Intrinsic Membrane Proteins Research Articles

The Arabidopsis PIP1;1 Aquaporin Represses Lateral Root Development and Nitrate Uptake Under Low Nitrate Availability.

Nitrate (NO3 -) deficiency decreases root water uptake and root hydraulic conductance. This adaptive response is correlated with reduced abundance and activity of plasma membrane intrinsic protein (PIP) aquaporins. We therefore screened changes in the root architecture of a complete set of Arabidopsis pip loss-of-function mutants grown under NO3 - deficiency to systematically approach the impact of PIPs under these conditions. NO3 - deprivation led to attenuated responses of specific pip single mutants compared to the strongly altered LR parameters of wild-type plants. In particular, pip1;1 exhibited a lower relative reduction in LR length and LR density, revealing that PIP1;1 represses LR development when NO3 - is scarce. Indeed, PIP1;1 compromises root and shoot NO3 - accumulation during early developmental stages. A fluorescent VENUS-PIP1;1 fusion revealed that PIP1;1 is specifically repressed in the pericycle, endodermis and at the flanks of emerging LRs upon NO3 - deficiency. Thus, LR plasticity and NO3 - uptake are affected by an interactive mechanism involving aquaporins (PIP1;1) and nitrate accumulation during seedling development under NO3 --deficient conditions.

Effects of foliar spraying with melatonin and chitosan Nano-encapsulated melatonin on tomato (Lycopersicon esculentum L. cv. Falcato) plants under salinity stress

Melatonin has been found to be crucial in the growth and development of plants under stress conditions. In this study, the effects of melatonin and nano melatonin regarding the growth and development of tomato plants, along with their photosynthetic pigment, phenol, and antioxidant activity, were investigated under saline conditions. The study was conducted using a completely randomized design with three replications, and the applied treatments were salt stress and foliar spraying of melatonin at a concentration of 0 (control), melatonin (Mel), and nano capsule-melatonin (Nano-Mel) at 500 µM. Salinity treatments included application of sodium chloride with two concentration of 0 mM NaCl (S1) and 50 mM NaCl (S2). Under saline conditions, Mel and Nano-Mel increased both shoot and root fresh and dry weights, improved relative water content (RWC), and enhanced antioxidant activity and phenolic content. Salinity elevated leaf ABA content, unaffected by Mel or Nano-Mel. Chlorophyll fluorescence and SPAD values demonstrated resilience to salinity with Mel and Nano-Mel applications. Nano-Mel notably mitigated Na + accumulation in leaves under salinity, helping maintain K + homeostasis. Proline levels rise due to salinity but decreased with Mel and Nano-Mel treatments. Electrolyte leakage (EL) increased under salinity but is significantly reduced by Mel, indicating enhanced membrane stability. The findings reveal that salinity stress significantly reduced plasma membrane intrinsic protein (PIP) expression in roots and leaves, whereas Mel and Nano-Mel treatments enhance PIP expression, particularly in roots. The study concludes that Mel and Nano-Mel effectively alleviate salinity-induced stress, promoting growth and maintaining physiological homeostasis in tomato plants.

A Plasma Membrane Intrinsic Protein Gene OfPIP2 Involved in Promoting Petal Expansion and Drought Resistance in Osmanthus fragrans

Osmanthus fragrans, a native to China, is renowned as a highly popular gardening plant. However, this plant faces significant challenges from drought stress, which can adversely affect its flowering. In this study, we found that the plasma membrane-localized gene OfPIP2 exhibited a substantial upregulation during the flowering stages and in response to drought stress. GUS staining has illustrated that the OfPIP2 promoter can drive GUS activity under drought conditions. The overexpression of OfPIP2 was found to enhance petal size by modulating epidermal cell dimensions in Petunia and tobacco. Moreover, this overexpression also bolstered drought tolerance, as evidenced by a reduction in stomatal aperture in both species. Furthermore, yeast one-hybrid (Y1H) and dual-luciferase (Dual-LUC) assays have indicated that the transcription factor OfMYB28 directly binds to the OfPIP2 promoter, thereby regulating its expression. Together, we speculated that a module of OfMYB28-OfPIP2 was not only involved in the enhancement of petal size but also conferred the improvement of drought tolerance in O. fragrans. These results contribute valuable insights into the molecular function of the OfPIP2 gene and lay a foundation for molecular breeding strategies in O. fragrans.

