RESUMO
Studies of the interactions between plants and their microbiome have been conducted worldwide in the search for growth-promoting representative strains for use as biological inputs for agriculture, aiming to achieve more sustainable agriculture practices. With a focus on the isolation of plant growth-promoting (PGP) bacteria with ability to alleviate N stress, representative strains that were found at population densities greater than 104 cells g-1 and that could grow in N-free semisolid media were isolated from soils under different management conditions and from the roots of tomato (Solanum lycopersicum) and lulo (Solanum quitoense) plants that were grown in those soils. A total of 101 bacterial strains were obtained, after which they were phylogenetically categorized and characterized for their basic PGP mechanisms. All strains belonged to the Proteobacteria phylum in the classes Alphaproteobacteria (61% of isolates), Betaproteobacteria (19% of isolates) and Gammaproteobacteria (20% of isolates), with distribution encompassing nine genera, with the predominant genus being Rhizobium (58.4% of isolates). Strains isolated from conventional horticulture (CH) soil composed three bacterial genera, suggesting a lower diversity for the diazotrophs/N scavenger bacterial community than that observed for soils under organic management (ORG) or secondary forest coverture (SF). Conversely, diazotrophs/N scavenger strains from tomato plants grown in CH soil comprised a higher number of bacterial genera than did strains isolated from tomato plants grown in ORG or SF soils. Furthermore, strains isolated from tomato were phylogenetically more diverse than those from lulo. BOX-PCR fingerprinting of all strains revealed a high genetic diversity for several clonal representatives (four Rhizobium species and one Pseudomonas species). Considering the potential PGP mechanisms, 49 strains (48.5% of the total) produced IAA (2.96-193.97 µg IAA mg protein-1), 72 strains (71.3%) solubilized FePO4 (0.40-56.00 mg l-1), 44 strains (43.5%) solubilized AlPO4 (0.62-17.05 mg l-1), and 44 strains produced siderophores (1.06-3.23). Further, 91 isolates (90.1% of total) showed at least one PGP trait, and 68 isolates (67.3%) showed multiple PGP traits. Greenhouse trials using the bacterial collection to inoculate tomato or lulo plants revealed increases in plant biomass (roots, shoots or both plant tissues) elicited by 65 strains (54.5% of the bacterial collection), of which 36 were obtained from the tomato rhizosphere, 15 were obtained from the lulo rhizosphere, and 14 originated from samples of soil that lacked plants. In addition, 18 strains showed positive inoculation effects on both Solanum species, of which 12 were classified as Rhizobium spp. by partial 16S rRNA gene sequencing. Overall, the strategy adopted allowed us to identify the variability in the composition of culturable diazotroph/N-scavenger representatives from soils under different management conditions by using two Solanum species as trap plants. The present results suggest the ability of tomato and lulo plants to enrich their belowground microbiomes with rhizobia representatives and the potential of selected rhizobial strains to promote the growth of Solanum crops under limiting N supply.
Assuntos
Bactérias/isolamento & purificação , Fixação de Nitrogênio/fisiologia , Rizosfera , Microbiologia do Solo , Solanum/microbiologia , Fenômenos Fisiológicos Bacterianos , Biodiversidade , Produtos Agrícolas/crescimento & desenvolvimento , Solanum lycopersicum/microbiologia , Microbiota , Nitrogênio/metabolismo , Desenvolvimento Vegetal , Proteobactérias/isolamento & purificação , Solanum/genética , Especificidade da EspécieRESUMO
This work aimed to study the inheritance of resistance to Phytophthora infestans in tomato plants, using the maximum likelihood estimation function. The susceptible cultivar Santa Clara (Solanum lycopersicum) was used as the female genitor and the P. infestans resistant S. habrochaites f. glabratum accession (BGH 6902) as the male genitor. F1 plants from the crossing were self-pollinated to produce F2 progenies, and also backcrossed with PR and PS to produce BC1:R and BC1:S generations, respectively. The tomato plants were inoculated 50 days after transplanting. Disease severity was evaluated via a diagrammatic scale. Comparison of the genetic models created using the maximum likelihood function revealed that the inheritance of resistance to P. infestans in S. habrochaites is conferred by a major gene with additive and dominance effects, polygenes with additive effects, plus the environmental effect. Vertical resistance can be explored using genes with major effects. Programs of recurrent and maker-assisted selection are considered efficient strategies with which to select genotypes that hold P. infestans resistance conferred by polygenes.
