Carotenoids play an important role in many physiological processes in plants and the phytoene desaturase gene （PDS3） encodes one of the important enzymes in the carotenoid biosynthesis pathway. Here we report the id...Carotenoids play an important role in many physiological processes in plants and the phytoene desaturase gene （PDS3） encodes one of the important enzymes in the carotenoid biosynthesis pathway. Here we report the identification and analysis of a T-DNA insertion mutant of PDS3 gene. Functional complementation confirmed that both the albino and dwarfphenotypes ofthepds3 mutant resulted from functional disruption of the PDS3 gene. Chloroplast development was arrested at the proplastid stage in thepds3 mutant. Further analysis showed that high level ofphytoene was accumulated in the pds3 mutant. Addition of exogenous GA3 could partially rescue the dwarf phenotype, suggesting that the dwarf phenotype ofthepds3 mutant might be due to GA deficiency. Microarray and RT-PCR analysis showed that disrupting PDS3 gene resulted in gene expression changes involved in at least 20 metabolic pathways, including the inhibition of many genes in carotenoid, chlorophyll, and GA biosynthesis pathways. Our data suggest that the accumulated phytoene in the pds3 mutant might play an important role in certain negative feedbacks to affect gene expression of diverse cellular pathways.展开更多
1-Deoxy-D-xylulose-5-phosphate reductoisomerase （DXR） is an important enzyme involved in the 2-C-methyi-D- erythritol-4-phosphate （MEP） pathway which provides the basic five-carbon units for isoprenoid biosynthesi...1-Deoxy-D-xylulose-5-phosphate reductoisomerase （DXR） is an important enzyme involved in the 2-C-methyi-D- erythritol-4-phosphate （MEP） pathway which provides the basic five-carbon units for isoprenoid biosynthesis. To investigate the role of the MEP pathway in plant development and metabolism, we carried out detailed analyses on a dxr mutant （GK_215C01） and two DXR transgenic co-suppression fines, OX-DXR-L2 and OX-DXR-L7. We found that the dxr mutant was albino and dwarf. It never bolted, had significantly reduced number of trichomes and most of the stomata could not close normally in the leaves. The two co-suppression lines produced more yellow inflorescences and albino sepals with no trichomes. The transcription levels of genes involved in tricbome initiation were found to be strongly affected, including GLABRA1, TRANSPARENT TESTA GLABROUS 1, TRIPTYCHON and SPINDLY, expression of which is regulated by gibberellic acids （GAs）. Exogenous application of GA3 could partially rescue the dwarf phenotype and the trichome initiation of dxr, whereas exogenous application of abscisic acid （ABA） could rescue the stomata closure defect, suggesting that lower levels of both GA and ABA contribute to the phenotype in the dxr mutants. We further found that genes involved in the biosynthetic pathways of GA and ABA were coordinately regulated. These results indicate that disruption of the plastidial MEP pathway leads to biosynthetic deficiency of photosynthetic pigments, GAs and ABA, and thus the developmental abnormalities, and that the flux from the cytoplasmic mevalonate pathway is not sufficient to rescue the deficiency caused by the blockage of the plastidial MEP pathway. These results reveal a critical role for the MEP biosynthetic pathway in controlling the biosynthesis of isoprenoids.展开更多
Pollen germination on the surface of compatible stigmatic tissues is an essential step for plant fertilization. Here we report that the Arabidopsis mutant bcll is male sterile as a result of the failure ofpollen germi...Pollen germination on the surface of compatible stigmatic tissues is an essential step for plant fertilization. Here we report that the Arabidopsis mutant bcll is male sterile as a result of the failure ofpollen germination. We show that the bcll mutant allele cannot be transmitted by male gametophytes and no homozygous bcll mutants were obtained. Analysis of pollen developmental stages indicates that the bcll mutation affects pollen germination but not pollen maturation. Molecular analysis demonstrates that the failure of pollen germination was caused by the disruption of AtBECLIN 1. AtBECLIN 1 is expressed predominantly in mature pollen and encodes a protein with significant homology to Beclin1/Atg6/Vps30 required for the processes of autophagy and vacuolar protein sorting （VPS） in yeast. We also show that AtBECLIN 1 is required for normal plant development, and that genes related to autophagy, VPS and the glycosylphosphatidylinositol anchor system, were affected by the deficiency of AtBECLIN 1.展开更多
