The CBL-interacting protein kinase OsCIPK1 phosphorylated by SAPK10 positively regulates responses to ABA and osmotic stress in rice

SubclassⅢsucrose nonfermenting1-related protein kinase 2s(SnRK2s)function in ABA and abiotic stress responses by unknown mechanisms.We found that osmotic stress/ABA-activated protein kinase 10(SAPK10),a member of rice SnRK2s,physically interacted with CBL-interacting protein kinase 1(OsCIPK1).OsCIPK1 expression was up-regulated by ABA and PEG treatment,and overexpression increased the ABA sensitivity of seed germination and root growth and plant osmotic stress tolerance.Osmotic stress or ABA-induced activation of OsCIPK1 is dependent on SAPK10.SAPK10 phosphorylates Thr-24 of OsCIPK1 in vitro,and this phosphorylation increases the activity of OsCIPK1 and positively regulates the function of OsCIPK1 in ABA responses and plant osmotic stress tolerance.This study suggests that OsCIPK1 is a direct phosphorylated substrate of SAPK10,and SAPK10-mediated phosphorylation of OsCIPK1 functions in ABA signaling and increases rice osmotic stress tolerance.

JA-mediated MYC2/LOX/AOS feedback loop regulates osmotic stress response in tea plant

Osmotic stress caused by low-temperature,drought and salinity was a prevalent abiotic stress in plant that severely inhibited plant development and agricultural yield,particularly in tea plant.Jasmonic acid(JA)is an important phytohormone involving in plant stress.However,underlying molecular mechanisms of JA modulated osmotic stress response remains unclear.In this study,high concentration of mannitol induced JA accumulation and increase of peroxidase activity in tea plant.Integrated transcriptome mined a JA signaling master,MYC2 transcription factor is shown as a hub regulator that induced by mannitol,expression of which positively correlated with JA biosynthetic genes(LOX and AOS)and peroxidase genes(PER).CsMYC2 was determined as a nuclei-localized transcription activator,furthermore,ProteinDNA interaction analysis indicated that CsMYC2 was positive regulator that activated the transcription of CsLOX7,CsAOS2,CsPER1 and CsPER3via bound with their promoters,respectively.Suppression of CsMYC2 expression resulted in a reduced JA content and peroxidase activity and osmotic stress tolerance of tea plant.Overexpression of CsMYC2 in Arabidopsis improved JA content,peroxidase activity and plants tolerance against mannitol stress.Together,we proposed a positive feedback loop mediated by CsMYC2,CsLOX7 and CsAOS2 which constituted to increase the tolerance of osmotic stress through fine-tuning the accumulation of JA levels and increase of POD activity in tea plant.

Overexpression of the transcription factor MdWRKY115 improves drought and osmotic stress tolerance by directly binding to the MdRD22 promoter in apple

Abiotic stress reduces plant yield and quality.WRKY transcription factors play key roles in abiotic stress responses in plants,but the molecular mechanisms by which WRKY transcription factors mediate responses to drought and osmotic stresses in apple(Malus×domestica Borkh.)remain unclear.Here,we functionally characterized the apple GroupⅢWRKY gene MdWRKY115.qRT-PCR analysis showed that MdWRKY115 expression was up-regulated by drought and osmotic stresses.GUS activity analysis revealed that the promoter activity of MdWRKY115 was enhanced under osmotic stress.Subcellular localization and transactivation assays indicated that MdWRKY115 was localized to the nucleus and had a transcriptional activity domain at the N-terminal region.Transgenic analysis revealed that the overexpression of MdWRKY115 in Arabidopsis plants and in apple callus markedly enhanced their tolerance to drought and osmotic stresses.DNA affinity purification sequencing showed that MdWRKY115 binds to the promoter of the stress-related gene MdRD22.This binding was further verified by an electrophoretic mobility shift assay.Collectively,these findings suggest that MdWRKY115 is an important regulator of osmotic and drought stress tolerance in apple.

