Although AGAMOUS-LIKE6 (AGL6) MADS-box genes are ancient with wide distributions in gymnosperms and angiosperms, their functions remain poorly understood. Here, we show the biological role of the AGL6-1ike gene, OsMAD...Although AGAMOUS-LIKE6 (AGL6) MADS-box genes are ancient with wide distributions in gymnosperms and angiosperms, their functions remain poorly understood. Here, we show the biological role of the AGL6-1ike gene, OsMADS6, in specifying floral organ and meristem identities in rice (Oryza sativa L.). OsMADS6 was strongly ex- pressed in the floral meristem at early stages. Subsequently, OsMADS6 transcripts were mainly detectable in paleas, lodicules, carpels and the integument of ovule, as well as in the receptacle. Compared to wild type plants, osmads6 mutants displayed altered palea identity, extra glume-like or mosaic organs, abnormal carpel development and loss of floral meristem determinacy. Strikingly, mutation of a SEPALLATA (SEP)-like gene, OsMADS1 (LHS1), enhanced the defect of osmads6 flowers, and no inner floral organs or glume-like structures were observed in whorls 2 and 3 of osmadsl-z osmads6-1 flowers. Furthermore, the osmadsl-z osmads6-1 double mutants developed severely indetermi- nate floral meristems. Our finding, therefore, suggests that the ancient OsMADS6 gene is able to specify "floral state" by determining floral organ and meristem identities in monocot crop rice together with OsMADS1.展开更多
The floral-organ-number mutant fon（t） was firstly discovered in the progeny of a cross between a diploid （Chunjiang 683） and a haploid （SARⅣ/-620-A） rice cultivar. The fon（t） mutant showed normal vegetative d...The floral-organ-number mutant fon（t） was firstly discovered in the progeny of a cross between a diploid （Chunjiang 683） and a haploid （SARⅣ/-620-A） rice cultivar. The fon（t） mutant showed normal vegetative development and produced normal inflorescence structures. Difference between the mutant and the wild type was observed when the stamen primordia began to form. The mature flowers offon（t） mutant showed open-hull phenotypes, which resulted in the exposure of stamens and stigmas. Normally, a single fon（t） floret consisted of six to nine stamens and one or two pistils. In addition, stamen/pistil-like structures and bulged tissues near ovaries were also observed in a few fon（t） florets. But homeotic transformation of lodicules into palea/lemma-like organs was observed almost in all the open-hull florets. The phenotypes offon（t） flowers also suggested thatfon（t） gene might affect flower organ identity in the inner whorls. Genetic analysis showed that thefon（t） mutant was controlled by a single recessive gene.展开更多
During organ development, many key regulators have been identified in plant genomes, which play a conserved role among plant species to control the organ identities and/or determine the organ size and shape. It is int...During organ development, many key regulators have been identified in plant genomes, which play a conserved role among plant species to control the organ identities and/or determine the organ size and shape. It is intriguing whether these key regulators can acquire diverse function and be integrated into different molecular pathways among different species, giving rise to the immense diversity of organ forms in nature. In this study, we have characterized and cloned LATHYROIDES （LATH）, a classical locus in pea, whose mutation displays pleiotropic alteration of lateral growth of organs and predominant effects on tendril and dorsal petal development. LATH encodes a WUSCHEL-related home- oboxl （WOX1） transcription factor, which has a conserved function in determining organ lateral growth among different plant species. Furthermore, we showed that LATH regulated the expression level of TENDRIL-LESS （TL）, a key factor in the control of tendril development in compound leaf, and LATH genetically interacted with LOBED STANDARD （LST）, a floral dorsal factor, to affect the dorsal petal identity. Thus, LATH plays multiple roles during organ development in pea： it maintains a conserved function controlling organ lateral outgrowth, and modulates organ identities in compound leaf and zygomorphic flower development, respectively. Our data indicated that a key regulator can play important roles in different aspects of organ development and dedicate to the complexity of the molecular mechanism in the control of organ development so as to create distinct organ forms in different species.展开更多
