RESUMO
A survey of the ecological variability within 52 populations of Schoenoplectus californicus (C.A. Mey.) Soják across its distributional range revealed that it is commonly found in nitrogen (N) limited areas, but rarely in phosphorus limited soils. We explored the hypothesis that S. californicus supplements its nitrogen demand by bacterial N2-fixation processes associated with its roots and rhizomes. We estimated N2-fixation of diazotrophs associated with plant rhizomes and roots from several locations throughout the species' range and conducted an experiment growing plants in zero, low, and high N additions. Nitrogenase activity in rhizomes and roots was measured using the acetylene reduction assay. The presence of diazotrophs was verified by the detection of the nifH gene. Nitrogenase activity was restricted to rhizomes and roots and it was two orders of magnitude higher in the latter plant organs (81 and 2032 nmol C2H4 g DW-1 d-1, respectively). Correspondingly, 40x more nifH gene copies were found on roots compared to rhizomes. The proportion of the nifH gene copies in total bacterial DNA was positively correlated with the nitrogenase activity. In the experiment, the contribution of fixed N to the plant N content ranged from 13.8% to 32.5% among clones from different locations. These are relatively high values for a non-cultivated plant and justify future research on the link between N-fixing bacteria and S. californicus production.
Assuntos
Cyperaceae/metabolismo , Fixação de Nitrogênio , Nitrogênio/metabolismo , Raízes de Plantas/metabolismo , Rizoma/metabolismo , Áreas Alagadas , Proteínas de Bactérias/metabolismo , Cyperaceae/microbiologia , Nitrogênio/química , Nitrogenase/metabolismo , América do Norte , Oxirredutases/metabolismo , Fósforo/química , Fósforo/metabolismo , Dispersão Vegetal , Proteínas de Plantas/metabolismo , Rizoma/microbiologia , Solo/química , América do Sul , Especificidade da EspécieRESUMO
Plant growth-promoting rhizobacteria stimulate plant growth and development via different mechanisms. In this study, we characterized the effect of volatiles from Bacillus methylotrophicus M4-96 isolated from the maize rhizosphere on root and shoot development, and auxin homeostasis in Arabidopsis thaliana. Phytostimulation occurred after 4 days of interaction between M4-96 and Arabidopsis grown on opposite sides of divided Petri plates, as revealed by enhanced primary root growth, root branching, leaf formation, and shoot biomass accumulation. Analysis of indole-3-acetic acid content revealed two- and threefold higher accumulation in the shoot and root of bacterized seedlings, respectively, compared to uninoculated plants, which was correlated with increased expression of the auxin response marker DR5::GUS. The auxin transport inhibitor 1-naphthylphthalamic acid inhibited primary root growth and lateral root formation in axenically grown seedlings and antagonized the plant growth-promoting effects of M4-96. Analysis of bacterial volatile compounds revealed the presence of four classes of compounds, including ten ketones, eight alcohols, one aldehyde, and two hydrocarbons. However, the abundance of ketones and alcohols represented 88.73 and 8.05%, respectively, of all airborne signals detected, with acetoin being the main compound produced. Application of acetoin had a different effect from application of volatiles, suggesting that either the entire pool or acetoin acting in concert with another unidentified compound underlies the strong phytostimulatory response. Taken together, our results show that B. methylotrophicus M4-96 generates bioactive volatiles that increase the active auxin pool of plants, stimulate the growth and formation of new organs, and reprogram root morphogenesis.
Assuntos
Arabidopsis/crescimento & desenvolvimento , Bacillus/fisiologia , Zea mays/microbiologia , Arabidopsis/metabolismo , Arabidopsis/microbiologia , Proteínas de Arabidopsis/genética , Proteínas de Arabidopsis/metabolismo , Bacillus/isolamento & purificação , Expressão Gênica , Regulação da Expressão Gênica de Plantas , Ácidos Indolacéticos/metabolismo , Rizoma/crescimento & desenvolvimento , Rizoma/metabolismo , Rizoma/microbiologia , Compostos Orgânicos Voláteis/metabolismoRESUMO
This study aimed to evaluate the biocontrol potential of bacteria isolated from different plant species and soils. The production of compounds related to phytopathogen biocontrol and/or promotion of plant growth in bacterial isolates was evaluated by measuring the production of antimicrobial compounds (ammonia and antibiosis) and hydrolytic enzymes (amylases, lipases, proteases, and chitinases) and phosphate solubilization. Of the 1219 bacterial isolates, 92% produced one or more of the eight compounds evaluated, but only 1% of the isolates produced all the compounds. Proteolytic activity was most frequently observed among the bacterial isolates. Among the compounds which often determine the success of biocontrol, 43% produced compounds which inhibit mycelial growth of Monilinia fructicola, but only 11% hydrolyzed chitin. Bacteria from different plant species (rhizosphere or phylloplane) exhibited differences in the ability to produce the compounds evaluated. Most bacterial isolates with biocontrol potential were isolated from rhizospheric soil. The most efficient bacteria (producing at least five compounds related to phytopathogen biocontrol and/or plant growth), 86 in total, were evaluated for their biocontrol potential by observing their ability to kill juvenile Mesocriconema xenoplax. Thus, we clearly observed that bacteria that produced more compounds related to phytopathogen biocontrol and/or plant growth had a higher efficacy for nematode biocontrol, which validated the selection strategy used.(AU)
Assuntos
Microbiologia do Solo , Rizoma/microbiologia , Rizosfera , Doenças das Plantas/prevenção & controle , Controle Biológico de VetoresRESUMO
Fusarium wilt of banana, which is caused by Fusarium oxysporum f. sp. cubense, is one of the most serious diseases of banana. F. oxysporum f. sp. cubense race 1 (Foc1) and race 4 (Foc4) are the most prevalent pathogens of banana cultivars in the world. To understand the differences in the infection processes between Foc1 and Foc4, green fluorescent protein-tagged strains of F. oxysporum f. sp. cubense tropical race 4 (FocTR4) and Foc1 were used to inoculate 'Brazil Cavendish' banana. At 2 days postinoculation (dpi), it was observed that the spores and hyphae of both Foc1 and Foc4 attached to the root hairs and root epidermis. At 3 dpi, the hyphae of both Foc1 and Foc4 were found in the vascular tissues of roots. However, Foc4 was observed in the parenchymal cells of banana root, whereas Foc1 was not found in parenchymal cells at 7 dpi. Furthermore, few Foc1 hyphae were observed in a few xylems whereas many more Foc4 hyphae were present in many xylems and phloems. Foc4 was observed in the vascular tissues of banana rhizomes, whereas no Foc1 was found in rhizomes 2 months after inoculation. The attachment process in F. oxysporum f. sp. cubense infection was further studied with scanning electron microscopy. Foc4 was observed to penetrate into banana roots from the intercellular space of the epidermis and wounds, whereas Foc1 mainly penetrated from the wounds but not from the intercellular space of the epidermis. Therefore, direct root penetration and rhizome vascular colonization by F. oxysporum f. sp. cubense are the key steps in the successful infection of Brazil Cavendish.
