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1.
Cell ; 170(1): 114-126.e15, 2017 Jun 29.
Artigo em Inglês | MEDLINE | ID: mdl-28666113

RESUMO

Rice feeds half the world's population, and rice blast is often a destructive disease that results in significant crop loss. Non-race-specific resistance has been more effective in controlling crop diseases than race-specific resistance because of its broad spectrum and durability. Through a genome-wide association study, we report the identification of a natural allele of a C2H2-type transcription factor in rice that confers non-race-specific resistance to blast. A survey of 3,000 sequenced rice genomes reveals that this allele exists in 10% of rice, suggesting that this favorable trait has been selected through breeding. This allele causes a single nucleotide change in the promoter of the bsr-d1 gene, which results in reduced expression of the gene through the binding of the repressive MYB transcription factor and, consequently, an inhibition of H2O2 degradation and enhanced disease resistance. Our discovery highlights this novel allele as a strategy for breeding durable resistance in rice.


Assuntos
Oryza/genética , Proteínas de Plantas/genética , Fatores de Transcrição/genética , Sequência de Bases , Cruzamento , Resistência à Doença , Técnicas de Inativação de Genes , Genoma de Planta , Estudo de Associação Genômica Ampla , Doenças das Plantas , Regiões Promotoras Genéticas
2.
Plant Physiol ; 187(4): 2852-2864, 2021 12 04.
Artigo em Inglês | MEDLINE | ID: mdl-34597396

RESUMO

Resistance (R) proteins are important components of plant innate immunity. Most known R proteins are nucleotide-binding site leucine-rich repeat (NLR) proteins. Although a number of signaling components downstream of NLRs have been identified, we lack a general understanding of the signaling pathways. Here, we used the interaction between rice (Oryza sativa) and Magnaporthe oryzae to study signaling of rice NLRs in response to blast infection. We found that in blast resistance mediated by the NLR PIRICULARIA ORYZAE RESISTANCE IN DIGU 3 (PID3), the guanine nucleotide exchange factor OsSPK1 works downstream of PID3. OsSPK1 activates the small GTPase OsRac1, which in turn transduces the signal to the transcription factor RAC IMMUNITY1 (RAI1). Further investigation revealed that the three signaling components also play important roles in disease resistance mediated by the distantly related NLR protein Pi9, suggesting that the OsSPK1-OsRac1-RAI1 signaling pathway could be conserved across rice NLR-induced blast resistance. In addition, we observed changes in RAI1 levels during blast infection, which led to identification of OsRPT2a, a subunit of the 19S regulatory particle of the 26S proteasome. OsRPT2a seemed to be responsible for RAI1 turnover in a 26S proteasome-dependent manner. Collectively, our results suggest a defense signaling route that might be common to NLR proteins in response to blast infection.


Assuntos
Magnaporthe/fisiologia , Proteínas NLR/genética , Oryza/genética , Doenças das Plantas/genética , Imunidade Vegetal/genética , Transdução de Sinais , Resistência à Doença/genética , Proteínas NLR/metabolismo , Oryza/microbiologia , Doenças das Plantas/microbiologia
3.
Proc Natl Acad Sci U S A ; 115(12): 3174-3179, 2018 03 20.
Artigo em Inglês | MEDLINE | ID: mdl-29432165

RESUMO

Crops carrying broad-spectrum resistance loci provide an effective strategy for controlling infectious disease because these loci typically confer resistance to diverse races of a pathogen or even multiple species of pathogens. Despite their importance, only a few crop broad-spectrum resistance loci have been reported. Here, we report the identification and characterization of the rice bsr-k1 (broad-spectrum resistance Kitaake-1) mutant, which confers broad-spectrum resistance against Magnaporthe oryzae and Xanthomonas oryzae pv oryzae with no major penalty on key agronomic traits. Map-based cloning reveals that Bsr-k1 encodes a tetratricopeptide repeats (TPRs)-containing protein, which binds to mRNAs of multiple OsPAL (OsPAL1-7) genes and promotes their turnover. Loss of function of the Bsr-k1 gene leads to accumulation of OsPAL1-7 mRNAs in the bsr-k1 mutant. Furthermore, overexpression of OsPAL1 in wild-type rice TP309 confers resistance to M. oryzae, supporting the role of OsPAL1 Our discovery of the bsr-k1 allele constitutes a significant conceptual advancement and provides a valuable tool for breeding broad-spectrum resistant rice.


Assuntos
Oryza/fisiologia , Doenças das Plantas/genética , Proteínas de Plantas/genética , Proteínas de Ligação a RNA/genética , Citoplasma/metabolismo , Resistência à Doença/genética , Regulação da Expressão Gênica de Plantas , Magnaporthe/patogenicidade , Mutação , Oryza/genética , Oryza/microbiologia , Doenças das Plantas/microbiologia , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/genética , Domínios Proteicos , RNA Mensageiro/metabolismo , Proteínas de Ligação a RNA/metabolismo , Sequências Repetitivas de Aminoácidos , Xanthomonas/patogenicidade
4.
New Phytol ; 223(2): 828-838, 2019 07.
Artigo em Inglês | MEDLINE | ID: mdl-30919975