Flower organ identity in rice is mainly determined by the A-,B-,C-and E-class genes,with the majority encoding MADS-box transcription factors.However,few studies have investigated how the expression of these floral or...Flower organ identity in rice is mainly determined by the A-,B-,C-and E-class genes,with the majority encoding MADS-box transcription factors.However,few studies have investigated how the expression of these floral organ identity genes is regulated during flower development.In this study,we identified a gene named SUPER WOMAN 2(SPW2),which is necessary for spikelet/floret development in rice by participating in the regulation of the expression of pistil identity genes such as OsMADS3,OsMADS13,OsMADS58 and DL.In the spw2 mutant,ectopic stigma/ovary-like tissues were observed in the non-pistil organs,including sterile lemma,lemma,palea,lodicule,and stamen,suggesting that the identities of these organs were severely affected by mutations in SPW2.SPW2 was shown to encode a plant-specific EMF1-like protein that is involved in H3K27me3 modification as an important component of the PRC2 complex.Expression analysis showed that the SPW2 mutation led to the ectopic expression of OsMADS3,OsMADS13,OsMADS58,and DL in non-pistil organs of the spikelet.The ChIP-qPCR results showed significant reductions in the levels of H3K27me3 modification on the chromatin of these genes.Thus,we demonstrated that SPW2 can mediate the process of H3K27me3 modification of pistil-related genes to regulate their expression in non-pistil organs of spikelets in rice.The results of this study expand our understanding of the molecular mechanism by which SPW2 regulates floral organ identity genes through epigenetic regulation.展开更多
During reproductive development, rice plants develop unique flower organs which determine the final grain yield. OsMADS1, one of SEPALLATA-like MADS-box genes, has been unraveled to play critical roles in rice floral ...During reproductive development, rice plants develop unique flower organs which determine the final grain yield. OsMADS1, one of SEPALLATA-like MADS-box genes, has been unraveled to play critical roles in rice floral organ identity specification and floral meristem determinacy. However, the molecular mechanisms underlying interactions of OsMADS1 with other floral homeotic genes in regulating flower development remains largely elusive. In this work, we studied the genetic interactions of OsMADS1 with B-, C-, and D-class genes along with physical interactions among their proteins. We show that the physical and genetic interactions between OsMADS1 and OsMADS3 are essential for floral meristem activity maintenance and organ identity specification; while OsMADS1 physically and genetically interacts with OsMADS58 in regu- lating floral meristem determinacy and suppressing spikelet meristem reversion. We provided important genetic evidence to support the neofunctionalization of two rice C-class genes （OsMADS3 and OsMADS58） during flower development. Gene expression profiling and quantitative RT-PCR analyses further revealed that OsMADS1 affects the expression of many genes involved in floral identity and hormone signaling, and chromatin immunoprecipitation （ChlP）-PCR assay further demonstrated that OsMADS17 is a direct target gene of OsMADS1. Taken together, these results reveal that OsMADS1 has diversified regulatory functions in specifying rice floral organ and meristem identity, probably through its genetic and physical interactions with different floral homeotic regulators.展开更多
Stamen is a unique plant organ wherein germ cells or microsporocytes that commit to meiosis are initiated from somatic cells during its early developmental process. While genes determining stamen identity are known ac...Stamen is a unique plant organ wherein germ cells or microsporocytes that commit to meiosis are initiated from somatic cells during its early developmental process. While genes determining stamen identity are known according to the ABC model of floral development, little information is available on how these genes affect germ cell initiation. By using the Affymetrix GeneChip Rice Genome Array to assess 51 279 tran- scripts, we established a dynamic gene expression profile （GEP） of the early developmental process of rice （Oryza sativa） stamen. Systematic analysis of the GEP data revealed novel expression patterns of some developmentally important genes including meiosis-, tapetum-, and phytohormone-related genes. Following the finding that a substantial amount of nuclear genes encoding photosynthetic proteins are ex- pressed at the low levels in early rice stamen, through the ChlP-seq analysis we found that a C-class MADS box protein, OsMADS58, binds many nuclear-encoded genes participated in photosystem and light reac- tions and the expression levels of most of them are increased when expression of OsMADS58 is downre- gulated in the osmads58 mutant. Furthermore, more pro-chloroplasts are observed and increased signals of reactive oxygen species are detected in the osmads58 mutant anthers. These findings implicate a novel link between stamen identity determination and hypoxia status establishment.展开更多