Coseismic Coulomb stress changes induced by a 2020-2021 M_(W)>7.0 Alaska earthquake sequence in and around the Shumagin gap and its influence on the Alaska-Aleutian subduction interface

Three M_(W)>7.0 earthquakes in 2020-2021 occurred in the Shumagin seismic gap and its adjacent area of the Alaska-Aleutian subduction zone,including the Mw7.8 Simeonof thrust earthquake on July 22,2020,the M_(W)7.6 Sand Point strike-slip earthquake on October 19,2020,and the M_(W)8.2 Chignik thrust earthquake on July 29,2021.The spatial and temporal proximity of these three earthquakes prompts us to probe stress-triggering effects among them.Here we examine the coseismic Coulomb stress change imparted by the three earthquakes and their influence on the subduction interface.Our results show that:(1)The Simeonof earthquake has strong loading effects on the subsequent Sand Point and Chignik earthquakes,with the Coulomb stress changes of 3.95 bars and 2.89 bars,respectively.The Coulomb stress change caused by the Sand Point earthquake at the hypocenter of the Chignik earthquake is merely around 0.01 bars,suggesting the negligible triggering effect on the latter earthquake;(2)The triggering effects of the Simeonof,Sand Point,and Chignik earthquakes on aftershocks within three months are not well pronounced because of the triggering rates of 38%,14%,and 43%respectively.Other factors may have played an important role in promoting the occurrence of these aftershocks,such as the roughness of the subduction interface,the complicated velocity structure of the lithosphere,and the heterogeneous prestress therein;(3)The three earthquakes caused remarkable coseismic Coulomb stress changes at the subduction interface nearby these mainshocks,with an average Coulomb stress change of 3.2 bars in the shallow region directly inwards the trench.

Exercise-induced modulation of miR-149-5p and MMP9 in LPS-triggered diabetic myoblast ER stress: licorice glycoside E as a potential therapeutic target

Background:This study explores the relationship between endoplasmic reticulum(ER)stress and diabetes,particularly focusing on the impact of physical exercise on ER stress mechanisms and identifying potential therapeutic drugs and targets for diabetes-related sepsis.The research also incorporates traditional physical therapy perspectives,emphasizing the genomic insights gained from exercise therapy in disease management and prevention.Methods:Gene analysis was conducted on the GSE168796 and GSE94717 datasets to identify ER stress-related genes.Gene interactions and immune cell correlations were mapped using GeneCard and STRING databases.A screening of 2,456 compounds from the TCMSP database was performed to identify potential therapeutic agents,with a focus on their docking potential.Techniques such as luciferase reporter gene assay and RNA interference were used to examine the interactions between microRNA-149-5p and MMP9.Results:The study identified 2,006 differentially expressed genes and 616 miRNAs.Key genes like MMP9,TNF-α,and IL1B were linked to an immunosuppressive state.Licorice glycoside E demonstrated high affinity for MMP9,suggesting its potential effectiveness in treating diabetes.The constructed miRNA network highlighted the regulatory roles of MMP9,IL1B,IFNG,and TNF-α.Experimental evidence confirmed the binding of microRNA-149-5p to MMP9,impacting apoptosis in diabetic cells.Conclusion:The findings highlight the regulatory role of microRNA-149-5p in managing MMP9,a crucial gene in diabetes pathophysiology.Licorice glycoside E emerges as a promising treatment option for diabetes,especially targeting MMP9 affected by ER stress.The study also underscores the significance of physical exercise in modulating ER stress pathways in diabetes management,bridging traditional physical therapy and modern scientific understanding.Our study has limitations.It focuses on the microRNA-149-5p-MMP9 network in sepsis,using cell-based methods without animal or clinical trials.Despite strong in vitro findings,in vivo studies are needed to confirm licorice glycoside E’s therapeutic potential and understand the microRNA-149-5p-MMP9 dynamics in real conditions.

Exosomes derived from microglia overexpressing miR-124-3p alleviate neuronal endoplasmic reticulum stress damage after repetitive mild traumatic brain injury