RESUMO

Plants depend on Resistance (R) genes, most of which encode nucleotide-binding site leucine-rich repeat (NLR) proteins, for pathogen race-specific disease resistance. However, only a few immediate downstream targets of R proteins have been characterized, and the signalling pathways for R-protein-induced immunity are largely unknown. In rice (Oryza sativa), NLR proteins serve as important immune receptors in the response to rice blast disease caused by the fungus Magnaporthe oryzae. We used site-directed mutagenesis to create an autoactive form of the NLR protein PID3 that confers blast resistance and used transgenic rice to test the resulting immunity and gene expression changes. We identified OsRac1, a known GTPase, as a signalling molecule in PID3-mediated blast resistance, implicating OsRac1 as a possible common factor downstream of rice NLR proteins. We also identified RAI1, a transcriptional activator, as a PID3 interactor required for PID3-mediated blast resistance and showed that RAI1 expression is induced by PID3 via a process mediated by OsRac1. This study describes a new signalling pathway for NLR protein-mediated blast resistance and shows that OsRac1 and RAI1 act together to play a critical role in this process.


Assuntos
Resistência à Doença , Nucleotídeos/metabolismo , Oryza/microbiologia , Doenças das Plantas/imunologia , Doenças das Plantas/microbiologia , Proteínas de Plantas/metabolismo , Proteínas/metabolismo , Transdução de Sinais , Sítios de Ligação , Resistência à Doença/genética , Regulação da Expressão Gênica de Plantas , Proteínas de Repetições Ricas em Leucina , Oryza/genética , Oryza/imunologia , Oryza/metabolismo , Doenças das Plantas/genética , Imunidade Vegetal , Proteínas de Plantas/genética , Ligação Proteica , Proteínas/genética , RNA Mensageiro/genética , RNA Mensageiro/metabolismo
5.
PLoS Genet ; 12(9): e1006311, 2016 09.
Artigo em Inglês | MEDLINE | ID: mdl-27618555

RESUMO

Previous studies have shown that multivesicular bodies (MVBs)/endosomes-mediated vesicular trafficking may play key roles in plant immunity and cell death. However, the molecular regulation is poorly understood in rice. Here we report the identification and characterization of a MVBs-localized AAA ATPase LRD6-6 in rice. Disruption of LRD6-6 leads to enhanced immunity and cell death in rice. The ATPase activity and homo-dimerization of LRD6-6 is essential for its regulation on plant immunity and cell death. An ATPase inactive mutation (LRD6-6E315Q) leads to dominant-negative inhibition in plants. The LRD6-6 protein co-localizes with the MVBs marker protein RabF1/ARA6 and interacts with ESCRT-III components OsSNF7 and OsVPS2. Further analysis reveals that LRD6-6 is required for MVBs-mediated vesicular trafficking and inhibits the biosynthesis of antimicrobial compounds. Collectively, our study shows that the AAA ATPase LRD6-6 inhibits plant immunity and cell death most likely through modulating MVBs-mediated vesicular trafficking in rice.


Assuntos
Adenosina Trifosfatases/biossíntese , Imunidade Celular/genética , Corpos Multivesiculares/genética , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas/genética , Adenosina Trifosfatases/genética , Arabidopsis/genética , Arabidopsis/crescimento & desenvolvimento , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Morte Celular/genética , Resistência à Doença/genética , Resistência à Doença/imunologia , Complexos Endossomais de Distribuição Requeridos para Transporte/genética , Complexos Endossomais de Distribuição Requeridos para Transporte/metabolismo , Endossomos/genética , Endossomos/metabolismo , Regulação da Expressão Gênica de Plantas , Corpos Multivesiculares/imunologia , Mutação , Oryza/genética , Oryza/crescimento & desenvolvimento , Doenças das Plantas/genética , Doenças das Plantas/imunologia , Folhas de Planta/genética , Folhas de Planta/imunologia , Plantas Geneticamente Modificadas/imunologia , Transporte Proteico/genética , Proteínas rab de Ligação ao GTP/genética
6.
Proc Natl Acad Sci U S A ; 113(41): E6026-E6035, 2016 10 11.
Artigo em Inglês | MEDLINE | ID: mdl-27663737

RESUMO

Hybrid rice is the dominant form of rice planted in China, and its use has extended worldwide since the 1970s. It offers great yield advantages and has contributed greatly to the world's food security. However, the molecular mechanisms underlying heterosis have remained a mystery. In this study we integrated genetics and omics analyses to determine the candidate genes for yield heterosis in a model two-line rice hybrid system, Liang-you-pei 9 (LYP9) and its parents. Phenomics study revealed that the better parent heterosis (BPH) of yield in hybrid is not ascribed to BPH of all the yield components but is specific to the BPH of spikelet number per panicle (SPP) and paternal parent heterosis (PPH) of effective panicle number (EPN). Genetic analyses then identified multiple quantitative trait loci (QTLs) for these two components. Moreover, a number of differentially expressed genes and alleles in the hybrid were mapped by transcriptome profiling to the QTL regions as possible candidate genes. In parallel, a major QTL for yield heterosis, rice heterosis 8 (RH8), was found to be the DTH8/Ghd8/LHD1 gene. Based on the shared allelic heterozygosity of RH8 in many hybrid rice cultivars, a common mechanism for yield heterosis in the present commercial hybrid rice is proposed.


