In the Medicago genus, triterpenic saponins are bioactive secondary metabolites constitutively synthesized in the aerial and subterranean parts of plants via the isoprenoid pathway. Exploitation of saponins as pharmac...In the Medicago genus, triterpenic saponins are bioactive secondary metabolites constitutively synthesized in the aerial and subterranean parts of plants via the isoprenoid pathway. Exploitation of saponins as pharmaceutics, agrochemicals and in the food and cosmetic industries has raised interest in identifying the enzymes involved in their synthesis. We have identified a cytochrome P450 （CYP72A67） involved in hemolytic sapogenin biosynthesis by a reverse genetic TILLING approach in a Medicago truncatula ethyl- methanesulfonate （EMS） mutagenized collection. Genetic and biochemical analyses, mutant complementation, and expression of the gene in a microsome yeast system showed that CYP72A67 is responsible for hydroxylation at the C-2 position downstream of oleanolic acid synthesis. The affinity of CYP72A67 for substrates with different substitutions at multiple carbon positions was investigated in the same in vitro yeast system, and in relation to two other CYP450s （CYP72A68） responsible for the production of medicagenic acid, the main sapogenin in M. truncatula leaves and roots. Full sib mutant and wild-type plants were compared for their sapogenin profile, expression patterns of the genes involved in sapogenin synthesis, and response to inoculation with Sinorhizobium meliloti. The results obtained allowed us to revise the hemolytic sapogenin pathway in M. truncatula and contribute to highlighting the tissue specificities （leaves/roots） of sapogenin synthesis.展开更多
The R2R3-MYB genes make up one of the largest transcription factor families in plants, and play regulatory roles in various biological processes such as development, metabolism and defense response. Although genome-wi...The R2R3-MYB genes make up one of the largest transcription factor families in plants, and play regulatory roles in various biological processes such as development, metabolism and defense response. Although genome-wide analyses of this gene family have been conducted in several species, R2R3-MYB genes have not been systematically analyzed in Medicago truncatula, a sequenced model legume plant. Here, we performed a comprehensive, genome-wide computational analysis of the structural characteristics, phylogeny, functions and expression patterns of M. truncatula R2R3-MYB genes. DNA binding domains are highly conserved among the 155 putative MtR2R3-MYB proteins that we identified. Chromosomal location analysis revealed that these genes were distributed across all eight chromosomes. Results showed that the expansion of the MtR2R3-MYB family was mainly attributable to segmental duplication and tandem duplication. A comprehensive classification was performed based on phylogenetic analysis of the R2R3-MYB gene families in M. truncatula, Arabidopsis thaliana and other plant species. Evolutionary relationships within clades were supported by clade-specific conserved motifs outside the MYB domain. Species-specific clades have been gained or lost during evolution, resulting in functional divergence. Also, tissue-specific expression patterns were investigated. The functions of stress response-related clades were further verified by the changes in transcript levels of representative R2R3-MYB genes upon treatment with abiotic and biotic stresses. This study is the first report on identification and characterization of R2R3-MYB gene family based on the genome of M. truncatula, and will facilitate functional analysis of this gene family in the future.展开更多
We identified de novo sucrose transporter （SUT） genes involved in long-distance transport of sucrose from photosynthetic source leaves towards sink organs in the model leguminous species Medicago truncatula. The ide...We identified de novo sucrose transporter （SUT） genes involved in long-distance transport of sucrose from photosynthetic source leaves towards sink organs in the model leguminous species Medicago truncatula. The iden- tification and functional analysis of sugar transporters provide key information on mechanisms that underlie carbon partitioning in plant-microorganism interactions. In that way, full-length sequences of the M. truncatula SUT （MtSUT） family were retrieved and biochemical characterization of MtSUT members was performed by heterologous expression in yeast. The MtSUT family now comprises six genes which distribute among Dicotyledonous clades. MtSUTI-1 and MtSUT4-1 are key members in regard to their expression profiles in source leaves and sink roots and were characterized as functional H~/sucrose transporters. Physiological and molecular responses to phosphorus supply and inoculation by the arbuscular mycorrhizal fungus （AMF） Glomus intraradices was studied by gene expression and sugar quantification analyses. Sucrose represents the main sugar transport form in M. truncatula and the expression profiles of MtSUTI-1, MtSUT2, and MtSUT4-1 highlight a fine-tuning regulation for beneficial sugar fluxes towards the fungal symbiont. Taken together, these results suggest distinct functions for proteins from the SUT1, SUT2, and SUT4 clades in plant and in bio- trophic interactions.展开更多