Permeation mechanisms of hydrogen peroxide and water through Plasma Membrane Intrinsic Protein aquaporins.

Hydrogen peroxide (H2O2) transport by aquaporins (AQP) is a critical feature for cellular redox signaling. However, the H2O2 permeation mechanism through these channels remains poorly understood. Through functional assays, two Plasma membrane Intrinsic Protein (PIP) AQP from Medicago truncatula, MtPIP2;2 and MtPIP2;3 have been identified as pH-gated channels capable of facilitating the permeation of both water (H2O) and H2O2. Employing a combination of unbiased and enhanced sampling molecular dynamics simulations, we investigated the key barriers and translocation mechanisms governing H2O2 permeation through these AQP in both open and closed conformational states. Our findings reveal that both H2O and H2O2 encounter their primary permeation barrier within the selectivity filter (SF) region of MtPIP2;3. In addition to the SF barrier, a second energetic barrier at the NPA (asparagine-proline-alanine) region that is more restrictive for the passage of H2O2 than for H2O, was found. This behavior can be attributed to a dissimilar geometric arrangement and hydrogen bonding profile between both molecules in this area. Collectively, these findings suggest mechanistic heterogeneity in H2O and H2O2 permeation through PIPs.

Aquaporin CmPIP2; 3 links H2O2 signal and antioxidation to modulate trehalose-induced cold tolerance in melon seedlings.

Cold stress severely restricts the growth and development of cold-sensitive crops. Trehalose (Tre), known as the 'sugar of life', plays key roles in regulating plant cold tolerance by triggering antioxidation. However, the relevant regulatory mechanism remains unclear. Here, we confirmed that Tre triggers apoplastic hydrogen peroxide (H2O2) production and thus plays key roles in improving the cold tolerance of melon (Cucumis melo var. makuwa Makino) seedlings. Moreover, Tre treatment can promote the transport of apoplastic H2O2 to the cytoplasm. This physiological process may depend on aquaporins. Further studies showed that a Tre-responsive plasma membrane intrinsic protein 2; 3 (CmPIP2; 3) had strong H2O2 transport function and that silencing CmPIP2; 3 significantly weakened apoplastic H2O2 transport and reduced the cold tolerance of melon seedlings. Yeast library and protein-DNA interaction technology were then used to screen two Tre-responsive transcription factors, abscisic acid responsive element (ABRE)-binding factor 2 (CmABF2) and abscisic acid responsive element (ABRE)-binding factor 3 (CmABF3), which can bind to the ABRE motif of the CmPIP2; 3 promoter and activate its expression. Silencing of CmABF2 and CmABF3 further dramatically increased the ratio of apoplastic H2O2/cytoplasm H2O2 and reduced the cold tolerance of melon seedlings. This study uncovered that Tre treatment induces CmABF2/3 to positively regulate CmPIP2; 3 expression. CmPIP2; 3 subsequently enhances the cold tolerance of melon seedlings by promoting the transport of apoplastic H2O2 into the cytoplasm for conducting redox signals and stimulating downstream antioxidation.

Phosphorylation of PIP2;7 by CPK28 or Phytophthora kinase effectors dampens pattern-triggered immunity in Arabidopsis

Plasma membrane intrinsic proteins (PIPs), a subclass of aquaporins, play an important role in plant immunity by acting as H2O2 transporters. Their homeostasis is mostly maintained by C-terminal serine phosphorylation. However, the kinases that phosphorylate PIPs and manipulate their turnover are largely unknown. Here, we found that Arabidopsis thaliana PIP2;7 positively regulates plant immunity by transporting H2O2. Arabidopsis CALCIUM-DEPENDENT PROTEIN KINASE 28 (CPK28) directly interacts with and phosphorylates PIP2;7 at Ser273/276 to induce its degradation. During pathogen infection, CPK28 dissociates from PIP2;7 and destabilizes, leading to PIP2;7 accumulation. As a countermeasure, oomycete pathogens produce conserved kinase effectors that stably bind to and mediate the phosphorylation of PIP2;7 to induce its degradation. Our study identifies PIP2;7 as a novel substrate of CPK28 and shows that its protein stability is negatively regulated by CPK28. Such phosphorylation could be mimicked by Phytophthora kinase effectors to promote infection. Accordingly, we developed a strategy to combat oomycete infection using a phosphorylation-resistant PIP2;7S273/276A mutant. The strategy only allows accumulation of PIP2;7S273/276A during infection to limit potential side effects on normal plant growth.

