ABSTRACT
Histone variants play crucial roles in gene expression, genome integrity, and chromosome segregation. We report that the four H2A variants in Arabidopsis define different genomic features, contributing to overall genomic organization. The histone variant H2A.W marks heterochromatin specifically and acts in synergy with heterochromatic marks H3K9me2 and DNA methylation to maintain transposon silencing. In vitro, H2A.W enhances chromatin condensation by promoting fiber-to-fiber interactions via its conserved C-terminal motif. In vivo, H2A.W is required for heterochromatin condensation, demonstrating that H2A.W plays critical roles in heterochromatin organization. Similarities in conserved motifs between H2A.W and another H2A variant in metazoans suggest that plants and animals share common mechanisms for heterochromatin condensation.
Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/metabolism , Chromatin Assembly and Disassembly , Heterochromatin/metabolism , Histones/metabolism , Amino Acid Sequence , Arabidopsis/genetics , Arabidopsis Proteins/chemistry , Arabidopsis Proteins/genetics , DNA Methylation , DNA Transposable Elements , Genome-Wide Association Study , Histones/chemistry , Histones/genetics , Molecular Sequence Data , Sequence AlignmentABSTRACT
Symbiotic interactions between rhizobia and legumes result in the formation of root nodules, which fix nitrogen that can be used for plant growth. Rhizobia usually invade legume roots through a plant-made tunnel-like structure called an infection thread (IT). RPG (Rhizobium-directed polar growth) encodes a coiled-coil protein that has been identified in Medicago truncatula as required for root nodule infection, but the function of RPG remains poorly understood. In this study, we identified and characterized RPG in Lotus japonicus and determined that it is required for IT formation. RPG was induced by Mesorhizobium loti or purified Nodulation factor and displayed an infection-specific expression pattern. Nodule inception (NIN) bound to the RPG promoter and induced its expression. We showed that RPG displayed punctate subcellular localization in L. japonicus root protoplasts and in root hairs infected by M. loti. The N-terminal predicted C2 lipid-binding domain of RPG was not required for this subcellular localization or for function. CERBERUS, a U-box E3 ligase which is also required for rhizobial infection, was found to be localized similarly in puncta. RPG co-localized and directly interacted with CERBERUS in the early endosome (TGN/EE) compartment and near the nuclei in root hairs after rhizobial inoculation. Our study sheds light on an RPG-CERBERUS protein complex that is involved in an exocytotic pathway mediating IT elongation.
Subject(s)
Lotus , Rhizobium , Rhizobium/genetics , Lotus/genetics , Lotus/metabolism , Ubiquitin-Protein Ligases/genetics , Ubiquitin-Protein Ligases/metabolism , Plant Proteins/genetics , Plant Proteins/metabolism , Symbiosis/genetics , Gene Expression Regulation, Plant , Root Nodules, Plant/genetics , Plant RootsABSTRACT
Linker H1 histones play an important role in animal and human pathogenesis, but their function in plant immunity is poorly understood. Here, we analyzed mutants of the three canonical variants of Arabidopsis H1 histones, namely H1.1, H1.2 and H1.3. We observed that double h1.1h1.2 and triple h1.1h1.2h1.3 (3h1) mutants were resistant to Pseudomonas syringae and Botrytis cinerea infections. Transcriptome analysis of 3h1 mutant plants showed H1s play a key role in regulating the expression of early and late defense genes upon pathogen challenge. Moreover, 3h1 mutant plants showed enhanced production of reactive oxygen species and activation of mitogen activated protein kinases upon pathogen-associated molecular pattern (PAMP) treatment. However, 3h1 mutant plants were insensitive to priming with flg22, a well-known bacterial PAMP which induces enhanced resistance in WT plants. The defective defense response in 3h1 upon priming was correlated with altered DNA methylation and reduced global H3K56ac levels. Our data place H1 as a molecular gatekeeper in governing dynamic changes in the chromatin landscape of defense genes during plant pathogen interaction.