OsMADS32 is a monocot specific MIKCc type MADS‐box gene that plays an important role in regulating rice floral meristem and organs identity, a crucial process for reproductive success and rice yield. However, its und...OsMADS32 is a monocot specific MIKCc type MADS‐box gene that plays an important role in regulating rice floral meristem and organs identity, a crucial process for reproductive success and rice yield. However, its underlying mechanism of action remains to be clarified. Here, we characterized a hypomorphic mutant allele of OsMADS32/CFO1, cfo1‐3 and identified its function in controlling rice flower development by bioinformatics and protein‐protein interaction analysis. The cfo1‐3 mutant produces defective flowers, including loss of lodicule identity, formation of ectopic lodicule or hull‐like organs and decreased stamen number, mimicking phenotypes related to the mutation of B class genes. Molecular characterization indicated that mis‐splicing of OsMADS32 transcripts in the cfo1‐3 mutant resulted in an extra eight amino acids in the K‐domain of OsMADS32 protein. By yeast two hybrid and bimolecular fluorescence comple-mentation assays, we revealed that the insertion of eight amino acids or deletion of the internal region in the K1 subdomain of OsMADS32 affects the interaction between OsMADS32 with PISTILLATA （PI）‐like proteins OsMADS2 and OsMADS4. This work provides new insight into the mecha-nism by which OsMADS32 regulates rice lodicule and stamen identity, by interaction with two PI‐like proteins via its K domain.展开更多
Current understanding of the classical ABC model of floral development has provided a new set of characters to evaluate floral evolution. However, what is still lacking is a clear assessment of this genetic program ac...Current understanding of the classical ABC model of floral development has provided a new set of characters to evaluate floral evolution. However, what is still lacking is a clear assessment of this genetic program across monocots. Here, to investigate the evolution of members of class A and B genes in monocots, we report the sequence characteristic and transcript expression of three new MADS-box genes in Alpinia oblongifolia Hayata. Sequence and phylogenetic analysis reveals that these genes are FUL-like and AP3-1ike. Therefore, they were termed AoFL1, AoFL2 and AoAP3. AoFL1 contains the FUL motif, but AoFL2 lacks this motif. Their expression revealed by in situ hybridization may reflect the ancestral function of FUL-like genes in the specification of inflorescence and floral meristems. The AoAP3 gene contains two conserved motifs, the PI-derived and paleoAP3 motifs. The AoAP3 transcripts located to the corolla and stamen, and hybridization signals were detected in the central whorl. These expression patterns suggest that the functions of homologous organ identity genes are diversified in A. oblongifolia. The implications of these findings on the conservation of homologous gene function are discussed.展开更多
Using PCR approach, three cDNA sequences, NTSQUA4, NTSQUA12 and NTSQUA15, were amplified from first_strand cDNAs of wild tobacco flower buds and identified as homologues for floral homeotic genes. All the three clones...Using PCR approach, three cDNA sequences, NTSQUA4, NTSQUA12 and NTSQUA15, were amplified from first_strand cDNAs of wild tobacco flower buds and identified as homologues for floral homeotic genes. All the three clones contained domains that a floral homeotic gene generally had, i.e. I domain, K domain and C_terminal domain except MADS_ box since the PCR primers were designed beyond this region. In addition, the amino acid sequences of them showed 50%-60% identity (70%-80% similarity) with the known floral organ identity class A gene AP1 and SQUA, possibly indicating that they are class A_like genes. NTSQUA4 and NTSQUA412 shared 95% identity in their amino acid sequence, while NTSQUA415 exhibited only 47% identity as compared with NTSQUA4 and NTSQUA12. Within tobacco flower, NTSQUA4 was expressed in sepals, petals and carpels, but not in stamens, while NTSQUA15 was expressed in every whorl of the flower. The possible functions of these genes are discussed.展开更多
基金We gratefully acknowledge B Han from National Center for Gene Research, Chinese Academy of Sciences (CAS) and Rice Genome Resource Center (RGRC) for providing BAC clone, cDNA clone and Tosl7 insertion line. We thank Z-J Luo and M-J Chen from Shanghai Jiao Tong University for mutant screening and generation of F2 populations, X-Y Gao from Institute of Plant Physiology and Ecology, SIBS, CAS, for SEM, H Yu from Nation- al University Of Singapore for critical reading of this manuscript and H Ma from Fudan University for helpful discussion. This work was supported by funds from the National Basic Research Program of China (2009CB941500, 2006CB 101700), the National Natural Science Foundation of China (30725022, 30830014 and 90717109) and the Shanghai Leading Academic Discipline Project (B205).