Assuntos
Phytophthora infestans/patogenicidade , Doenças das Plantas/microbiologia , Solanum/microbiologia , Genótipo , Solanum/genéticaRESUMO
In 1976, a virus with flexuous, filamentous virions typical of the family Potyviridae was isolated from symptomatic pepino (Solanum muricatum) plants growing in two valleys in Peru's coastal desert region. In 2014, a virus with similar-shaped virions was isolated from asymptomatic fruits obtained from pepino plants growing in six coastal valleys and a valley in Peru's Andean highlands. Both were identified subsequently as Wild potato mosaic virus (WPMV) by serology or high-throughput sequencing (HTS). The symptoms caused by two old and seven new isolates from pepino were examined in indicator plants. Infected solanaceous hosts varied considerably in their sensitivities to infection and individual isolates varied greatly in virulence. All seven new isolates caused quick death of infected Nicotiana benthamiana plants and more than half of them killed infected plants of Physalis floridana and S. chancayense. These three species were the most sensitive to infection. The most virulent isolate was found to be BA because it killed five of eight solanaceous host species whereas CA was the least severe because it only killed N. benthamiana. Using HTS, complete genomic sequences of six isolates were obtained, with one isolate (FE) showing evidence of recombination. The distances between individual WPMV isolates in phylogenetic trees and the geographical distances between their collection sites were found to be unrelated. The individual WPMV isolates displayed nucleotide sequence identities of 80.9-99.8%, whereas the most closely related virus, Potato virus V (PVV), was around 75% identical to WPMV. WPMV, PVV, and Peru tomato virus formed clusters of similar phylogenetic diversity, and were found to be distinct but related viruses within the overall Potato virus Y lineage. WPMV infection seems widespread and of likely economic significance to pepino producers in Peru's coastal valleys. Because it constitutes the fifth virus found infecting pepino and this crop is entirely vegetatively propagated, development of healthy pepino stock programs is advocated.
Assuntos
Genoma Viral , Potyvirus , Solanum , Genoma Viral/genética , Peru , Filogenia , Potyvirus/classificação , Potyvirus/genética , Solanum/microbiologia , Especificidade da EspécieRESUMO
The aim of this work was the production of bioactive metabolites by submerged fermentation from the fungus Diaporthe schini, followed by their extraction, separation and characterization. Different solvents (methanol, dichloromethane and hexane) were used for the extraction of metabolites from the fermentation broth and the extracts obtained were evaluated by in vitro antibacterial and antifungal activity. The separation and characterization of the extract from the hexane extraction was performed by column chromatography and GC-MS, respectively. The extracts had a great inhibitory action on the Gram-positive bacteria Staphylococcus epidermidis and Staphylococcus aureus, on the Gram-negative bacteria Enterobacter aerogenes and Klebsiella pneumoniae and on the fungus Candida krusei. The main metabolites produced were: 13-docosenamide, (Z)-; 2-hexadecene, 3,7,11,15-tetramethyl; 9-octadecenamide and 11-octadecenoic acid. Studies related to the antibacterial and antifungal activities of metabolites extracted from microorganisms are found in the literature. However, works about the identification of metabolites produced by submerged fermentation from Diaporthe schini were not found until the present moment. This work is an initial study where the conditions of the process can be optimized by looking for the production of a specific compound and can be a promising source for obtaining new drugs.
Assuntos
Antibacterianos/farmacologia , Antifúngicos/farmacologia , Ascomicetos/metabolismo , Antibacterianos/isolamento & purificação , Antifúngicos/isolamento & purificação , Ascomicetos/genética , Bactérias/efeitos dos fármacos , Bactérias/crescimento & desenvolvimento , Candida/efeitos dos fármacos , Candida/crescimento & desenvolvimento , DNA Fúngico/análise , Fermentação , Testes de Sensibilidade Microbiana , Solanum/microbiologiaRESUMO
The oomycete Phytophthora infestans, causal agent of the tomato and potato late blight, generates important economic and environmental losses worldwide. As current control strategies are becoming less effective, there is a need for studies on oomycete metabolism to help identify promising and more effective targets for chemical control. The pyrimidine pathways are attractive metabolic targets to combat tumors, virus and parasitic diseases but have not yet been studied in Phytophthora. Pyrimidines are involved in several critical cellular processes and play structural, metabolic and regulatory functions. Here, we used genomic and transcriptomic information to survey the pyrimidine metabolism during the P. infestans life cycle. After assessing the putative gene machinery for pyrimidine salvage and de novo synthesis, we inferred genealogies for each enzymatic domain in the latter pathway, which displayed a mosaic origin. The last two enzymes of the pathway, orotate phosphoribosyltransferase and orotidine-5-monophosphate decarboxylase, are fused in a multi-domain enzyme and are duplicated in some P. infestans strains. Two splice variants of the third gene (dihydroorotase) were identified, one of them encoding a premature stop codon generating a non-functional truncated protein. Relative expression profiles of pyrimidine biosynthesis genes were evaluated by qRT-PCR during infection in Solanum phureja. The third and fifth genes involved in this pathway showed high up-regulation during biotrophic stages and down-regulation during necrotrophy, whereas the uracil phosphoribosyl transferase gene involved in pyrimidine salvage showed the inverse behavior. These findings suggest the importance of de novo pyrimidine biosynthesis during the fast replicative early infection stages and highlight the dynamics of the metabolism associated with the hemibiotrophic life style of pathogen.