Radicle emergence and reserves mobilization are two distinct programmes that are thought to control germination. Both programs are influenced by abscissic acid （ABA） but how this hormone controls seed germination is...Radicle emergence and reserves mobilization are two distinct programmes that are thought to control germination. Both programs are influenced by abscissic acid （ABA） but how this hormone controls seed germination is still poorly known. Phenotypic and microscopic observations of the embryo axis of Medicago truncatula during germination in mitotic inhibition condition triggered by 10 μM oryzalin showed that cell division was not required to allow radicle emergence. A suppressive subtractive hybridization showed that more than 10% of up-regulated genes in the embryo axis encoded proteins related to cell-wall biosynthesis. The expression of α-expansins, pectin-esterase, xylogucan-endotransglycosidase, cellulose synthase, and extensins was monitored in the embryo axis of seeds germinated on water, constant and transitory ABA. These genes were overexpressed before completion of germination in the control and strongly inhibited by ABA. The expression was re-established in the ABA transitory-treatment after the seeds were transferred back on water and proceeded to germination. This proves these genes as contributors to the completion of germination and strengthen the idea that cell-wall loosening and remodeling in relation to cell expansion in the embryo axis is a determinant feature in germination. Our results also showed that ABA controls germination through the control of radicle emergence, namely by inhibiting cell-wall loosening and expansion.展开更多
Synonymous codon usage pattern presumably reflects gene expression optimization as a result of molecular evolution. Though much attention has been paid to various model organisms ranging from prokaryotes to eukaryotes...Synonymous codon usage pattern presumably reflects gene expression optimization as a result of molecular evolution. Though much attention has been paid to various model organisms ranging from prokaryotes to eukaryotes, codon usage has yet been extensively investigated for model legume Medicago truncatula. In present study, 39 531 available coding sequences （CDSs） from M. truncatula were examined for codon usage bias （CUB）. Based on analyses including neutrality plots, effective number of codons plots, and correlations between optimal codons frequency and codon adaptation index, we conclude that natural selection is a major driving force in M. truncatula CUB. We have identified 30 optimal codons encoding 18 amino acids based on relative synonymous codon usage. These optimal codons characteristically end with A or T, except for AGG and TTG encoding arginine and leucine respectively. Optimal codon usage is positively correlated with the GC content at three nucleotide positions of codons and the GC content of CDSs. The abundance of expressed sequence tag is a proxy for gene expression intensity in the legume, but has no relatedness with either CDS length or GC content. Collectively, we unravel the synonymous codon usage pattern in M. truncatula, which may serve as the valuable information on genetic engineering of the model legume and forage crop.展开更多
In the present study, the phosphorus-absorption capacity of transgenic Arabidopsis plants ectopically ex- pressing a novel phytase gene from Medicago truncatula Barrel Medic was evaluated. A full-length cDNA encoding ...In the present study, the phosphorus-absorption capacity of transgenic Arabidopsis plants ectopically ex- pressing a novel phytase gene from Medicago truncatula Barrel Medic was evaluated. A full-length cDNA encoding an extracellular form of phytase was isolated from the model legume M. truncatula. The phytase gene （MtPHY1） has an open reading frame of I 632 bp predicted to encode 543 amino acids, including an N- terminal signal peptide of 27 amino acids. The genomic sequence of the MtPHY1 gene is 5 151 bp, containing seven exons interrupted by six introns. Under high-Pi （2 mmol/L） growth conditions, higher levels of MtPHY1 transcripts accumulated in the leaf and stem than in the root. The transcript level was reduced in the stem and increased in the root, with no obvious changes in the hybridization signal detected in the leaf under IowPi （10 pmol/L） conditions. Chimeric transgenes were constructed by placing MtPHY1 under the control of the constitutive CaMV35S promoter and the root-specific MtPT1 promoter. Phytase activities in root apoplast of transgenic Arabidopsis were 12.3- to 16.2-fold of that in control plants. The phytase expressed was secreted into the rhizosphere, as demonstrated by HPLC analysis of phytate degradation by root exudates. Ectopic expression of MtPHY1 in Arabidopsis, leading to significant improvement in organic phosphorus absorption and plant growth, indicated that MtPHY1 has great potential for improving plant phosphorus absorption and phytoremediation.展开更多
Legume rhizobium symbiosis is initiated upon perception of bacterial secreted lipo-chitooligosaccharides （LCOs）. Perception of these signals by the plant initiates a signaling cascade that leads to nodule formation....Legume rhizobium symbiosis is initiated upon perception of bacterial secreted lipo-chitooligosaccharides （LCOs）. Perception of these signals by the plant initiates a signaling cascade that leads to nodule formation. Several studies have implicated a function for cytokinin in this process. However, whether cytokinin accu- mulation and subsequent signaling are an integral part of rhizobium LCO signaling remains elusive. Here, we show that cytokinin signaling is required for the majority of transcriptional changes induced by rhizo- bium LCOs. In addition, we demonstrate that several cytokinins accumulate in the root susceptible zone 3 h after rhizobium LCO application, including the biologically most active cytokinins, trans-zeatin and iso- pentenyl adenine. These responses are dependent on calcium- and calmodulin-dependent protein kinase （CCaMK）, a key protein in rhizobial LCO-induced signaling. Analysis of the ethylene-insensitive Mtein21 Mtsickle mutant showed that LCO-induced cytokinin accumulation is negatively regulated by ethylene. Together with transcriptional induction of ethylene biosynthesis genes, it suggests a feedback loop negatively regulating LCO signaling and subsequent cytokinin accumulation. We argue that cytokinin accumulation is a key step in the pathway leading to nodule organogenesis and that this is tightly controlled by feedback loops.展开更多