Subject(s)
Arabidopsis Proteins , Arabidopsis , Histones , Host-Pathogen Interactions , Plant Diseases , Plant Immunity , Arabidopsis/genetics , Arabidopsis/immunology , Arabidopsis/metabolism , Arabidopsis/microbiology , Arabidopsis Proteins/genetics , Arabidopsis Proteins/metabolism , Bacterial Proteins/immunology , Chromatin/chemistry , Chromatin/genetics , Chromatin/metabolism , DNA Methylation , Gene Expression Regulation, Plant , Histones/genetics , Histones/metabolism , Mitogen-Activated Protein Kinases/metabolism , Mutation , Pathogen-Associated Molecular Pattern Molecules/immunology , Pathogen-Associated Molecular Pattern Molecules/metabolism , Plant Diseases/immunology , Plant Diseases/microbiology , Plant Immunity/genetics , Plant Immunity/immunology , Pseudomonas syringae/immunology , Pseudomonas syringae/metabolism , Reactive Oxygen Species/metabolismABSTRACT
In Arabidopsis, CHG DNA methylation is controlled by the H3K9 methylation mark through a self-reinforcing loop between DNA methyltransferase CHROMOMETHYLASE3 (CMT3) and H3K9 histone methyltransferase KRYPTONITE/SUVH4 (KYP). We report on the structure of KYP in complex with methylated DNA, substrate H3 peptide, and cofactor SAH, thereby defining the spatial positioning of the SRA domain relative to the SET domain. The methylated DNA is bound by the SRA domain with the 5mC flipped out of the DNA, while the H3(1-15) peptide substrate binds between the SET and post-SET domains, with the ε-ammonium of K9 positioned adjacent to bound SAH. These structural insights, complemented by functional data on key mutants of residues lining the 5mC and H3K9-binding pockets within KYP, establish how methylated DNA recruits KYP to the histone substrate. Together, the structures of KYP and previously reported CMT3 complexes provide insights into molecular mechanisms linking DNA and histone methylation.
Subject(s)
Arabidopsis Proteins/chemistry , Arabidopsis Proteins/metabolism , Arabidopsis/genetics , DNA Methylation , DNA, Plant/chemistry , DNA, Plant/genetics , Histone-Lysine N-Methyltransferase/chemistry , Histone-Lysine N-Methyltransferase/metabolism , Histones/physiology , Arabidopsis/chemistry , Arabidopsis/metabolism , Binding Sites/genetics , Epigenesis, Genetic , Gene Expression Regulation, Plant , Genome, Plant , Models, Molecular , S-Adenosylhomocysteine/metabolism , X-Ray DiffractionABSTRACT
DNA methylation is a mechanism of epigenetic gene regulation and genome defense conserved in many eukaryotic organisms. In Arabidopsis, the DNA methyltransferase domains rearranged methylase 2 (DRM2) controls RNA-directed DNA methylation in a pathway that also involves the plant-specific RNA Polymerase V (Pol V). Additionally, the Arabidopsis genome encodes an evolutionarily conserved but catalytically inactive DNA methyltransferase, DRM3. Here, we show that DRM3 has moderate effects on global DNA methylation and small RNA abundance and that DRM3 physically interacts with Pol V. In Arabidopsis drm3 mutants, we observe a lower level of Pol V-dependent noncoding RNA transcripts even though Pol V chromatin occupancy is increased at many sites in the genome. These findings suggest that DRM3 acts to promote Pol V transcriptional elongation or assist in the stabilization of Pol V transcripts. This work sheds further light on the mechanism by which long noncoding RNAs facilitate RNA-directed DNA methylation.