VPS 15 protein is a component of the phosphatidylinositol 3-kinase complex which plays a pivotal role in the development of yeast and mammalian cells. The knowledge about the function of its homologue in plants remain...VPS 15 protein is a component of the phosphatidylinositol 3-kinase complex which plays a pivotal role in the development of yeast and mammalian cells. The knowledge about the function of its homologue in plants remains limited. Here we report that AtVPS15, a homologue of yeast VPS15p in Arabidopsis, plays an essential role in pollen germination. Homozygous T-DNA insertion mutants of AtVPS15 could not be obtained from the progenies of self-pollinated heterozygous mutants. Reciprocal crosses between atvps15 mutants and wild-type Arabidopsis revealed that the T-DNA insertion was not able to be transmitted by male gametophytes. DAPI staining, Alexander＇s stain and scanning electron microscopic analysis showed that atvps15 heterozygous plants produced pollen grains that were morphologically indistinguishable from wild-type pollen, whereas in vitro germination experiments revealed that germination of the pollen grains was defective. GUS staining analysis of transgenic plants expressing the GUS reporter gene driven by the AtVPS15 promoter showed that AtVPS15 was mainly expressed in pollen grains. Finally, DUALmembrane yeast two-hybrid analysis demonstrated that AtVPS15 might interact directly with AtVPS34. These results suggest that AtVPS15 is very important for pollen germination, possibly through modulation of the activity of PI3-kinase.展开更多
Complex I （the NADH：ubiquinone oxidoreductase） of the mitochondrial respiratory chain is a complicated, multi-subunit, membrane- bound assembly and contains more than 40 different proteins in higher plants. In this...Complex I （the NADH：ubiquinone oxidoreductase） of the mitochondrial respiratory chain is a complicated, multi-subunit, membrane- bound assembly and contains more than 40 different proteins in higher plants. In this paper, we characterize the Arabidopsis homologue （designated as AtCIB22） of the B22 subunit of eukaryotic mitochondriai Complex I. AtCIB22 is a single-copy gene and is highly con- served throughout eukaryotes. AtCIB22 protein is located in mitochondria and the AtC1B22 gene is widely expressed in different tissues. Mutant Arabidopsis plants with a disrupted AtC1B22 gene display pleiotropic phenotypes including shorter roots, smaller plants and de- layed flowering. Stress analysis indicates that the AtC1B22 mutants＇ seed germination and early seedling growth are severely inhibited by sucrose deprivation stress but more tolerant to ethanol stress. Molecular analysis reveals that in moderate knockdown AtCIB22 mutants, genes including cell redox proteins and stress related proteins are significantly up-regulated, and that in severe knockdown AtCIB22 mu- tants, the alternative respiratory pathways including NDA1, NDB2, AOXla and AtPUMP1 are remarkably elevated. These data demon- strate that AtCIB22 is essential for plant development and mitochondrial electron transport chains in Arabidopsis. Our findings also en- hance our understanding about the physiological role of Complex I in plants.展开更多
In plants, the meristem has to maintain a separate population of pluripotent cells that serve two main tasks, i.e., self-maintenance and organ initiation, which are separated spatially in meristem. Prior to our study,...In plants, the meristem has to maintain a separate population of pluripotent cells that serve two main tasks, i.e., self-maintenance and organ initiation, which are separated spatially in meristem. Prior to our study, WUS and WUS.like WOX genes had been reported as essential for the development of the SAM. In this study, the consequences of gain of WOX1 function are described. Here we report the identification of an Arabidopsis gain-of-function mutant woxl-D, in which the expression level of the WOX1 （WUSCHEL HOMEOBOX 1） was elevated and subtle defects in meristem development were