Root nodule symbiosis(RNs)between legumes and rhizobia is a major source of nitrogen in agricultural systems.Effective symbiosis requires precise regulation of plant defense responses.The role of the defense hormone j...Root nodule symbiosis(RNs)between legumes and rhizobia is a major source of nitrogen in agricultural systems.Effective symbiosis requires precise regulation of plant defense responses.The role of the defense hormone jasmonic acid(JA)in the immune response has been extensively studied.Current research shows that JA can play either a positive or negative regulatory role in RNS depending on its concentration,but the molecular mechanisms remain to be elucidated.In this study,we found that inoculation with the rhizobia Sm1021 induces the JA pathway in Medicago truncatula,and blocking the JA pathway significantly reduces the number of infection threads.Mutations in the MtMYc2 gene,which encodes a JA signaling master transcription factor,significantly inhibited rhizobia infection,terminal differentiation,and symbiotic cell formation.Combining RNA sequencing and chromatin immunoprecipitation sequencing,we discovered that MtMYc2 regulates the expression of nodule-specific MtDNF2,MtNAD1,and MtSymCRK to suppress host defense,while it activates MtDNF1 expression to regulate the maturation of MtNCRs,which in turn promotes bacteroid formation.More importantly,MtMYC2 participates in symbiotic signal transduction by promoting the expression of MtiPD3.Notably,the MtMYC2-MtiPD3 transcriptional regulatory module is specifically present in legumes,and the Mtmyc2 mutants are susceptible to the infection by the pathogen Rhizoctonia solani.Collectively,these findings reveal the molecular mechanisms of how the JA pathway regulates RNS,broadening our understanding of the roles of JA in plant-microbe interactions.展开更多
Plants are capable of regulating their shoot architecture in response to diverse internal and external environments.The circadian clock is an adaptive mechanism that integrates information from internal and ambient co...Plants are capable of regulating their shoot architecture in response to diverse internal and external environments.The circadian clock is an adaptive mechanism that integrates information from internal and ambient conditions to help plants cope with recurring environmental fluctuations.Despite the current understanding of plant circadian clock and genetic framework underlying plant shoot architecture,the intricate connection between these two adaptive mechanisms remains largely unclear.In this study,we elucidated how the core clock gene LUX ARRHYTHMO(LUX)regulates shoot architecture in the model legume plant Medicago truncatula.We show that mtlux mutant displays increased main stem height,reduced lateral shoot length,and decreased the number of lateral branches and biomass yield.Gene expression analysis revealed that Mt LUX regulated shoot architecture by repressing the expression of strigolactone receptor MtD14 and MtTB1/MtTCP1A,a TCP gene that functions centrally in modulating shoot architecture.In vivo and in vitro experiments showed that Mt LUX directly binds to a cis-element in the promoter of MtTB1/MtTCP1A,suggesting that Mt LUX regulates branching by rhythmically suppressing MtTB1/MtTCP1A.This work demonstrates the regulatory effect of the circadian clock on shoot architecture,offering a new understanding underlying the genetic basis towards the flexibility of plant shoot architecture.展开更多
Nodulation is an energy-expensive behavior driven by legumes by providing carbon sources to bacteroids and obtaining nitrogen sources in return.The energy sensor sucrose nonfermenting 1-related protein kinase 1(SnRK1)...Nodulation is an energy-expensive behavior driven by legumes by providing carbon sources to bacteroids and obtaining nitrogen sources in return.The energy sensor sucrose nonfermenting 1-related protein kinase 1(SnRK1)is the hub of energy regulation in eukaryotes.However,the molecular mechanism by which SnRK1 coordinates the allocation of energy and substances during symbiotic nitrogen fixation(SNF)remains unknown.In this study,we identified the novel legume-specific SnRK1α4,a member of the SnRK1 family that positively regulates SNF.Phenotypic analysis showed that nodule size and nitrogenase activity increased in SnRK1α4-overexpressing plants and decreased significantly in snrk1α4 mutants.We demonstrated that a key upstream kinase involved in nodulation,Does Not Make Infection 2(DMI2),can phosphorylate SnRK1α4 at Thr175 to cause its activation.Further evidence clarified that SnRK1α4 phosphorylates the malate dehydrogenases MDH1/2 to promote malate production in the cytoplasm,supplying carbon sources to bacteroids.Therefore,our findings reveal an essential role of the DMI2–SnRK1α4–MDH pathway in supplying carbon sources to bacteroids for SNF and provide a new module for constructing cereal crops with SNF.展开更多
