Changes in ambient temperature profoundly affect plant growth and performance.Therefore,the molecu-larbasis of plant acclimation to temperature fluctuation is of great interest.In this study,we discovered that GLYCINE...Changes in ambient temperature profoundly affect plant growth and performance.Therefore,the molecu-larbasis of plant acclimation to temperature fluctuation is of great interest.In this study,we discovered that GLYCINE-RICH RNA-BINDING PROTEIN 7(GRP7)contributes to cold and heat tolerance in Arabidopsis thaliana.We found that exposure to a warm temperature rapidly induces GRP7 condensates in planta,which can be reversed by transfer to a lower temperature.Cell biology and biochemical assays revealed that GRP7 undergoes liquid-liquid phase separation(LLPS)in vivo and in vitro.LLPS of GRP7 in the cyto-plasm contributes to the formation of stress granules that recruit RNA,along with the translation machinery component eukaryotic initiation factor 4E1(elF4E1)and the mRNA chaperones COLD SHOCK PROTEIN 1(CSP1)and CSP3,to inhibit translation.Moreover,natural variations in GRP7 affecting the residue phos-phorylated by the receptorkinase FERONIA alter its capacity to undergo LLPS and correlate with the adap-tation of some Arabidopsis accessions to a widertemperature range.Taken together,ourfindings illustrate the role of translational control mediated by GRP7 LLPS to confer plants with temperature resilience.展开更多
Abiotic and biotic stressors adversely affect plant survival,biomass generation,and crop yields.As the global availability of arable land declines and the impacts of global warming intensify,such stressors may have in...Abiotic and biotic stressors adversely affect plant survival,biomass generation,and crop yields.As the global availability of arable land declines and the impacts of global warming intensify,such stressors may have increasingly pronounced effects on agricultural productivity.Currently,researchers face the overarching challenge of comprehensively enhancing plant resilience to abiotic and biotic stressors.The secondary cell wall plays a crucial role in bolstering the stress resistance of plants.To increase plant resistance to stress through genetic manipulation of the secondary cell wall,we cloned a cell wall protein designated glycine-rich protein-like(GhGRPL)from cotton fibers,and found that it is specifically expressed during the period of secondary cell wall biosynthesis.Notably,this protein differs from its Arabidopsis homolog,AtGRP,since its glycine-rich domain is deficient in glycine residues.GhGRPL is involved in secondary cell wall deposition.Upregulation of GhGRPL enhances lignin accumulation and,consequently,the thickness of the secondary cell walls,thereby increasing the plant’s resistance to abiotic stressors,such as drought and salinity,and biotic threats,including Verticillium dahliae infection.Conversely,interference with GhGRPL expression in cotton reduces lignin accumulation and compromises that resistance.Taken together,our findings elucidate the role of GhGRPL in regulating secondary cell wall development through its influence on lignin deposition,which,in turn,reinforces cell wall robustness and impermeability.These findings highlight the promising near-future prospect of adopting GhGRPL as a viable,effective approach for enhancing plant resilience to abiotic and biotic stress factors.展开更多
Rice(Oryza sativa L.)production is threatened by global warming associated with extreme high temperatures,and rice heat sensitivity is differed when stress occurs between daytime and nighttime.However,the underlying m...Rice(Oryza sativa L.)production is threatened by global warming associated with extreme high temperatures,and rice heat sensitivity is differed when stress occurs between daytime and nighttime.However,the underlying molecular mechanism are largely unknown.We show here that two glycine-rich RNA binding proteins,Os GRP3 and Os GRP162,are required for thermotolerance in rice,especially at nighttime.The rhythmic expression of Os GRP3/Os GRP162 peaks at midnight,and at these coincident times,is increased by heat stress.This is largely dependent on the evening complex component Os ELF3-2.We next found that the double mutant of Os GRP3/Os GRP162 is strikingly more sensitive to heat stress in terms of survival rate and seed setting rate when comparing to the wild-type plants.Interestingly,the defect in thermotolerance is more evident when heat stress occurred in nighttime than that in daytime.Upon heat stress,the double mutant of Os GRP3/Os GRP162 displays globally reduced expression of heat-stress responsive genes,and increases of m RNA alternative splicing dominated by exon-skipping.This study thus reveals the important role of Os GRP3/Os GRP162 in thermotolerance in rice,and unravels the mechanism on how Os GRP3/Os GRP162 regulate thermotolerance in a diurnal manner.展开更多