Superior osmotic stress tolerance in oilseed rape transformed with wild-type Rhizobium rhizogenes

Key messageNatural transformation with R. rhizogenes enhances osmotic stress tolerance in oilseed rape through increasing osmoregulation capacity, enhancing maintenance of hydraulic integrity and total antioxidant capacity.Transformation of plants using wild strains of agrobacteria is termed natural transformation and is not covered by GMO legislation in, e.g., European Union and Japan. In this study, offspring lines of Rhizobium rhizogenes naturally transformed oilseed rape (Brassica napus), i.e., A11 and B3 (termed root-inducing (Ri) lines), were investigated for osmotic stress resilience. Under polyethylene glycol 6000 (PEG) 10% (w/v)-induced osmotic stress, the Ri lines, particularly A11, had less severe leaf wilting, higher stomatal conductance (8.2 times more than WT), and a stable leaf transpiration rate (about 2.9 mmol m−2 s−1). Although the leaf relative water content and leaf water potential responded similarly to PEG treatment between the Ri lines and WT, a significant reduction of the turgid weight to dry weight ratio in A11 and B3 indicated a greater capacity of osmoregulation in the Ri lines. Moreover, the upregulation of plasma membrane intrinsic proteins genes (PIPs) in roots and downregulation of these genes in leaves of the Ri lines implied a better maintenance of hydraulic integrity in relation to the WT. Furthermore, the Ri lines had greater total antioxidant capacity (TAC) than the WT under PEG stress. Collectively, the enhanced tolerance of the Ri lines to PEG-induced osmotic stress could be attributed to the greater osmoregulation capacity, better maintenance of hydraulic integrity, and greater TAC than the WT. In addition, Ri-genes (particularly rolA and rolD) play roles in response to osmotic stress in Ri oilseed rape. This study reveals the potential of R. rhizogenes transformation for application in plant drought resilience.

Variations in Sweet Pepper Growth, Physiology, and Gene Expression in Shock and Gradual Salt Stress

ABSTRACT Salinity is a major abiotic stressor that can significantly impact plant growth and productivity. To evaluate the physiological responses of sweet pepper (Capsicum annuum L. Ps301) to gradual salinity stress (SLS) and shock salinity stress (STS), an experiment was conducted using a factorially based completely randomized design (CRD). The sodium chloride (NaCl) induced salinity treatments were administered in the nutrient solution in two forms: STS and SLS. The SLS was applied for 8 days, with stress levels increasing every 2 days; STS was administered for 48 h. The signaling response was activated more rapidly and abruptly in response to STS; however, after the various adaptive responses to salinity, these signals were more activated in response to SLS. The STS led to plant growth a greater decrease than SLS. The electrolyte leakage (EL) increased under both SLS and STS by 60% and 61% compared to the control, respectively. Other stress indices, such as phenol and abscisic acid (ABA) content, increased by 26% and 71% under SLS compared to STS. Salinity increased the leaf water potential (LWP) in two salt regimes, and the root water potential (RWP) in STS conditions only. Salinity stress decreased plasma membrane intrinsic protein gene expression in roots, especially in STS compared to SLS. The STS increased plasma membrane intrinsic protein (PIP) expression in leaves more than SLS. It was concluded that salinity impairs the growth and physiological characteristics of pepper; however, the signaling and stress indices respond differently to STS and SLS application.

The role of light intensity in water transport and homeostasis across different vapor pressure deficit conditions