Elevated concentrations of atmospheric CO2 can alter plant secondary metabolites,which play important roles in the interactions among plants,herbivorous insects and natural enemies.However,few studies have examined th...Elevated concentrations of atmospheric CO2 can alter plant secondary metabolites,which play important roles in the interactions among plants,herbivorous insects and natural enemies.However,few studies have examined the cascading effects of host plant secondary metabolites on tri-trophic interactions under elevated CO2(eCO2).In this study,we determined the effects of eCO2 on the growth and foliar phenolics of Medicago truncatula and the cascading effects on two color genotypes oiAcyrthosiphon pisum(pink vs.green)and their parasitoid Aphidius avenae in the field open-top chambers.Our results showed that eCO2 increased photosynthetic rate,nodule number,yield and the total phenolic content of M.truncatula.eCO2 had contrasting effects on two genotypes of A.pisum;the green genotype demonstrated increased population abundance,fecundity,growth and feeding efficiency,while the pink genotype showed decreased fitness and these were closely associated with the foliar genstein content.Furthermore,eCO2 decreased the parasitic rate of A.avenae independent of aphid genotypes.eCO2 prolonged the emergence time and reduced the emergence rate and percentage of females when associated with the green genotype,but little difference,except for increased percentage of females,was observed in A.avenae under eCO2 when associated with the pink genotype,indicating that parasitoids can perceive and discriminate the qualities of aphid hosts.We concluded that eCO2 altered plant phenolics and thus the performance of aphids and parasitoids.Our results indicate that plant phenolics vary by different abiotic and biotic stimuli and could potentially deliver the cascading effects of eCO2 to the higher trophic levels.Our results also suggest that the green genotype is expected to perform better in future eCO2 because of decreased plant resistance after its infestation and decreased parasitic rate.展开更多
Maintaining genomic integrity and stability is particularly important for stem cells,which are at the top of the cell lineage origin.Here,we discovered that the plant-specific histone methyltransferase SUVR2 maintains...Maintaining genomic integrity and stability is particularly important for stem cells,which are at the top of the cell lineage origin.Here,we discovered that the plant-specific histone methyltransferase SUVR2 maintains the genome integrity of the root tip stem cells through chromatin remodeling and liquid-liquid phase separation(LLPS)when facing DNA double-strand breaks(DSBs).The histone methyltransferase SUVR2(MtSUVR2)has histone methyltransferase activity and catalyzes the conversion of histone H3 lysine 9 monomethylation(H3K9me1)to H3K9me2/3 in vitro and in Medicago truncatula.Under DNA damage,the proportion of heterochromatin decreased and the level of DSB damage marker y-H2AX increased in suvr2 mutants,indicating that MtSUVR2 promotes the compaction of the chromatin structure through H3K9 methylation modification to protect DNA from damage.Interestingly,MtSUVR2 was induced by DSBs to phase separate and form droplets to localize at the damage sites,and this was confirmed by immunofluorescence and fluorescence recovery after photobleaching experiments.The IDR1 and lowcomplexity domain regions of MtSUVR2 determined its phase separation in the nucleus,whereas the IDR2 region determined the interaction with the homologous recombinase MtRAD51.Furthermore,we found that MtSUVR2 drove the phase separation of MtRAD51 to form"DNA repair bodies,"which could enhance the stability of MtRAD51 proteins to facilitate error-free homologous recombination repair of stem cells.Taken together,our study reveals that chromatin remodeling-associated proteins participate in DNA repair through LLPS.展开更多
Medicago truncatula is a chosen model for legumes towards deciphering fundamental legume biology,especially symbiotic nitrogen fixation.Current genomic resources for M.truncatula include a completed whole genome seque...Medicago truncatula is a chosen model for legumes towards deciphering fundamental legume biology,especially symbiotic nitrogen fixation.Current genomic resources for M.truncatula include a completed whole genome sequence information for R108 and Jemalong A17 accessions along with the sparse draft genome sequences for other 226 M.truncatula accessions.These genomic resources are complemented by the availability of mutant resources such as retrotransposon(Tnt1)insertion mutants in R108 and fast neutron bombardment(FNB)mutants in A17.In addition,several M.truncatula databases such as small secreted peptides(SSPs)database,transporter protein database,gene expression atlas,proteomic atlas,and metabolite atlas are available to the research community.This review describes these resources and provide information regarding how to access these resources.展开更多
Acute ozone is a model abiotic elicitor of oxidative stress and a useful tool for understanding biochemical and molecular events during oxidative signaling. Two Medicago truncatula accessions with contrasting response...Acute ozone is a model abiotic elicitor of oxidative stress and a useful tool for understanding biochemical and molecular events during oxidative signaling. Two Medicago truncatula accessions with contrasting responses to ozone were used to examine translational regulation during ozone stress. In ozone-resistant JE154, significant reduction in ri- bosome loading was observed within one hour of ozone treatment, suggesting energy homeostasis as a vital factor for oxidative stress management. Polysomal RNA-based expression profiling with Affymetrix arrays revealed extensive changes in the translatomes of both accessions. Messenger RNAs with low GC content in their 5＇ and 3＇-UTRs were pref- erentially associated with polysomes during oxidative stress. Genebins analysis revealed extensive changes in various gene ontologies in both accessions. Extensive changes in nicotinate and nicotinamide metabolism genes were corrobo- rated with increased levels of NAD＋ and NADH in JE154. The significantly lower NAD＋：NADH redox status in JE154, in conjunction with higher ATP amounts, provided a cellular milieu conducive for overcoming oxidative stress. Low levels of ATP, NADH, and suppression of antioxidant defense responses, abet build-up of ozone-derived ROS and ultimately lead to oxidative cell death in Jemalong.展开更多
