RESUMO
BACKGROUND: Rice sheath blight (caused by Rhizoctonia solani) and tobacco mosaic virus are very important plant diseases, causing a huge loss in global crop production. Paenibacillus kribbensis PS04 is a broad-spectrum biocontrol agent, used for controlling these diseases. Previously, extracellular polysaccharides (EPS) from P. kribbensis PS04 had been purified and their structure was inferred to be fructosan. This study aimed to evaluate the effects of exogenous EPS treatment on plantpathogen interactions. RESULTS: Plant defense genes such as phenylalanine ammonia-lyase, catalase, chitinase, allene oxide synthase, and PR1a proteins were significantly induced by exogenous EPS treatment. Moreover, subsequent challenge of EPSpretreated plants with the pathogens (R. solani or tobacco mosaic virus) resulted in higher expression of defenseassociated genes. Increased activities of defense-associated enzymes, total phenols, and flavonoids were also observed in EPS pretreated plants. The contents of malondialdehyde in plants, which act as indicator of lipid peroxidation, were reduced by EPS treatment. CONCLUSIONS: This study comprehensively showed that EPS produced from P. kribbensis PS04 enhances disease resistance in plants by the activation of defense-associated genes as well as through the enhancement of activities of defense-related enzymes.
Assuntos
Doenças das Plantas/imunologia , Rhizoctonia/patogenicidade , Vírus do Mosaico do Tabaco/patogenicidade , Paenibacillus/imunologia , Doenças das Plantas/microbiologia , Polissacarídeos Bacterianos , Controle Biológico de Vetores , Interações Hospedeiro-Patógeno , Paenibacillus/genética , Resistência à Doença/genética , Reação em Cadeia da Polimerase em Tempo Real , Frutose/análogos & derivadosRESUMO
Trichoderma spp. are filamentous fungi that colonize plant roots conferring beneficial effects to plants, either indirectly through the induction of their defense systems or directly through the suppression of phytopathogens in the rhizosphere. Transcriptomic analyses of Trichoderma spp. emerged as a powerful method for identifying the molecular events underlying the establishment of this beneficial relationship. Here, we focus on the transcriptomic response of Trichoderma virens during its interaction with Arabidopsis seedlings. The main response of T. virens to cocultivation with Arabidopsis was the repression of gene expression. The biological processes of transport and metabolism of carbohydrates were downregulated, including a set of cell wall-degrading enzymes putatively relevant for root colonization. Repression of such genes reached their basal levels at later times in the interaction, when genes belonging to the biological process of copper ion transport were induced, a necessary process providing copper as a cofactor for cell wall-degrading enzymes with the auxiliary activities class. RNA-Seq analyses showed the induction of a member of the SNF2 family of chromatin remodelers/helicase-related proteins, which was named IPA-1 (increased protection of Arabidopsis-1). Sequence analyses of IPA-1 showed its closest relatives to be members of the Rad5/Rad16 and SNF2 subfamilies; however, it grouped into a different clade. Although deletion of IPA-1 in T. virens did not affect its growth, the antibiotic activity of Δipa-1 culture filtrates against Rhizoctonia solani diminished but it remained unaltered against Botrytis cinerea. Triggering of the plant defense genes in plants treated with Δipa-1 was higher, showing enhanced resistance against Pseudomonas syringae but not against B. cinerea as compared with the wild type.
Assuntos
Antibiose , Arabidopsis/microbiologia , Montagem e Desmontagem da Cromatina , Resistência à Doença , Rhizoctonia/patogenicidade , Trichoderma/fisiologia , Humanos , Doenças das Plantas/microbiologia , Raízes de Plantas , TranscriptomaRESUMO
Snakin-1 is a cysteine-rich antimicrobial peptide (AMP) isolated from potato tubers, with broad-spectrum activity. It belongs to the Snakin/GASA family, whose members have been studied because of their diverse roles in important plant processes, including defense. To analyze if this defensive function may lead to disease tolerance in lettuce, one of the most worldwide consumed leafy vegetable, we characterized three homozygous transgenic lines overexpressing Snakin-1. They were biologically assessed by the inoculation with the fungal pathogens Rhizoctonia solani and Sclerotinia sclerotiorum both in vitro and in planta at the greenhouse. When in vitro assays were performed with R. solani on Petri dishes containing crude plant extracts it was confirmed that the expressed Snakin-1 protein has antimicrobial activity. Furthermore, transgenic lines showed a better response than wild type in in vivo challenges against R. solani both in chamber and in greenhouse. In addition, two of these lines showed significant in vivo protection against the pathogen S. sclerotiorum in challenge assays on adult plants. Our results show that Snakin-1 is an interesting candidate gene for the selection/breeding of lettuce plants with increased fungal tolerance.