We previously reported that miR-124-3p is markedly upregulated in microglia-derived exosomes following repetitive mild traumatic brain injury.However,its impact on neuronal endoplasmic reticulum stress following repetitive mild traumatic brain injury remains unclear.In this study,we first used an HT22 scratch injury model to mimic traumatic brain injury,then co-cultured the HT22 cells with BV2 microglia expressing high levels of miR-124-3p.We found that exosomes containing high levels of miR-124-3p attenuated apoptosis and endoplasmic reticulum stress.Furthermore,luciferase reporter assay analysis confirmed that miR-124-3p bound specifically to the endoplasmic reticulum stress-related protein IRE1α,while an IRE1αfunctional salvage experiment confirmed that miR-124-3p targeted IRE1αand reduced its expression,thereby inhibiting endoplasmic reticulum stress in injured neurons.Finally,we delivered microglia-derived exosomes containing miR-124-3p intranasally to a mouse model of repetitive mild traumatic brain injury and found that endoplasmic reticulum stress and apoptosis levels in hippocampal neurons were significantly reduced.These findings suggest that,after repetitive mild traumatic brain injury,miR-124-3 can be transferred from microglia-derived exosomes to injured neurons,where it exerts a neuroprotective effect by inhibiting endoplasmic reticulum stress.Therefore,microglia-derived exosomes containing miR-124-3p may represent a novel therapeutic strategy for repetitive mild traumatic brain injury.

Adaptive strategy of Nitraria sibirica to transient salt,alkali and osmotic stresses via the alteration of Na+/K+fluxes around root tips

Nitraria sibirica Pall.is an important shrub with a strong salt-alkali tolerance,but the mechanism underlying this tolerance remains obscure.In this study,N.sibirica,with salt-sensitive Vigna radiata(Linn.)Wilczek as the control,was subjected to transient salt stress(100 mM NaCl),alkali stress(50 mM Na_(2)CO_(3)),and osmotic stress(175 mM mannitol).The ionic fluxes of Na^(+)and K^(+)in the root apical region were measured.Results show that,under salt and alkali stress,N.sibirica roots exhibited higher capacities to limit Na+influx and reduce K+efflux,thereby resulting in lower Na^(+)/K^(+)ratios compared with V.radiata roots.Alkali stress induced stronger Na^(+)influx and K+efflux in the root salt stress treatment;Na^(+)influx was mainly observed in the root cap,while K^(+)efflux was mainly observed in the elongation zone.While under osmotic stress,N.sibirica roots showed stronger Na+efflux and weaker K+efflux than V.radiata roots.Na+efflux was mainly observed in the root elongation zone,while K+efflux was in the root cap.These results reveal the ionic strategy of N.sibirica in response to transient salt,alkali,and osmotic stresses through the regulation of Na+/K+flux homeostasis.

Relationship between ATPase activity and conjugated polyamines in mito-chondrial membrane from wheat seedling roots under osmotic stress

The effects of osmotic stress on the ATPase activity, the contents of —SH group and conjugated polyamines in mitochondrial membrane from wheat seedling [Triticum aestivum L. cv. Yumai No.18(drought-tolerant) and cv. Yumai No.9(drought-sensitive)] roots were investigated. The results showed that ATPase activity and —SH group content decreased with polyethylene glycol(PEG) 6000(-0.55 MPa) treatment for 7 d, in concert with the decrease of the ratio of noncovalently conjugated spermidine(NCC-Spd)/noncovalently conjugated putrescine(NCC-Put) and increase of the covalently conjugated putrescine(CC-Put). Osmotic stress injury to Yangmai No.9 seedlings was alleviated greatly with 1 mmol/L exogenous spermidine(Spd), in concert with marked increases of the ratio of NCC-Spd/NCC-Put, —SH group contents and ATPase activity in mitochondrial membrane. Under osmotic stress, the concomitant treatment of Yumai No.18 seedlings with methylglyoxyl bis(guanylhydrazone) (MGBG), an inhibitor of S-adenosyl methionine decarboxylase(SAMDC), and phenanthrolin (o-Phen), an inhibitor of transglutaminase(TGase), caused a significant decrease of the ratio of NCC-Spd / NCC-Put, CC-Put contents, respectively, in concert with the marked decreases of ATPase activity, —SH group content and its tolerance to osmotic stress. All the results above suggested that osmotic stress tolerance of wheat seedlings was associated with the ATPase activity, the contents of —SH group, NCC-Spd and CC-Put in mitochondrial membrane.