Phytosulfokine-α(PSK-α),a sulfated pentapeptide with the sequence YIYTQ,is encoded by a small precursor gene family in Arabidopsis.PSK-αregulates multiple growth and developmental processes as a novel peptide hormo...Phytosulfokine-α(PSK-α),a sulfated pentapeptide with the sequence YIYTQ,is encoded by a small precursor gene family in Arabidopsis.PSK-αregulates multiple growth and developmental processes as a novel peptide hormone.Despite its importance,functions of PSK-αin M.truncatula growth remains unknown.In this study,we identified five genes to encode PSK-αprecursors in M.truncatula.All of these precursors possess conserved PSK-αsignature motif.Expression pattern analysis of these MtPSK genes revealed that each gene was expressed in a tissue-specific or ubiquitous pattern and three of them were remarkably expressed in root.Treatment of M.truncatula seedlings with synthetic PSK-αpeptide significantly promoted root elongation.In addition,expression analysis of downstream genes by RNA-seq and qRT-PCR assays suggested that PSK-αsignaling might regulate cell wall structure via PMEI-PME module to promote root cell growth.Taken together,our results shed light on the mechanism by which PSK-αpromotes root growth in M.truncatula,providing a new resource for improvement of root growth in agriculture.展开更多
文摘In the Medicago genus, triterpenic saponins are bioactive secondary metabolites constitutively synthesized in the aerial and subterranean parts of plants via the isoprenoid pathway. Exploitation of saponins as pharmaceutics, agrochemicals and in the food and cosmetic industries has raised interest in identifying the enzymes involved in their synthesis. We have identified a cytochrome P450 （CYP72A67） involved in hemolytic sapogenin biosynthesis by a reverse genetic TILLING approach in a Medicago truncatula ethyl- methanesulfonate （EMS） mutagenized collection. Genetic and biochemical analyses, mutant complementation, and expression of the gene in a microsome yeast system showed that CYP72A67 is responsible for hydroxylation at the C-2 position downstream of oleanolic acid synthesis. The affinity of CYP72A67 for substrates with different substitutions at multiple carbon positions was investigated in the same in vitro yeast system, and in relation to two other CYP450s （CYP72A68） responsible for the production of medicagenic acid, the main sapogenin in M. truncatula leaves and roots. Full sib mutant and wild-type plants were compared for their sapogenin profile, expression patterns of the genes involved in sapogenin synthesis, and response to inoculation with Sinorhizobium meliloti. The results obtained allowed us to revise the hemolytic sapogenin pathway in M. truncatula and contribute to highlighting the tissue specificities （leaves/roots） of sapogenin synthesis.
基金supported by the National Natural Science Foundation of China(31372362)
文摘The R2R3-MYB genes make up one of the largest transcription factor families in plants, and play regulatory roles in various biological processes such as development, metabolism and defense response. Although genome-wide analyses of this gene family have been conducted in several species, R2R3-MYB genes have not been systematically analyzed in Medicago truncatula, a sequenced model legume plant. Here, we performed a comprehensive, genome-wide computational analysis of the structural characteristics, phylogeny, functions and expression patterns of M. truncatula R2R3-MYB genes. DNA binding domains are highly conserved among the 155 putative MtR2R3-MYB proteins that we identified. Chromosomal location analysis revealed that these genes were distributed across all eight chromosomes. Results showed that the expansion of the MtR2R3-MYB family was mainly attributable to segmental duplication and tandem duplication. A comprehensive classification was performed based on phylogenetic analysis of the R2R3-MYB gene families in M. truncatula, Arabidopsis thaliana and other plant species. Evolutionary relationships within clades were supported by clade-specific conserved motifs outside the MYB domain. Species-specific clades have been gained or lost during evolution, resulting in functional divergence. Also, tissue-specific expression patterns were investigated. The functions of stress response-related clades were further verified by the changes in transcript levels of representative R2R3-MYB genes upon treatment with abiotic and biotic stresses. This study is the first report on identification and characterization of R2R3-MYB gene family based on the genome of M. truncatula, and will facilitate functional analysis of this gene family in the future.
文摘We identified de novo sucrose transporter （SUT） genes involved in long-distance transport of sucrose from photosynthetic source leaves towards sink organs in the model leguminous species Medicago truncatula. The iden- tification and functional analysis of sugar transporters provide key information on mechanisms that underlie carbon partitioning in plant-microorganism interactions. In that way, full-length sequences of the M. truncatula SUT （MtSUT） family were retrieved and biochemical characterization of MtSUT members was performed by heterologous expression in yeast. The MtSUT family now comprises six genes which distribute among Dicotyledonous clades. MtSUTI-1 and MtSUT4-1 are key members in regard to their expression profiles in source leaves and sink roots and were characterized as functional H~/sucrose transporters. Physiological and molecular responses to phosphorus supply and inoculation by the arbuscular mycorrhizal fungus （AMF） Glomus intraradices was studied by gene expression and sugar quantification analyses. Sucrose represents the main sugar transport form in M. truncatula and the expression profiles of MtSUTI-1, MtSUT2, and MtSUT4-1 highlight a fine-tuning regulation for beneficial sugar fluxes towards the fungal symbiont. Taken together, these results suggest distinct functions for proteins from the SUT1, SUT2, and SUT4 clades in plant and in bio- trophic interactions.