observed. The woxl-D mutant phenotype is dwarfed and slightly bushy, with a smaller shoot apex. The woxl-D mutant also produced small and dark green leaves, and exhibited a failure in anther dehiscence and male sterility. Molecular evidences showed that the transcription of the stem cell marker gene CLV3 was down-regulated in the meristem of woxl-D but accumulated in the other regions, i.e., in the root-hypocotyl junction and at the sites for lateral root initiation. The fact that the organ size and cell size in leaves of woxl-D are smaller than those in wild type suggests that cell expansion is possibly affected in order to have partially retarded the development of lateral organs, possibly through alteration of CLV3 expression pattern in the meristem. An S-adenosylmethionine decarboxylase （SAMDC） protein, SAMDC1, was found able to interact with WOX1 by yeast two-hybrid and pull-down assays in vitro. HPLC analysis revealed a significant reduction of polyamine content in woxl-D. Our results suggest that WOX1 plays an important role in meristem development in Arabidopsis, possibly via regulation of SAMDC activity and polyamine homeostasis, and/or by regulating CLV3 expression.展开更多
Plant architecture is an important factor for crop production. Some members of microRNA156 (miR156) and their target genes SQUAMOSA Promoter-Binding Protein-Like (SPL) were identified to play essential roles in the es...Plant architecture is an important factor for crop production. Some members of microRNA156 (miR156) and their target genes SQUAMOSA Promoter-Binding Protein-Like (SPL) were identified to play essential roles in the establishment of plant architecture. However, the roles and regulation of miR156 is not well understood yet. Here, we identified a T-DNA insertion mutant Osmtd1 (Oryza sativa multi-tillering and dwarf mutant). Osmtd1 produced more tillers and displayed short stature phenotype. We determined that the dramatic morphological changes were caused by a single T-DNA insertion in Osmtd1. Further analysis revealed that the T-DNA insertion was located in the gene Os08g34258 encoding a putative inhibitor I family protein. Os08g34258 was knocked out and OsmiR156f was significantly upregulated in Osmtd1. Overexpression of Os08g34258 in Osmtd1 complemented the defects of the mutant architecture, while overexpression of OsmiR156f in wild-type rice phenocopied Osmtd1. We showed that the expression of OsSPL3, OsSPL12, and OsSPL14 were significantly downregulated in Osmtd1 or OsmiR156f overexpressed lines, indicating that OsSPL3, OsSPL12, and OsSPL14 were possibly direct target genes of OsmiR156f. Our results suggested that OsmiR156f controlled plant architecture by mediating plant stature and tiller outgrowth and may be regulated by an unknown protease inhibitor I family protein.展开更多
Green petals pose a challenge for pollinators to distinguish flowers from leaves,but they are valuable as a specialty flower trait.However,little is understood about the molecular mechanisms that underlie the developm...Green petals pose a challenge for pollinators to distinguish flowers from leaves,but they are valuable as a specialty flower trait.However,little is understood about the molecular mechanisms that underlie the development of green petals.Here,we report that CINCINNATA(CIN)-like TEOSINTE BRANCHED 1/CYCLOIDEA/PCF(TCP)proteins play key roles in the control of petal color.The septuple tcp2/3/4/5/10/13/17 mutant produced flowers with green petals due to chlorophyll accumulation.Expression of TCP4 complemented the petal phenotype of tcp2/3/4/5/10/13/17.We found that chloroplasts were converted into leucoplasts in the distal parts of wild-type petals but not in the proximal parts during flower development,whereas plastid conversion was compromised in the distal parts of tcp2/3/4/5/10/13/17 petals.TCP4 and most CIN-like TCPs were predominantly expressed in distal petal regions,consistent with the green–white pattern in wild-type petals and the petal greening observed in the distal parts of tcp2/3/4/5/10/13/17 petals.RNA-sequencing data revealed that most chlorophyll biosynthesis genes were downregulated in the white distal parts of wild-type petals,but these genes had elevated expression in the distal green parts of tcp2/3/4/5/10/13/17 petals and the green proximal parts of wild-type petals.We revealed that TCP4 repressed chlorophyll biosynthesis by directly binding to the promoters of PROTOCHLOROPHYLLIDE REDUCTASE(PORB),DIVINYL REDUCTASE(DVR),and SUPPRESSOR OF OVEREXPRESSION OF CO 1(SOC1),which are known to promote petal greening.We found that the conversion of chloroplasts to leucoplasts and the green coloration in the proximal parts of petals appeared to be conserved among plant species.Our findings uncover a major molecular mechanism that underpins the formation of petal color patterns and provide a foundation for the breeding of plants with green flowers.展开更多