Subject(s)
Arabidopsis/enzymology , DNA Methylation/physiology , DNA-Directed RNA Polymerases/genetics , Methyltransferases/physiology , RNA, Messenger/genetics , Arabidopsis/genetics , Genes, Plant , Methyltransferases/genetics , RNA, Messenger/metabolismABSTRACT
DNA methylation in Arabidopsis thaliana is maintained by at least four different enzymes: DNA methyltransferase1 (MET1), chromomethylase3 (CMT3), domains rearranged methyltransferase2 (DRM2), and chromomethylase2 (CMT2). However, DNA methylation is established exclusively by the enzyme DRM2, which acts in the RNA-directed DNA methylation (RdDM) pathway. Some RdDM components belong to gene families and have partially redundant functions, such as the endoribonucleases dicer-like 2, 3, and 4, and involved in de novo2 (IDN2) interactors IDN2-like 1 and 2. Traditional mutagenesis screens usually fail to detect genes if they are redundant, as the loss of one gene can be compensated by a related gene. In an effort to circumvent this issue, we used coexpression data to identify closely related genes that are coregulated with genes in the RdDM pathway. Here we report the discovery of two redundant proteins, SNF2-ring-helicase-like1 and -2 (FRG1 and -2) that are putative chromatin modifiers belonging to the SNF2 family of helicase-like proteins. Analysis of genome-wide bisulfite sequencing shows that simultaneous mutations of FRG1 and -2 cause defects in methylation at specific RdDM targeted loci. We also show that FRG1 physically associates with Su(var)3-9-related SUVR2, a known RdDM component, in vivo. Combined, our results identify FRG1 and FRG2 as previously unidentified components of the RdDM machinery.
Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/genetics , Chromatin/chemistry , Chromosomal Proteins, Non-Histone/metabolism , DNA Methylation , RNA, Plant/chemistry , Arabidopsis/metabolism , Chromatin/metabolism , Chromatin Assembly and Disassembly , Epigenesis, Genetic , Gene Expression Regulation, Plant , Gene Silencing , Mutagenesis , Mutation , RNA Interference , RNA, Small Interfering/geneticsABSTRACT
In eukaryotic cells, environmental and developmental signals alter chromatin structure and modulate gene expression. Heterochromatin constitutes the transcriptionally inactive state of the genome and in plants and mammals is generally characterized by DNA methylation and histone modifications such as histone H3 lysine 9 (H3K9) methylation. In Arabidopsis thaliana, DNA methylation and H3K9 methylation are usually colocated and set up a mutually self-reinforcing and stable state. Here, in contrast, we found that SUVR5, a plant Su(var)3-9 homolog with a SET histone methyltransferase domain, mediates H3K9me2 deposition and regulates gene expression in a DNA methylation-independent manner. SUVR5 binds DNA through its zinc fingers and represses the expression of a subset of stimulus response genes. This represents a novel mechanism for plants to regulate their chromatin and transcriptional state, which may allow for the adaptability and modulation necessary to rapidly respond to extracellular cues.
Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/genetics , Arabidopsis/metabolism , DNA Methylation , Gene Expression Regulation, Plant , Gene Silencing , Histone-Lysine N-Methyltransferase/metabolism , Histones/metabolism , Arabidopsis Proteins/chemistry , Base Sequence , Binding Sites , DNA-Binding Proteins/chemistry , DNA-Binding Proteins/metabolism , Histone Demethylases/metabolism , Histone-Lysine N-Methyltransferase/chemistry , Nucleotide Motifs , Promoter Regions, Genetic , Protein Binding , Protein Interaction Domains and Motifs , Zinc FingersABSTRACT
Arbuscular mycorrhiza (AM) fungi form nutrient-acquiring symbioses with the majority of higher plants. Nutrient exchange occurs via arbuscules, highly branched hyphal structures that are formed within root cortical cells. With a view to identifying host genes involved in AM development, we isolated Lotus japonicus AM-defective mutants via a microscopic screen of an ethyl methanesulfonate-mutagenized population. A standardized mapping procedure was developed that facilitated positioning of the defective loci on the genetic map of L. japonicus, and, in five cases, allowed identification of mutants of known symbiotic genes. Two additional mutants representing independent loci did not form mature arbuscules during symbiosis with two divergent AM fungal species, but exhibited signs of premature arbuscule arrest or senescence. Marker gene expression patterns indicated that the two mutants are affected in distinct steps of arbuscule development. Both mutants formed wild-type-like root nodules upon inoculation with Mesorhizobium loti, indicating that the mutated loci are essential during AM but not during root nodule symbiosis.