Assuntos
Genoma de Planta , Vigor Híbrido/genética , Hibridização Genética , Oryza/genética , Oryza/metabolismo , Fenótipo , Locos de Características Quantitativas , Transcriptoma , Alelos , Mapeamento Cromossômico , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Genes de Plantas , Ligação Genética , Genômica/métodos , Genótipo , Polimorfismo de Nucleotídeo Único , Característica Quantitativa Herdável
7.
Biochem Biophys Res Commun ; 495(1): 487-492, 2018 01 01.
Artigo em Inglês | MEDLINE | ID: mdl-29122595

RESUMO

The cereal crops (such as rice and maize) which belong to the grass family, are the most important grain crops for human beings, and the development of their flower and inflorescence architecture has attracted extensive attention. Although multiple genes involved in the regulation of floral and inflorescence organogenesis have been identified, the underlying molecular mechanisms are largely unknown. Previously, we identified rice depressed palea1 (dp1) mutants with defects in main structure of palea and its enhancer RETARDED PALEA1 (REP1). DP1 is an AT-hook protein while REP1 is a TCP transcription factor, both of which are important regulators of palea development. However, the relationship of these two proteins has not been elucidated yet. Here, we demonstrated that DP1 interacts physically with REP1 both in yeast and in rice protoplasts. Considering the close phylogenetic relationship between maize and rice, we further hypothesize that their orthologs in maize, BARREN STALK FASTIGIATE (BAF1) and BRANCH ANGLE DEFECTIVE 1 (BAD1), may interact physically. Subsequently, we verified their physical interaction, indicating that the interaction between AT-hook proteins and TCP proteins is conserved in rice and maize. Our findings may reveal a novel molecular mechanism of floral and inflorescence development in grasses.


Assuntos
Oryza/metabolismo , Proteínas de Plantas/metabolismo , Mapas de Interação de Proteínas , Protoplastos/metabolismo , Fatores de Transcrição/metabolismo , Regulação da Expressão Gênica de Plantas , Oryza/genética , Filogenia , Proteínas de Plantas/análise , Proteínas de Plantas/genética , Fatores de Transcrição/análise , Fatores de Transcrição/genética , Zea mays/genética
8.
BMC Plant Biol ; 18(1): 157, 2018 Aug 06.
Artigo em Inglês | MEDLINE | ID: mdl-30081823

RESUMO

BACKGROUND: Flowering time is a key trait for regional adaption and seed production in rice (Oryza sativa L.). Forward and reverse genetic studies have characterized a number of flowering-time genes. However, co-expression analysis has not been used to identify the flowering-time genes. RESULTS: We predicted a G2-like family transcription factor, OsPHL3, by co-expression networks analysis with photoperiodic flowering pathway genes. OsPHL3 contains a MYB-CC domain, and was localized in the nucleus with transcriptional activation potential. OsPHL3 was mainly expressed in the leaves and exhibited a circadian rhythmic expression pattern. Rice lines overexpressing OsPHL3 showed a delayed flowering time in the genetic background of TP309 under both long-day (Beijing) and short-day (Hainan) conditions. By contrast, the knockout rice lines of OsPHL3 by CRISPR/Cas9 technology promoted flowering time regardless of genetic backgrounds (i.e. Nipponbare and TP309) or day length. Further analysis indicated that OsPHL3 delayed flowering time by down-regulating the expression of Hd3a and RFT1 through promoting Hd1 under long-day conditions (LDs), or suppressing Ehd1/Hd1 under short-day conditions (SDs). CONCLUSIONS: Our results suggested that co-expression analysis is a useful strategy for identifying novel flowering-time genes in rice.


Assuntos
Flores/genética , Oryza/genética , Proteínas de Plantas/genética , Fatores de Transcrição/genética , Flores/crescimento & desenvolvimento , Técnicas de Silenciamento de Genes , Oryza/crescimento & desenvolvimento , Oryza/metabolismo , Fotoperíodo , Filogenia , Proteínas de Plantas/fisiologia , Reação em Cadeia da Polimerase em Tempo Real , Genética Reversa , Homologia de Sequência do Ácido Nucleico , Fatores de Transcrição/fisiologia , Transcriptoma
9.
New Phytol ; 218(2): 774-788, 2018 04.
Artigo em Inglês | MEDLINE | ID: mdl-29411384

RESUMO

Natural antisense long noncoding RNAs (lncRNAs) are widespread in many organisms. However, their biological functions remain largely unknown, particularly in plants. We report the identification and characterization of an endogenous lncRNA, TWISTED LEAF (TL), which is transcribed from the opposite strand of the R2R3 MYB transcription factor gene locus, OsMYB60, in rice (Oryza sativa). TL and OsMYB60 were found to be coexpressed in many different tissues, and the expression level of TL was higher than that of OsMYB60. Downregulation of TL by RNA interference (RNAi) and overexpression of OsMYB60 resulted in twisted leaf blades in transgenic rice. The expression level of OsMYB60 was significantly increased in TL-RNAi transgenic plants. This suggests that TL may play a cis-regulatory role on OsMYB60 in leaf morphological development. We also determined that the antisense transcription suppressed the sense gene expression by mediating chromatin modifications. We further discovered that a C2H2 transcription factor, OsZFP7, is an OsMYB60 binding partner and involved in leaf development. Taken together, these findings reveal that the cis-natural antisense lncRNA plays a critical role in maintaining leaf blade flattening in rice. Our study uncovers a regulatory mechanism of lncRNA in plant leaf development.