Increases in recorded high temperatures around the world are causing plant thermomorphogenesis and decreasing crop productivity.PHYTOCHROME INTERACTING FACTOR 4(PIF4)is a central positive regulator of plant thermomorp...Increases in recorded high temperatures around the world are causing plant thermomorphogenesis and decreasing crop productivity.PHYTOCHROME INTERACTING FACTOR 4(PIF4)is a central positive regulator of plant thermomorphogenesis.However,the molecular mechanisms underlying PIF4-regulated thermomorphogenesis remain largely unclear.In this study,we identified ABNORMAL THERMOMORPHOGENESIS 1(ABT1)as an important negative regulator of PIF4 and plant thermomorphogenesis.Overexpression of ABT1 in the activation tagging mutant abt1-D caused shorter hypocotyls and petioles under moderately high temperature(HT).ABT1 encodes WRKY14,which belongs to subgroup II of the WRKY transcription factors.Overexpression of ABT1/WRKY14 or its close homologs,including ABT2/WRKY35,ABT3/WRKY65,and ABT4/WRKY69in transgenic plants caused insensitivity to HT,whereas the quadruple mutant abt1 abt2 abt3 abt4 exhibited greater sensitivity to HT.ABTs were expressed in hypocotyls,cotyledons,shoot apical meristems,and leaves,but their expression were suppressed by HT.Biochemical assays showed that ABT1 can interact with TCP5,a known positive regulator of PIF4,and interrupt the formation of the TCP5-PIF4 complex and repress its transcriptional activation activity.Genetic analysis showed that ABT1 functioned antagonistically with TCP5,BZR1,and PIF4 in plant thermomorphogenesis.Taken together,our results identify ABT1/WRKY14 as a critical repressor of plant thermomorphogenesis and suggest that ABT1/WRKY14,TCP5,and PIF4 may form a sophisticated regulatory module to fine-tune PIF4 activity and temperature-dependent plant growth.展开更多
Salicylic acid methyltransferase （SAMT）, benzoic acid methyltransferase （BAMT） and theobromine methyltransferase （TH） （henceforth, SABATH） family proteins belong to a unique class of mehtyltransferase that can...Salicylic acid methyltransferase （SAMT）, benzoic acid methyltransferase （BAMT） and theobromine methyltransferase （TH） （henceforth, SABATH） family proteins belong to a unique class of mehtyltransferase that can methylate small molecular compounds Including indole-3-acidic acid （IAA）, salicylic acid （SA） and jasmonic acid （JA）, in plants. Here we report that the GAMT2 protein, which has 34.2% similarity with IAMT1 in the amino acid sequence, can methylate gibberellic acid （GA）. Biolnformatics analysis suggests that GAMT2 may be able to methylate one molecule larger than SA. GAMT2 is predominantly expressed in the developing seed embryo and endosperm in Arabidopsis. During seed germination, the expression of GAMT2 decreases until the cotyledons expand out of the seed coat. Overexpression of GAMT2 in Arabidopsis resulted in multiple phenotypes, including dwarfism, retarded growth, late flowering, and reduced fertility, which are similar to the phenotypes of GA-deficient mutants. Seed germination assay showed that GAMT2 overexpression in plants was hypersensitive to GA biosynthesis inhibitor （ancymidol） and abscisic acid （ABA） treatments, whereas the GAMT2 null mutant （SALK_075450） was slightly Insensitive to such treatments, suggesting that GAMT2 may methylate GA or ABA. Enzyme activity analysis indicated that GAMT2 was able to methylate GA3 into Methyi-GA3 in vitro, but could not methylate ABA. Microarray analysis on GAMT2 overexpression plants suggested that Methyl-GA may be an Inactive form of GA in Arabidopsis. These data suggest that GAMT2 Is Involved in seed maturation and germination by modulating GA activity.展开更多
基金the National Natural Science Foundation of China （Grant No. 30470172）.