Assuntos
Lactuca/genética , Doenças das Plantas/prevenção & controle , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Ascomicetos/patogenicidade , Resistência à Doença , Lactuca/crescimento & desenvolvimento , Lactuca/microbiologia , Doenças das Plantas/microbiologia , Folhas de Planta/genética , Folhas de Planta/metabolismo , Plantas Geneticamente Modificadas/crescimento & desenvolvimento , Plantas Geneticamente Modificadas/microbiologia , Espécies Reativas de Oxigênio/metabolismo , Rhizoctonia/patogenicidadeRESUMO
Eighty-one Rhizoctonia-like isolates were identified based on morphology and nuclei-staining methods from natural and agricultural soils of the Cerrado (Brazilian savanna). The nucleotide similarity analysis of ITS1-5.8S-ITS2 regions identified 14 different taxa, with 39.5% of isolates assigned to Waitea circinata (zeae, oryzae, and circinata varieties), while 37.0% belonged to Thanatephorus cucumeris anastomosis groups (AGs) AG1-IB, AG1-ID, AG1-IE, AG4-HGI, and AG4-HGIII. Ceratobasidium spp. AG-A, AG-F, AG-Fa, AG-P, and AG-R comprised 23.5%. Rhizoctonia zeae (19.8%), R. solani AG1-IE (18.6%), and binucleate Rhizoctonia AG-A (8.6%) were the most frequent anamorphic states found. Root rot severity caused by the different taxa varied from low to high on common beans, and tended to be low to average in maize. Twenty-two isolates were pathogenic to both hosts, suggesting difficulties in managing Rhizoctonia root rots with crop rotation. These results suggest that cropping history affects the geographical arrangement of AGs, with a prevalence of AG1 in the tropical zone from central to north Brazil while the AG4 group was most prevalent from central to subtropical south. W. circinata var. zeae was predominant in soils under maize production. To our knowledge, this is the first report on the occurrence of W. circinata var. circinata in Brazil.
Assuntos
Produtos Agrícolas/microbiologia , Doenças das Plantas/microbiologia , Rhizoctonia/genética , Rhizoctonia/patogenicidade , Brasil , FilogeniaRESUMO
The fungus Rhizoctonia oryzae-sativae is an important pathogen that causes the aggregated sheath spot disease on rice. In this study, we investigated the genetic structure of rice-adapted populations of R. oryzae-sativae sampled from traditional rice-cropping areas from the Paraíba Valley, São Paulo, Brazil, and from Meta, in the Colombian Llanos, in South America. We used five microsatellite loci to measure population differentiation and infer the pathogen's reproductive system. Gene flow was detected among the three populations of R. oryzae-sativae from lowland rice in Brazil and Colombia. In contrast, a lack of gene flow was observed between the lowland and the upland rice populations of the pathogen. Evidence of sexual reproduction including low clonality, Hardy-Weinberg equilibrium within loci and gametic equilibrium between loci, indicated the predominance of a mixed reproductive system in all populations. In addition, we assessed the adaptive potential of the Brazilian populations of R. oryzae-sativae to emerge as a pathogen to Urochloa spp. (signalgrass) based on greenhouse aggressiveness assays. The Brazilian populations of R. oryzae-sativae were probably only incipiently adapted as a pathogen to Urochloa spp. Comparison between RST and QST showed the predominance of diversifying selection in the divergence between the two populations of R. oryzae-sativae from Brazil.
Assuntos
Genética Populacional , Oryza/microbiologia , Doenças das Plantas/microbiologia , Poaceae/microbiologia , Rhizoctonia/genética , Brasil , Fluxo Gênico , Genótipo , Geografia , Repetições de Microssatélites/genética , Rhizoctonia/isolamento & purificação , Rhizoctonia/patogenicidadeRESUMO
The fungus Rhizoctonia solani anastomosis group (AG)-1 IA emerged in the early 1990s as an important pathogen causing foliar blight and collar rot on pastures of the genus Urochloa (signalgrass) in South America. We tested the hypothesis that this pathogen emerged following a host shift or jump as a result of geographical overlapping of host species. The genetic structure of host and regional populations of R. solani AG-1 IA infecting signalgrass, rice, and soybean in Colombia and Brazil was analyzed using nine microsatellite loci in 350 isolates to measure population differentiation and infer the pathogen reproductive system. Phylogeographical analyses based on the microsatellite loci and on three DNA sequence loci were used to infer historical migration patterns and test hypotheses about the origin of the current pathogen populations. Cross pathogenicity assays were conducted to measure the degree of host specialization in populations sampled from different hosts. The combined analyses indicate that the pathogen populations currently infecting Urochloa in Colombia and Brazil most likely originated from a population that originally infected rice. R. solani AG-1 IA populations infecting Urochloa exhibit a mixed reproductive system including both sexual reproduction and long-distance dispersal of adapted clones, most likely on infected seed. The pathogen population on Urochloa has a genetic structure consistent with a high evolutionary potential and showed evidence for host specialization.