Assuntos
Padronização Corporal/genética , Genes de Plantas , Oryza/genética , Folhas de Planta/genética , RNA Antissenso/genética , RNA Longo não Codificante/genética , Cromatina/metabolismo , Regulação da Expressão Gênica de Plantas , Técnicas de Silenciamento de Genes , Loci Gênicos , Fases de Leitura Aberta/genética , Fenótipo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas , Ligação Proteica , Interferência de RNA , RNA Antissenso/metabolismo , RNA Longo não Codificante/metabolismo , Fatores de Transcrição/metabolismo , Transcrição Gênica
10.
BMC Plant Biol ; 17(1): 166, 2017 Oct 19.
Artigo em Inglês | MEDLINE | ID: mdl-29052517

RESUMO

BACKGROUND: Dof (DNA binding with one finger) proteins, a class of plant-specific transcription factors which contain a conserved C2-C2-type zinc finger domain, are involved in many fundamental processes. In the Arabidopsis photoperiod response pathway, CDF (CYCLING DOF FACTOR) proteins have a primary role as acting via transcriptional repression of the direct FLOWERING LOCUS T (FT) activator CONSTANS (CO). Our previous study indicated that one of CDF homologs, OsDOf12, was involved in photoperiodic flowering. However, the functional characterization of other rice CDF like genes is still in progress. Here, we characterized the function of OsDof4 in rice. RESULTS: Phylogenic analysis indicated that OsDof4 is closely clustered into the same subgroup with CDFs and OsDof12. The subcellular localization experiment and transcriptional activity assay suggested that OsDof4 may function as a transcription factor. The diurnal expression pattern indicated that OsDof4 was regulated by endogenous circadian clock. Overexpression of OsDof4 led to earlier flowering under natural long-day field conditions (NLDs) and late flowering under natural short-day field conditions (NSDs), respectively. We compared the expression level of key floral genes in vector line and OsDof4-ox lines grown under long-day conditions (LDs) and short-day conditions (SDs). Real-time q-PCR results demonstrated that under LDs, Hd3a, RFT1 and Ehd1 were up-regulated whereas under SDs they were down-regulated. Hd1 was down-regulated at dusk period independent of photoperiods. CONCLUSIONS: Taken these results together, we may speculate that the abnormal flowering responses in OsDof4-ox plants under LDs and SDs might be mediated by Ehd1 and Hd1.


Assuntos
Flores/crescimento & desenvolvimento , Oryza/fisiologia , Proteínas de Plantas/fisiologia , Dedos de Zinco/fisiologia , Regulação da Expressão Gênica de Plantas/fisiologia , Genes de Plantas/genética , Genes de Plantas/fisiologia , Oryza/metabolismo , Fotoperíodo , Filogenia , Proteínas de Plantas/metabolismo
11.
J Integr Plant Biol ; 58(6): 600-9, 2016 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-26356550

RESUMO

Jasmonates (JAs) are rapidly induced after wounding and act as key regulators for wound induced signaling pathway. However, what perceives the wound signal and how that triggers JA biosynthesis remains poorly understood. To identify components involved in Arabidopsis wound and JA signaling pathway, we screened for mutants with abnormal expression of a luciferase reporter, which is under the control of a wound-responsive promoter of an ethylene response factor (ERF) transcription factor gene, RAP2.6 (Related to APetala 2.6). The rea1 (RAP2.6 expresser in shoot apex) mutant constitutively expressed the RAP2.6-LUC reporter gene in young leaves. Along with the typical JA phenotypes including shorter petioles, loss of apical dominance, accumulation of anthocyanin pigments and constitutive expression of JA response gene, rea1 plants also displayed cell death and accumulated high levels of JA in response to wounding. The phenotype of rea1 mutant is caused by a gain-of-function mutation in the C-terminus of a mechanosensitive ion channel MscS-like 10 (MSL10). MSL10 is localized in the plasma membrane and is expressed predominantly in root tip, shoot apex and vascular tissues. These results suggest that MSL10 is involved in the wound-triggered early signal transduction pathway and possibly in regulating the positive feedback synthesis of JA.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/metabolismo , Morte Celular/fisiologia , Ciclopentanos/metabolismo , Proteínas de Membrana/metabolismo , Mutação/genética , Oxilipinas/metabolismo , Arabidopsis/genética , Proteínas de Arabidopsis/genética , Morte Celular/genética , Regulação da Expressão Gênica de Plantas/genética , Proteínas de Membrana/genética , Transdução de Sinais/genética , Transdução de Sinais/fisiologia
12.
BMC Plant Biol ; 15: 49, 2015 Feb 13.
Artigo em Inglês | MEDLINE | ID: mdl-25849162