文摘Carotenoids play an important role in many physiological processes in plants and the phytoene desaturase gene （PDS3） encodes one of the important enzymes in the carotenoid biosynthesis pathway. Here we report the identification and analysis of a T-DNA insertion mutant of PDS3 gene. Functional complementation confirmed that both the albino and dwarfphenotypes ofthepds3 mutant resulted from functional disruption of the PDS3 gene. Chloroplast development was arrested at the proplastid stage in thepds3 mutant. Further analysis showed that high level ofphytoene was accumulated in the pds3 mutant. Addition of exogenous GA3 could partially rescue the dwarf phenotype, suggesting that the dwarf phenotype ofthepds3 mutant might be due to GA deficiency. Microarray and RT-PCR analysis showed that disrupting PDS3 gene resulted in gene expression changes involved in at least 20 metabolic pathways, including the inhibition of many genes in carotenoid, chlorophyll, and GA biosynthesis pathways. Our data suggest that the accumulated phytoene in the pds3 mutant might play an important role in certain negative feedbacks to affect gene expression of diverse cellular pathways.
基金Acknowledgments This work was supported by the National Natural Science Foundation of China （NSFC Grant 90717003 to L-J Qu）.
文摘1-Deoxy-D-xylulose-5-phosphate reductoisomerase （DXR） is an important enzyme involved in the 2-C-methyi-D- erythritol-4-phosphate （MEP） pathway which provides the basic five-carbon units for isoprenoid biosynthesis. To investigate the role of the MEP pathway in plant development and metabolism, we carried out detailed analyses on a dxr mutant （GK_215C01） and two DXR transgenic co-suppression fines, OX-DXR-L2 and OX-DXR-L7. We found that the dxr mutant was albino and dwarf. It never bolted, had significantly reduced number of trichomes and most of the stomata could not close normally in the leaves. The two co-suppression lines produced more yellow inflorescences and albino sepals with no trichomes. The transcription levels of genes involved in tricbome initiation were found to be strongly affected, including GLABRA1, TRANSPARENT TESTA GLABROUS 1, TRIPTYCHON and SPINDLY, expression of which is regulated by gibberellic acids （GAs）. Exogenous application of GA3 could partially rescue the dwarf phenotype and the trichome initiation of dxr, whereas exogenous application of abscisic acid （ABA） could rescue the stomata closure defect, suggesting that lower levels of both GA and ABA contribute to the phenotype in the dxr mutants. We further found that genes involved in the biosynthetic pathways of GA and ABA were coordinately regulated. These results indicate that disruption of the plastidial MEP pathway leads to biosynthetic deficiency of photosynthetic pigments, GAs and ABA, and thus the developmental abnormalities, and that the flux from the cytoplasmic mevalonate pathway is not sufficient to rescue the deficiency caused by the blockage of the plastidial MEP pathway. These results reveal a critical role for the MEP biosynthetic pathway in controlling the biosynthesis of isoprenoids.
基金The work was supported by the National Natural Science Foundation of China(Grant No.30470172 and 30221120261)the National Special Projects for R&D of Transgenic Plants(J99-A-001).
文摘Pollen germination on the surface of compatible stigmatic tissues is an essential step for plant fertilization. Here we report that the Arabidopsis mutant bcll is male sterile as a result of the failure ofpollen germination. We show that the bcll mutant allele cannot be transmitted by male gametophytes and no homozygous bcll mutants were obtained. Analysis of pollen developmental stages indicates that the bcll mutation affects pollen germination but not pollen maturation. Molecular analysis demonstrates that the failure of pollen germination was caused by the disruption of AtBECLIN 1. AtBECLIN 1 is expressed predominantly in mature pollen and encodes a protein with significant homology to Beclin1/Atg6/Vps30 required for the processes of autophagy and vacuolar protein sorting （VPS） in yeast. We also show that AtBECLIN 1 is required for normal plant development, and that genes related to autophagy, VPS and the glycosylphosphatidylinositol anchor system, were affected by the deficiency of AtBECLIN 1.