文摘Radicle emergence and reserves mobilization are two distinct programmes that are thought to control germination. Both programs are influenced by abscissic acid （ABA） but how this hormone controls seed germination is still poorly known. Phenotypic and microscopic observations of the embryo axis of Medicago truncatula during germination in mitotic inhibition condition triggered by 10 μM oryzalin showed that cell division was not required to allow radicle emergence. A suppressive subtractive hybridization showed that more than 10% of up-regulated genes in the embryo axis encoded proteins related to cell-wall biosynthesis. The expression of α-expansins, pectin-esterase, xylogucan-endotransglycosidase, cellulose synthase, and extensins was monitored in the embryo axis of seeds germinated on water, constant and transitory ABA. These genes were overexpressed before completion of germination in the control and strongly inhibited by ABA. The expression was re-established in the ABA transitory-treatment after the seeds were transferred back on water and proceeded to germination. This proves these genes as contributors to the completion of germination and strengthen the idea that cell-wall loosening and remodeling in relation to cell expansion in the embryo axis is a determinant feature in germination. Our results also showed that ABA controls germination through the control of radicle emergence, namely by inhibiting cell-wall loosening and expansion.
基金supported by the National Basic Research Program of China (2014CB138702)the National Natural Science Foundation of China (31502001)
文摘Synonymous codon usage pattern presumably reflects gene expression optimization as a result of molecular evolution. Though much attention has been paid to various model organisms ranging from prokaryotes to eukaryotes, codon usage has yet been extensively investigated for model legume Medicago truncatula. In present study, 39 531 available coding sequences （CDSs） from M. truncatula were examined for codon usage bias （CUB）. Based on analyses including neutrality plots, effective number of codons plots, and correlations between optimal codons frequency and codon adaptation index, we conclude that natural selection is a major driving force in M. truncatula CUB. We have identified 30 optimal codons encoding 18 amino acids based on relative synonymous codon usage. These optimal codons characteristically end with A or T, except for AGG and TTG encoding arginine and leucine respectively. Optimal codon usage is positively correlated with the GC content at three nucleotide positions of codons and the GC content of CDSs. The abundance of expressed sequence tag is a proxy for gene expression intensity in the legume, but has no relatedness with either CDS length or GC content. Collectively, we unravel the synonymous codon usage pattern in M. truncatula, which may serve as the valuable information on genetic engineering of the model legume and forage crop.
基金Supported by the Samuel Roberts Noble Foundation and the Hebei Provincial Natural Science Foundation of China （300112）.
文摘In the present study, the phosphorus-absorption capacity of transgenic Arabidopsis plants ectopically ex- pressing a novel phytase gene from Medicago truncatula Barrel Medic was evaluated. A full-length cDNA encoding an extracellular form of phytase was isolated from the model legume M. truncatula. The phytase gene （MtPHY1） has an open reading frame of I 632 bp predicted to encode 543 amino acids, including an N- terminal signal peptide of 27 amino acids. The genomic sequence of the MtPHY1 gene is 5 151 bp, containing seven exons interrupted by six introns. Under high-Pi （2 mmol/L） growth conditions, higher levels of MtPHY1 transcripts accumulated in the leaf and stem than in the root. The transcript level was reduced in the stem and increased in the root, with no obvious changes in the hybridization signal detected in the leaf under IowPi （10 pmol/L） conditions. Chimeric transgenes were constructed by placing MtPHY1 under the control of the constitutive CaMV35S promoter and the root-specific MtPT1 promoter. Phytase activities in root apoplast of transgenic Arabidopsis were 12.3- to 16.2-fold of that in control plants. The phytase expressed was secreted into the rhizosphere, as demonstrated by HPLC analysis of phytate degradation by root exudates. Ectopic expression of MtPHY1 in Arabidopsis, leading to significant improvement in organic phosphorus absorption and plant growth, indicated that MtPHY1 has great potential for improving plant phosphorus absorption and phytoremediation.
文摘Legume rhizobium symbiosis is initiated upon perception of bacterial secreted lipo-chitooligosaccharides （LCOs）. Perception of these signals by the plant initiates a signaling cascade that leads to nodule formation. Several studies have implicated a function for cytokinin in this process. However, whether cytokinin accu- mulation and subsequent signaling are an integral part of rhizobium LCO signaling remains elusive. Here, we show that cytokinin signaling is required for the majority of transcriptional changes induced by rhizo- bium LCOs. In addition, we demonstrate that several cytokinins accumulate in the root susceptible zone 3 h after rhizobium LCO application, including the biologically most active cytokinins, trans-zeatin and iso- pentenyl adenine. These responses are dependent on calcium- and calmodulin-dependent protein kinase （CCaMK）, a key protein in rhizobial LCO-induced signaling. Analysis of the ethylene-insensitive Mtein21 Mtsickle mutant showed that LCO-induced cytokinin accumulation is negatively regulated by ethylene. Together with transcriptional induction of ethylene biosynthesis genes, it suggests a feedback loop negatively regulating LCO signaling and subsequent cytokinin accumulation. We argue that cytokinin accumulation is a key step in the pathway leading to nodule organogenesis and that this is tightly controlled by feedback loops.
基金National Natural Science Foundation of China(32370253,and 32070272)the National Key Research Development Program of China(2022YFA0912100,and 2023YFF1001400).