RESUMO

BACKGROUND: Rice blast disease is one of the most destructive diseases of rice worldwide. We previously cloned the rice blast resistance gene Pid2, which encodes a transmembrane receptor-like kinase containing an extracellular B-lectin domain and an intracellular serine/threonine kinase domain. However, little is known about Pid2-mediated signaling. RESULTS: Here we report the functional characterization of the U-box/ARM repeat protein OsPUB15 as one of the PID2-binding proteins. We found that OsPUB15 physically interacted with the kinase domain of PID2 (PID2K) in vitro and in vivo and the ARM repeat domain of OsPUB15 was essential for the interaction. In vitro biochemical assays indicated that PID2K possessed kinase activity and was able to phosphorylate OsPUB15. We also found that the phosphorylated form of OsPUB15 possessed E3 ligase activity. Expression pattern analyses revealed that OsPUB15 was constitutively expressed and its encoded protein OsPUB15 was localized in cytosol. Transgenic rice plants over-expressing OsPUB15 at early stage displayed cell death lesions spontaneously in association with a constitutive activation of plant basal defense responses, including excessive accumulation of hydrogen peroxide, up-regulated expression of pathogenesis-related genes and enhanced resistance to blast strains. We also observed that, along with plant growth, the cell death lesions kept spreading over the whole seedlings quickly resulting in a seedling lethal phenotype. CONCLUSIONS: These results reveal that the E3 ligase OsPUB15 interacts directly with the receptor-like kinase PID2 and regulates plant cell death and blast disease resistance.


Assuntos
Morte Celular , Regulação da Expressão Gênica de Plantas , Oryza/fisiologia , Proteínas de Plantas/genética , Proteínas Quinases/genética , Ubiquitina-Proteína Ligases/genética , Sequência de Aminoácidos , Resistência à Doença , Imunidade Inata , Magnaporthe/fisiologia , Oryza/enzimologia , Oryza/genética , Oryza/imunologia , Filogenia , Doenças das Plantas/genética , Doenças das Plantas/imunologia , Doenças das Plantas/microbiologia , Imunidade Vegetal/fisiologia , Proteínas de Plantas/química , Proteínas de Plantas/metabolismo , Plantas Geneticamente Modificadas/genética , Plantas Geneticamente Modificadas/imunologia , Plantas Geneticamente Modificadas/fisiologia , Proteínas Quinases/metabolismo , Ubiquitina-Proteína Ligases/química , Ubiquitina-Proteína Ligases/metabolismo
13.
Plant Cell ; 24(6): 2562-77, 2012 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-22685166

RESUMO

In Arabidopsis thaliana, the GSK3/SHAGGY-like kinase BRASSINOSTEROID-INSENSITIVE2 (BIN2) plays a critical role in the brassinosteroid (BR) signaling pathway by negatively regulating the activities of bri1-EMS-SUPPRESSOR1/BRASSINAZOLE-RESISTANT1 family transcription factors that regulate the expression of downstream BR-responsive genes. In this study, we analyzed the function of a rice (Oryza sativa) GSK3/SHAGGY-like kinase (GSK2), which is one of the orthologs of BIN2. Overexpression of GSK2 (Go) led to plants with typical BR loss-of-function phenotypes, and suppression of GSK2 resulted in enhanced BR signaling phenotypes. DWARF AND LOW-TILLERING (DLT) is a positive regulator that mediates several BR responses in rice. Suppression of DLT can enhance the phenotypes of BR receptor mutant d61-1, and overexpression of DLT obviously suppressed the BR loss-of-function phenotypes of both d61-1 and Go, suggesting that DLT functions downstream of GSK2 to modulate BR responses. Indeed, GSK2 can interact with DLT and phosphorylate DLT. Moreover, brassinolide treatment can induce the dephosphorylation of DLT, leading to the accumulation of dephosphorylated DLT protein. In GSK2 transgenic plants, the DLT phosphorylation level is dictated by the GSK2 level. These results demonstrate that DLT is a GSK2 substrate, further reinforcing that the BIN2/GSK2 kinase has multiple substrates that carry out various BR responses.


Assuntos
Brassinosteroides/metabolismo , Oryza/metabolismo , Proteínas de Plantas/metabolismo , Proteínas Quinases/metabolismo , Proteínas de Arabidopsis/metabolismo , Brassinosteroides/farmacologia , Núcleo Celular/metabolismo , Proteínas de Ligação a DNA , Regulação da Expressão Gênica de Plantas , Dados de Sequência Molecular , Mutação , Proteínas Nucleares/metabolismo , Oryza/efeitos dos fármacos , Oryza/genética , Oryza/crescimento & desenvolvimento , Fenótipo , Fosforilação , Desenvolvimento Vegetal , Proteínas de Plantas/genética , Plantas Geneticamente Modificadas , Proteínas Quinases/genética , Transdução de Sinais/genética , Esteroides Heterocíclicos/farmacologia
14.
BMC Plant Biol ; 14: 158, 2014 Jun 06.
Artigo em Inglês | MEDLINE | ID: mdl-24906444

RESUMO

BACKGROUND: The shape of grass leaves possesses great value in both agronomy and developmental biology research. Leaf rolling is one of the important traits in rice (Oryza sativa L.) breeding. MYB transcription factors are one of the largest gene families and have important roles in plant development, metabolism and stress responses. However, little is known about their functions in rice. RESULTS: In this study, we report the functional characterization of a rice gene, OsMYB103L, which encodes an R2R3-MYB transcription factor. OsMYB103L was localized in the nucleus with transactivation activity. Overexpression of OsMYB103L in rice resulted in a rolled leaf phenotype. Further analyses showed that expression levels of several cellulose synthase genes (CESAs) were significantly increased, as was the cellulose content in OsMYB103L overexpressing lines. Knockdown of OsMYB103L by RNA interference led to a decreased level of cellulose content and reduced mechanical strength in leaves. Meanwhile, the expression levels of several CESA genes were decreased in these knockdown lines. CONCLUSIONS: These findings suggest that OsMYB103L may target CESA genes for regulation of cellulose synthesis and could potentially be engineered for desirable leaf shape and mechanical strength in rice.