基金supported by the National Basic Research Program of China(No.2009CB941502)
文摘VPS 15 protein is a component of the phosphatidylinositol 3-kinase complex which plays a pivotal role in the development of yeast and mammalian cells. The knowledge about the function of its homologue in plants remains limited. Here we report that AtVPS15, a homologue of yeast VPS15p in Arabidopsis, plays an essential role in pollen germination. Homozygous T-DNA insertion mutants of AtVPS15 could not be obtained from the progenies of self-pollinated heterozygous mutants. Reciprocal crosses between atvps15 mutants and wild-type Arabidopsis revealed that the T-DNA insertion was not able to be transmitted by male gametophytes. DAPI staining, Alexander＇s stain and scanning electron microscopic analysis showed that atvps15 heterozygous plants produced pollen grains that were morphologically indistinguishable from wild-type pollen, whereas in vitro germination experiments revealed that germination of the pollen grains was defective. GUS staining analysis of transgenic plants expressing the GUS reporter gene driven by the AtVPS15 promoter showed that AtVPS15 was mainly expressed in pollen grains. Finally, DUALmembrane yeast two-hybrid analysis demonstrated that AtVPS15 might interact directly with AtVPS34. These results suggest that AtVPS15 is very important for pollen germination, possibly through modulation of the activity of PI3-kinase.
基金supported by the National Basic Research Program of China (No. 2009CB941503)
文摘Complex I （the NADH：ubiquinone oxidoreductase） of the mitochondrial respiratory chain is a complicated, multi-subunit, membrane- bound assembly and contains more than 40 different proteins in higher plants. In this paper, we characterize the Arabidopsis homologue （designated as AtCIB22） of the B22 subunit of eukaryotic mitochondriai Complex I. AtCIB22 is a single-copy gene and is highly con- served throughout eukaryotes. AtCIB22 protein is located in mitochondria and the AtC1B22 gene is widely expressed in different tissues. Mutant Arabidopsis plants with a disrupted AtC1B22 gene display pleiotropic phenotypes including shorter roots, smaller plants and de- layed flowering. Stress analysis indicates that the AtC1B22 mutants＇ seed germination and early seedling growth are severely inhibited by sucrose deprivation stress but more tolerant to ethanol stress. Molecular analysis reveals that in moderate knockdown AtCIB22 mutants, genes including cell redox proteins and stress related proteins are significantly up-regulated, and that in severe knockdown AtCIB22 mu- tants, the alternative respiratory pathways including NDA1, NDB2, AOXla and AtPUMP1 are remarkably elevated. These data demon- strate that AtCIB22 is essential for plant development and mitochondrial electron transport chains in Arabidopsis. Our findings also en- hance our understanding about the physiological role of Complex I in plants.
基金supported by the National Natural Science Foundation of China(90717003 and 30625002 to L.-J. Q.)partially by the 111 Project
文摘In plants, the meristem has to maintain a separate population of pluripotent cells that serve two main tasks, i.e., self-maintenance and organ initiation, which are separated spatially in meristem. Prior to our study, WUS and WUS.like WOX genes had been reported as essential for the development of the SAM. In this study, the consequences of gain of WOX1 function are described. Here we report the identification of an Arabidopsis gain-of-function mutant woxl-D, in which the expression level of the WOX1 （WUSCHEL HOMEOBOX 1） was elevated and subtle defects in meristem development were observed. The woxl-D mutant phenotype is dwarfed and slightly bushy, with a smaller shoot apex. The woxl-D mutant also produced small and dark green leaves, and exhibited a failure in anther dehiscence and male sterility. Molecular evidences showed that the transcription of the stem cell marker gene CLV3 was down-regulated in the meristem of woxl-D but accumulated in the other regions, i.e., in the root-hypocotyl junction and at the sites for lateral root initiation. The fact that the organ size and cell size in leaves of woxl-D are smaller than those in wild type suggests that cell expansion is possibly affected in order to have partially retarded the development of lateral organs, possibly through alteration of CLV3 expression pattern in the meristem. An S-adenosylmethionine decarboxylase （SAMDC） protein, SAMDC1, was found able to interact with WOX1 by yeast two-hybrid and pull-down assays in vitro. HPLC analysis revealed a significant reduction of polyamine content in woxl-D. Our results suggest that WOX1 plays an important role in meristem development in Arabidopsis, possibly via regulation of SAMDC activity and polyamine homeostasis, and/or by regulating CLV3 expression.