Subject(s)
Fungi/physiology , Gene Expression Regulation, Plant , Lotus/genetics , Mesorhizobium/physiology , Mycorrhizae/genetics , Chromosome Mapping , Ethyl Methanesulfonate/pharmacology , Fungi/growth & development , Fungi/ultrastructure , Genetic Loci , Hyphae , Lotus/growth & development , Lotus/microbiology , Lotus/ultrastructure , Mutation , Mycorrhizae/growth & development , Mycorrhizae/ultrastructure , Phenotype , Plant Proteins/genetics , Plant Proteins/metabolism , Plant Root Nodulation , Plant Roots/genetics , Plant Roots/microbiology , Plant Roots/ultrastructure , Root Nodules, Plant , Sequence Analysis, DNA , SymbiosisABSTRACT
Legumes form symbioses with arbuscular mycorrhiza (AM) fungi and nitrogen fixing root nodule bacteria. Intracellular root infection by either endosymbiont is controlled by the activation of the calcium and calmodulin-dependent kinase (CCaMK), a central regulatory component of the plant's common symbiosis signaling network. We performed a microscopy screen for Lotus japonicus mutants defective in AM development and isolated a mutant, nena, that aborted fungal infection in the rhizodermis. NENA encodes a WD40 repeat protein related to the nucleoporins Sec13 and Seh1. Localization of NENA to the nuclear rim and yeast two-hybrid experiments indicated a role for NENA in a conserved subcomplex of the nuclear pore scaffold. Although nena mutants were able to form pink nodules in symbiosis with Mesorhizobium loti, root hair infection was not observed. Moreover, Nod factor induction of the symbiotic genes NIN, SbtM4, and SbtS, as well as perinuclear calcium spiking, were impaired. Detailed phenotypic analyses of nena mutants revealed a rhizobial infection mode that overcame the lack of rhizodermal responsiveness and carried the hallmarks of crack entry, including a requirement for ethylene. CCaMK-dependent processes were only abolished in the rhizodermis but not in the cortex of nena mutants. These data support the concept of tissue-specific components for the activation of CCaMK.
Subject(s)
Lotus/metabolism , Mycorrhizae/pathogenicity , Plant Proteins/physiology , Rhizobium/pathogenicity , Symbiosis , Cloning, Molecular , Genetic Complementation Test , Molecular Sequence Data , Mutation , Reverse Transcriptase Polymerase Chain Reaction , Two-Hybrid System TechniquesABSTRACT
Plants are extremely plastic organisms. They continuously receive and integrate environmental information and adjust their growth and development to favour fitness and survival. When this integration of information affects subsequent life stages or the development of subsequent generations, it can be considered an environmental memory. Thus, plant memory is a relevant mechanism by which plants respond adaptively to different environments. If the cost of maintaining the response is offset by its benefits, it may influence evolutionary trajectories. As such, plant memory has a sophisticated underlying molecular mechanism with multiple components and layers. Nonetheless, when mathematical modelling is combined with knowledge of ecological, physiological, and developmental effects as well as molecular mechanisms as a tool for understanding plant memory, the combined potential becomes unfathomable for the management of plant communities in natural and agricultural ecosystems. In this review, we summarize recent advances in the understanding of plant memory, discuss the ecological requirements for its evolution, outline the multilayered molecular network and mechanisms required for accurate and fail-proof plant responses to variable environments, point out the direct involvement of the plant metabolism and discuss the tremendous potential of various types of models to further our understanding of the plant's environmental memory. Throughout, we emphasize the use of plant memory as a tool to unlock the secrets of the natural world.
