered and noncompacted (control), well-watered and compacted; unwatered and noncompacted, and unwatered and compacted. In L. glutinosa, rooting depth and root elongation were severely restricted when soil bulk density increased from around 1.12 to 1.62 g cm(-3), whereas soil compaction had little effect on these parameters in A. confusa. As soil drying progressed, root water potential and osmotic potential declined more slowly in L. glutinosa than in A. confusa. Both the soil drying and compaction treatments significantly stimulated the accumulation of root abscisic acid (ABA) in both species. Soil drying damaged the root cell membrane of A. confusa, but had little influence on the root cell membrane of L. glutinosa. Soil drying had a greater effect on root hydraulic conductivity (L(p)) in L. glutinosa than in A. confusa, whereas the effect of soil compaction on L(p) was less in L. glutinosa than in A. confusa. Soil drying enhanced the effects of soil compaction on root L(p). We conclude that soil drying and compaction have large species-specific effects on the distribution, growth and physiology of roots. The relationships of these root properties to the species' ability to tolerate unfavorable soil conditions were examined.