RESUMEN
Phytophthora sojae is an oomycete and a pathogen of soybean that causes root rot. During infection P. sojae delivers effector proteins into host cells to foster disease. However, effector-triggered immunity (ETI) results when pathogen factors are recognized by host resistance (R) proteins. We have now identified the P. sojae Avr1d gene, which encodes a predicted effector protein with the amino acid motif Arg-X-Leu-Arg (RXLR). Genetic mapping of 16 different P. sojae isolates and of a segregating F2 population of 40 individuals shows that the predicted RXLR effector gene Avh6 precisely cosegregates with the Avr1d phenotype. Transient expression assays confirm that Avr1d triggers cell death specifically in Rps1d soybean plants. The Avr1d gene is present in P. sojae strains that are avirulent on Rps1d, whereas the gene is deleted from the genome of virulent strains. Two sequence variants of the Avr1d gene encoding different protein products occur in P. sojae strains, but both are recognized by Rps1d and cause ETI. Liposome binding assays show that Avr1d has affinity for phosphatidylinositol 4-phosphate and that binding can be disrupted by mutation of lysine residues in the carboxy-terminal effector domain of the protein. The identification of Avr1d aids pathogen diagnostics and soybean cultivar development.
Asunto(s)
Glycine max/microbiología , Phytophthora/metabolismo , Phytophthora/patogenicidad , Enfermedades de las Plantas/parasitología , Eliminación de Gen , Regulación de la Expresión Génica , Haplotipos , Phytophthora/genética , Proteínas Recombinantes/genética , Proteínas Recombinantes/metabolismo , VirulenciaRESUMEN
Plants have evolved pathogen-associated molecular pattern (PAMP)-triggered immunity (PTI) and effector-triggered immunity (ETI) to protect themselves from infection by diverse pathogens. Avirulence (Avr) effectors that trigger plant ETI as a result of recognition by plant resistance (R) gene products have been identified in many plant pathogenic oomycetes and fungi. However, the virulence functions of oomycete and fungal Avr effectors remain largely unknown. Here, we combined bioinformatics and genetics to identify Avr3b, a new Avr gene from Phytophthora sojae, an oomycete pathogen that causes soybean root rot. Avr3b encodes a secreted protein with the RXLR host-targeting motif and C-terminal W and Nudix hydrolase motifs. Some isolates of P. sojae evade perception by the soybean R gene Rps3b through sequence mutation in Avr3b and lowered transcript accumulation. Transient expression of Avr3b in Nicotiana benthamiana increased susceptibility to P. capsici and P. parasitica, with significantly reduced accumulation of reactive oxygen species (ROS) around invasion sites. Biochemical assays confirmed that Avr3b is an ADP-ribose/NADH pyrophosphorylase, as predicted from the Nudix motif. Deletion of the Nudix motif of Avr3b abolished enzyme activity. Mutation of key residues in Nudix motif significantly impaired Avr3b virulence function but not the avirulence activity. Some Nudix hydrolases act as negative regulators of plant immunity, and thus Avr3b might be delivered into host cells as a Nudix hydrolase to impair host immunity. Avr3b homologues are present in several sequenced Phytophthora genomes, suggesting that Phytophthora pathogens might share similar strategies to suppress plant immunity.