Assuntos
Phytophthora infestans/genética , Phytophthora infestans/metabolismo , Phytophthora infestans/patogenicidade , Pirimidinas/biossíntese , Processamento Alternativo , Clonagem Molecular , Di-Hidro-Orotase/genética , Di-Hidro-Orotase/metabolismo , Orotato Fosforribosiltransferase/genética , Orotato Fosforribosiltransferase/metabolismo , Orotidina-5'-Fosfato Descarboxilase/genética , Orotidina-5'-Fosfato Descarboxilase/metabolismo , Filogenia , Pirimidinas/metabolismo , Solanum/microbiologiaRESUMO
The diversity and antimicrobial activity of endophytic fungi associated with the Brazilian medicinal plant Solanum cernuum Vell. were studied during summer and winter seasons. A total of 246 fungal isolates were obtained, including 225 filamentous fungi and 21 yeasts. They were identified by morphological, physiological, and molecular methods. Fifty-five different taxa represented by the phyla Ascomycota (33 taxa), Basidiomycota (21 taxa), and Zygomycota (one taxon) were identified. The most abundant taxa were closely related to Arthrobotrys foliicola , Colletotrichum gloeosporioides , Coprinellus radians , Glomerella acutata , Diatrypella frostii , Phoma glomerata , Mucor sp., Phlebia subserialis , Phoma moricola , Phanerochaete sordida , and Colletotrichum sp. A total of 265 fungal extracts were screened and 64 (26.01%) displayed antimicrobial activities. Among these extracts, 18 (28.12%) presented antibacterial and antifungal activities, 42 (65.62%) displayed selective antibacterial activity, and four (6.25%) exhibited only antifungal activity. The best values of minimum inhibitory concentration were obtained from extracts of Cryptococcus rajasthanensis , Glomerella acutata, Leptosphaeria sp., and Phoma glomerata ranging from 7.8 to 15.62 µg/mL. This study is the first survey of the endophytic fungi community associated with S. cernuum, and our results show that they can represent a promising source of bioactive compounds.
Assuntos
Anti-Infecciosos/farmacologia , Bactérias/efeitos dos fármacos , Biodiversidade , Fungos/efeitos dos fármacos , Fungos/fisiologia , Plantas Medicinais/microbiologia , Solanum/microbiologia , Anti-Infecciosos/isolamento & purificação , Brasil , Misturas Complexas/farmacologia , DNA Espaçador Ribossômico/genética , Endófitos , Fungos/química , Fungos/classificação , Fungos/genética , Fungos/isolamento & purificação , Genes de RNAr/genética , Testes de Sensibilidade Microbiana , Dados de Sequência Molecular , FilogeniaRESUMO
Two Solanum genotypes, a wild relative of cultivated potato S. cajamarquense (Cjm) and an advanced tetraploid clone B3C1 (B3), were inoculated with two Phytophthora infestans isolates and leaves were sampled at 72 and 96 h after inoculation. Gene expression in the inoculated versus noninoculated samples was monitored using the Institute of Genomic Research (TIGR) 10K potato array and real-time reverse transcriptase-polymerase chain reaction (RT-PCR). The current experiment is study number 83 of the TIGR expression profiling service project, and all data are publicly available in the Solanaceae Gene Expression Database (SGED) at ftp://ftp.tigr.org/pub/data/s_tuberosum/SGED. Differentially regulated cDNA clones were selected separately for each isolate-time point interaction by significant analysis of microarray (SAM), and differentially regulated clones were classified into functional categories by MapMan. The results show that the genes activated in B3 and Cjm have largely the same biological functions and are commonly activated when plants respond to pathogen attack. The genes activated within biological function categories were considerably different between the genotypes studied, suggesting that the defence pathways activated in B3 and Cjm during the tested conditions may involve unique genes. However, as indicated by real-time RT-PCR, some of the genes thought to be genotype specific may be activated across genotypes at other time points during disease development.