文摘Although AGAMOUS-LIKE6 (AGL6) MADS-box genes are ancient with wide distributions in gymnosperms and angiosperms, their functions remain poorly understood. Here, we show the biological role of the AGL6-1ike gene, OsMADS6, in specifying floral organ and meristem identities in rice (Oryza sativa L.). OsMADS6 was strongly ex- pressed in the floral meristem at early stages. Subsequently, OsMADS6 transcripts were mainly detectable in paleas, lodicules, carpels and the integument of ovule, as well as in the receptacle. Compared to wild type plants, osmads6 mutants displayed altered palea identity, extra glume-like or mosaic organs, abnormal carpel development and loss of floral meristem determinacy. Strikingly, mutation of a SEPALLATA (SEP)-like gene, OsMADS1 (LHS1), enhanced the defect of osmads6 flowers, and no inner floral organs or glume-like structures were observed in whorls 2 and 3 of osmadsl-z osmads6-1 flowers. Furthermore, the osmadsl-z osmads6-1 double mutants developed severely indetermi- nate floral meristems. Our finding, therefore, suggests that the ancient OsMADS6 gene is able to specify "floral state" by determining floral organ and meristem identities in monocot crop rice together with OsMADS1.
基金This work was supported by the Program for Changjiang Scholars and Innovative Research Team in University (No. IRTO453)
文摘The floral-organ-number mutant fon（t） was firstly discovered in the progeny of a cross between a diploid （Chunjiang 683） and a haploid （SARⅣ/-620-A） rice cultivar. The fon（t） mutant showed normal vegetative development and produced normal inflorescence structures. Difference between the mutant and the wild type was observed when the stamen primordia began to form. The mature flowers offon（t） mutant showed open-hull phenotypes, which resulted in the exposure of stamens and stigmas. Normally, a single fon（t） floret consisted of six to nine stamens and one or two pistils. In addition, stamen/pistil-like structures and bulged tissues near ovaries were also observed in a few fon（t） florets. But homeotic transformation of lodicules into palea/lemma-like organs was observed almost in all the open-hull florets. The phenotypes offon（t） flowers also suggested thatfon（t） gene might affect flower organ identity in the inner whorls. Genetic analysis showed that thefon（t） mutant was controlled by a single recessive gene.
基金National Natural Science Foundation of China，Science and Technology Planning Project of Guangdong Province,China
文摘During organ development, many key regulators have been identified in plant genomes, which play a conserved role among plant species to control the organ identities and/or determine the organ size and shape. It is intriguing whether these key regulators can acquire diverse function and be integrated into different molecular pathways among different species, giving rise to the immense diversity of organ forms in nature. In this study, we have characterized and cloned LATHYROIDES （LATH）, a classical locus in pea, whose mutation displays pleiotropic alteration of lateral growth of organs and predominant effects on tendril and dorsal petal development. LATH encodes a WUSCHEL-related home- oboxl （WOX1） transcription factor, which has a conserved function in determining organ lateral growth among different plant species. Furthermore, we showed that LATH regulated the expression level of TENDRIL-LESS （TL）, a key factor in the control of tendril development in compound leaf, and LATH genetically interacted with LOBED STANDARD （LST）, a floral dorsal factor, to affect the dorsal petal identity. Thus, LATH plays multiple roles during organ development in pea： it maintains a conserved function controlling organ lateral outgrowth, and modulates organ identities in compound leaf and zygomorphic flower development, respectively. Our data indicated that a key regulator can play important roles in different aspects of organ development and dedicate to the complexity of the molecular mechanism in the control of organ development so as to create distinct organ forms in different species.