Asunto(s)
Fosforilasas/metabolismo , Phytophthora/enzimología , Inmunidad de la Planta , Factores de Virulencia/metabolismo , Adenosina Difosfato Ribosa/metabolismo , Alelos , Genotipo , Datos de Secuencia Molecular , Mutación , NAD/metabolismo , Fosforilasas/química , Fosforilasas/genética , Phytophthora/genética , Phytophthora/patogenicidad , Enfermedades de las Plantas/parasitología , Pirofosfatasas/química , Especies Reactivas de Oxígeno/metabolismo , Glycine max/inmunología , Glycine max/parasitología , Nicotiana/inmunología , Nicotiana/metabolismo , Nicotiana/parasitología , Factores de Virulencia/biosíntesis , Hidrolasas NudixRESUMEN
Transglutaminases (TGases) are ubiquitous enzymes that catalyze selective cross-linking between protein-bound glutamine and lysine residues; the resulting isopeptide bond confers high resistance to proteolysis. Phytophthora sojae, a pathogen of soybean, secretes a Ca(2+)-dependent TGase (GP42) that is activating defense responses in both host and non-host plants. A GP42 fragment of 13 amino acids, termed Pep-13, was shown to be absolutely indispensable for both TGase and elicitor activity. GP42 does not share significant primary sequence similarity with known TGases from mammals or bacteria. This suggests that GP42 has evolved novel structural and catalytic features to support enzymatic activity. We have solved the crystal structure of the catalytically inactive point mutant GP42 (C290S) at 2.95 Å resolution and identified residues involved in catalysis by mutational analysis. The protein comprises three domains that assemble into an elongated structure. Although GP42 has no structural homolog, its core region displays significant similarity to the catalytic core of the Mac-1 cysteine protease from Group A Streptococcus, a member of the papain-like superfamily of cysteine proteases. Proteins that are taxonomically related to GP42 are only present in plant pathogenic oomycetes belonging to the order of the Peronosporales (e.g. Phytophthora, Hyaloperonospora, and Pythium spp.) and in marine Vibrio bacteria. This suggests that a lateral gene transfer event may have occurred between bacteria and oomycetes. Our results offer a basis to design and use highly specific inhibitors of the GP42-like TGase family that may impair the growth of important oomycete and bacterial pathogens.
Asunto(s)
Oomicetos/metabolismo , Phytophthora/genética , Vibrio/metabolismo , Secuencia de Aminoácidos , Dominio Catalítico , Cristalografía por Rayos X/métodos , Análisis Mutacional de ADN , Evolución Molecular , Inmunidad Innata , Modelos Moleculares , Datos de Secuencia Molecular , Mutagénesis Sitio-Dirigida , Petroselinum/microbiología , Filogenia , Proteínas Recombinantes/metabolismo , Homología de Secuencia de Aminoácido , Solanum tuberosum/microbiología , Transglutaminasas/metabolismo , Microbiología del AguaRESUMEN
Necrosis- and ethylene-inducing-like proteins (NLP) are widely distributed in eukaryotic and prokaryotic plant pathogens and are considered to be important virulence factors. We identified, in total, 70 potential Phytophthora sojae NLP genes but 37 were designated as pseudogenes. Sequence alignment of the remaining 33 NLP delineated six groups. Three of these groups include proteins with an intact heptapeptide (Gly-His-Arg-His-Asp-Trp-Glu) motif, which is important for necrosis-inducing activity, whereas the motif is not conserved in the other groups. In total, 19 representative NLP genes were assessed for necrosis-inducing activity by heterologous expression in Nicotiana benthamiana. Surprisingly, only eight genes triggered cell death. The expression of the NLP genes in P. sojae was examined, distinguishing 20 expressed and 13 nonexpressed NLP genes. Real-time reverse-transcriptase polymerase chain reaction results indicate that most NLP are highly expressed during cyst germination and infection stages. Amino acid substitution ratios (Ka/Ks) of 33 NLP sequences from four different P. sojae strains resulted in identification of positive selection sites in a distinct NLP group. Overall, our study indicates that expansion and pseudogenization of the P. sojae NLP family results from an ongoing birth-and-death process, and that varying patterns of expression, necrosis-inducing activity, and positive selection suggest that NLP have diversified in function.
Asunto(s)
Regulación del Desarrollo de la Expresión Génica/genética , Familia de Multigenes/genética , Nicotiana/parasitología , Phytophthora/genética , Enfermedades de las Plantas/parasitología , Secuencias de Aminoácidos , Secuencia de Aminoácidos , Sustitución de Aminoácidos , Secuencia de Bases , Biblioteca de Genes , Modelos Moleculares , Datos de Secuencia Molecular , Necrosis , Filogenia , Phytophthora/metabolismo , Phytophthora/patogenicidad , Seudogenes , Reacción en Cadena en Tiempo Real de la Polimerasa , Alineación de Secuencia , Análisis de Secuencia de ADNRESUMEN
The use of resistance genes in elite soybean cultivars is one of the most widely used methods to manage Phytophthora sojae. This method relies on effector-triggered immunity, where a Resistant to P. sojae (Rps) gene product from the plant recognizes a specific effector from the pathogen, encoded by an avirulence (Avr) gene. Many Avr genes from P. sojae have been identified in the last decade, allowing a better exploitation of this type of resistance. The objective of the present study was to identify the Avr gene triggering immunity derived from the soybean resistance gene Rps8. The analysis of a segregating F2 progeny coupled with a genotyping-by-sequencing approach led to the identification of a putative Avr8 locus. The investigation of this locus using whole-genome sequencing data from 31 isolates of P. sojae identified Avr3a as the likely candidate for Avr8. Long-read sequencing also revealed that P. sojae isolates can carry up to five copies of the Avr3a gene, compared to the four previously reported. Haplotype and transcriptional analyses showed that amino acid changes and absence of Avr3a transcripts from P. sojae isolates caused changes in virulence towards Rps8. Functional analyses using CRISPR/Cas9 knockout and constitutive expression demonstrated that Rps8 interacted with Avr3a. We also showed that a specific allele of Avr3a is recognized by Rps3a but not Rps8. While Rps3a and Rps8 have been previously described as closely linked, this is the first report of a clear distinction hitherto undefined between these two resistance genes.