Assuntos
Fluxo Gênico , Glycine max/microbiologia , Especificidade de Hospedeiro , Oryza/microbiologia , Rhizoctonia/genética , Brasil , Colômbia , Variação Genética , Repetições de Microssatélites , Filogeografia , Doenças das Plantas , Rhizoctonia/patogenicidadeRESUMO
Rhizoctonia solani Kühn, es un patógeno de suelo, que causa enfermedades en un amplio rango de hospedantes de cultivos agrícolas. Las necesidades de reducir el uso indiscriminado de fungicidas químicos conllevan al estudio de otras alternativas menos agresivas al medio ambiente. Los monoterpenos presentes en los aceites esenciales de plantas, presentan una marcada actividad biocida y son compuestos que resultan menos perjudiciales desde el punto de vista ambiental. Se prepararon cuatro mezclas de monoterpenos (timol-entol, timolalcanfor, timol-citronelal y timol-1,8 cineol) con el objetivo de evaluar la actividad antifúngica in vitro sobre R. solani, mediante su efecto sinérgico. Se empleó el método de adición al medio de cultivo de cada una de las mezclas a concentraciones de: 0,5; 0,1; 0,05; 0,03 y 0,01 por ciento, partiendo de una solución madre al 10 por ciento en dimetilsufóxido (DMSO) al 5 por ciento. Se calcularon los porcentajes de inhibición y los resultados se analizaron estadísticamente. Se determinaron las dosis inhibitorias medias y se clasificaron la toxicidad de cada una de las mezclas en: inocua, ligera, moderada y tóxica a cada concentración. Las cuatro mezclas mostraron 100 por ciento de inhibición a las concentraciones de: 0, 5; 0, 1; 0,05 y 0,03 por ciento. A la menor concentración las mezclas de timol-mentol y timol-cineol fueron las de mayor efecto inhibitorio del crecimiento micelial de R. solani.
Rhizoctonia solani Kühn is a soil pathogen that causes diseases in a wide host range from agricultural crops. The need to reduce the indiscriminate use of chemical fungicide it has led to the study of other less aggressive environment alternatives. Monoterpenes, common components in plants essential oils, shows a maked biocide activity and are compounds less harmful from an environmental point of view. With the objective of evaluate the in vitro antifungal activity against R. solani, four binary monoterpenes mixtures (thymol-menthol, thymol-camphor, thymol-citronellal and thymol-1,8 cineol) with the objective of evaluate the in vitro antifungal activity against R. solani, four monoterpenes mixtures were prepared. The method of addition to culture media of each of the mixtures in concentrations of: 0, 5; 0, 1; 0, 05; 0, 03 and 0, 01 percent, from a stock solution to 10 percent in dimethylsulfoxide (DMSO) to 5 percent were used. The inhibition percentages were calculated and the results were statistically analyzed. Median inhibitory dose was determined and the toxicity of each mixture were classified in: harmless, light, moderate and toxic in each concentration. The four mixtures showed 100 percent of micelial growth inhibition in concentrations of 0, 5; 0, 1; 0,05 and 0,03 percent. The thymol-cineol and thymol-menthol mixtures had the higher inhibitory effect on micelial growth of R. solani at the lower concentration.