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...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.展开更多
The RNA‐binding glycine‐rich protein（RB‐GRP）family is characterized by the presence of a glycine‐rich domain arranged in（Gly）n‐X repeats and an RNA‐recognition motif（RRM）. RB‐GRPs participate in varied ph...The RNA‐binding glycine‐rich protein（RB‐GRP）family is characterized by the presence of a glycine‐rich domain arranged in（Gly）n‐X repeats and an RNA‐recognition motif（RRM）. RB‐GRPs participate in varied physiological and biochemical processes especially in the stress response of plants. In this study, a total of 23 RB‐GRPs distributed on 10 chromosomes were identified in maize（Zea mays L.）, and they were divided into four subgroups according to their conserved domain architecture. Five pairs of paralogs were identified,while none of them was located on the same chromosomal region, suggesting that segmental duplication is predominant in the duplication events of the RB‐GRPs in maize. Comparative analysis of RB‐GRPs in maize, Arabidopsis（Arabidopsis thaliana L.）, rice（Oryza sativa L.）, and wheat（Triticum aestivum）revealed that two exclusive subgroups were only identified in maize. Expression of eight ZmRB‐GRPs was significantly regulated by at least two kinds of stresses. In addition, cis‐elements predicted in the promoter regions of the ZmRB‐GRPs also indicated that these ZmRB‐GRPs would be involved in stress response of maize. The preliminary genome‐wide analysis of the RB‐GRPs in maize would provide useful information for further study on the function of the ZmRB‐GRPs.展开更多
BACKGROUND： As a member of the LIM protein family Ⅱ, cysteine- and glycine-rich protein-2 （CRP2） has been demonstrated to play a role in the regulation of growth and differentiation of eukaryotic cells. Our previo...BACKGROUND： As a member of the LIM protein family Ⅱ, cysteine- and glycine-rich protein-2 （CRP2） has been demonstrated to play a role in the regulation of growth and differentiation of eukaryotic cells. Our previous study has demonstrated that CRP2 can be detected in the embryonic rat inner ear but not in the adult rat inner ear. However, at present, the expression of LIM protein family H members in stem or precursor cells has not been described. OBJECTIVE： To determine the expression and sub-cellular localization of CRP2 in olfactory stem cells. DESIGN, TIME AND SETTING： An experiment with repeated measures was performed in the Laboratory of Otorhinolaryngology, Head and Neck Surgery, Xijing Hospital, the Fourth Military Medical University from February 2008 to April 2008. MATERIALS： Olfactory stem cells, and rabbit-anti-CRP2 polyclonal antibody were prepared and kept in our laboratory. METHODS： Reverse transcription polymerase chain reaction and Western blot analysis were used to detect expression of CRP2 in olfactory stem cells. Immunocytochemistry was also used to localize CRP2 in olfactory stem cells. MAIN OUTCOME MEASURES： The expression and sub-cellular localization of CRP2 in rat olfactory stem cells. RESULTS： CRP2 expression was found in olfactory stem cells, and CRP2 was distributed in both the nucleus and the cytoplasm. CONCLUSION： Confirmation of the expression and distribution of CRP2 in olfactory stem cells.展开更多
To better understand the mechanism of sugar signaling in rice cell, the suspension-cultured rice cells were transferred from sucrose-containing (+S) to sucrose-free (-S) of MS culture medium, we found that ribosomal R...To better understand the mechanism of sugar signaling in rice cell, the suspension-cultured rice cells were transferred from sucrose-containing (+S) to sucrose-free (-S) of MS culture medium, we found that ribosomal RNAs (rRNAs) were degraded progressively. This suggests that carbon, nitrogen, and phosphate were recycled in this process and the reduction in cellular rRNAs might lead to decreased translation to save energy in response to sugar starvation. Differential screening revealed that two groups of genes, sugar-starvation-repressed (SSR) and