Assuntos
Oryza/metabolismo , Oryza/fisiologia , Folhas de Planta/fisiologia , Proteínas de Plantas/metabolismo , Fatores de Transcrição/metabolismo , Sequência de Aminoácidos , Fenômenos Biomecânicos , Celulose/metabolismo , Regulação para Baixo/genética , Perfilação da Expressão Gênica , Regulação da Expressão Gênica de Plantas , Técnicas de Silenciamento de Genes , Genes de Plantas , Glucosiltransferases/genética , Glucosiltransferases/metabolismo , Proteínas de Fluorescência Verde/metabolismo , Dados de Sequência Molecular , Especificidade de Órgãos/genética , Oryza/genética , Fenótipo , Proteínas de Plantas/química , Plantas Geneticamente Modificadas , Transporte Proteico , Alinhamento de Sequência , Frações Subcelulares/metabolismo , Fatores de Transcrição/química , Ativação Transcricional , Regulação para Cima/genética
15.
Plant Physiol ; 161(3): 1375-91, 2013 Mar.
Artigo em Inglês | MEDLINE | ID: mdl-23292790

RESUMO

MicroRNA319 (miR319) is one of the first characterized and conserved microRNA families in plants and has been demonstrated to target TCP (for TEOSINTE BRANCHED/CYCLOIDEA/PROLIFERATING CELL FACTORS [PCF]) genes encoding plant-specific transcription factors. MiR319 expression is regulated by environmental stimuli, suggesting its involvement in plant stress response, although experimental evidence is lacking and the underlying mechanism remains elusive. This study investigates the role that miR319 plays in the plant response to abiotic stress using transgenic creeping bentgrass (Agrostis stolonifera) overexpressing a rice (Oryza sativa) miR319 gene, Osa-miR319a. We found that transgenic plants overexpressing Osa-miR319a displayed morphological changes and exhibited enhanced drought and salt tolerance associated with increased leaf wax content and water retention but reduced sodium uptake. Gene expression analysis indicated that at least four putative miR319 target genes, AsPCF5, AsPCF6, AsPCF8, and AsTCP14, and a homolog of the rice NAC domain gene AsNAC60 were down-regulated in transgenic plants. Our results demonstrate that miR319 controls plant responses to drought and salinity stress. The enhanced abiotic stress tolerance in transgenic plants is related to significant down-regulation of miR319 target genes, implying their potential for use in the development of novel molecular strategies to genetically engineer crop species for enhanced resistance to environmental stress.


Assuntos
Adaptação Fisiológica/genética , Agrostis/genética , Secas , Regulação da Expressão Gênica de Plantas/efeitos dos fármacos , MicroRNAs/genética , Desenvolvimento Vegetal/genética , Cloreto de Sódio/farmacologia , Adaptação Fisiológica/efeitos dos fármacos , Agrostis/anatomia & histologia , Agrostis/efeitos dos fármacos , Agrostis/crescimento & desenvolvimento , Sequência de Bases , Membrana Celular/efeitos dos fármacos , Membrana Celular/fisiologia , Regulação da Expressão Gênica no Desenvolvimento/efeitos dos fármacos , Genes de Plantas/genética , Pleiotropia Genética/efeitos dos fármacos , MicroRNAs/metabolismo , Dados de Sequência Molecular , Tamanho do Órgão/efeitos dos fármacos , Tamanho do Órgão/genética , Oryza/efeitos dos fármacos , Oryza/genética , Fenótipo , Fotossíntese/efeitos dos fármacos , Fotossíntese/genética , Desenvolvimento Vegetal/efeitos dos fármacos , Folhas de Planta/anatomia & histologia , Folhas de Planta/efeitos dos fármacos , Plantas Geneticamente Modificadas , Salinidade , Tolerância ao Sal/efeitos dos fármacos , Tolerância ao Sal/genética , Sódio/metabolismo , Estresse Fisiológico/efeitos dos fármacos , Estresse Fisiológico/genética , Água/metabolismo , Ceras/metabolismo
16.
Plant Cell Environ ; 36(12): 2207-18, 2013 Dec.
Artigo em Inglês | MEDLINE | ID: mdl-23651319

RESUMO

MicroRNA319 (miR319) family is one of the conserved microRNA (miRNA) families among diverse plant species. It has been reported that miR319 regulates plant development in dicotyledons, but little is known at present about its functions in monocotyledons. In rice (Oryza sativa L.), the MIR319 gene family comprises two members, Osa-MIR319a and Osa-MIR319b. Here, we report an expression pattern analysis and a functional characterization of the two Osa-MIR319 genes in rice. We found that overexpressing Osa-MIR319a and Osa-MIR319b in rice both resulted in wider leaf blades. Leaves of osa-miR319 overexpression transgenic plants showed an increased number of longitudinal small veins, which probably accounted for the increased leaf blade width. In addition, we observed that overexpressing osa-miR319 led to enhanced cold tolerance (4 °C) after chilling acclimation (12 °C) in transgenic rice seedlings. Notably, under both 4 and 12 °C low temperatures, Osa-MIR319a and Osa-MIR319b were down-regulated while the expression of miR319-targeted genes was induced. Furthermore, genetically down-regulating the expression of either of the two miR319-targeted genes, OsPCF5 and OsPCF8, in RNA interference (RNAi) plants also resulted in enhanced cold tolerance after chilling acclimation. Our findings in this study demonstrate that miR319 plays important roles in leaf morphogenesis and cold tolerance in rice.