Vapor pressure deficit (VPD) directly affects the driving force of plant water movement by altering the water potential gradient between the atmosphere and plants and indirectly influences the resistance to water movement by regulating plant structure. Concurrently, light intensity modulates both the driving force and resistance to water movement by regulating plant morphology and nonstructural carbohydrate synthesis. Despite significant advances in the understanding of the regulatory effects of VPD on water absorption and transport in tomatoes, the effect of light intensity regulation under varying VPDs on water transport and homeostasis remains to be clarified. Here, we investigated the effects of two light intensities (L300; 300 µmol m–2 s–1, L600; 600 µmol m–2 s–1) on plant anatomy, physiological traits, hydraulic properties, and expression of plasma membrane intrinsic proteins (PIPs) and tonoplast intrinsic proteins (TIPs) in tomatoes subjected to long-term high and low VPDs. In addition, we analysed the correlations and path coefficients of these indicators. These results indicate that higher light intensity reduces resistance to water movement by enhancing root morphology, vessel parameters in roots and stems, leaf vein density, stomatal morphology, physiological traits, and expression of SlTIPs and SlPIPs in both roots and leaves. Concurrently, increased light intensity boosts the driving force of water movement by amplifying the water potential difference and transpiration under low VPD. However, under high VPD, elevated light intensities create a larger water potential difference, prompting plants to reduce this excessive force by decreasing transpiration and stomatal conductance, thereby maintaining water homeostasis. These findings suggest that light intensity can effectively regulate water homeostasis by dynamically optimising plant structure, hydraulic properties, and the expression of SlTIPs and SlPIPs across different VPDs, providing a theoretical foundation for practical light intensity management in agriculture.

Early Diversification of Membrane Intrinsic Proteins (MIPs) in Eukaryotes.

Membrane intrinsic proteins (MIPs), including aquaporins (AQPs) and aquaglyceroporins (GLPs), form an ancient family of transporters for water and small solutes across biological membranes. The evolutionary history and functions of MIPs have been extensively studied in vertebrates and land plants, but their widespread presence across the eukaryotic tree of life suggests both a more complex evolutionary history and a broader set of functions than previously thought. That said, the early evolution of MIPs remains obscure. The presence of one GLP and four AQP clades across both bacteria and archaea suggests that the first eukaryotes could have possessed up to five MIPs. Here, we report on a previously unknown richness in MIP diversity across all major eukaryotic lineages, including unicellular eukaryotes, which make up the bulk of eukaryotic diversity. Three MIP clades have likely deep evolutionary origins, dating back to the last eukaryotic common ancestor (LECA), and support the presence of a complex MIP repertoire in early eukaryotes. Overall, our findings highlight the growing complexity of the reconstructed LECA genome: the dynamic evolutionary history of MIPs was set in motion when eukaryotes were in their infancy followed by radiative bursts across all main eukaryotic lineages.

PIP family-based association studies uncover ZmPIP1;6 involved in Pb accumulation and water absorption in maize roots

Excessive lead (Pb) in the soil affects crop growth and development, thus threatening human beings via food chains. Plasma membrane intrinsic proteins (PIPs) facilitate the transport of substrates across cell membranes. Herein, we characterized maize PIPs and identified eight Pb accumulation-associated PIP genes using association studies. Among these, ZmPIP1;6 was simultaneously correlated with root Pb concentrations under various Pb treatment stages. Significant correlations were observed between the ZmPIP1;6 expression abundance and Pb accumulation in maize roots. Ectopic expression in yeast showed that ZmPIP1;6 conferred Pb accumulation in the cells and affected Pb tolerance in yeast. Overexpression in maize demonstrated that ZmPIP1;6 altered the Pb concentration performance and root moisture content under Pb stress. Meanwhile, protein interaction analyses suggested that ZmPIP1; 6 and three PIP2 members formed isoforms and facilitate water uptake in maize roots. However, ZmPIP1; 6 improved Pb absorption in maize roots probably by interacting with CASP-like protein 2C3 and/or another metal transporter. Moreover, the significant variants in the ZmPIP1;6 promoter caused the variations in ZmPIP1;6 expression level and Pb accumulation among various maize germplasms. Our study will contribute to understanding of PIP family-mediated Pb accumulation in crops and bioremediation of Pb-polluted soils.

Genetic variability of aquaporin expression in maize: From eQTLs to a MITE insertion regulating PIP2;5 expression.