基金supported by the Chongqing Modern Agricultural Industry Technology System,China(CQMAITS202301)the National Natural Science Foundation of China(32100287 and 31971919)+2 种基金the Natural Science Foundation of Chongqing,China(cstc2020jcyj-jq X0020 and cstc2021ycjh-bgzxm0066)the China Postdoctoral Science Foundation Funded Project(2020M683219)the Fundamental Research Funds for the Central Universities,China(SWU-XDJH202315)。
文摘Flower organ identity in rice is mainly determined by the A-,B-,C-and E-class genes,with the majority encoding MADS-box transcription factors.However,few studies have investigated how the expression of these floral organ identity genes is regulated during flower development.In this study,we identified a gene named SUPER WOMAN 2(SPW2),which is necessary for spikelet/floret development in rice by participating in the regulation of the expression of pistil identity genes such as OsMADS3,OsMADS13,OsMADS58 and DL.In the spw2 mutant,ectopic stigma/ovary-like tissues were observed in the non-pistil organs,including sterile lemma,lemma,palea,lodicule,and stamen,suggesting that the identities of these organs were severely affected by mutations in SPW2.SPW2 was shown to encode a plant-specific EMF1-like protein that is involved in H3K27me3 modification as an important component of the PRC2 complex.Expression analysis showed that the SPW2 mutation led to the ectopic expression of OsMADS3,OsMADS13,OsMADS58,and DL in non-pistil organs of the spikelet.The ChIP-qPCR results showed significant reductions in the levels of H3K27me3 modification on the chromatin of these genes.Thus,we demonstrated that SPW2 can mediate the process of H3K27me3 modification of pistil-related genes to regulate their expression in non-pistil organs of spikelets in rice.The results of this study expand our understanding of the molecular mechanism by which SPW2 regulates floral organ identity genes through epigenetic regulation.
文摘During reproductive development, rice plants develop unique flower organs which determine the final grain yield. OsMADS1, one of SEPALLATA-like MADS-box genes, has been unraveled to play critical roles in rice floral organ identity specification and floral meristem determinacy. However, the molecular mechanisms underlying interactions of OsMADS1 with other floral homeotic genes in regulating flower development remains largely elusive. In this work, we studied the genetic interactions of OsMADS1 with B-, C-, and D-class genes along with physical interactions among their proteins. We show that the physical and genetic interactions between OsMADS1 and OsMADS3 are essential for floral meristem activity maintenance and organ identity specification; while OsMADS1 physically and genetically interacts with OsMADS58 in regu- lating floral meristem determinacy and suppressing spikelet meristem reversion. We provided important genetic evidence to support the neofunctionalization of two rice C-class genes （OsMADS3 and OsMADS58） during flower development. Gene expression profiling and quantitative RT-PCR analyses further revealed that OsMADS1 affects the expression of many genes involved in floral identity and hormone signaling, and chromatin immunoprecipitation （ChlP）-PCR assay further demonstrated that OsMADS17 is a direct target gene of OsMADS1. Taken together, these results reveal that OsMADS1 has diversified regulatory functions in specifying rice floral organ and meristem identity, probably through its genetic and physical interactions with different floral homeotic regulators.
文摘Stamen is a unique plant organ wherein germ cells or microsporocytes that commit to meiosis are initiated from somatic cells during its early developmental process. While genes determining stamen identity are known according to the ABC model of floral development, little information is available on how these genes affect germ cell initiation. By using the Affymetrix GeneChip Rice Genome Array to assess 51 279 tran- scripts, we established a dynamic gene expression profile （GEP） of the early developmental process of rice （Oryza sativa） stamen. Systematic analysis of the GEP data revealed novel expression patterns of some developmentally important genes including meiosis-, tapetum-, and phytohormone-related genes. Following the finding that a substantial amount of nuclear genes encoding photosynthetic proteins are ex- pressed at the low levels in early rice stamen, through the ChlP-seq analysis we found that a C-class MADS box protein, OsMADS58, binds many nuclear-encoded genes participated in photosystem and light reac- tions and the expression levels of most of them are increased when expression of OsMADS58 is downre- gulated in the osmads58 mutant. Furthermore, more pro-chloroplasts are observed and increased signals of reactive oxygen species are detected in the osmads58 mutant anthers. These findings implicate a novel link between stamen identity determination and hypoxia status establishment.