Effects of osmotic stress on antioxidant enzymes activities in leaf discs of P_(SAG12)-IPT modified gerbera

Leaf senescence is often caused by water deficit and the chimeric gene PSA612-1PT is an auto-regulated gene delaying leaf senescence. Using in vitro leaf discs culture system, the changes of contents of chlorophylls, carotenoids, soluble protein and thiobarbituric acid reactive substance （TBARS） and antioxidant enzymes activities were investigated during leaf senescence of PSA612-1PT modified gerbera induced by osmotic stress compared with the control plant （wild type）. Leaf discs were incubated in 20%, 40% （w/v） polyethylene glycol （PEG） 6 000 nutrient solution for 20 h under continuous light [ 130 μmol/（m^2·s）]. The results showed that the contents of chlorophylls, carotenoids and soluble protein were decreased by osmotic stress with the decrease being more pronounced at 40% PEG, but that, at the same PEG concentration the decrease in the transgenic plants was significantly lower than that in the control plant. The activities of superoxide dismutase （SOD）, catalases （CAT）, ascorbate peroxidase （APX）, guaiacol peroxidase （GPX） and dehydroascorbate reductase （DHAR） were stimulated by PEG treatment. However, the increases were higher in PSA612-IPT transgenic plants than in the control plants, particularly at 40% PEG treatment. Lipid peroxidation （TBARS content） was increased by PEG treatment with the increase being much lower in transgenic plant than in the control plant. It could be concluded that the increases in the activities ofantioxidant enzymes including SOD, CAT, APX, GPX and DHAR were responsible for the delay of leaf senescence induced by osmotic stress.

Proteomic analysis of salt and osmotic-drought stress in alfalfa seedlings

Alfalfa is widely grown and is one of the most important forage crops in the world, but its growth and biomass production are markedly reduced under salt and drought stress, particularly during the early seedling stage. To identify the mechanisms behind salt and drought responsiveness at the alfalfa seedling stage, the proteins expressed were analyzed under no-treatment, 200 mol L–1 Na Cl and 180 g L–1 PEG treatment conditions during the seedling stage. Out of more than 800 protein spots detected on two-dimensional electrophoresis（2-DE） g els, 35 proteins showed statistically significant responses（P〈0.05） to Na Cl and PEG stress, which were selected for tandem mass spectrometric identification, owing to their good resolution and abundance levels, and 32 proteins were positively identified. The identified proteins were divided into seven functional categories： photosynthetic metabolism, protein biosynthesis, folding and assembly, carbohydrate metabolism-associated proteins, stress defense related protein, metabolism of nucleic acid, other function categories and unknown proteins. Our results suggested that these proteins may play roles in alfalfa adaptation to salt and drought stress. Further study of these proteins will provide insights into the molecular mechanisms of abiotic stress and the discovery of new candidate markers in alfalfa.

Interactive effect of shade and PEG-induced osmotic stress on physiological responses of soybean seedlings

Intensively farmed crops used to experience numerous environmental stresses.Among these,shade and drought significantly influence the morpho-physiological and biochemical attributes of plants.However,the interactive effect of shade and drought on the growth and development of soybean under dense cropping systems has not been reported yet.This study investigated the interactive effect of PEG-induced osmotic stress and shade on soybean seedlings.The soybean cultivar viz.,C-103 was subjected to PEG-induced osmotic stress from polyethylene glycol 6000(PEG-6000)under shading and non-shading conditions.PEG-induced osmotic stress significantly reduced the relative water contents,morphological parameters,carbohydrates and chlorophyll contents under both light environments.A significant increase was observed in osmoprotectants,reactive oxygen species and antioxidant enzymes in soybean seedlings.Henceforth,the findings revealed that,seedlings grown under non-shading conditions produced more malondialdehyde and hydrogen peroxide contents as compared to the shade-treated plants when subjected to PEG-induced osmotic stress.Likewise,the shaded plants accumulated more sugars and proline than non-shaded ones under drought stress.Moreover,it was found that nonshaded grown plants were more sensitive to PEG-induced osmotic stress than those exposed to shading conditions,which suggested that shade could boost the protective mechanisms against osmotic stress or at least would not exaggerate the adverse effects of PEG-induced osmotic stress in soybean seedlings.