Plant aquaporins are involved in numerous physiological processes, such as cellular homeostasis, tissue hydraulics, transpiration, and nutrient supply, and are key players of the response to environmental cues. While varying expression patterns of aquaporin genes have been described across organs, developmental stages, and stress conditions, the underlying regulation mechanisms remain elusive. Hence, this work aimed to shed light on the expression variability of 4 plasma membrane intrinsic protein (PIP) genes in maize (Zea mays) leaves, and its genetic causes, through expression quantitative trait locus (eQTL) mapping across a 252-hybrid diversity panel. Significant genetic variability in PIP transcript abundance was observed to different extents depending on the isoforms. The genome-wide association study mapped numerous eQTLs, both local and distant, thus emphasizing the existing natural diversity of PIP gene expression across the studied panel and the potential to reveal regulatory actors and mechanisms. One eQTL associated with PIP2;5 expression variation was characterized. Genomic sequence comparison and in vivo reporter assay attributed, at least partly, the local eQTL to a transposon-containing polymorphism in the PIP2;5 promoter. This work paves the way to the molecular understanding of PIP gene regulation and its possible integration into larger networks regulating physiological and stress adaptation processes.

Variation in photosynthetic capacity of Salvia przewalskii along elevational gradients on the eastern Qinghai-Tibetan Plateau, China

Elevational variation in plant growing environment drives diversification of photosynthetic capacity, however, the mechanism behind this reaction is poorly understood. We measured leaf gas exchange, chlorophyll fluorescence, anatomical characteristics, and biochemical traits of Salvia przewalskii at elevations ranging from 2400 m to 3400 m above sea level (a.s.l) on the eastern Qinghai-Tibetan Plateau, China. We found that photosynthetic capacity showed an initial increase and then a decrease with rising elevation, and the best state observed at 2800 m a.s.l. Environmental factors indirectly regulated photosynthetic capacity by affecting stomatal conductance (gs), mesophyll conductance (gm), maximum velocity of carboxylation (Vc max), and maximum capacity for photosynthetic electron transport (Jmax). The average temperature (T) and total precipitation (P) during the growing season had the highest contribution to the variation of photosynthetic capacity of S. przewalskii in subalpine areas, which were 25% and 24%, respectively. Photosynthetic capacity was mainly affected by diffusional limitations (71%–89%), and mesophyll limitation (lm) played a leading role. The variation of gm was attributed to the effects of environmental factors on the volume fraction of intercellular air space (fias), the thickness of cell wall (Tcw), the surface of mesophyll cells and chloroplasts exposed to intercellular airspace (Sm, Sc), and plasma membrane intrinsic protein (PIPs, PIP1, PIP2), independent of carbonic anhydrase (CA). Optimization of leaf tissue structure and adaptive physiological responses enabled plants to efficiently cope with variable climate conditions of high-elevation areas, and the while maintaining high levels of carbon assimilation.

Aquaporins: Multifunctional Players in Plant Growth, Development and Stress Responses

Aquaporins are small membrane proteins belonging to the group of Major Intrinsic proteins (MIPs) that facilitate the transport of water and small molecules across biological membranes. They were initially identified as water channels and later it has become clear that they also play important roles in several other physiological processes in plants. Plants have different types of aquaporins in terms of their location, gating properties, and solute specificity and can mainly divide into four major subfamilies. They are tonoplast intrinsic proteins (TIPs), nodulin26-like intrinsic proteins (NIPs), plasma membrane intrinsic proteins (PIPs), and the small basic intrinsic proteins (SIPs). All these aquaporins play a variety of physiological activities at all stages of plant growth and development. Also, they are believed to play a significant part in the plant’s defense mechanisms against biotic and abiotic stresses. This review comprehensively addresses the roles of diverse aquaporin homologs in growth and development and their response to different environmental factors.

Improved Waterlogging Tolerance in Roots of Cucumber Plants after Inoculation with Arbuscular Mycorrhizal Fungi

Mycorrhizal symbiosis enhances host plant resistance to various unfavorable environmental stresses, but whether and how it also enhances waterlogging tolerance in cucumber plants is not known. The objective of this study was to analyze the effect of Paraglomus occultum inoculation on biomass production, osmolyte levels, and the expression of 12 heat shock protein 70 (Hsp70) genes and 14 plasma membrane intrinsic protein (PIP) genes in the roots of cucumber plants under a short-term waterlogging stress (WS) (5 days) condition. Although the short-term WS treatment significantly inhibited the arbuscular mycorrhizal fungal colonization of roots, the inoculation with arbuscular mycorrhizal fungi (AMFs) significantly increased leaf, stem, and root biomass under WS. AMF inoculation also significantly increased root glucose, sucrose, betaine, and proline contents, along with decreased fructose levels, compared with the uninoculated control. More CsHsp70 and CsPIP genes were up-regulated in AMF-inoculated plants than in AMF-uninoculated plants in response to WS. AMF inoculation showed no significant effect on the expression of any of the examined CsHsp70 genes under no-waterlogging stress, but it did raise the expression of 11 of 12 CsHsp70 genes under WS. AMF colonization also down-regulated or had no effect on CsPIP expression under no-waterlogging stress, whereas it up-regulated the expression of 12 of the 14 CsPIP genes under WS. It is concluded that AMF inoculation enhances waterlogging tolerance in cucumber plants by increasing osmolyte levels and stress-responsive gene (CsPIP and CsHsp70) expression.