ABSTRACT
We have established tools for forward and reverse genetic analysis of the legume Lotus (Lotus japonicus). A structured population of M2 progeny of 4,904 ethyl methanesulfonate-mutagenized M1 embryos is available for single nucleotide polymorphism mutation detection, using a TILLING (for Targeting Induced Local Lesions IN Genomes) protocol. Scanning subsets of this population, we identified a mutation load of one per 502 kb of amplified fragment. Moreover, we observed a 1:10 ratio between homozygous and heterozygous mutations in the M2 progeny. This reveals a clear difference in germline genetics between Lotus and Arabidopsis (Arabidopsis thaliana). In addition, we assembled M2 siblings with obvious phenotypes in overall development, starch accumulation, or nitrogen-fixing root nodule symbiosis in three thematic subpopulations. By screening the nodulation-defective population of M2 individuals for mutations in a set of 12 genes known to be essential for nodule development, we identified large allelic series for each gene, generating a unique data set that combines genotypic and phenotypic information facilitating structure-function studies. This analysis revealed a significant bias for replacements of glycine (Gly) residues in functionally defective alleles, which may be explained by the exceptional structural features of Gly. Gly allows the peptide chain to adopt conformations that are no longer possible after amino acid replacement. This previously unrecognized vulnerability of proteins at Gly residues could be used for the improvement of algorithms that are designed to predict the deleterious nature of single nucleotide polymorphism mutations. Our results demonstrate the power, as well as the limitations, of ethyl methanesulfonate mutagenesis for forward and reverse genetic studies. (Original mutant phenotypes can be accessed at http://data.jic.bbsrc.ac.uk/cgi-bin/lotusjaponicus Access to the Lotus TILLING facility can be obtained through http://www.lotusjaponicus.org or http://revgenuk.jic.ac.uk).
Subject(s)
Ethyl Methanesulfonate/pharmacology , Lotus/genetics , Mutagenesis , Plant Root Nodulation/genetics , Symbiosis/genetics , Alleles , Arabidopsis/genetics , DNA, Plant/genetics , Databases, Genetic , Gene Expression Regulation, Plant , Genes, Plant , Genome, Plant , Molecular Sequence Data , Mutation , PhenotypeABSTRACT
BACKGROUND: One of the fascinating aspects of epigenetic regulation is that it provides means to rapidly adapt to environmental change. This is particularly relevant in the plant kingdom, where most species are sessile and exposed to increasing habitat fluctuations due to global warming. Although the inheritance of epigenetically controlled traits acquired through environmental impact is a matter of debate, it is well documented that environmental cues lead to epigenetic changes, including chromatin modifications, that affect cell differentiation or are associated with plant acclimation and defense priming. Still, in most cases, the mechanisms involved are poorly understood. An emerging topic that promises to reveal new insights is the interaction between epigenetics and metabolism. SCOPE OF REVIEW: This study reviews the links between metabolism and chromatin modification, in particular histone acetylation, histone methylation, and DNA methylation, in plants and compares them to examples from the mammalian field, where the relationship to human diseases has already generated a larger body of literature. This study particularly focuses on the role of reactive oxygen species (ROS) and nitric oxide (NO) in modulating metabolic pathways and gene activities that are involved in these chromatin modifications. As ROS and NO are hallmarks of stress responses, we predict that they are also pivotal in mediating chromatin dynamics during environmental responses. MAJOR CONCLUSIONS: Due to conservation of chromatin-modifying mechanisms, mammals and plants share a common dependence on metabolic intermediates that serve as cofactors for chromatin modifications. In addition, plant-specific non-CG methylation pathways are particularly sensitive to changes in folate-mediated one-carbon metabolism. Finally, reactive oxygen and nitrogen species may fine-tune epigenetic processes and include similar signaling mechanisms involved in environmental stress responses in plants as well as animals.