Asunto(s)
Glycine max , Phytophthora infestans , Alelos , Haplotipos/genética , Phytophthora infestans/genética , Enfermedades de las Plantas , Glycine max/genética , Virulencia/genéticaRESUMEN
Two Soybean mosaic virus (SMV) G2 isolates, L and L-RB, sharing high-sequence similarly but differing in ability to break Rsv4-mediated resistance in soybean, were investigated. Infectious clones corresponding to these two isolates and their chimeric clones resulting from swapping different regions of genomic cDNA between L and L-RB were constructed. Only L-RB or chimeras containing the middle fragment of L-RB cDNA showed virulence on Rsv4-genotype soybean. Sequence comparison analysis revealed that the middle genomic region of L and L-RB encodes four different amino acids. Point mutagenesis demonstrated that a single amino acid substitution (Q1033K) in the P3 protein determined virulence toward Rsv4 resistance. In addition, six new SMV Rsv4 resistance-breaking isolates, variants of the second passage on Williams 82 infected with the chimeras or mutants noninfectious on soybean carrying Rsv4, were obtained. Sequencing data indicated that these new isolates contain either the Q1033K mutation or a new substitution (G1054R) in P3. Site-directed mutagenesis confirmed the virulence role of the G1054R mutation on Rsv4-genotype soybean. Taken together, these data suggest that P3 of the SMV G2 strain is an avirulent determinant for Rsv4 and one single nucleotide mutation in P3 may be sufficient to compromise its elicitor function.
Asunto(s)
Glycine max/virología , Virus del Mosaico/genética , Sustitución de Aminoácidos , Secuencia de Bases , Quimera/genética , Clonación Molecular , ADN Complementario/genética , ADN Viral/genética , Datos de Secuencia Molecular , Virus del Mosaico/aislamiento & purificación , Virus del Mosaico/patogenicidad , Mutagénesis Sitio-Dirigida , Mutación , Enfermedades de las Plantas/virología , Alineación de Secuencia , Homología de Secuencia de Aminoácido , Homología de Secuencia de Ácido Nucleico , Glycine max/genética , VirulenciaRESUMEN
The transcriptome of the oomycete plant pathogen Phytophthora sojae was profiled at ten different developmental and infection stages based on a 3'-tag digital gene-expression protocol. More than 90 million clean sequence tags were generated and compared with the P. sojae genome and its 19,027 predicted genes. A total of 14,969 genes were detected, of which 10,044 were deemed reliable because they mapped to unambiguous tags. A comparison of the whole-library genes' expression patterns suggested four groups: i) mycelia and zoosporangia, ii) zoospores and cysts, iii) germinating cysts, and iv) five infection site libraries (IF1.5 to IF24h). The libraries from the different groups showed major transitional shifts in gene expression. From the ten libraries, 722 gene expression?pattern clusters were obtained and the top 16 clusters, containing more than half of the genes, comprised enriched genes with different functions including protein localization, triphosphate metabolism, signaling process, and noncoding RNA metabolism. An evaluation of the average expression level of 30 pathogenesis-related gene families revealed that most were infection induced but with diverse expression patterns and levels. A web-based server named the Phytophthora Transcriptional Database has been established.