sugar-starvation-activated (SSA) genes, were regulated by sugar in an opposing manner. Northern-blot analysis showed that two major hybridization signals of 0.8 and 1.9 kb were induced strongly under sugar starvation. The two populations of genes corresponded with homologs of α-amylases (1.9 kb) and the glycine-rich proteins (GRPs) gene family (0.8 kb), and all were SSA genes. Expression of GRP genes was strongly induced in sugar-starved cells, which suggests that GRPs may help to protect cells against nutritional stress. Treatment of +S and -S cells with the protein kinase (PK) inhibitor staurosporine (St) and the serine/theronine phosphoprotein phosphatases 1 (PP1) and 2A (PP2A) inhibitor okadaic acid (OA) revealed that PP1 and PP2A (PPs) might be involved in increasing SSR gene expression in +S cells, and that activation of the majority of the SSA genes in -S cells might be due to PKs activity. These results suggested that PKs and PPs might be involved in the sugar regulation of SSR and SSA gene expression. An in-gel PK activity assay demonstrated that the activity of two classes of PKs (50 and 66 kDa) may be induced rapidly after transfer of +S cells to -S medium. Following transfer of -S cells to +S medium, a novel class of 38 kDa PK was induced rapidly and showed high activity. The 38 kDa PK might play a role in sugar sensing, and the 50 and 66 kDa PKs might play roles in signal sensing under sugar starvation in rice cells. These results provide valuable informat展开更多
Based on the transcriptomic sequencing, three genes of Alexandrium pacificum encoding ubiquitin(UBI), telomerase(TEL) and glycine-rich protein(GRP) relating to cell division were isolated and characterized. The full-l...Based on the transcriptomic sequencing, three genes of Alexandrium pacificum encoding ubiquitin(UBI), telomerase(TEL) and glycine-rich protein(GRP) relating to cell division were isolated and characterized. The full-length cD NA of GRP was obtained through approach of rapid amplification of c DNA ends(RACE). Four conserved domains, including DNA-and RNAbinding sites or motifs, cold shock domain at the N-terminal, and zinc-finger structure of CCCH type at the C-terminal were idenrtified. Phylogenetic analysis revealed that the deduced amino acid sequence of GRP tends to cluster together with proteins harboring cold shock domain. The expressions of these three genes were analyzed with quantitative PCR(qPCR). It was found that the expressions of these three genes at the logarithmic growth phase and induced logarithmic growth phase were all higher than those at lagging growth phase(P < 0.05). The expression patterns of these three genes were coincident with the proliferative capacity of the algae, e.g., displaying increased expression level at log and induced growth phases. The functions of these genes and their possible roles during harmful algal blooms(HAB) were discussed.展开更多
基金supported by grants from National Natural Science Foundation of China(NSFC-32000208 and NSFC-32070769)National Key R&D Program of China(2023YFD1401100)+1 种基金China Postdoctoral Science Foundation funded project(2020M672475)the Science and Technology Innovation Program of Hunan Province(Nonos.2021JJ10015,2021JJ40060,2023ZJ1080,and 2021JJ40056).
文摘Changes in ambient temperature profoundly affect plant growth and performance.Therefore,the molecu-larbasis of plant acclimation to temperature fluctuation is of great interest.In this study,we discovered that GLYCINE-RICH RNA-BINDING PROTEIN 7(GRP7)contributes to cold and heat tolerance in Arabidopsis thaliana.We found that exposure to a warm temperature rapidly induces GRP7 condensates in planta,which can be reversed by transfer to a lower temperature.Cell biology and biochemical assays revealed that GRP7 undergoes liquid-liquid phase separation(LLPS)in vivo and in vitro.LLPS of GRP7 in the cyto-plasm contributes to the formation of stress granules that recruit RNA,along with the translation machinery component eukaryotic initiation factor 4E1(elF4E1)and the mRNA chaperones COLD SHOCK PROTEIN 1(CSP1)and CSP3,to inhibit translation.Moreover,natural variations in GRP7 affecting the residue phos-phorylated by the receptorkinase FERONIA alter its capacity to undergo LLPS and correlate with the adap-tation of some Arabidopsis accessions to a widertemperature range.Taken together,ourfindings illustrate the role of translational control mediated by GRP7 LLPS to confer plants with temperature resilience.