Subject(s)
Chromatin/metabolism , Plants/genetics , Plants/metabolism , Chromatin/genetics , DNA Methylation/genetics , Epigenesis, Genetic/genetics , Epigenomics/methods , Histones/genetics , Reactive Oxygen Species/metabolism , Signal Transduction , Stress, PhysiologicalABSTRACT
Small RNAs regulate chromatin modifications such as DNA methylation and gene silencing across eukaryotic genomes. In plants, RNA-directed DNA methylation (RdDM) requires 24-nucleotide small interfering RNAs (siRNAs) that bind to ARGONAUTE 4 (AGO4) and target genomic regions for silencing. RdDM also requires non-coding RNAs transcribed by RNA polymerase V (Pol V) that probably serve as scaffolds for binding of AGO4-siRNA complexes. Here, we used a modified global nuclear run-on protocol followed by deep sequencing to capture Pol V nascent transcripts genome-wide. We uncovered unique characteristics of Pol V RNAs, including a uracil (U) common at position 10. This uracil was complementary to the 5' adenine found in many AGO4-bound 24-nucleotide siRNAs and was eliminated in a siRNA-deficient mutant as well as in the ago4/6/9 triple mutant, suggesting that the +10 U signature is due to siRNA-mediated co-transcriptional slicing of Pol V transcripts. Expression of wild-type AGO4 in ago4/6/9 mutants was able to restore slicing of Pol V transcripts, but a catalytically inactive AGO4 mutant did not correct the slicing defect. We also found that Pol V transcript slicing required SUPPRESSOR OF TY INSERTION 5-LIKE (SPT5L), an elongation factor whose function is not well understood. These results highlight the importance of Pol V transcript slicing in RNA-mediated transcriptional gene silencing, which is a conserved process in many eukaryotes.
Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/metabolism , DNA Methylation , DNA-Directed RNA Polymerases/metabolism , Gene Silencing , RNA, Guide, Kinetoplastida/metabolism , RNA, Small Nuclear/metabolism , Argonaute Proteins/metabolism , Genome-Wide Association Study , Transcription, GeneticABSTRACT
DNA methylation generally functions as a repressive transcriptional signal, but it is also known to activate gene expression. In either case, the downstream factors remain largely unknown. By using comparative interactomics, we isolated proteins in Arabidopsis thaliana that associate with methylated DNA. Two SU(VAR)3-9 homologs, the transcriptional antisilencing factor SUVH1, and SUVH3, were among the methyl reader candidates. SUVH1 and SUVH3 bound methylated DNA in vitro, were associated with euchromatic methylation in vivo, and formed a complex with two DNAJ domain-containing homologs, DNAJ1 and DNAJ2. Ectopic recruitment of DNAJ1 enhanced gene transcription in plants, yeast, and mammals. Thus, the SUVH proteins bind to methylated DNA and recruit the DNAJ proteins to enhance proximal gene expression, thereby counteracting the repressive effects of transposon insertion near genes.
Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/genetics , DNA Methylation , Gene Expression Regulation, Plant , HSP40 Heat-Shock Proteins/metabolism , Histone-Lysine N-Methyltransferase/metabolism , Methyltransferases/metabolism , Transcription, Genetic , Arabidopsis/enzymology , HSP40 Heat-Shock Proteins/chemistry , Protein DomainsABSTRACT
Proteases catalyze the hydrolysis of peptide bonds in proteins/peptides inside or outside of cells. They play important roles in development and responses to environmental stresses. In arbuscular mycorrhiza (AM), symbiosis-induced protease genes were found by large-scale transcriptome analyses in different plant species, suggesting that proteolytic processes are implicated in AM. In legumes, some of these were also transcriptionally activated during the root nodule symbiosis. However, the precise function of these symbiosis-induced proteases remains unknown. Here we present a compilation of the symbiosis-induced proteases identified so far and discuss their possible roles in symbiosis.