Assuntos
Adaptação Fisiológica/genética , Temperatura Baixa , Regulação da Expressão Gênica de Plantas , MicroRNAs/genética , Morfogênese/genética , Oryza/genética , Folhas de Planta/crescimento & desenvolvimento , Sequência de Bases , Regulação para Baixo/genética , Perfilação da Expressão Gênica , Genes de Plantas , MicroRNAs/metabolismo , Dados de Sequência Molecular , Oryza/fisiologia , Fenótipo , Folhas de Planta/anatomia & histologia , Folhas de Planta/genética , Plantas Geneticamente Modificadas , Estresse Fisiológico
17.
J Exp Bot ; 64(14): 4389-402, 2013 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-24085578

RESUMO

Proper branching and successful reproductive growth is of great importance for rice productivity. Substantial progress has been made in uncovering the molecular mechanisms underlying tillering control and spikelet sterility. However, rice tillering is developmentally controlled, and how it is regulated coordinately with reproductive growth remains unclear. This study characterized a rice mutant, the most obvious phenotypes of which are high tillering, reduced height, and infertile spikelets (named this1). Similarly to the high tiller number and dwarf mutants in rice, the increased tiller number of this1 plants is ascribed to the release of tiller bud outgrowth rather than to increased tiller bud formation. In the this1 mutant, however, the accelerated rate of branching was delayed until the stem elongation stage, while other mutants lost the ability to control branching at all developmental stages. The seed-setting rate of this1 was less than half that of the wild type, owing to defects in pollen maturation, anther dehiscence, and flower opening. Histological analyses showed that the mutation in this1 resulted in anisotropic cell expansion and cell division. Using a map-based cloning approach, This1 was found to encode a class III lipase. Homology searches revealed that THIS1 is conserved in both monocots and eudicots, suggesting that it plays fundamental role in regulating branch and spikelet fertility, as well as other aspects of developmental control. The relative change in expression of marker genes highlighted the possibility that This1 is involved in phytohormone signalling pathways, such as those for strigolactone and auxin. Thus, This1 provides joint control between shoot branching and reproductive development.


Assuntos
Flores/fisiologia , Lipase/metabolismo , Oryza/enzimologia , Oryza/crescimento & desenvolvimento , Proteínas de Plantas/metabolismo , Sequência de Aminoácidos , Anisotropia , Sequência de Bases , Contagem de Células , Divisão Celular , Proliferação de Células , Forma Celular , Clonagem Molecular , Citoplasma/enzimologia , Fertilidade , Regulação da Expressão Gênica de Plantas , Ácidos Indolacéticos/metabolismo , Lactonas/metabolismo , Lipase/química , Dados de Sequência Molecular , Mutação/genética , Oryza/anatomia & histologia , Oryza/genética , Filogenia , Infertilidade das Plantas , Proteínas de Plantas/química , Caules de Planta/crescimento & desenvolvimento , Pólen/crescimento & desenvolvimento
18.
Plant Cell ; 22(11): 3692-709, 2010 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-21045165

RESUMO

Recent identification of the Arabidopsis thaliana tyrosylprotein sulfotransferase (TPST) and a group of Tyr-sulfated peptides known as root meristem growth factors (RGFs) highlights the importance of protein Tyr sulfation in plant growth and development. Here, we report the action mechanism of TPST in maintenance of the root stem cell niche, which in the Arabidopsis root meristem is an area of four mitotically inactive quiescent cells plus the surrounding mitotically active stem cells. Mutation of TPST leads to defective maintenance of the root stem cell niche, decreased meristematic activity, and stunted root growth. We show that TPST expression is positively regulated by auxin and that mutation of this gene affects auxin distribution by reducing local expression levels of several PIN genes and auxin biosynthetic genes in the stem cell niche region. We also show that mutation of TPST impairs basal- and auxin-induced expression of the PLETHORA (PLT) stem cell transcription factor genes and that overexpression of PLT2 rescues the root meristem defects of the loss-of-function mutant of TPST. Together, these results support that TPST acts to maintain root stem cell niche by regulating basal- and auxin-induced expression of PLT1 and PLT2. TPST-dependent sulfation of RGFs provides a link between auxin and PLTs in regulating root stem cell niche maintenance.