Assuntos
Fusarium , Musa , Raízes de Plantas , Rizoma , Fusarium/fisiologia , Musa/microbiologia , Raízes de Plantas/microbiologia , Rizoma/microbiologiaRESUMO
Endophyte microorganisms live inside plants without causing them any apparent damage. Recently, endophytic microorganisms have attracted attention because they can produce bioactive compounds of biotechnological interest. The endophytic microorganisms in Paris polyphylla var. yunnanensis (Liliaceae) - a species used since antiquity in traditional Chinese medicine - are under scrutiny because they may be responsible for producing the bioactive metabolites associated with the plant. The levels of bioactive metabolites in the rhizomes of P. polyphylla increase with rhizome age. To elucidate the roles played by endophytes in the accumulation of bioactive metabolites, we investigated the community structure and diversity of the endophytic microorganisms in P. polyphylla rhizomes of different ages (4, 6, and 8 years) using 16S rRNA and internal transcribed spacer (ITS) sequence analysis. 16S rDNA amplicon pyrosequencing revealed that the number of operational taxonomic units was lower in the 8-year-old samples than in the other samples. A total of 28 phyla were observed in the P. polyphylla samples and the predominant bacteria were of the Cyanobacteria and Proteobacteria phyla. Moreover, the percentage of Cyanobacteria increased with rhizome age. Similarly, ITS1 amplicon pyrosequencing identified developmental changes in the most abundant fungal classes; some classes were more prevalent in the 8-year-old rhizomes than in younger rhizomes, indicating the importance in secondary metabolism in older rhizomes. Our study showed that endophyte microorganism diversity and prevalence depend on P. polyphylla rhizome age. There was also an indication that some endophyte microorganisms contribute to the higher saponin content in older P. polyphylla specimens.
Assuntos
Cianobactérias/genética , Liliaceae/genética , Proteobactérias/genética , RNA Ribossômico 16S/genética , Rizoma/microbiologia , Bactérias/genética , Cianobactérias/isolamento & purificação , Endófitos/genética , Liliaceae/microbiologia , Medicina Tradicional Chinesa , Proteobactérias/isolamento & purificação , Rizoma/genéticaRESUMO
The screening for hydrolases-producing, halotolerant, and spore-forming gram-positive bacteria from the root, rhizosphere, and non-rhizosphere soil of Blutaparon portulacoides, a plant found in the Restinga de Jurubatiba located at the northern region of Rio de Janeiro State, Brazil, resulted in the isolation of 22 strains. These strains were identified as Halobacillus blutaparonensis (n = 2), Oceanobacillus picturae (n = 5), and Oceanobacillus iheyensis (n = 15), and all showed the ability to produce different extracellular enzymes. A total of 20 isolates (90.9 %) showed activity for protease, 5 (22.7 %) for phytase, 3 (13.6 %) for cellulase, and 2 (9.1 %) for amylase. Some bacterial strains were capable of producing three (13.6 %) or two (9.1 %) distinct hydrolytic enzymes. However, no bacterial strain with ability to produce esterase and DNase was observed. The isolate designated M9, belonging to the species H. blutaparonensis, was the best producer of protease and also yielded amylase and phytase. This strain was chosen for further studies regarding its protease activity. The M9 strain produced similar amounts of protease when grown either without or with different NaCl concentrations (from 0.5 to 10 %). A simple inspection of the cell-free culture supernatant by gelatin-sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE) revealed the presence of three major alkaline proteases of 40, 50, and 70 kDa, which were fully inhibited by phenylmethylsulfonyl fluoride (PMSF) and tosyl-L-phenylalanine chloromethyl ketone (TPCK) (two classical serine protease inhibitors). The secreted proteases were detected in a wide range of temperature (from 4 to 45 °C) and their hydrolytic activities were stimulated by NaCl (up to 10 %). The serine proteases produced by the M9 strain cleaved gelatin, casein, albumin, and hemoglobin, however, in different extensions. Collectively, these results suggest the potential use of the M9 strain in biotechnological and/or industrial processes.