Subject(s)
Peptide Hydrolases/physiology , Plant Roots/enzymology , Symbiosis/physiology , Enzyme Induction , Gene Expression Regulation, PlantABSTRACT
DNA methylation is an epigenetic mechanism that has important functions in transcriptional silencing and is associated with repressive histone methylation (H3K9me). To further investigate silencing mechanisms, we screened a mutagenized Arabidopsis thaliana population for expression of SDCpro-GFP, redundantly controlled by DNA methyltransferases DRM2 and CMT3. Here, we identify the hypomorphic mutant mthfd1-1, carrying a mutation (R175Q) in the cytoplasmic bifunctional methylenetetrahydrofolate dehydrogenase/methenyltetrahydrofolate cyclohydrolase (MTHFD1). Decreased levels of oxidized tetrahydrofolates in mthfd1-1 and lethality of loss-of-function demonstrate the essential enzymatic role of MTHFD1 in Arabidopsis. Accumulation of homocysteine and S-adenosylhomocysteine, genome-wide DNA hypomethylation, loss of H3K9me and transposon derepression indicate that S-adenosylmethionine-dependent transmethylation is inhibited in mthfd1-1. Comparative analysis of DNA methylation revealed that the CMT3 and CMT2 pathways involving positive feedback with H3K9me are mostly affected. Our work highlights the sensitivity of epigenetic networks to one-carbon metabolism due to their common S-adenosylmethionine-dependent transmethylation and has implications for human MTHFD1-associated diseases.
Subject(s)
Arabidopsis Proteins/metabolism , Arabidopsis/genetics , DNA Methylation/genetics , Methenyltetrahydrofolate Cyclohydrolase/metabolism , Methylenetetrahydrofolate Dehydrogenase (NADP)/metabolism , Arabidopsis Proteins/genetics , Cytoplasm/drug effects , Cytoplasm/metabolism , DNA Demethylation , Epigenesis, Genetic , Folic Acid/metabolism , Gene Expression Regulation, Plant/drug effects , Gene Silencing , Green Fluorescent Proteins/metabolism , Histones/metabolism , Homeostasis/drug effects , Lysine/metabolism , Methenyltetrahydrofolate Cyclohydrolase/genetics , Methionine/pharmacology , Methylenetetrahydrofolate Dehydrogenase (NADP)/genetics , Models, Biological , Mutation/genetics , Protein Transport/drug effects , S-Adenosylmethionine/metabolism , Tetrahydrofolates/pharmacologyABSTRACT
Bugs such as Nezara viridula (Linnaeus) and Dichelops melacanthus (Dallas) are considered the main insect pests of wheat crop in Brazil. The use of the entomopathogenic fungus Metarhizium anisopliae (Metschnikoff) may be an alternative for the management of these insects in the crop. The objective of this work was to verify the pathogenicity of different isolates of M. anisopliae on adults of N. viridula and D. melacanthus under laboratory and greenhouse conditions. In the laboratory, isolates 05RA, 11RA, 08RA and 02RA were obtained from N. viridula and D. melacanthus infested with M. anisopliae. Also, a high pathogenicity (100% of mortality) of both species was recorded in a bioassay of the topical application 8 Days After Application (DAA). However, compared to the other isolates, the 08RA isolate showed the highest pathogenicity in a shorter time interval for N. viridula (Mean Time "MT" = 2.8 days) and D. melacanthus (MT = 4.0 days). Under greenhouse conditions, the 08RA isolate provided a mortality of 44.9% (N. viridula) and 35.7% (D. melacanthus) in the same evaluation period. However, at 14 DAA, the mortality was 100% for both species, with the MT values of N. viridula and D. melacanthus being obtained at 8 days and 10 days, respectively. The fungus M. anisopliae is a promising alternative for the control of adult N. viridula and D. melacanthus in wheat crop.(AU)