Asunto(s)
Perfilación de la Expresión Génica/métodos , Phytophthora/genética , Enfermedades de las Plantas/microbiología , Transcriptoma/genética , Secuencia de Bases , Análisis por Conglomerados , ADN Complementario/genética , Bases de Datos Genéticas , Etiquetas de Secuencia Expresada , Expresión Génica/genética , Biblioteca de Genes , Sitios Genéticos/genética , Genoma/genética , Internet , Anotación de Secuencia Molecular , Datos de Secuencia Molecular , Phytophthora/crecimiento & desarrollo , Phytophthora/fisiología , ARN Mensajero/genética , Análisis de Secuencia de ADN , Factores de TiempoRESUMEN
Whether a seed coat of a soybean (Glycine max L. Mer.) seed is permeable or non-permeable is governed by a number of quantitative trait loci further influenced by environmental factors. In soybean seeds, water loss is controlled by a thin, inconspicuous outer cuticle. When intact, the outer cuticle constitutes a barrier to water passage; however, the presence of minute cracks in the cuticle results in the ready passage of water. We explored the timing of cuticular development in soybean seeds by measuring the deposition of the cutin in relation to seed growth and cell viability. Cutin deposition occurred early in the development and ceased just prior to the final stage of rapid seed expansion. Cracks in the cuticle appeared after cutin synthesis ceased while the seed continued to grow. In permeable seeds (regardless of genotype) the resistance of the cuticle to water passage increased steadily during development until seed expansion was maximal and cracks appeared in the cuticle. Once cracks formed, they became the primary site of water passage and the cuticle lost its ability to control the process. In non-permeable seeds, no cracks appeared at this critical point and the cuticle continued to restrict water passage. Microarray analysis of gene expression during seed coat development revealed a complex transcriptome with many genes uniquely expressed in the seed coat. However, the expression patterns were remarkably similar between permeable and non-permeable types, in keeping with the complexity of the underlying genetics of seed coat permeability.
Asunto(s)
Glycine max/crecimiento & desarrollo , Epidermis de la Planta/crecimiento & desarrollo , Semillas/crecimiento & desarrollo , Análisis de Varianza , Arabidopsis/enzimología , Supervivencia Celular , Perfilación de la Expresión Génica , Regulación del Desarrollo de la Expresión Génica , Regulación de la Expresión Génica de las Plantas , Genes de Plantas/genética , Lípidos de la Membrana/metabolismo , Análisis de Secuencia por Matrices de Oligonucleótidos , Permeabilidad , Epidermis de la Planta/genética , Epidermis de la Planta/metabolismo , Epidermis de la Planta/ultraestructura , Semillas/genética , Semillas/metabolismo , Semillas/ultraestructura , Glycine max/genética , Glycine max/metabolismo , Glycine max/ultraestructura , Ácidos Sulfónicos/metabolismo , Factores de Tiempo , Agua/metabolismoRESUMEN
Phytophthora sojae is the causal agent of root and stem rot of soybean (Glycine max). Various cultivars with partial resistance to the pathogen have been developed to mitigate this damage. Herein, two contrasting genotypes, the cultivar Conrad (with strong partial resistance) and the line OX760-6 (with weak partial resistance), were compared regarding their amounts of preformed and induced suberin components, and to early events during the P. sojae infection process. To colonize the root, hyphae grew through the suberized middle lamellae between epidermal cells. This took 2 to 3 h longer in Conrad than in OX760-6, giving Conrad plants more time to establish their chemical defenses. Subsequent growth of hyphae through the endodermis was also delayed in Conrad. This cultivar had more preformed aliphatic suberin than the line OX760-6 and was induced to form more aliphatic suberin several days prior to that of OX760-6. However, the induced suberin was formed subsequent to the initial infection process. Eventually, the amount of induced suberin (measured 8 days postinoculation) was the same in both genotypes. Preformed root epidermal suberin provides a target for selection and development of new soybean cultivars with higher levels of expression of partial resistance to P. sojae.
Asunto(s)
Glycine max/microbiología , Lípidos/análisis , Phytophthora/fisiología , Enfermedades de las Plantas/microbiología , Raíces de Plantas/microbiología , Interacciones Huésped-Patógeno , Inmunidad Innata , Microscopía Fluorescente , Raíces de Plantas/metabolismo , Glycine max/metabolismoRESUMEN
A new genetic locus mediating avirulence in the potato late blight pathogen Phytophthora infestans has been discovered. The Avr3b-Avr10-Avr11 locus is recognized by three different potato resistance genes, and is different from other Avr loci that have been identified thus far. This locus encodes a large protein with a WD40 domain and sequence similarities to transcription factors. Multiple, truncated copies of this gene have arisen by gene amplification and are characteristic of avirulent strains of P. infestans. Here, we describe the new avirulence locus and discuss the importance of this finding.