基金supported by the National Natural Science Foundation of China (no. 91317312 and 91117006)Open Foundation Project for Hunan Provincial Higher Institutional Innovation Platform (no. 09K052)Hunan Provincial Key Laboratory for Crop Germplasm Innovation and Utilization (no. 12KFXM05)
文摘Plant architecture is an important factor for crop production. Some members of microRNA156 (miR156) and their target genes SQUAMOSA Promoter-Binding Protein-Like (SPL) were identified to play essential roles in the establishment of plant architecture. However, the roles and regulation of miR156 is not well understood yet. Here, we identified a T-DNA insertion mutant Osmtd1 (Oryza sativa multi-tillering and dwarf mutant). Osmtd1 produced more tillers and displayed short stature phenotype. We determined that the dramatic morphological changes were caused by a single T-DNA insertion in Osmtd1. Further analysis revealed that the T-DNA insertion was located in the gene Os08g34258 encoding a putative inhibitor I family protein. Os08g34258 was knocked out and OsmiR156f was significantly upregulated in Osmtd1. Overexpression of Os08g34258 in Osmtd1 complemented the defects of the mutant architecture, while overexpression of OsmiR156f in wild-type rice phenocopied Osmtd1. We showed that the expression of OsSPL3, OsSPL12, and OsSPL14 were significantly downregulated in Osmtd1 or OsmiR156f overexpressed lines, indicating that OsSPL3, OsSPL12, and OsSPL14 were possibly direct target genes of OsmiR156f. Our results suggested that OsmiR156f controlled plant architecture by mediating plant stature and tiller outgrowth and may be regulated by an unknown protease inhibitor I family protein.
基金supported by the National Science Fund for Distinguished Young Scholars of China(grant 31725005)the Science Fund for the Creative Research Groups of the National Natural Science Foundation of China(grant 31621001)the National Key R&D Program of China(2018YFE0204700).
文摘Green petals pose a challenge for pollinators to distinguish flowers from leaves,but they are valuable as a specialty flower trait.However,little is understood about the molecular mechanisms that underlie the development of green petals.Here,we report that CINCINNATA(CIN)-like TEOSINTE BRANCHED 1/CYCLOIDEA/PCF(TCP)proteins play key roles in the control of petal color.The septuple tcp2/3/4/5/10/13/17 mutant produced flowers with green petals due to chlorophyll accumulation.Expression of TCP4 complemented the petal phenotype of tcp2/3/4/5/10/13/17.We found that chloroplasts were converted into leucoplasts in the distal parts of wild-type petals but not in the proximal parts during flower development,whereas plastid conversion was compromised in the distal parts of tcp2/3/4/5/10/13/17 petals.TCP4 and most CIN-like TCPs were predominantly expressed in distal petal regions,consistent with the green–white pattern in wild-type petals and the petal greening observed in the distal parts of tcp2/3/4/5/10/13/17 petals.RNA-sequencing data revealed that most chlorophyll biosynthesis genes were downregulated in the white distal parts of wild-type petals,but these genes had elevated expression in the distal green parts of tcp2/3/4/5/10/13/17 petals and the green proximal parts of wild-type petals.We revealed that TCP4 repressed chlorophyll biosynthesis by directly binding to the promoters of PROTOCHLOROPHYLLIDE REDUCTASE(PORB),DIVINYL REDUCTASE(DVR),and SUPPRESSOR OF OVEREXPRESSION OF CO 1(SOC1),which are known to promote petal greening.We found that the conversion of chloroplasts to leucoplasts and the green coloration in the proximal parts of petals appeared to be conserved among plant species.Our findings uncover a major molecular mechanism that underpins the formation of petal color patterns and provide a foundation for the breeding of plants with green flowers.