GmGPDH12,a mitochondrial FAD-GPDH from soybean, increases salt and osmotic stress resistance by modulating redox state and respiration

In plants,glycerol-3-phosphate dehydrogenase(GPDH)catalyzes the interconversion of glycerol-3-phosphate(G3P)and dihydroxyacetone phosphate(DHAP)coupled to the reduction/oxidation of the nicotinamide adenine dinucleotide(NADH)pool,and plays a central role in glycerolipid metabolism and stress response.Previous studies have focused mainly on the NAD+-dependent GPDH isoforms,neglecting the role of flavin adenine dinucleotide(FAD)-dependent GPDHs.We isolated and characterized three mitochondrialtargeted FAD-GPDHs in soybean,of which one isoform(GmGPDH12)showed a significant transcriptional response to NaCl and mannitol treatments,suggesting the existence of a major FAD-GPDH isoform acting in soybean responses to salt and osmotic stress.An enzyme kinetic assay showed that the purified GmGPDH12 protein possessed the capacity to oxidize G3P to DHAP in the presence of FAD.Overexpression and RNA interference of GmGPDH12 in soybean hairy roots resulted in elevated tolerance and sensitivity to salt and osmotic stress,respectively.G3P contents were significantly lower in GmGPDH12-overexpressing hair roots and higher in knockdown hair roots,indicating that GmGPDH12 was essential for G3P catabolism.A significant perturbation in redox status of NADH,ascorbic acid(ASA)and glutathione(GSH)pools was observed in GmGPDH12-knockdown plants under stress conditions.The impaired redox balance was manifested by higher reactive oxygen species generation and consequent cell damage or death;however,overexpressing plants showed the opposite results for these traits.GmGPDH12 overexpression contributed to maintaining constant respiration rates under salt or osmotic stress by regulating mRNA levels of key mitochondrial respiratory enzymes.This study provides new evidence for the roles of mitochondria-localized GmGPDH12 in conferring resistance to salt or osmotic stress by maintaining cellular redox homeostasis,protecting cells and respiration from oxidative injury.

Effects of short-term osmotic stress on leaf hydraulic conductivity and ZmPIPs mRNA accumulation in maize seedlings

Plants maintain water balance by varying hydraulic properties, and plasma membrane intrinsic proteins（PIPs） may be involved in this process. Leaf xylem and root hydraulic conductivity and the m RNA contents of four highly expressed Zm PIP genes（Zm PIP1;1, Zm PIP1;2, Zm PIP2;2, and Zm PIP2;5） in maize（Zea mays） seedlings were investigated. Under well-watered conditions, leaf hydraulic conductivity（K_（leaf）） varied diurnally and was correlated with whole-plant hydraulic conductivity. Similar diurnal rhythms of leaf transpiration rate（E）, K_（leaf） and root hydraulic conductivity（K_（root）） in well-watered plants are important for maintaining whole-plant water balance. After 2 h of osmotic stress treatment induced by 10% polyethylene glycol 6000, the K_（root） of stressed plants decreased but K_（leaf） increased, compared with well-watered plants. The m RNA contents of four Zm PIPs were significantly up-regulated in the leaves of stressed plants, especially for Zm PIP1;2. Meanwhile, Zm PIP2;5 was significantly down-regulated in the roots of stressed plants. After 4 h of osmotic stress treatment, the E and leaf xylem water potentials of stressed plants unexpectedly increased. The increase in K_（leaf） and a partial recovery of K_（root） may have contributed to this process. The m RNA content of Zm PIP1;2 but not of the other three genes was up-regulated in roots at this time. In summary, the m RNA contents of these four Zm PIPs associated with K_（leaf） and K_（root） change in maize seedlings during short-term osmotic stress, especially for Zm PIP1;2 and Zm PIP2;5, which may help to further reveal the hydraulic resistance adjustment role of Zm PIPs.

Osmotic Stress Resistance of Transgenic Tobacco Expressed Soybean Lysine-rich Protein Gene

PM2 gene （accession number： M80664） with high lysine content from soybean （Glycine max） was found in GenBank by changing three BLASTp parameters. Amino acid composition analysis of PM2 showed that Lys content was on the high level of 18.22%. Protein encoded by PM＇2 also belonged to the family of late embryogenesis abundant （LEA） proteins, which was considered that it had a strong relation with the abiotic stress resistance. In this experiment, PM2 gene was obtained from dry soybean seeds by RT-PCR, plant expression vector pEMTPM2 was constructed, and then transformed into tobacco by using agrobacterium-mediated method. Eight salt and drought tolerant lines were obtained from 31 differentiated lines. Real-time PCR showed that PM2 gene overexpressed in all four PCR positive lines with the osmotic stress resistance. These results confirmed that the overexpression of PM2 gene enhanced the osmotic stress resistance of transgenic tobacco.

