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1.
Sci Total Environ ; 772: 145594, 2021 Jun 10.
Artigo em Inglês | MEDLINE | ID: mdl-33770866

RESUMO

Biological nitrogen fixation (BNF) in sediments is an important source of bioavailable nitrogen in aquatic systems. However, the effect of habitat change caused by eutrophication on nitrogen fixation within sediments is still unclear. In this study, nitrogen fixation rates and diazotroph diversities in sediments with heterogeneous ecological status in one eutrophic lake were investigated by using an isotope tracer method and sequencing of nitrogen-fixing (nif) genes. The results showed that both nitrogenase activity (NA) and nifH abundance in sediments of blooms area were higher than those in vegetation-dominated habitats. Correlation analysis showed that NA was correlated closely to nifH abundance, dissolved sulfide, and iron. The diazotrophic assemblage contained mainly Proteobacterial sequences belonging to Cluster I and III, and the variations of diazotrophic community could be explained by total nitrogen content, total phosphorus content, organic matters, sulfides, ammonium and iron content. Moreover, the co-occurrence network analysis showed the Alphaproteobacteria shaped the major interactions in diazotrophic community, and sediment properties had stronger effect on diazotrophic community in cyanobacteria-dominated habitat. This study revealed that habitat heterogeneity in eutrophic lakes shaped different succession of BNF in sediments and cyanobacterial blooms significantly improved the nitrogen-fixing activity in sediments, which broadened our understanding of nitrogen cycle and nutrient management in eutrophic freshwater lakes.


Assuntos
Lagos , Fixação de Nitrogênio , China , Ecossistema , Eutrofização , Sedimentos Geológicos , Nitrogênio/análise , Fósforo/análise
2.
Mar Pollut Bull ; 165: 112126, 2021 Apr.
Artigo em Inglês | MEDLINE | ID: mdl-33667934

RESUMO

Benthic nitrogen fixation in the tropical estuaries plays a major role in marine nitrogen cycle, its contribution to nitrogen budget and players behind process is not well understood. The present study was estimated the benthic nitrogen fixation rate in a tropical estuary (Cochin) and also evaluated the contribution of various diazotrophic bacterial communities. Nitrogen fixation was detected throughout year (0.1-1.11 nmol N g-1 h-1); higher activity was observed in post-monsoon. The nifH gene abundance was varied from 0.8 × 104 to 0.6 × 108 copies g-1dry sediment; highest was detected in post-monsoon. The Cluster I and Cluster III were the dominant diazotrophs. Sulfur reducing bacterial phylotypes (Deltaproteobacteria) contributed up to 2-72% of total nitrogen fixation. These bacteria may provide new nitrogen to these systems, counteracting nitrogen loss via denitrification and anammox. Overall, the study explained the importance of benthic nitrogen fixation and role of diazotrophs in a monsoon influenced tropical estuarine environments.


Assuntos
Estuários , Fixação de Nitrogênio , Bactérias , Nitrogênio/análise , Enxofre
3.
Sci Total Environ ; 773: 145066, 2021 Jun 15.
Artigo em Inglês | MEDLINE | ID: mdl-33582326

RESUMO

Biological nitrogen fixation is a key process for the maintenance of natural ecosystems productivity. In tropical forests, the contribution of asymbiotic nitrogen fixation (ANF) to the nitrogen (N) input has been underestimated, even though few studies have shown that ANF may be as important as symbiotic nitrogen fixation in such environments. The inputs and abiotic modulators of ANF in the Amazon forest are not completely understood. Here, we determined ANF rates and estimated the N inputs from ANF in the phyllosphere, litter and rhizospheric soil of nine tree species in the Amazon forest over time, including an extreme drought period induced by the El Niño-Southern Oscillation. Our data showed that ANF rates in the phyllosphere were 2.8- and 17.6-fold higher than in the litter and rhizospheric soil, respectively, and was highly dependent on tree taxon. Sampling time was the major factor modulating ANF in all forest compartments. At the driest period, ANF rates were approximately 1.8-fold and 13.1-fold higher than at periods with higher rainfall, before and after the extreme drought period, respectively. Tree species was a key modulator of ANF in the phyllosphere, as well as N and Vanadium concentrations. Carbon, molybdenum and vanadium concentrations were significant modulators of ANF in the litter. Based on ANF rates at the three sampling times, we estimated that the N input in the Amazon forest through ANF in the phyllosphere, litter and rhizospheric soil, was between 0.459 and 0.714 kg N ha-1 yr-1. Our results highlight the importance of ANF in the phyllosphere for the N input in the Amazon forest, and suggest that changes in the patterns of ANF driven by large scale climatic events may impact total N inputs and likely alter forest productivity.