Assuntos
Phytophthora infestans/fisiologia , Solanum tuberosum/genética , Solanum tuberosum/microbiologia , Solanum/genética , Solanum/microbiologia , Genótipo , Análise de Sequência com Séries de Oligonucleotídeos , Proteínas de Plantas/genética , Reação em Cadeia da Polimerase Via Transcriptase ReversaRESUMO
Phylogenetic relationships of Phytophthora infestans sensu lato in the Andean highlands of South America were examined. Three clonal lineages (US-1, EC-1, EC-3) and one heterogeneous lineage (EC-2) were found in association with different host species in genus Solanum. The EC-2 lineage includes two mitochondrial (mtDNA) haplotypes, Ia and Ic. Isolates of P. infestans sensu lato EC-2 fit the morphological description of P. infestans but are different from any genotypes of P. infestans described to date. All isolates of P. infestans sensu lato from Ecuador were amplified by a P. infestans specific primer (PINF), and restriction fragment length patterns were identical in isolates amplified with ITS primers 4 and 5. The EC-1 clonal lineage of P. infestans sensu lato from S. andreanum, S. columbianum, S. paucijugum, S. phureja, S. regularifolium, S. tuberosum and S. tuquerense was confirmed to be P. infestans based on sequences of the cytochrome oxidase I (cox I) gene and intron 1 of ras gene. The EC-2 isolates with the Ic haplotype formed a distinct branch in the same clade with P. infestans and P. mirabilis, P. phaseoli and P. ipomoeae for both cox I and ras intron 1 phylogenies and were identified as the newly described species P. andina. Ras intron 1 sequence data suggests that P. andina might have arisen via hybridization between P. infestans and P. mirabilis.
Assuntos
Filogenia , Phytophthora/classificação , Phytophthora/genética , Doenças das Plantas/microbiologia , Solanum/microbiologia , DNA Mitocondrial/genética , Equador , Complexo IV da Cadeia de Transporte de Elétrons/genética , Proteínas Fúngicas/genética , Dados de Sequência Molecular , Phytophthora/isolamento & purificação , Polimorfismo de Fragmento de Restrição , Proteínas ras/genéticaRESUMO
Late blight caused by Phytophthora infestans is one of the most limiting diseases in solanaceous crops in the world. This pathogen is a main constraint in the highland Andes, where these plants are grown under high humidity conditions and continuous cropping. The aim of this research was to increase the available information on the biology of P. infestans, specifically on its level of genetic variation in south-western Colombia, an area where various solanaceous crops susceptible to this pathogen converge. The study was carried out by using AFLP molecular markers with the restriction enzymes EcoRI and MseI and different primer combinations. Results indicated a low level of genetic variation among the 26 isolates evaluated, with only 18 polymorphic bands out of 135 amplicons obtained (13.43%), a Nei's genetic diversity index of 0.04, and a Shannon's information index of 0.06.
Assuntos
Variação Genética , Phytophthora infestans/genética , Colômbia , Phytophthora infestans/isolamento & purificação , Solanum/microbiologiaRESUMO
The central highlands of Mexico are considered to be a center of genetic diversity for both the potato late blight pathogen and for tuber-bearing Solanum spp. Recent work conducted in Mexico and South America sheds new light on the biology and evolution of Phytophthora infestans and other related Phytophthora pathogens. It now appears that Mexican Solanum species, which coevolved with P. infestans and were previously known for providing a source of R-genes, also provide a source of quantitative, rate-reducing resistance that is highly effective, stable, and durable. It is now apparent that Mexico is the center of origin not only of the potato late blight pathogen P. infestans, but also of several related Phytophthora species including P. mirabilis, P. ipomoeae, and possibly P. phaseoli. We close with the hypothesis that these Phytophthora species evolved sympatrically from one ancestral host through adaptive radiation onto their respective four host families.
Assuntos
Phytophthora/genética , Phytophthora/fisiologia , México , Phytophthora/classificação , Doenças das Plantas/microbiologia , Solanum/microbiologiaRESUMO
Genetic variability in Crinipellis perniciosa, the causal organism of witches' broom disease in Theobroma cacao, was determined in strains originating from T. cacao and other susceptible host species Heteropterys acutifolia and Solanum lycocarpum in Brazil, in order to clarify host specificity and geographical variability. RFLP analysis of the ribosomal DNA ITS regions (rDNA ITS), and the mitochondrial DNA small subunit ribosomal DNA gene (mtDNA SSU rDNA) did not reveal any genetic variability in 120 tested strains, possibly serving only as species level markers. Genetic variability was observed in the ribosomal DNA IGS spacer region, in terms of IGS size, RFLPs and sequence data. Phylogenetic analyses (using CLUSTAL W, PHYLIP and TREEVIEW) indicated considerable differences between C. perniciosa strains from T. cacao and those from H. acutifolia (85-86%) and S. lycocarpum (95-96%). Sequence differences also indicated that C. perniciosa from T. cacao in Bahia is less variable (98%) when compared to the pathogen on T. cacao in Amazonas (97-98%), perhaps reflecting a recent introduction to T. cacao in Bahia.