Asunto(s)
Phytophthora/genética , Solanum tuberosum/inmunología , Modelos Genéticos , Phytophthora/patogenicidad , Enfermedades de las Plantas/inmunología , Enfermedades de las Plantas/microbiología , Solanum tuberosum/microbiología , Virulencia/genética , Factores de Virulencia/genéticaRESUMEN
Six unique expressed sequence tag (EST) libraries were generated from four developmental stages of Phytophthora sojae P6497. RNA was extracted from mycelia, swimming zoospores, germinating cysts, and soybean (Glycine max (L.) Merr.) cv. Harosoy tissues heavily infected with P. sojae. Three libraries were created from mycelia growing on defined medium, complex medium, and nutrient-limited medium. The 26,943 high-quality sequences obtained clustered into 7,863 unigenes composed of 2,845 contigs and 5,018 singletons. The total number of P. sojae unigenes matching sequences in the genome assembly was 7,412 (94%). Of these unigenes, 7,088 (90%) matched gene models predicted from the P. sojae sequence assembly, but only 2,047 (26%) matched P. ramorum gene models. Analysis of EST frequency from different growth conditions and morphological stages revealed genes that were specific to or highly represented in particular growth conditions and life stages. Additionally, our results indicate that, during infection, the pathogen derives most of its carbon and energy via glycolysis of sugars in the plant. Sequences identified with putative roles in pathogenesis included avirulence homologs possessing the RxLR motif, elicitins, and hydrolytic enzymes. This large collection of P. sojae ESTs will serve as a valuable public genomic resource.
Asunto(s)
Etiquetas de Secuencia Expresada , Perfilación de la Expresión Génica , Genes Fúngicos , Phytophthora/genética , Análisis por Conglomerados , Biblioteca de Genes , Datos de Secuencia Molecular , Phytophthora/crecimiento & desarrollo , Análisis de Secuencia de ADN , Glycine max/microbiologíaRESUMEN
Cyclophilins (CYPs) belong to the immunophilin superfamily with peptidyl-prolyl cis-trans isomerase (PPIase) activity. They catalyze the interconversion of the cis- and trans-rotamers of the peptidyl-prolyl amide bond of peptides. A yeast-two-hybrid screening using the isoflavonoid regulator GmMYB176 as bait identified GmCYP1 as one of the interacting proteins in soybean embryos. GmCYP1 localizes both in the nucleus and cytoplasm, and interacts in planta with GmMYB176, in the nucleus, and with SGF14l (a soybean 14-3-3 protein) in the nucleus and the cytoplasm. GmCYP1 contains a single cyclophilin-like domain and displays a high sequence identity with other plant CYPs that are known to have stress-specific function. Tissue-specific expression of GmCYP1 revealed higher expression in developing seeds compared to other vegetative tissues, suggesting their seed-specific role. Furthermore, GmCYP1 transcript level was reduced in response to stress. Since isoflavonoids are involved in plant stress resistance against biotic and abiotic factors, the interaction of GmCYP1 with the isoflavonoid regulators GmMYB176 and 14-3-3 protein suggests its role in defense in soybean.
Asunto(s)
Ciclofilinas/metabolismo , Glycine max/metabolismo , Proteínas de Soja/metabolismo , Proteínas 14-3-3/metabolismo , Núcleo Celular/química , Núcleo Celular/metabolismo , Citoplasma/química , Citoplasma/metabolismo , Proteínas de Soja/aislamiento & purificación , Estrés FisiológicoRESUMEN
The process of RNA splicing influences many physiological processes, including plant immunity. However, how plant parasites manipulate host RNA splicing process remains unknown. Here we demonstrate that PsAvr3c, an avirulence effector from oomycete plant pathogen Phytophthora sojae, physically binds to and stabilizes soybean serine/lysine/arginine-rich proteins GmSKRPs. The SKRPs are novel proteins that associate with a complex that contains plant spliceosome components, and are negative regulators of plant immunity. Analysis by RNA-seq data indicates that alternative splicing of pre-mRNAs from 401 soybean genes, including defense-related genes, is altered in GmSKRP1 and PsAvr3c overexpressing lines compared to control plants. Representative splicing events mediated by GmSKRP1 and PsAvr3c are tested by infection assays or by transient expression in soybean plants. Our results show that plant pathogen effectors can reprogram host pre-mRNA splicing to promote disease, and we propose that pathogens evolved such strategies to defeat host immune systems.