基金National Science Fund for Distinguished Young Scholars of China(grant no.31725005)National Natural Science Foundation of China(grant no.31970194)National Key Research and Development Program of China(2017YFA0503800).
文摘Increases in recorded high temperatures around the world are causing plant thermomorphogenesis and decreasing crop productivity.PHYTOCHROME INTERACTING FACTOR 4(PIF4)is a central positive regulator of plant thermomorphogenesis.However,the molecular mechanisms underlying PIF4-regulated thermomorphogenesis remain largely unclear.In this study,we identified ABNORMAL THERMOMORPHOGENESIS 1(ABT1)as an important negative regulator of PIF4 and plant thermomorphogenesis.Overexpression of ABT1 in the activation tagging mutant abt1-D caused shorter hypocotyls and petioles under moderately high temperature(HT).ABT1 encodes WRKY14,which belongs to subgroup II of the WRKY transcription factors.Overexpression of ABT1/WRKY14 or its close homologs,including ABT2/WRKY35,ABT3/WRKY65,and ABT4/WRKY69in transgenic plants caused insensitivity to HT,whereas the quadruple mutant abt1 abt2 abt3 abt4 exhibited greater sensitivity to HT.ABTs were expressed in hypocotyls,cotyledons,shoot apical meristems,and leaves,but their expression were suppressed by HT.Biochemical assays showed that ABT1 can interact with TCP5,a known positive regulator of PIF4,and interrupt the formation of the TCP5-PIF4 complex and repress its transcriptional activation activity.Genetic analysis showed that ABT1 functioned antagonistically with TCP5,BZR1,and PIF4 in plant thermomorphogenesis.Taken together,our results identify ABT1/WRKY14 as a critical repressor of plant thermomorphogenesis and suggest that ABT1/WRKY14,TCP5,and PIF4 may form a sophisticated regulatory module to fine-tune PIF4 activity and temperature-dependent plant growth.
基金Supported by the National Natural Science Foundation of China （GN 30625002） to LJ Qu.
文摘Salicylic acid methyltransferase （SAMT）, benzoic acid methyltransferase （BAMT） and theobromine methyltransferase （TH） （henceforth, SABATH） family proteins belong to a unique class of mehtyltransferase that can methylate small molecular compounds Including indole-3-acidic acid （IAA）, salicylic acid （SA） and jasmonic acid （JA）, in plants. Here we report that the GAMT2 protein, which has 34.2% similarity with IAMT1 in the amino acid sequence, can methylate gibberellic acid （GA）. Biolnformatics analysis suggests that GAMT2 may be able to methylate one molecule larger than SA. GAMT2 is predominantly expressed in the developing seed embryo and endosperm in Arabidopsis. During seed germination, the expression of GAMT2 decreases until the cotyledons expand out of the seed coat. Overexpression of GAMT2 in Arabidopsis resulted in multiple phenotypes, including dwarfism, retarded growth, late flowering, and reduced fertility, which are similar to the phenotypes of GA-deficient mutants. Seed germination assay showed that GAMT2 overexpression in plants was hypersensitive to GA biosynthesis inhibitor （ancymidol） and abscisic acid （ABA） treatments, whereas the GAMT2 null mutant （SALK_075450） was slightly Insensitive to such treatments, suggesting that GAMT2 may methylate GA or ABA. Enzyme activity analysis indicated that GAMT2 was able to methylate GA3 into Methyi-GA3 in vitro, but could not methylate ABA. Microarray analysis on GAMT2 overexpression plants suggested that Methyl-GA may be an Inactive form of GA in Arabidopsis. These data suggest that GAMT2 Is Involved in seed maturation and germination by modulating GA activity.