文摘Root nodule symbiosis(RNs)between legumes and rhizobia is a major source of nitrogen in agricultural systems.Effective symbiosis requires precise regulation of plant defense responses.The role of the defense hormone jasmonic acid(JA)in the immune response has been extensively studied.Current research shows that JA can play either a positive or negative regulatory role in RNS depending on its concentration,but the molecular mechanisms remain to be elucidated.In this study,we found that inoculation with the rhizobia Sm1021 induces the JA pathway in Medicago truncatula,and blocking the JA pathway significantly reduces the number of infection threads.Mutations in the MtMYc2 gene,which encodes a JA signaling master transcription factor,significantly inhibited rhizobia infection,terminal differentiation,and symbiotic cell formation.Combining RNA sequencing and chromatin immunoprecipitation sequencing,we discovered that MtMYc2 regulates the expression of nodule-specific MtDNF2,MtNAD1,and MtSymCRK to suppress host defense,while it activates MtDNF1 expression to regulate the maturation of MtNCRs,which in turn promotes bacteroid formation.More importantly,MtMYC2 participates in symbiotic signal transduction by promoting the expression of MtiPD3.Notably,the MtMYC2-MtiPD3 transcriptional regulatory module is specifically present in legumes,and the Mtmyc2 mutants are susceptible to the infection by the pathogen Rhizoctonia solani.Collectively,these findings reveal the molecular mechanisms of how the JA pathway regulates RNS,broadening our understanding of the roles of JA in plant-microbe interactions.
基金supported by Laboratory of Lingnan Modern Agriculture Project(NZ2021001)State Key Laboratory for Conservation and Utilization of Subtropical Agrobioresources(SKICUSA-a202007)Natural Science Foundation of Guangdong Province(2022A1515011027,2019A1515012009)。
文摘Plants are capable of regulating their shoot architecture in response to diverse internal and external environments.The circadian clock is an adaptive mechanism that integrates information from internal and ambient conditions to help plants cope with recurring environmental fluctuations.Despite the current understanding of plant circadian clock and genetic framework underlying plant shoot architecture,the intricate connection between these two adaptive mechanisms remains largely unclear.In this study,we elucidated how the core clock gene LUX ARRHYTHMO(LUX)regulates shoot architecture in the model legume plant Medicago truncatula.We show that mtlux mutant displays increased main stem height,reduced lateral shoot length,and decreased the number of lateral branches and biomass yield.Gene expression analysis revealed that Mt LUX regulated shoot architecture by repressing the expression of strigolactone receptor MtD14 and MtTB1/MtTCP1A,a TCP gene that functions centrally in modulating shoot architecture.In vivo and in vitro experiments showed that Mt LUX directly binds to a cis-element in the promoter of MtTB1/MtTCP1A,suggesting that Mt LUX regulates branching by rhythmically suppressing MtTB1/MtTCP1A.This work demonstrates the regulatory effect of the circadian clock on shoot architecture,offering a new understanding underlying the genetic basis towards the flexibility of plant shoot architecture.
基金the National Key Research Development Program of China(2022YFA0912100)the National Natural Science Foundation of China(NFSC+1 种基金32070272)the major demonstration project"The Open Competition"for seed industry science and technology innovation in Inner Mongolia(2022JBGS0016).
文摘Nodulation is an energy-expensive behavior driven by legumes by providing carbon sources to bacteroids and obtaining nitrogen sources in return.The energy sensor sucrose nonfermenting 1-related protein kinase 1(SnRK1)is the hub of energy regulation in eukaryotes.However,the molecular mechanism by which SnRK1 coordinates the allocation of energy and substances during symbiotic nitrogen fixation(SNF)remains unknown.In this study,we identified the novel legume-specific SnRK1α4,a member of the SnRK1 family that positively regulates SNF.Phenotypic analysis showed that nodule size and nitrogenase activity increased in SnRK1α4-overexpressing plants and decreased significantly in snrk1α4 mutants.We demonstrated that a key upstream kinase involved in nodulation,Does Not Make Infection 2(DMI2),can phosphorylate SnRK1α4 at Thr175 to cause its activation.Further evidence clarified that SnRK1α4 phosphorylates the malate dehydrogenases MDH1/2 to promote malate production in the cytoplasm,supplying carbon sources to bacteroids.Therefore,our findings reveal an essential role of the DMI2–SnRK1α4–MDH pathway in supplying carbon sources to bacteroids for SNF and provide a new module for constructing cereal crops with SNF.
文摘Elevated concentrations of atmospheric CO2 can alter plant secondary metabolites,which play important roles in the interactions among plants,herbivorous insects and natural enemies.However,few studies have examined the cascading effects of host plant secondary metabolites on tri-trophic interactions under elevated CO2(eCO2).In this study,we determined the effects of eCO2 on the growth and foliar phenolics of Medicago truncatula and the cascading effects on two color genotypes oiAcyrthosiphon pisum(pink vs.green)and their parasitoid Aphidius avenae in the field open-top chambers.Our results showed that eCO2 increased photosynthetic rate,nodule number,yield and the total phenolic content of M.truncatula.eCO2 had contrasting effects on two genotypes of A.pisum;the green genotype demonstrated increased population abundance,fecundity,growth and feeding efficiency,while the pink genotype showed decreased fitness and these were closely associated with the foliar genstein content.Furthermore,eCO2 decreased the parasitic rate of A.avenae independent of aphid genotypes.eCO2 prolonged the emergence time and reduced the emergence rate and percentage of females when associated with the green genotype,but little difference,except for increased percentage of females,was observed in A.avenae under eCO2 when associated with the pink genotype,indicating that parasitoids can perceive and discriminate the qualities of aphid hosts.We concluded that eCO2 altered plant phenolics and thus the performance of aphids and parasitoids.Our results indicate that plant phenolics vary by different abiotic and biotic stimuli and could potentially deliver the cascading effects of eCO2 to the higher trophic levels.Our results also suggest that the green genotype is expected to perform better in future eCO2 because of decreased plant resistance after its infestation and decreased parasitic rate.