Assuntos
Humanos , Antifúngicos , Monoterpenos , Rhizoctonia/isolamento & purificação , Rhizoctonia/patogenicidade , Timol , Óleos VoláteisRESUMO
Twenty-one isolates of Rhizoctonia solani were categorized into three anastomosis groups consisting of AG-4-HG-I (eight isolates), AG-2-2 (nine isolates) and AG-5 (four isolates). Their pathogenic capacities were tested on cotton cultivar Giza 86. Pre-emergence damping-off varied in response to the different isolates; however, the differences were not significant. Soluble proteins of the fungal isolates were electrophoresed using SDS-PAGE and gel electrophoreses. A dendrogram of the protein banding patterns by the UPGMA of arithmetic means placed the fungal isolates into distinct groups. There was no evidence of a relationship between protein dendrogram, anastomosis grouping or level of virulence or geographic origin. The dendrogram generated from these isolates based on PCR analysis with five RAPD-PCR primers showed high levels of genetic similarity among the isolates from the same geographical locations. There was partially relationship between the genetic similarity and AGs or level of virulence or geographic origin based on RAPD dendrogram. These results demonstrate that RAPD technique is a useful tool in determining the genetic characterization among isolates of R. solani.
Assuntos
Proteínas Fúngicas/genética , Gossypium/microbiologia , Técnica de Amplificação ao Acaso de DNA Polimórfico/métodos , Rhizoctonia/genética , Rhizoctonia/isolamento & purificação , Primers do DNA/metabolismo , Egito , Eletroforese em Gel de Poliacrilamida , Geografia , Técnicas de Tipagem Micológica , Filogenia , Doenças das Plantas/microbiologia , Rhizoctonia/classificação , Rhizoctonia/patogenicidade , Plântula/microbiologiaRESUMO
ABSTRACT The basidiomycetous fungus Rhizoctonia solani anastomosis group (AG)-1 IA is a major pathogen in Latin America causing sheath blight (SB) of rice. Particularly in Venezuela, the fungus also causes banded leaf and sheath blight (BLSB) on maize, which is considered an emerging disease problem where maize replaced traditional rice-cropping areas or is now planted in adjacent fields. Our goals in this study were to elucidate (i) the effects of host specialization on gene flow between sympatric and allopatric rice and maize-infecting fungal populations and (ii) the reproductive mode of the fungus, looking for evidence of recombination. In total, 375 isolates of R. solani AG1 IA sampled from three sympatric rice and maize fields in Venezuela (Portuguesa State) and two allopatric rice fields from Colombia (Meta State) and Panama (Chiriquí State) were genotyped using 10 microsatellite loci. Allopatric populations from Venezuela, Colombia, and Panama were significantly differentiated (Phi(ST) of 0.16 to 0.34). Partitioning of the genetic diversity indicated differentiation between sympatric populations from different host species, with 17% of the total genetic variation distributed between hosts while only 3 to 6% was distributed geographically among the sympatric Venezuelan fields. We detected symmetrical historical migration between the rice- and the maize-infecting populations from Venezuela. Rice- and maize-derived isolates were able to infect both rice and maize but were more aggressive on their original hosts, consistent with host specialization. Because the maize- and rice-infecting populations are still cross-pathogenic, we postulate that the genetic differentiation was relatively recent and mediated via a host shift. An isolation with migration analysis indicated that the maize-infecting population diverged from the rice-infecting population between 40 and 240 years ago. Our findings also suggest that maize-infecting populations have a mainly recombining reproductive system whereas the rice-infecting populations have a mixed reproductive system in Latin America.
Assuntos
Interações Hospedeiro-Patógeno , Oryza/microbiologia , Rhizoctonia/genética , Zea mays/microbiologia , Frequência do Gene , Especiação Genética , Genótipo , América Latina , Repetições de Microssatélites , Dinâmica Populacional , Rhizoctonia/patogenicidadeRESUMO
The chit1 gene from the entomopathogenic fungus Metarhizium anisopliae, encoding the endochitinase CHIT42, was placed under the control of the CaMV 35S promoter, and the resulting construct was transferred to tobacco. Seventeen kanamycin-resistant transgenic lines were recovered, and the presence of the transgene was confirmed by polymerase chain reactions and Southern blot hybridization. The number of chit1 copies was determined to be varying from one to four. Copy number had observable effects neither on plant growth nor development. Substantial heterogeneity concerning production of the recombinant chitinase, and both general and specific chitinolytic activities were detected in leaf extracts from primary transformants. The highest chitinase activities were found in plants harboring two copies of chit1 inserts at different loci. Progeny derived from self-pollination of the primary transgenics revealed a stable inheritance pattern, with transgene segregation following a mendelian dihybrid ratio. Two selected plants expressing high levels of CHIT42 were consistently resistant to the soilborne pathogen Rhizoctonia solani, suggesting a direct relationship between enzyme activity and reduction of foliar area affected by fungal lesions. To date, this is the first report of resistance to fungal attack in plants mediated by a recombinant chitinase from an entomopathogenic and acaricide fungus.