RESUMO
The coordinated expression of highly related homoeologous genes in polyploid species underlies the phenotypes of many of the world's major crops. Here we combine extensive gene expression datasets to produce a comprehensive, genome-wide analysis of homoeolog expression patterns in hexaploid bread wheat. Bias in homoeolog expression varies between tissues, with ~30% of wheat homoeologs showing nonbalanced expression. We found expression asymmetries along wheat chromosomes, with homoeologs showing the largest inter-tissue, inter-cultivar, and coding sequence variation, most often located in high-recombination distal ends of chromosomes. These transcriptionally dynamic genes potentially represent the first steps toward neo- or subfunctionalization of wheat homoeologs. Coexpression networks reveal extensive coordination of homoeologs throughout development and, alongside a detailed expression atlas, provide a framework to target candidate genes underpinning agronomic traits in wheat.
Assuntos
Regulação da Expressão Gênica de Plantas , Poliploidia , Transcrição Gênica , Triticum/genética , Pão , Perfilação da Expressão Gênica , Regulação da Expressão Gênica no Desenvolvimento , Genoma de Planta , RNA de Plantas/genética , Análise de Sequência de RNA , Triticum/crescimento & desenvolvimentoRESUMO
During pathogenesis, fungal pathogens are exposed to a variety of fungitoxic compounds. This may be particularly relevant to Botrytis cinerea, a plant pathogen that has a broad host range and, consequently, is subjected to exposure to many plant defense compounds. In practice, the pathogen is controlled with fungicides belonging to different chemical groups. ATP-binding cassette (ABC) transporters might provide protection against plant defense compounds and fungicides by ATP-driven efflux mechanisms. To test this hypothesis, we cloned BcatrB, an ABC transporter-encoding gene from B. cinerea. This gene encodes a 1,439 amino acid protein with nucleotide binding fold (NBF) and transmembrane (TM) domains in a [NBF-TM6]2 topology. The amino acid sequence has 31 to 67% identity with ABC transporters from various fungi. The expression of BcatrB is up regulated by treatment of B. cinerea germlings with the grapevine phytoalexin resveratrol and the fungicide fenpiclonil. BcatrB replacement mutants are not affected in saprophytic growth on different media but are more sensitive to resveratrol and fenpiclonil than the parental isolate. Furthermore, virulence of deltaBcatrB mutants on grapevine leaves was slightly reduced. These results indicate that BcatrB is a determinant in sensitivity of B. cinerea to plant defense compounds and fungicides.
Assuntos
Transportadores de Cassetes de Ligação de ATP/fisiologia , Botrytis/efeitos dos fármacos , Pirróis/farmacologia , Estilbenos/farmacologia , Transportadores de Cassetes de Ligação de ATP/química , Transportadores de Cassetes de Ligação de ATP/genética , Sequência de Aminoácidos , Botrytis/metabolismo , Botrytis/patogenicidade , Clonagem Molecular , Dados de Sequência Molecular , Resveratrol , Homologia de Sequência de Aminoácidos , VirulênciaRESUMO
This study demonstrates that the ATP-binding cassette (ABC) transporter BcatrB from Botrytis cinerea influences the activity of phenlpyrrole fungicides against the pathogen. This conclusion is based on toxicity assays and northern analysis experiments which show that BcatrB replacement mutants, which do not express the BcatrB gene, show an increased sensitivity to the phenylpyrrole fungicides fludioxonil and fenpiclonil. Mutants overexpressing BcatrB exhibit a decreased sensitivity to these fungicides. In addition, accumulation of fludioxonil by BcatrB replacement mutants was higher than by wild-type isolates. For mutants overexpressing BcatrB the reverse was observed. Additional ABC and major facilitator superfamily (MFS) transporter genes were identified in an expressed sequence tag (EST) database, suggesting that B cinerea has gene families of ABC and MFS transporters. Corresponding fragments of ten ABC (BcatrC-BcatrN) and three MFS transporter genes (Bcmfs1-4) were cloned and characterised. Fludioxonil affected the transcript level of some members of these gene families in germlings during a short treatment with the fungicide at sub-lethal concentrations. Hence, other ABC and MFS transporters may affect the activity of phenylpyrrole fungicides as well. Other fungicides such as the anilinopyrimidine fungicide cyprodinil, the azole fungicide tebuconazole, the dicarboximide fungicide iprodione and the strobilurin fungicide trifloxystrobin also induced transcription of some of the ABC and MFS transporter genes identified. Therefore, we propose that various ABC and MFS transporters function in protection of the fungus against fungicides and are involved in multi-drug resistance development.
Assuntos
Transportadores de Cassetes de Ligação de ATP/genética , Botrytis/genética , Dioxóis/farmacologia , Resistência a Múltiplos Medicamentos/genética , Fungicidas Industriais/farmacologia , Genes Fúngicos , Pirróis/farmacologia , Transportadores de Cassetes de Ligação de ATP/metabolismo , Transporte Biológico Ativo , Northern Blotting , Botrytis/efeitos dos fármacos , Botrytis/metabolismo , Clonagem Molecular , Proteínas Fúngicas/química , Proteínas Fúngicas/genética , Regulação Fúngica da Expressão Gênica , Genes de RNAr , Testes de Sensibilidade Microbiana , Mutação , Doenças das Plantas/microbiologia , RNA Mensageiro/metabolismo , Análise de Sequência de RNARESUMO
Survival of microorganisms in natural environments is favored by the capacity to produce compounds toxic to competing organisms and the ability to resist the effects of such toxic compounds. Both factors contribute to a competitive advantage of organisms in ecosystems. All organisms have evolved active transport mechanisms by which endogenous and exogenous toxicants can be secreted. Two major classes of transporter proteins are the ATP-binding cassette (ABC) and the major facilitator superfamily (MFS) transporters. Members of both classes can have broad and overlapping substrate specificities for natural toxic compounds and can be regarded as a "first-line defense barrier" in survival mechanisms. In plant pathogens, these transporters can play an essential role in protection against plant defense compounds during pathogenesis. Also, some transporters actively secrete host-specific and non-host-specific toxins. Remarkably, ABC and MFS transporters can also play a major role in fungicide sensitivity and resistance. Their role in multidrug resistance of Aspergillus nidulans, Candida albicans, and Saccharomyces cerevisiae to azoles and other fungitoxic compounds is well established. Knowledge of ABC and MFS transporters opens possibilities of developing novel strategies for controlling plant diseases, either by modulation of transporter activity or by transgenic expression of transporter genes in plants.