Assuntos
Ecossistema , Fixação de Nitrogênio , Florestas , Nitrogênio , Solo , Árvores
4.
Int J Mol Sci ; 22(3)2021 Feb 02.
Artigo em Inglês | MEDLINE | ID: mdl-33540521

RESUMO

Application of diazotrophs (N2-fixing microorganisms) can decrease the overuse of nitrogen (N) fertilizer. Until now, there are few studies on the effects of diazotroph application on microbial communities of major crops. In this study, the diazotrophic and endospore-forming Paenibacillus triticisoli BJ-18 was inoculated into maize soils containing different N levels. The effects of inoculation on the composition and abundance of the bacterial, diazotrophic and fungal communities in the rhizosphere and root/shoot endosphere of maize were evaluated by sequencing the 16S rRNA, nifH gene and ITS (Inter Transcribed Spacer) region. P. triticisoli BJ-18 survived and propagated in all the compartments of the maize rhizosphere, root and shoot. The abundances and diversities of the bacterial and diazotrophic communities in the rhizosphere were significantly higher than in both root and shoot endospheres. Each compartment of the rhizosphere, root and shoot had its specific bacterial and diazotrophic communities. Our results showed that inoculation reshaped the structures of the bacterial, diazotrophic and fungal communities in the maize rhizosphere and endosphere. Inoculation reduced the interactions of the bacteria and diazotrophs in the rhizosphere and endosphere, while it increased the fungal interactions. After inoculation, the abundances of Pseudomonas, Bacillus and Paenibacillus in all three compartments, Klebsiella in the rhizosphere and Paenibacillus in the root and shoot were significantly increased, while the abundances of Fusarium and Giberella were greatly reduced. Paenibacillus was significantly correlated with plant dry weight, nitrogenase, N2-fixing rate, P solubilization and other properties of the soil and plant.


Assuntos
Código de Barras de DNA Taxonômico , Microbiota , Paenibacillus/fisiologia , Rizosfera , Microbiologia do Solo , Zea mays/microbiologia , Bactérias/isolamento & purificação , Bactérias/metabolismo , Fungos/isolamento & purificação , Micobioma , Fixação de Nitrogênio , Paenibacillus/metabolismo , Raízes de Plantas/microbiologia
5.
Microbiome ; 9(1): 53, 2021 02 23.
Artigo em Inglês | MEDLINE | ID: mdl-33622403

RESUMO

BACKGROUND: Mosses in high-latitude ecosystems harbor diverse bacterial taxa, including N2-fixers which are key contributors to nitrogen dynamics in these systems. Yet the relative importance of moss host species, and environmental factors, in structuring these microbial communities and their N2-fixing potential remains unclear. We studied 26 boreal and tundra moss species across 24 sites in Alaska, USA, from 61 to 69° N. We used cultivation-independent approaches to characterize the variation in moss-associated bacterial communities as a function of host species identity and site characteristics. We also measured N2-fixation rates via 15N2 isotopic enrichment and identified potential N2-fixing bacteria using available literature and genomic information. RESULTS: Host species identity and host evolutionary history were both highly predictive of moss microbiome composition, highlighting strong phylogenetic coherence in these microbial communities. Although less important, light availability and temperature also influenced composition of the moss microbiome. Finally, we identified putative N2-fixing bacteria specific to some moss hosts, including potential N2-fixing bacteria outside well-studied cyanobacterial clades. CONCLUSIONS: The strong effect of host identity on moss-associated bacterial communities demonstrates mosses' utility for understanding plant-microbe interactions in non-leguminous systems. Our work also highlights the likely importance of novel bacterial taxa to N2-fixation in high-latitude ecosystems. Video Abstract.