Asunto(s)
Regulación de la Expresión Génica de las Plantas/inmunología , Glycine max/parasitología , Interacciones Huésped-Patógeno/genética , Phytophthora/patogenicidad , Inmunidad de la Planta/genética , Empalme Alternativo , Interacciones Huésped-Patógeno/inmunología , Phytophthora/fisiología , Enfermedades de las Plantas/genética , Enfermedades de las Plantas/inmunología , Precursores del ARN/genética , ARN de Planta/genética , Análisis de Secuencia de ARN , Proteínas de Soja/genética , Proteínas de Soja/metabolismo , Glycine max/genética , Glycine max/inmunología , Empalmosomas/genética , Empalmosomas/metabolismo , Factores de Virulencia/metabolismoRESUMEN
The year 2004 was an exciting one for the Phytophthora research community. The United States Department of Energy Joint Genome Institute (JGI) completed the draft genome sequence of two Phytophthora species, Phytophthora sojae and Phytophthora ramorum. In August of that year over 50 people gathered at JGI in Walnut Creek, California, for an annotation jamboree and searched for the secrets and surprises that the two genomes have in petto. This culminated in a paper in Science in September of this year describing the highlights of the sequencing project and emphasizing the power of having the genome sequences of two closely related organisms. This MPMI Focus issue on Phytophthora genomics contains a number of more specialized manuscripts centered on gene annotation and genome organization, and complemented with manuscripts that rely on genomics resources.
Asunto(s)
Genoma , Phytophthora/genética , Proteínas Algáceas/química , Proteínas Algáceas/clasificación , Proteínas Algáceas/genética , Secuencias de Aminoácidos , Evolución Biológica , Genómica , Filogenia , Phytophthora/clasificación , Phytophthora/patogenicidadRESUMEN
BACKGROUND: Hydrophobic protein from soybean (HPS) is an 8 kD cysteine-rich polypeptide that causes asthma in persons allergic to soybean dust. HPS is synthesized in the pod endocarp and deposited on the seed surface during development. Past evidence suggests that the protein may mediate the adherence or dehiscence of endocarp tissues during maturation and affect the lustre, or glossiness of the seed surface. RESULTS: A comparison of soybean germplasm by genomic DNA blot hybridization shows that the copy number and structure of the Hps locus is polymorphic among soybean cultivars and related species. Changes in Hps gene copy number were also detected by comparative genomic DNA hybridization using cDNA microarrays. The Hps copy number polymorphisms co-segregated with seed lustre phenotype and HPS surface protein in a cross between dull- and shiny-seeded soybeans. In soybean cultivar Harosoy 63, a minimum of 27 +/- 5 copies of the Hps gene were estimated to be present in each haploid genome. The isolation and analysis of genomic clones indicates that the core Hps locus is comprised of a tandem array of reiterated units, with each 8.6 kb unit containing a single HPS open reading frame. CONCLUSION: This study shows that polymorphisms at the Hps locus arise from changes in the gene copy number via gene amplification. We present a model whereby Hps copy number modulates protein expression levels and seed lustre, and we suggest that gene amplification may result from selection pressures imposed on crop plants.
Asunto(s)
Amplificación de Genes/genética , Dosificación de Gen/genética , Glycine max/genética , Proteínas de Plantas/genética , ADN de Plantas/genética , Análisis de Secuencia por Matrices de Oligonucleótidos , Fenotipo , Polimorfismo Genético/genética , SemillasRESUMEN
An emerging group of proteins found in many plant pathogens are related to their ability to cause plant cell death. These proteins may be identified by the presence of a common NPP1 (necrosis-inducing Phytophthora protein) domain, and have collectively been named NLPs (Nep1-like proteins). The NLPs are distinguished by their wide distribution across taxa and their broad spectrum of activity against dicotyledonous plants. The function of NLPs is not known but there is strong evidence that they may act as positive virulence factors, accelerating disease and pathogen growth in plant hosts. Interest in NLPs is gaining momentum as more members of this protein family are discovered in more species of plant pathogens.