Effects of Polyamine on Leaf Physiological and Biochemical Indexes of Maize Seedlings under Osmotic Stress

[Objective]The research aimed to probe into the mechanism of polyamine enhancing the tolerance of plants to drought stress and provide theoretical basis for application of polyamine in process of maize drought resistance. [Method]With PEG-6000 simulating natural drought,the change in content of soluble protein and relative water content were investigated in seedling leaves of two maize cultivars,Nongda 108 and Yedan 13 under osmotic stress with exogenous Spd treatment. [Result]On the 7th day,leaf relative water content and the content of soluble protein decreased more significantly in leaves of Yedan 13 （drought-sensitive） than in Nongda 108 （drought-tolerant）. Exogenous Spd treatment not only obviously inhibited the decrease of leaf relative water content,but also increased the content of soluble protein. [Conclusion]Exogenous Spd treatment could enhance the tolerance of maize seedlings to osmotic stress,via improving the content of soluble protein in seedling leaves.

Effects of Nitric Oxide on the Germination of Wheat Seeds and Its Reactive Oxygen Species Metabolisms Under Osmotic Stress

Effects of sodium nitroprusside (SNP), a nitric oxide (NO) donor, on the germination and metabolism of reactive oxygen species were surveyed in wheat (Triticum aestivum L.) seeds. Germination of wheat seeds and even the elongation of radicle and plumule were dramatically promoted by SNP treatments during the germination under osmotic stress. Meanwhile, activities of amylase and EP were enhanced, thus leading to the degradation of storage reserve in seeds. After osmotic stress was removed, higher viability of wheat seeds was also maintained. In addition, the activities of CAT, APX and the content of proline were increased by SNP treatment simultaneously, but activities of LOX were inhibited, and both of which were beneficial for improving the antioxidant capacity during the germination of wheat seeds under osmotic stress. It was also shown that the increase of the activity of amylase induced by SNP in embryoless half-seeds of wheat in the beginning period of germination (6 h) might be indirectly related to GA(3).

Effect of Osmotic Stress with PEG6000 on Osmotic Adjustment Solute in Seedling Leaves of Chenopodium album L.

[ Objective] The purpose was to discuss drought resistance mechanism of Chenopodium album L. and supply theoretical basis and practical guidance for artificial cultivation and popularization of C. album. [ Method] C. album seedlings grown to 6th leaf stage were conducted osmotic stress treatment with PEG6000 osmotic whose concentration was set up as 0, 5%, 10% and 20% and the various physiological indices of the 3rd -5th function leaves in upper plant were determined after being treated for 0, 1,3, 5, 7 and 9 d. [ Result] Under osmotic stress with 5% PGE, the relative water content （RWC） of C. album reduced less. Under osmotic stress with 10%, the RWC in seedling leaves of C. album decreased to 62% on the fifth day and the leaves began to wither. Under osmotic stress with 20%, the RWC in seedling leaves of C. album decreased to 61.9% on the third day and the leaves appeared withering, and the RWC decreased to 48.6% on the 7th day and the leaves were dry and yellow. Proline contents in seedling leaves of C. album under osmotic stress with 5%, 10% and 20% PEG were 7.64, 10.9 and 29.4 times of CK on the 7th day. [ Conclusion] C. album hed some adaptability to moderate osmotic stress, but the PEG osmotic stress with high concentration and long time might lead to severe damage on C. album.

Effects of Drought Stress on Osmotic Regulation Substances of Five Catalpa bungei Clones