基金the Key Projects in Science and Technology of Inner Mongolia(2021ZD0031)the Opening Fund of the Key Laboratory of Forage and Endemic Crop Biology,the Ministry of Education(FECBOF2021001)+1 种基金the National Natural Science Foundation of China(NFSC32070272)for supporting this work.
文摘Maintaining genomic integrity and stability is particularly important for stem cells,which are at the top of the cell lineage origin.Here,we discovered that the plant-specific histone methyltransferase SUVR2 maintains the genome integrity of the root tip stem cells through chromatin remodeling and liquid-liquid phase separation(LLPS)when facing DNA double-strand breaks(DSBs).The histone methyltransferase SUVR2(MtSUVR2)has histone methyltransferase activity and catalyzes the conversion of histone H3 lysine 9 monomethylation(H3K9me1)to H3K9me2/3 in vitro and in Medicago truncatula.Under DNA damage,the proportion of heterochromatin decreased and the level of DSB damage marker y-H2AX increased in suvr2 mutants,indicating that MtSUVR2 promotes the compaction of the chromatin structure through H3K9 methylation modification to protect DNA from damage.Interestingly,MtSUVR2 was induced by DSBs to phase separate and form droplets to localize at the damage sites,and this was confirmed by immunofluorescence and fluorescence recovery after photobleaching experiments.The IDR1 and lowcomplexity domain regions of MtSUVR2 determined its phase separation in the nucleus,whereas the IDR2 region determined the interaction with the homologous recombinase MtRAD51.Furthermore,we found that MtSUVR2 drove the phase separation of MtRAD51 to form"DNA repair bodies,"which could enhance the stability of MtRAD51 proteins to facilitate error-free homologous recombination repair of stem cells.Taken together,our study reveals that chromatin remodeling-associated proteins participate in DNA repair through LLPS.
基金supported by the National Science Foundation USA,Plant Genome Program grants (DBI 0703285,IOS-1127155,and IOS-1733470)in part by Noble Research Institute,LLC.
文摘Medicago truncatula is a chosen model for legumes towards deciphering fundamental legume biology,especially symbiotic nitrogen fixation.Current genomic resources for M.truncatula include a completed whole genome sequence information for R108 and Jemalong A17 accessions along with the sparse draft genome sequences for other 226 M.truncatula accessions.These genomic resources are complemented by the availability of mutant resources such as retrotransposon(Tnt1)insertion mutants in R108 and fast neutron bombardment(FNB)mutants in A17.In addition,several M.truncatula databases such as small secreted peptides(SSPs)database,transporter protein database,gene expression atlas,proteomic atlas,and metabolite atlas are available to the research community.This review describes these resources and provide information regarding how to access these resources.
文摘Acute ozone is a model abiotic elicitor of oxidative stress and a useful tool for understanding biochemical and molecular events during oxidative signaling. Two Medicago truncatula accessions with contrasting responses to ozone were used to examine translational regulation during ozone stress. In ozone-resistant JE154, significant reduction in ri- bosome loading was observed within one hour of ozone treatment, suggesting energy homeostasis as a vital factor for oxidative stress management. Polysomal RNA-based expression profiling with Affymetrix arrays revealed extensive changes in the translatomes of both accessions. Messenger RNAs with low GC content in their 5＇ and 3＇-UTRs were pref- erentially associated with polysomes during oxidative stress. Genebins analysis revealed extensive changes in various gene ontologies in both accessions. Extensive changes in nicotinate and nicotinamide metabolism genes were corrobo- rated with increased levels of NAD＋ and NADH in JE154. The significantly lower NAD＋：NADH redox status in JE154, in conjunction with higher ATP amounts, provided a cellular milieu conducive for overcoming oxidative stress. Low levels of ATP, NADH, and suppression of antioxidant defense responses, abet build-up of ozone-derived ROS and ultimately lead to oxidative cell death in Jemalong.
基金This work is supported by the National Natural Science Foundation of China(31500197).
文摘Phytosulfokine-α(PSK-α),a sulfated pentapeptide with the sequence YIYTQ,is encoded by a small precursor gene family in Arabidopsis.PSK-αregulates multiple growth and developmental processes as a novel peptide hormone.Despite its importance,functions of PSK-αin M.truncatula growth remains unknown.In this study,we identified five genes to encode PSK-αprecursors in M.truncatula.All of these precursors possess conserved PSK-αsignature motif.Expression pattern analysis of these MtPSK genes revealed that each gene was expressed in a tissue-specific or ubiquitous pattern and three of them were remarkably expressed in root.Treatment of M.truncatula seedlings with synthetic PSK-αpeptide significantly promoted root elongation.In addition,expression analysis of downstream genes by RNA-seq and qRT-PCR assays suggested that PSK-αsignaling might regulate cell wall structure via PMEI-PME module to promote root cell growth.Taken together,our results shed light on the mechanism by which PSK-αpromotes root growth in M.truncatula,providing a new resource for improvement of root growth in agriculture.