Os percevejos Nezaraviridula (Linnaeus) e o Dichelops melacanthus (Dallas) são considerados os principais insetos-pragas na cultura do trigo no Brasil. A utilização do fungo entomopatogênico Metarhiziumanisopliae (Metschnikoff) pode ser uma alternativa para o manejo destes insetos na cultura. O objetivo do trabalho foi verificar a patogenicidade de diferentes isolados de M.anisopliae sobre adultos de N. viridula ou D. melacanthus em condições de laboratório e casa de vegetação. Em laboratório, os isolados 05RA, 11RA, 08RA e 02RA provenientes de adultos de N. viridula ou D. melacanthus, infestados a campo pelo fungo M. anisopliae, apresentaram elevada patogenicidade (100% de mortalidade) de ambas as espécies em bioensaio de aplicação tópica 8 dias após aplicação (DAA). Entretanto, o isolado 08RA apresentou a maior patogenicidade em um menor intervalo de tempo para N. viridula (Tempo Médio "TM" = 2,8 dias) e D. melacanthus (TM = 4,0 dias) em relação aos demais isolados. Em casa de vegetação, o isolado 08RA proporcionou mortalidade de 44,9% (N. viridula) e 35,7% (D. melacanthus) no mesmo período de avaliação. Contudo, aos 14 DAA, a mortalidade foi de 100% para ambas as espécies, com valores de TM de 8 dias e 10 dias para N. viridula e D. melacanthus, respectivamente. O fungo M. anisopliae constitui uma alternativa promissora de controle de adultos de N. viridula ou D. melacanthus na cultura do trigo.(AU)
Subject(s)
Triticum , Pest Control, Biological , Cimicidae , Fungi , Insect ControlABSTRACT
Studies combining comparative genomics and information on biochemical pathways have revealed that protein evolution can be affected by the amount of pleiotropy associated with a particular gene. The amount of pleiotropy, in turn, can be a function of the position at which a gene operates in a pathway and the pathway structure. Genes that serve as convergence points and have several partners (so-called hubs) often show the greatest constraint and hence the slowest rate of protein evolution. In this article, we have studied five genes (Pto, Fen, Rin4, Prf and Pfi) in a defence signalling network in a wild tomato species, Solanum peruvianum. These proteins operate together and contribute to bacterial resistance in tomato. We predicted that Prf (and possibly Pfi), which serves as a convergence point for upstream signals, should show greater evolutionary constraint. However, we found instead that two of the genes which potentially interact with pathogen ligands, Rin4 and Fen, have evolved under strong evolutionary constraint, whereas Prf and Pfi, which probably function further downstream in the network, show evidence of balancing selection. This counterintuitive observation may be probable in pathogen defence networks, because pathogens may target positions throughout resistance networks to manipulate or nullify host resistance, thereby leaving a molecular signature of host-parasite co-evolution throughout a single network.
Subject(s)
Disease Resistance/physiology , Plant Proteins/metabolism , Solanum lycopersicum/metabolism , Disease Resistance/genetics , Evolution, Molecular , Genetic Variation/genetics , Solanum lycopersicum/genetics , Solanum lycopersicum/microbiology , Solanum lycopersicum/parasitology , Plant Proteins/geneticsABSTRACT
The brush mutant of Lotus japonicus exhibits a temperature-dependent impairment in nodule, root, and shoot development. At 26 degrees C, brush formed fewer nodules, most of which were not colonized by rhizobia bacteria. Primary root growth was retarded and the anatomy of the brush root apical meristem revealed distorted cellular organization and reduced cell expansion. Reciprocal grafting of brush with wild-type plants indicated that this genotype only affected the root and that the shoot phenotype was a secondary effect. The root and nodulation phenotype cosegregated as a single Mendelian trait and the BRUSH gene could be mapped to the short arm of chromosome 2. At 18 degrees C, the brush root anatomy was rescued and similar to the wild type, and primary root length, number of infection threads, and nodule formation were partially rescued. Superficially, the brush root phenotype resembled the ethylene-related thick short root syndrome. However, treatment with ethylene inhibitor did not recover the observed phenotypes, although brush primary roots were slightly longer. The defects of brush in root architecture and infection thread development, together with intact nodule architecture and complete absence of symptoms from shoots, suggest that BRUSH affects cellular differentiation in a tissue-dependent way.