基金supported by the Special Fund for the Youth Team of the Southwest Universities,China(SWUXJPY 202306)the Fundamental Research Funds for the Central Universities,China(SWU-KR23009)the National Natural Sciences Foundation of China(U2003209 and 31871539)。
文摘Abiotic and biotic stressors adversely affect plant survival,biomass generation,and crop yields.As the global availability of arable land declines and the impacts of global warming intensify,such stressors may have increasingly pronounced effects on agricultural productivity.Currently,researchers face the overarching challenge of comprehensively enhancing plant resilience to abiotic and biotic stressors.The secondary cell wall plays a crucial role in bolstering the stress resistance of plants.To increase plant resistance to stress through genetic manipulation of the secondary cell wall,we cloned a cell wall protein designated glycine-rich protein-like(GhGRPL)from cotton fibers,and found that it is specifically expressed during the period of secondary cell wall biosynthesis.Notably,this protein differs from its Arabidopsis homolog,AtGRP,since its glycine-rich domain is deficient in glycine residues.GhGRPL is involved in secondary cell wall deposition.Upregulation of GhGRPL enhances lignin accumulation and,consequently,the thickness of the secondary cell walls,thereby increasing the plant’s resistance to abiotic stressors,such as drought and salinity,and biotic threats,including Verticillium dahliae infection.Conversely,interference with GhGRPL expression in cotton reduces lignin accumulation and compromises that resistance.Taken together,our findings elucidate the role of GhGRPL in regulating secondary cell wall development through its influence on lignin deposition,which,in turn,reinforces cell wall robustness and impermeability.These findings highlight the promising near-future prospect of adopting GhGRPL as a viable,effective approach for enhancing plant resilience to abiotic and biotic stress factors.
基金the State Key Project of Research and Development Plan of China(2022YFF1001603 and2021YFF1000404)the Natural Science Foundation of Zhejiang,China(LD21C020001)。
文摘Rice(Oryza sativa L.)production is threatened by global warming associated with extreme high temperatures,and rice heat sensitivity is differed when stress occurs between daytime and nighttime.However,the underlying molecular mechanism are largely unknown.We show here that two glycine-rich RNA binding proteins,Os GRP3 and Os GRP162,are required for thermotolerance in rice,especially at nighttime.The rhythmic expression of Os GRP3/Os GRP162 peaks at midnight,and at these coincident times,is increased by heat stress.This is largely dependent on the evening complex component Os ELF3-2.We next found that the double mutant of Os GRP3/Os GRP162 is strikingly more sensitive to heat stress in terms of survival rate and seed setting rate when comparing to the wild-type plants.Interestingly,the defect in thermotolerance is more evident when heat stress occurred in nighttime than that in daytime.Upon heat stress,the double mutant of Os GRP3/Os GRP162 displays globally reduced expression of heat-stress responsive genes,and increases of m RNA alternative splicing dominated by exon-skipping.This study thus reveals the important role of Os GRP3/Os GRP162 in thermotolerance in rice,and unravels the mechanism on how Os GRP3/Os GRP162 regulate thermotolerance in a diurnal manner.
基金funded by the National Natural Science Foundation of China (30560062)the Natural Science Foundation of Yunnan Province, China (2003C0342M)the Science-Technology Foundation of Tobacco Company of Yunnan Province, China (06A02)
文摘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.
基金financially supported by the National Natural Science Foundation of China (31171565 and 31371635)
文摘The RNA‐binding glycine‐rich protein（RB‐GRP）family is characterized by the presence of a glycine‐rich domain arranged in（Gly）n‐X repeats and an RNA‐recognition motif（RRM）. RB‐GRPs participate in varied physiological and biochemical processes especially in the stress response of plants. In this study, a total of 23 RB‐GRPs distributed on 10 chromosomes were identified in maize（Zea mays L.）, and they were divided into four subgroups according to their conserved domain architecture. Five pairs of paralogs were identified,while none of them was located on the same chromosomal region, suggesting that segmental duplication is predominant in the duplication events of the RB‐GRPs in maize. Comparative analysis of RB‐GRPs in maize, Arabidopsis（Arabidopsis thaliana L.）, rice（Oryza sativa L.）, and wheat（Triticum aestivum）revealed that two exclusive subgroups were only identified in maize. Expression of eight ZmRB‐GRPs was significantly regulated by at least two kinds of stresses. In addition, cis‐elements predicted in the promoter regions of the ZmRB‐GRPs also indicated that these ZmRB‐GRPs would be involved in stress response of maize. The preliminary genome‐wide analysis of the RB‐GRPs in maize would provide useful information for further study on the function of the ZmRB‐GRPs.