Assuntos
Briófitas/microbiologia , Fixação de Nitrogênio , Bactérias Fixadoras de Nitrogênio/classificação , Bactérias Fixadoras de Nitrogênio/metabolismo , Alaska , Filogenia
6.
Int J Mol Sci ; 22(1)2021 Jan 04.
Artigo em Inglês | MEDLINE | ID: mdl-33406726

RESUMO

Iron is an essential nutrient for the legume-rhizobia symbiosis and nitrogen-fixing bacteroids within root nodules of legumes have a very high demand for the metal. Within the infected cells of nodules, the bacteroids are surrounded by a plant membrane to form an organelle-like structure called the symbiosome. In this review, we focus on how iron is transported across the symbiosome membrane and accessed by the bacteroids.


Assuntos
Bacteroides/fisiologia , Fabaceae/metabolismo , Ferro/metabolismo , Nitrogênio/metabolismo , Organelas/metabolismo , Proteínas de Plantas/metabolismo , Simbiose , Transporte Biológico , Fabaceae/microbiologia , Fixação de Nitrogênio , Organelas/microbiologia , Nódulos Radiculares de Plantas/metabolismo , Nódulos Radiculares de Plantas/microbiologia
7.
Environ Sci Technol ; 55(2): 1310-1318, 2021 01 19.
Artigo em Inglês | MEDLINE | ID: mdl-33389989

RESUMO

Biological nitrogen fixation (BNF) represents the natural pathway by which mosses meet their demands for bioavailable/reactive nitrogen (Nr) in peatlands. However, following intensification of nitrogen fertilizer and fossil fuel use, atmospheric Nr deposition has increased exposing peatlands to Nr loading often above the ecological threshold. As BNF is energy intensive, therefore, it is unclear whether BNF shuts down when Nr availability is no longer a rarity. We studied the response of BNF under a gradient of Nr deposition extending over decades in three peatlands in the U.K., and at a background deposition peatland in Sweden. Experimental nitrogen fertilization plots in the Swedish site were also evaluated for BNF activity. In situ BNF activity of peatlands receiving Nr deposition of 6, 17, and 27 kg N ha-1 yr-1 was not shut down but rather suppressed by 54, 69, and 74%, respectively, compared to the rates under background Nr deposition of ∼2 kg N ha-1 yr-1. These findings were corroborated by similar BNF suppression at the fertilization plots in Sweden. Therefore, contribution of BNF in peatlands exposed to chronic Nr deposition needs accounting when modeling peatland's nitrogen pools, given that nitrogen availability exerts a key control on the carbon capture of peatlands, globally.


Assuntos
Briófitas , Fixação de Nitrogênio , Carbono , Nitrogênio/análise , Suécia
8.
Sci Total Environ ; 767: 144252, 2021 May 01.
Artigo em Inglês | MEDLINE | ID: mdl-33429279

RESUMO

Tillage has a considerable effect on the soil ecosystem and its services, including microbial communities. Harnessing beneficial microbes is a sustainable way to optimizing crop management and agricultural production. Although diazotrophs play a major role in global biological nitrogen fixation, the effects of tillage on diazotrophic communities in the rhizosphere are not fully understood. In the present study, we investigated the diazotrophic community in wheat rhizosphere soil under different tillage treatments in a long-term experiment, i.e., plow tillage (considered as conventional tillage), chisel plow tillage (considered as conservation tillage), and zero tillage (considered as conservation tillage). Tillage led to a divergent distribution in the rhizosphere diazotrophic community and significant changes in community structure. Tillage caused specific responses from members/modules of the rhizosphere diazotrophic community co-occurrence network, and the relative abundance of keystone taxa was higher under conservation tillage than under conventional tillage. The increased abundance of tillage-sensitive modules under conservation tillage had a broad and significant positive correlation with rhizosphere nutrient availability, whereas the opposite was true for conventional tillage. Differences in nutrients under different tillage practices may lead to different assembly processes of diazotrophs. Overall, our findings indicate that tillage significantly affects the assembly and composition of the rhizosphere diazotrophic community, emphasizing the importance of improved substrate availability for rhizosphere diazotrophic modules under conservation tillage. This knowledge could deepen our understanding of the rhizosphere functional microbial community (e.g., biological nitrogen fixation).