Assuntos
Proteínas de Arabidopsis/metabolismo , Arabidopsis/enzimologia , Arabidopsis/crescimento & desenvolvimento , Ácidos Indolacéticos/metabolismo , Raízes de Plantas/citologia , Raízes de Plantas/crescimento & desenvolvimento , Nicho de Células-Tronco , Sulfotransferases/metabolismo , Fatores de Transcrição/metabolismo , Arabidopsis/anatomia & histologia , Proteínas de Arabidopsis/genética , Regulação da Expressão Gênica de Plantas , Meristema/anatomia & histologia , Meristema/crescimento & desenvolvimento , Meristema/metabolismo , Mutação , Filogenia , Raízes de Plantas/metabolismo , Transdução de Sinais/fisiologia , Sulfotransferases/classificação , Sulfotransferases/genética , Fatores de Transcrição/genética
19.
Theor Appl Genet ; 126(5): 1257-72, 2013 May.
Artigo em Inglês | MEDLINE | ID: mdl-23423653

RESUMO

Sheath blight (SB), caused by Rhizoctonia solani kühn, is one of the most serious global rice diseases. No major resistance genes to SB have been identified so far. All discovered loci are quantitative resistance to rice SB. The qSB-11(LE) resistance quantitative trait locus (QTL) has been previously reported on chromosome 11 of Lemont (LE). In this study, we report the precise location of qSB-11 (LE) . We developed a near isogenic line, NIL-qSB11(TQ), by marker-assisted selection that contains susceptible allele(s) from Teqing (TQ) at the qSB-11 locus in the LE genetic background. NIL-qSB11(TQ) shows higher susceptibility to SB than LE in both field and greenhouse tests, suggesting that this region of LE contains a QTL contributing to SB resistance. In order to eliminate the genetic background effects and increase the accuracy of phenotypic evaluation, a total of 112 chromosome segment substitution lines (CSSLs) with the substituted segment specific to the qSB-11 (LE) region were produced as the fine mapping population. The genetic backgrounds and morphological characteristics of these CSSLs are similar to those of the recurrent parent LE. The donor TQ chromosomal segments in these CSSL lines contiguously overlap to bridge the qSB-11 (LE) region. Through artificial inoculation, all CSSLs were evaluated for resistance to SB in the field in 2005. For the recombinant lines, their phenotypes were evaluated in the field for another 3 years and during the final year were also evaluated in a controlled greenhouse environment, showing a consistent phenotype in SB resistance across years and conditions. After comparing the genotypic profile of each CSSL with its phenotype, we are able to localize qSB-11 (LE) to the region defined by two cleaved-amplified polymorphic sequence markers, Z22-27C and Z23-33C covering 78.871 kb, based on the rice reference genome. Eleven putative genes were annotated within this region and three of them were considered the most likely candidates. The results of this study will greatly facilitate the cloning of the genes responsible for qSB-11 (LE) and marker-assisted breeding to incorporate qSB-11 (LE) into other rice cultivars.


Assuntos
Mapeamento Cromossômico , Resistência à Doença/genética , Oryza/genética , Doenças das Plantas/genética , Locos de Características Quantitativas , Rhizoctonia/patogenicidade , Cromossomos de Plantas , DNA de Plantas/genética , Genes de Plantas , Ligação Genética , Marcadores Genéticos , Imunidade Inata/genética , Oryza/imunologia , Oryza/microbiologia , Fenótipo , Doenças das Plantas/microbiologia , Rhizoctonia/genética , Rhizoctonia/imunologia
20.
Nature ; 449(7159): 243-7, 2007 Sep 13.
Artigo em Inglês | MEDLINE | ID: mdl-17694048

RESUMO

Pathogenic microbes use effectors to enhance susceptibility in host plants. However, plants have evolved a sophisticated immune system to detect these effectors using cognate disease resistance proteins, a recognition that is highly specific, often elicits rapid and localized cell death, known as a hypersensitive response, and thus potentially limits pathogen growth. Despite numerous genetic and biochemical studies on the interactions between pathogen effector proteins and plant resistance proteins, the structural bases for such interactions remain elusive. The direct interaction between the tomato protein kinase Pto and the Pseudomonas syringae effector protein AvrPto is known to trigger disease resistance and programmed cell death through the nucleotide-binding site/leucine-rich repeat (NBS-LRR) class of disease resistance protein Prf. Here we present the crystal structure of an AvrPto-Pto complex. Contrary to the widely held hypothesis that AvrPto activates Pto kinase activity, our structural and biochemical analyses demonstrated that AvrPto is an inhibitor of Pto kinase in vitro. The AvrPto-Pto interaction is mediated by the phosphorylation-stabilized P+1 loop and a second loop in Pto, both of which negatively regulate the Prf-mediated defences in the absence of AvrPto in tomato plants. Together, our results show that AvrPto derepresses host defences by interacting with the two defence-inhibition loops of Pto.


Assuntos
Proteínas de Bactérias/química , Proteínas de Bactérias/imunologia , Inibidores de Proteínas Quinases/química , Proteínas Quinases/química , Proteínas Quinases/metabolismo , Pseudomonas syringae/fisiologia , Solanum lycopersicum/imunologia , Cristalografia por Raios X , Solanum lycopersicum/enzimologia , Solanum lycopersicum/microbiologia , Fosforilação , Doenças das Plantas/imunologia , Doenças das Plantas/microbiologia , Inibidores de Proteínas Quinases/imunologia , Pseudomonas syringae/química , Pseudomonas syringae/imunologia
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