The structural basis for light harvesting in organisms producing phycobiliproteins.

Cyanobacteria, red algae, and cryptophytes produce 2 classes of proteins for light harvesting: water-soluble phycobiliproteins (PBP) and membrane-intrinsic proteins that bind chlorophylls (Chls) and carotenoids. In cyanobacteria, red algae, and glaucophytes, phycobilisomes (PBS) are complexes of brightly colored PBP and linker (assembly) proteins. To date, 6 structural classes of PBS have been described: hemiellipsoidal, block-shaped, hemidiscoidal, bundle-shaped, paddle-shaped, and far-red-light bicylindrical. Two additional antenna complexes containing single types of PBP have also been described. Since 2017, structures have been reported for examples of all of these complexes except bundle-shaped PBS by cryogenic electron microscopy. PBS range in size from about 4.6 to 18 mDa and can include ∼900 polypeptides and bind >2000 chromophores. Cyanobacteria additionally produce membrane-associated proteins of the PsbC/CP43 superfamily of Chl a/b/d-binding proteins, including the iron-stress protein IsiA and other paralogous Chl-binding proteins (CBP) that can form antenna complexes with Photosystem I (PSI) and/or Photosystem II (PSII). Red and cryptophyte algae also produce CBP associated with PSI but which belong to the Chl a/b-binding protein superfamily and which are unrelated to the CBP of cyanobacteria. This review describes recent progress in structure determination for PBS and the Chl proteins of cyanobacteria, red algae, and cryptophytan algae.

Plasma membrane intrinsic proteins SlPIP2;5 gene regulates tolerance to high VPD in tomato

Plasma membrane intrinsic proteins (PIPs) play important roles in plant growth, development, and abiotic stress responses. However, the functions and regulatory mechanisms of PIPs under high vapor pressure deficit (VPD) stress are not yet well understood. In this study, we used tobacco rattle virus (TRV) - mediated virus-induced gene silencing (VIGS) technology to discover that SlPIP2;5 is involved in the acclimation process of tomato plants to high VPD. Under high VPD, the TRV-SlPIP2;5 strain exhibited improved growth compared to the TRV strain, and silencing SlPIP2;5 increased the enzyme activities of the antioxidant system of tomato plants and reduced the sensitivity of plants to atmospheric drought stress by eliminating reactive oxygen species produced by the plants. Additionally, leaf structure was improved, with decreased thickness of the epidermis and spongy tissue, leading to enhanced water transport efficiency. Furthermore, the TRV-SlPIP2;5 strain had increased stomatal length and individual stomatal area, which changed net assimilation rate and stomatal conductance to air drought. In addition, SlPIP2;5 was localized on the plasma membrane, and its promoter region contained numerous core elements and V-myb avian myeloblastosis viral oncogene homolog (MYB) binding sites. Yeast One-Hybrid Assay (Y1H) and Dual Luciferase (LUC) assays showed that the transcription factor SlMYB110 can bind to SlPIP2;5 and regulate its expression. qRT-PCR analysis revealed that high VPD inhibited the expression of SlMYB110, consistent with the expression pattern of SlPIP2;5. Therefore, we speculate that under high VPD, SlMYB110 negatively regulates the expression of SlPIP2;5, thereby modulating the tolerance of tomato plants to atmospheric drought. These findings provide a theoretical basis for improving tomato response to high VPD, and SlPIP2;5 was added to the high VPD-responsive gene pool as a valuable gene that could be used to improve high VPD tolerance in tomato through genetic engineering.

AtPIP1;4 and AtPIP2;4 Cooperatively Mediate H2O2 Transport to Regulate Plant Growth and Disease Resistance.