Assuntos
Rizosfera , Triticum , Agricultura , Fixação de Nitrogênio , Solo , Microbiologia do Solo
9.
Oecologia ; 195(2): 489-497, 2021 Feb.
Artigo em Inglês | MEDLINE | ID: mdl-33394128

RESUMO

Cloud forests have been found to lose more nitrogen in stream discharge than they gain from atmospheric deposition. They also support a large diversity and biomass of tree epiphytes, predominately composed of cryptogams. Since cryptogam epiphytes harbor nitrogen fixing cyanobacteria, they may help make up for the nitrogen loss from ecosystems. We assessed cryptogam biomass on the ground, boles and branches in Quercus costaricensis dominated stands near the tree line in the Cordillera de Talamanca, Costa Rica. Nitrogen fixation was assayed using 15N2 uptake. Total cryptogam biomass was 2 977 kg ha-1, with 67% being found on the lower branches. Bryophytes and chlorolichens made up 53% and 44%, respectively, of the biomass. Half of the bryophyte mass was composed of the liverwort Plagiochila heterophylla, and 66% of the chlorolichen of Lobariella pallida. There were no significant differences in nitrogen fixation rates between the cryptogam species, with a mean rate of 5.04 µg N g-1 day-1 during the predominantly wet condition in the forest. The overall nitrogen input from fixation was 6.1 kg N ha-1 year-1, of which 78% came from bryophytes, 18% from chlorolichens, and 4% from cyanolichens. Only 2.0% of the fixation occurred in cryptogams on the ground, whereas 67%, 24%, and 7% occurred on the lower branches, boles, and upper branches, respectively. These results show that tree epiphytes constitute a significant source of nitrogen for these forests, due to the trees' large surface area, and can make up for the nitrogen lost from these ecosystems.


Assuntos
Líquens , Fixação de Nitrogênio , Ascomicetos , Biomassa , Costa Rica , Ecossistema , Florestas , Nitrogênio/análise , Árvores
10.
Artigo em Inglês | MEDLINE | ID: mdl-33439117

RESUMO

Here, we describe three endosymbiotic bacterial strains isolated from the gills of the shipworm, Bankia setacea (Teredinidae: Bivalvia). These strains, designated as Bs08T, Bs12T and Bsc2T, are Gram-stain-negative, microaerobic, gammaproteobacteria that grow on cellulose and a variety of substrates derived from lignocellulose. Phenotypic characterization, phylogeny based on 16S rRNA gene and whole genome sequence data, amino acid identity and percentage of conserved proteins analyses, show that these strains are novel and may be assigned to the genus Teredinibacter. The three strains may be differentiated and distinguished from other previously described Teredinibacter species based on a combination of four characteristics: colony colour (Bs12T, purple; others beige to brown), marine salt requirement (Bs12T, Bsc2T and Teredinibacter turnerae strains), the capacity for nitrogen fixation (Bs08T and T. turnerae strains) and the ability to respire nitrate (Bs08T). Based on these findings, we propose the names Teredinibacter haidensis sp. nov. (type strain Bs08T=ATCC TSD-121T=KCTC 62964T), Teredinibacter purpureus sp. nov. (type strain Bs12T=ATCC TSD-122T=KCTC 62965T) and Teredinibacter franksiae sp. nov. (type strain Bsc2T=ATCC TSD-123T=KCTC 62966T).


Assuntos
Bivalves/microbiologia , Gammaproteobacteria/classificação , Brânquias/microbiologia , Filogenia , Animais , Técnicas de Tipagem Bacteriana , Composição de Bases , DNA Bacteriano/genética , Ácidos Graxos/química , Gammaproteobacteria/isolamento & purificação , Fixação de Nitrogênio , Oceano Pacífico , Pigmentação , RNA Ribossômico 16S/genética , Análise de Sequência de DNA , Washington , Madeira
11.
Planta ; 253(1): 7, 2021 Jan 02.
Artigo em Inglês | MEDLINE | ID: mdl-33387090