文摘BACKGROUND： As a member of the LIM protein family Ⅱ, cysteine- and glycine-rich protein-2 （CRP2） has been demonstrated to play a role in the regulation of growth and differentiation of eukaryotic cells. Our previous study has demonstrated that CRP2 can be detected in the embryonic rat inner ear but not in the adult rat inner ear. However, at present, the expression of LIM protein family H members in stem or precursor cells has not been described. OBJECTIVE： To determine the expression and sub-cellular localization of CRP2 in olfactory stem cells. DESIGN, TIME AND SETTING： An experiment with repeated measures was performed in the Laboratory of Otorhinolaryngology, Head and Neck Surgery, Xijing Hospital, the Fourth Military Medical University from February 2008 to April 2008. MATERIALS： Olfactory stem cells, and rabbit-anti-CRP2 polyclonal antibody were prepared and kept in our laboratory. METHODS： Reverse transcription polymerase chain reaction and Western blot analysis were used to detect expression of CRP2 in olfactory stem cells. Immunocytochemistry was also used to localize CRP2 in olfactory stem cells. MAIN OUTCOME MEASURES： The expression and sub-cellular localization of CRP2 in rat olfactory stem cells. RESULTS： CRP2 expression was found in olfactory stem cells, and CRP2 was distributed in both the nucleus and the cytoplasm. CONCLUSION： Confirmation of the expression and distribution of CRP2 in olfactory stem cells.
文摘To better understand the mechanism of sugar signaling in rice cell, the suspension-cultured rice cells were transferred from sucrose-containing (+S) to sucrose-free (-S) of MS culture medium, we found that ribosomal RNAs (rRNAs) were degraded progressively. This suggests that carbon, nitrogen, and phosphate were recycled in this process and the reduction in cellular rRNAs might lead to decreased translation to save energy in response to sugar starvation. Differential screening revealed that two groups of genes, sugar-starvation-repressed (SSR) and sugar-starvation-activated (SSA) genes, were regulated by sugar in an opposing manner. Northern-blot analysis showed that two major hybridization signals of 0.8 and 1.9 kb were induced strongly under sugar starvation. The two populations of genes corresponded with homologs of α-amylases (1.9 kb) and the glycine-rich proteins (GRPs) gene family (0.8 kb), and all were SSA genes. Expression of GRP genes was strongly induced in sugar-starved cells, which suggests that GRPs may help to protect cells against nutritional stress. Treatment of +S and -S cells with the protein kinase (PK) inhibitor staurosporine (St) and the serine/theronine phosphoprotein phosphatases 1 (PP1) and 2A (PP2A) inhibitor okadaic acid (OA) revealed that PP1 and PP2A (PPs) might be involved in increasing SSR gene expression in +S cells, and that activation of the majority of the SSA genes in -S cells might be due to PKs activity. These results suggested that PKs and PPs might be involved in the sugar regulation of SSR and SSA gene expression. An in-gel PK activity assay demonstrated that the activity of two classes of PKs (50 and 66 kDa) may be induced rapidly after transfer of +S cells to -S medium. Following transfer of -S cells to +S medium, a novel class of 38 kDa PK was induced rapidly and showed high activity. The 38 kDa PK might play a role in sugar sensing, and the 50 and 66 kDa PKs might play roles in signal sensing under sugar starvation in rice cells. These results provide valuable informat
基金supported by the National Natural Science Foundation of China (Nos. 41676091 41176098)Shandong Province Natural Science Foundation (No. ZR 2011DZ002)
文摘Based on the transcriptomic sequencing, three genes of Alexandrium pacificum encoding ubiquitin(UBI), telomerase(TEL) and glycine-rich protein(GRP) relating to cell division were isolated and characterized. The full-length cD NA of GRP was obtained through approach of rapid amplification of c DNA ends(RACE). Four conserved domains, including DNA-and RNAbinding sites or motifs, cold shock domain at the N-terminal, and zinc-finger structure of CCCH type at the C-terminal were idenrtified. Phylogenetic analysis revealed that the deduced amino acid sequence of GRP tends to cluster together with proteins harboring cold shock domain. The expressions of these three genes were analyzed with quantitative PCR(qPCR). It was found that the expressions of these three genes at the logarithmic growth phase and induced logarithmic growth phase were all higher than those at lagging growth phase(P < 0.05). The expression patterns of these three genes were coincident with the proliferative capacity of the algae, e.g., displaying increased expression level at log and induced growth phases. The functions of these genes and their possible roles during harmful algal blooms(HAB) were discussed.