The rapid production of hydrogen peroxide (H2O2) is a hallmark of plants' successful recognition of pathogen infection and plays a crucial role in innate immune signaling. Aquaporins (AQPs) are membrane channels that facilitate the transport of small molecular compounds across cell membranes. In plants, AQPs from the plasma membrane intrinsic protein (PIP) family are utilized for the transport of H2O2, thereby regulating various biological processes. Plants contain two PIP families, PIP1s and PIP2s. However, the specific functions and relationships between these subfamilies in plant growth and immunity remain largely unknown. In this study, we explore the synergistic role of AtPIP1;4 and AtPIP2;4 in regulating plant growth and disease resistance in Arabidopsis. We found that in plant cells treated with H2O2, AtPIP1;4 and AtPIP2;4 act as facilitators of H2O2 across membranes and the translocation of externally applied H2O2 from the apoplast to the cytoplasm. Moreover, AtPIP1;4 and AtPIP2;4 collaborate to transport bacterial pathogens and flg22-induced apoplastic H2O2 into the cytoplasm, leading to increased callose deposition and enhanced defense gene expression to strengthen immunity. These findings suggest that AtPIP1;4 and AtPIP2;4 cooperatively mediate H2O2 transport to regulate plant growth and immunity.

Comparative differences in maintaining membrane fluidity and remodeling cell wall between Glycine soja and Glycine max leaves under drought

Water shortage is one of the most important environmental factors limiting crop yield. In this study, we used wild soybean (Glycine soja Sieb. et Zucc.) and soybean (Glycinemax (L.) Merr.) seedlings as experimental materials, simulated drought stress using soil gravimetry, measured growth and physiological parameters, and analyzed differentially expressed genes and metabolites in the leaves of seedling by integrated transcriptomics and metabolomics techniques. The results indicate that under water deficit, Glycine soja maintained stable photosynthate by accumulating Mg2+, Fe3+, Mn2+, Zn2+ and B3+, and improved water absorption by increasing root growth. Notably, Glycine soja enhanced linoleic acid metabolism and plasma membrane intrinsic protein (PIP1) gene expression to maintain membrane fluidity, and increased pentose, glucuronate and galactose metabolism and thaumatin protein genes expression to remodel the cell wall, thereby increasing water-absorption to better tolerate to drought stress. In addition, it was found that secondary phenolic metabolism, such as phenylpropane biosynthesis, flavonoid biosynthesis and ascobate and aldarate metabolism were weakened, resulting in the collapse of the antioxidant system, which was the main reason for the sensitivity of Glycine max to drought stress. These results provide new insights into plant adaptation to water deficit and offer a theoretical basis for breeding soybean varieties with drought tolerance.

CmoPIP1-4 confers drought tolerance in pumpkin by altering hydrogen sulfide signaling

Drought is a major limiting factor for the growth and development of pumpkins. Plasma membrane intrinsic proteins (PIPs) are major water channels that play a crucial role in the regulation of cellular water status and solute trafficking during drought conditions. CmoPIP1-4 is a plasma membrane-localized protein that is significantly upregulated in roots and leaves under drought-stress conditions. In this study, the overexpression of CmoPIP1-4 enhances drought resistance in yeast. In contrast, CRISPR-mediated CmoPIP1-4 knockout in pumpkin roots increased drought sensitivity. This increased drought sensitivity of CmoPIP1-4 knockout plants is associated with a decline in the levels of hydrogen sulfide (H2S) and abscisic acid (ABA), accompanied by an increase in water loss caused by greater levels of transpiration and stomatal conductance. In addition, the sensitivity of CmoPIP1-4 CRISPR plants is further aggravated by reduced antioxidative enzyme activity, decreased proline and sugar contents, and extensive root damage. Furthermore, expression profiles of genes such as CmoHSP70s, CmoNCED3, CmoNCED4, and others involved in metabolic activities were markedly reduced in CmoPIP1-4 CRISPR plants. Moreover, we also discovered an interaction between the drought-responsive gene CmoDCD and CmoPIP1-4, indicating their potential role in activating H2S-mediated signaling in pumpkin, which could confer drought tolerance. The findings of our study collectively demonstrate CmoPIP1-4 plays a crucial role in the regulation of H2S-mediated signaling, influencing stomatal density and aperture in pumpkin plants, and thereby enhancing their drought tolerance.