基金supported by the Funds from National Natural Science Foundation of China (30971739,31270222,31230051,and J1210047)Key Project on Basic Research from Science and Technology Commission of Shanghai (14JC1403900)the Innovation Program of Shanghai Municipal Education Commission (13ZZ018)
文摘OsMADS32 is a monocot specific MIKCc type MADS‐box gene that plays an important role in regulating rice floral meristem and organs identity, a crucial process for reproductive success and rice yield. However, its underlying mechanism of action remains to be clarified. Here, we characterized a hypomorphic mutant allele of OsMADS32/CFO1, cfo1‐3 and identified its function in controlling rice flower development by bioinformatics and protein‐protein interaction analysis. The cfo1‐3 mutant produces defective flowers, including loss of lodicule identity, formation of ectopic lodicule or hull‐like organs and decreased stamen number, mimicking phenotypes related to the mutation of B class genes. Molecular characterization indicated that mis‐splicing of OsMADS32 transcripts in the cfo1‐3 mutant resulted in an extra eight amino acids in the K‐domain of OsMADS32 protein. By yeast two hybrid and bimolecular fluorescence comple-mentation assays, we revealed that the insertion of eight amino acids or deletion of the internal region in the K1 subdomain of OsMADS32 affects the interaction between OsMADS32 with PISTILLATA （PI）‐like proteins OsMADS2 and OsMADS4. This work provides new insight into the mecha-nism by which OsMADS32 regulates rice lodicule and stamen identity, by interaction with two PI‐like proteins via its K domain.
基金Supported by the National Natural Science Foundation of China (30225007)the Natural Science Foundation of Yunnan Province,China (2005C0057M)
文摘Current understanding of the classical ABC model of floral development has provided a new set of characters to evaluate floral evolution. However, what is still lacking is a clear assessment of this genetic program across monocots. Here, to investigate the evolution of members of class A and B genes in monocots, we report the sequence characteristic and transcript expression of three new MADS-box genes in Alpinia oblongifolia Hayata. Sequence and phylogenetic analysis reveals that these genes are FUL-like and AP3-1ike. Therefore, they were termed AoFL1, AoFL2 and AoAP3. AoFL1 contains the FUL motif, but AoFL2 lacks this motif. Their expression revealed by in situ hybridization may reflect the ancestral function of FUL-like genes in the specification of inflorescence and floral meristems. The AoAP3 gene contains two conserved motifs, the PI-derived and paleoAP3 motifs. The AoAP3 transcripts located to the corolla and stamen, and hybridization signals were detected in the central whorl. These expression patterns suggest that the functions of homologous organ identity genes are diversified in A. oblongifolia. The implications of these findings on the conservation of homologous gene function are discussed.
文摘Using PCR approach, three cDNA sequences, NTSQUA4, NTSQUA12 and NTSQUA15, were amplified from first_strand cDNAs of wild tobacco flower buds and identified as homologues for floral homeotic genes. All the three clones contained domains that a floral homeotic gene generally had, i.e. I domain, K domain and C_terminal domain except MADS_ box since the PCR primers were designed beyond this region. In addition, the amino acid sequences of them showed 50%-60% identity (70%-80% similarity) with the known floral organ identity class A gene AP1 and SQUA, possibly indicating that they are class A_like genes. NTSQUA4 and NTSQUA412 shared 95% identity in their amino acid sequence, while NTSQUA415 exhibited only 47% identity as compared with NTSQUA4 and NTSQUA12. Within tobacco flower, NTSQUA4 was expressed in sepals, petals and carpels, but not in stamens, while NTSQUA15 was expressed in every whorl of the flower. The possible functions of these genes are discussed.