Asunto(s)
Proteínas Bacterianas/aislamiento & purificación , Proteínas Fúngicas/aislamiento & purificación , Plantas/microbiología , Bacterias/patogenicidad , Proteínas Bacterianas/química , Proteínas Fúngicas/química , Hongos/patogenicidad , Genoma Fúngico , Filogenia , VirulenciaRESUMEN
The Phytophthora sojae avirulence gene Avr3a encodes an effector that is capable of triggering immunity on soybean plants carrying the resistance gene Rps3a. P. sojae strains that express Avr3a are avirulent to Rps3a plants, while strains that do not are virulent. To study the inheritance of Avr3a expression and virulence towards Rps3a, genetic crosses and self-fertilizations were performed. A cross between P. sojae strains ACR10 X P7076 causes transgenerational gene silencing of Avr3a allele, and this effect is meiotically stable up to the F5 generation. However, test-crosses of F1 progeny (ACR10 X P7076) with strain P6497 result in the release of silencing of Avr3a. Expression of Avr3a in the progeny is variable and correlates with the phenotypic penetrance of the avirulence trait. The F1 progeny from a direct cross of P6497 X ACR10 segregate for inheritance for Avr3a expression, a result that could not be explained by parental imprinting or heterozygosity. Analysis of small RNA arising from the Avr3a gene sequence in the parental strains and hybrid progeny suggests that the presence of small RNA is necessary but not sufficient for gene silencing. Overall, we conclude that inheritance of the Avr3a gene silenced phenotype relies on factors that are variable among P. sojae strains.
Asunto(s)
Silenciador del Gen/fisiología , Phytophthora/genética , Cruzamientos Genéticos , Regulación de la Expresión Génica , Genotipo , Phytophthora/patogenicidad , Phytophthora/fisiología , Enfermedades de las Plantas/parasitología , Especificidad de la Especie , Factores de Virulencia/genética , Factores de Virulencia/fisiologíaRESUMEN
The interaction between soybean and the phytopathogenic oomycete Phytophthora sojae is controlled by host resistance (Rps) genes and pathogen avirulence (Avr) genes. We have mapped the Avr1a locus in F(2) populations derived from four different P. sojae races. Four RAPD and nine AFLP markers linked to Avr1a were initially identified. Nine markers were used to compare genetic linkage maps of the Avr1a locus in two distinct F(2) populations. Distorted segregation ratios favoring homozygous genotypes were noted in both crosses. Segregation analysis of all the markers in one F(2) population of 90 progeny generated a map of 113.2 cM encompassing Avr1a, with one marker cosegregating with the gene. The cosegregating DNA marker was used to isolate P. sojae BAC clones and construct a physical map covering 170 kb, from which additional DNA markers were developed. Three markers occurring within the BAC contig were mapped in an enlarged population of 486 F(2) progeny. Avr1a was localized to a 114-kb interval, and an average physical to genetic distance ratio of 391 kb/cM was calculated for this region. This work provides a basis for the positional cloning of Avr1a.
Asunto(s)
Proteínas Algáceas/genética , Mapeo Cromosómico , Genes Fúngicos , Phytophthora/genética , Cromosomas Artificiales Bacterianos , Cruzamientos Genéticos , Proteínas Fúngicas/genética , Genes Dominantes , Marcadores Genéticos , Phytophthora/patogenicidad , VirulenciaRESUMEN
Hydrophobic protein from soybean (HPS) is present in soybean dust and is an allergen (Gly m 1) that causes asthma in allergic individuals. Past studies have shown that HPS occurs on the seed surface. To determine the microscopic localization of HPS during seed development, monoclonal antibodies to HPS were used to visualize the protein by fluorescence and transmission electron microscopy. Seed coat and endocarp sections were also examined for pectin, cellulose, callose, starch, and protein by histochemical staining. HPS is present in the endocarp epidermal cells at 18 to 28 days post anthesis. At later stages of seed development, HPS occurs in extracellular secretions that accumulate unevenly on the endocarp epidermis and seed surface. HPS is synthesized by the endocarp epidermis and deposited on the seed surface as part of a heterogeneous matrix.