[Objective] This study aimed to investigate the effects of drought stress on osmotic regulation substances of Catalpa bungei clones. [Method] Potting experimental was carried out with plastic film sealing method, to differentiate the degrees of drought stress based on changes in soil water content and leaf water potential and investigate variations in contents of osmotic regulation substances（free proline,soluble sugar and soluble protein） in leaves of five Catalpa bungei clones under different degrees of drought stresses. [Result] According to changes in soil water content and leaf water potential, the whole process of drought stress was divided into normal level（CK, with soil water content of 97.49% and leaf water potential of-0.54MPa）, light drought（LD with soil water content of 59.96% and leaf water potential of-1.28 MPa）, mediate drought（MD with soil water content of 34.19% and leaf water potential of-2.32 MPa） and severe drought（SD with soil water content of 14.52%and leaf water potential of-2.99 MPa）. The soil water content and leaf water potential of five Catalpa bungei clones reached the highest correlation in exponential fitting, with an average R2of 0.989 3（P0.001）. The free proline content in leaves of five Catalpa bungei clones increased rapidly with the increasing degree of drought stress（P 0.001）. To be specific, free proline contents of Catalpa bungei clones015-1 and 7080 were 34.39 and 33.41 times of the normal level under severe drought conditions, which reached an extremely significant level（P0.001）; the free proline content of Catalpa bungei clone 1-3 rapidly increased to（855.46±227.52） μg/g Fw under light drought conditions. The soluble protein content in leaves showed different variation trends. To be specific, the soluble protein content of Catalpa bungei clone 7080 was the lowest at various drought stages and reached（1.644 ±0.137）mg/g Fw under normal conditions; the soluble protein content of Catalpa bungei clone 1-3 was relatively high under normal conditions and was reduced rapidly under light drought conditions, showing different response patterns. [Conclusion] Osmotic regulation substances in leaves of five Catalpa bungei clones all have certain response to drought stress. Free proline is the most important osmotic regulation substance, followed by soluble protein, while soluble sugar makes no significant contribution. According to changes in content of osmotic regulation substances, Catalpa bungei clone 7080 has relatively high capacity and good drought resistance, while Catalpa bungei clone 1-3 has the rapidest response to drought stress.

OSMOTIC STRESS DECREASES THE ACTIVITY OF ATPASE ASSOCIATED WITH THE ROOT CAP PLASMODESMATAIN ZEA MAYS

With light and electron microscopy the substructural change and the ATPase activity of corn (Zea mays L.) root cap cells after short-term osmotic stress were studied. Some spoke-like fine strands originating from the departed periplasm and stretching towards cell wall could be observed even after plasmolysis. By observing the precipitation of ATPase activity product (lead phosphate) at plasma membrane and plasmodesmata, it was found that the fine strands were plasma membrane-lined channels surrounding the cytoplasm and that they still firmly connected to the plasmodesmata during plasmolysis. Compared with the control (unstressed), a sharp decrease of ATPase activity in the plasmodesmata of the stressed cells was observed. Inhibition of energy metabolism in these limited locales would affect the physiological activity, maybe including the regulation of permeability and the change of size exclusion limit (SEL) of plasmodesmata.

Characterization and Expression Analysis of Four Glycine-Rich RNA-Binding Proteins Involved in Osmotic Response in Tobacco (Nicotiana tabacum cv. Xanthi)

Plants have developed many signals and specific genes＇ regulations at both transcriptional and post-transcriptional levels in order to tolerate and adapt to various environmental stresses. RNA-binding proteins （RBPs） play crucial roles in the post- transcriptional regulation via mRNA splicing, polyadenylation, sequence editing, transport, mRNA stability, mRNA localization, and translation. In this paper, four cDNAs of glycine-rich RNA-binding proteins （GR-RBPs）, named NtRGP-la, -lb, -2, and -3, were isolated from Nicotiana tabacum by RT-PCR analysis, and special emphases were given to the sequences alignment, phylogenetic analysis and gene expression. Sequences alignment revealed minor difference of cDNA sequences, but no difference of deduced proteins between N. sylvestris and N. tabacum. Phylogenetic alignment revealed that four cDNAs in tobacco were clustered into two different groups. NtRGP-2 and -3 were evolutionarily closest to Arabidopsis GR-RBPs genes and related to animal GR-RBPs genes, while NtRGP-la and -lb were closest to Gramineae GR-RBPs genes. The expression analyses of these four NtRGPs in response to different abiotic stresses revealed the similar expression pattern. Moreover, the four NtRGPs, especially NtRGP-la and NtRGP-3, were strongly induced by stresses including water, wound, cold, and high temperature, weakly induced by PEG, drought and SA, while reduced by NaC1 and unaffected by ABA treatment. The fact that all of these abiotic stresses included in our experiments affected the water balance and resulted in osmotic stress on cellular level, suggests that NtRGPs in tobacco should be a family of crucial osmosis-related proteins, and may play a key role in signal transduction with ABA-independent pathway under abiotic stresses.