RESUMO

MAIN CONCLUSION: ST1 and ST6 are possibly involved in primary and lateral root and symbiotic nodule development, but only ST6 participates in the interaction with hemibiotrophic fungi. Specific tissue (ST) proteins have been shown to be involved in several processes related to plant nutritional status, development, and responses to biotic agents. In particular, ST1 and ST6 are mainly expressed in roots throughout plant development. Here, we analyze where and how the expression of the genes encoding both proteins are modulated in the legume model plant Medicago truncatula in response to the plant developmental program, nodulation induced by a beneficial nitrogen-fixing bacterium (Sinorhizobium meliloti) and the defense response triggered by a pathogenic hemibiotrophic fungus (Fusarium oxysporum). Gene expression results show that ST1 and ST6 participate in the vasculature development of both primary and lateral roots, although only ST6 is related to meristem activity. ST1 and ST6 clearly display different roles in the biotic interactions analyzed, where ST1 is activated in response to a N2-fixing bacterium and ST6 is up-regulated after inoculation with F. oxysporum. The role of ST1 and ST6 in the nodulation process may be related to nodule organogenesis rather than to the establishment of the interaction itself, and an increase in ST6 correlates with the activation of the salicylic acid signaling pathway during the infection and colonization processes. These results further support the role of ST6 in response to hemibiotrophic fungi. This research contributes to the understanding of the complex network that controls root biology and strengthens the idea that ST proteins are involved in several processes such as primary and lateral root development, nodule organogenesis, and the plant-microbe interaction.


Assuntos
Fusarium , Medicago truncatula , Proteínas de Plantas , Raízes de Plantas , Sinorhizobium meliloti , Simbiose , Fusarium/fisiologia , Regulação da Expressão Gênica de Plantas , Medicago truncatula/genética , Medicago truncatula/crescimento & desenvolvimento , Medicago truncatula/microbiologia , Fixação de Nitrogênio , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo , Raízes de Plantas/genética , Raízes de Plantas/crescimento & desenvolvimento , Nódulos Radiculares de Plantas/metabolismo , Nódulos Radiculares de Plantas/microbiologia , Sinorhizobium meliloti/fisiologia
12.
Sci Total Environ ; 754: 142202, 2021 Feb 01.
Artigo em Inglês | MEDLINE | ID: mdl-33254844

RESUMO

Biological nitrogen fixation (BNF) is a fundamental part of nitrogen cycling in tropical forests, yet little is known about the contribution made by free-living nitrogen fixers inhabiting the often-extensive forest canopy. We used the acetylene reduction assay, calibrated with 15N2, to measure free-living BNF on forest canopy leaves, vascular epiphytes, bryophytes and canopy soil, as well as on the forest floor in leaf litter and soil. We used a combination of calculated and published component densities to upscale free-living BNF rates to the forest level. We found that bryophytes and leaves situated in the canopy in particular displayed high mass-based rates of free-living BNF. Additionally, we calculated that nearly 2 kg of nitrogen enters the forest ecosystem through free-living BNF every year, 40% of which was fixed by the various canopy components. Our results reveal that in the studied tropical lowland forest a large part of the nitrogen input through free-living BNF stems from the canopy, but also that the total nitrogen inputs by free-living BNF are lower than previously thought and comparable to the inputs of reactive nitrogen by atmospheric deposition.


Assuntos
Fixação de Nitrogênio , Solo , Ecossistema , Florestas , Nitrogênio , Árvores , Clima Tropical
13.
Ambio ; 50(1): 203-214, 2021 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-32314265

RESUMO

Dense blooms of diazotrophic filamentous cyanobacteria are formed every summer in the Baltic Sea. We estimated their contribution to nitrogen fixation by combining two decades of cyanobacterial biovolume monitoring data with recently measured genera-specific nitrogen fixation rates. In the Bothnian Sea, estimated nitrogen fixation rates were 80 kt N year-1, which has doubled during recent decades and now exceeds external loading from rivers and atmospheric deposition of 69 kt year-1. The estimated contribution to the Baltic Proper was 399 kt N year-1, which agrees well with previous estimates using other approaches and is greater than the external input of 374 kt N year-1. Our approach can potentially be applied to continuously estimate nitrogen loads via nitrogen fixation. Those estimates are crucial for ecosystem adaptive management since internal nitrogen loading may counteract the positive effects of decreased external nutrient loading.


Assuntos
Cianobactérias , Fixação de Nitrogênio , Países Bálticos , Ecossistema , Nitrogênio/análise , Água do Mar
14.
Postepy Biochem ; 66(1): 49-61, 2020 03 31.
Artigo em Polonês | MEDLINE | ID: mdl-33320473

RESUMO

Lipopolysaccharides synthesized by rhizobia have a various structure. Differences are observed in lipid A (considered as the most conservative part of LPS), in the core region, and in the O-specific polysaccharide. Lipids A may have different compositions of the sugar backbone and the acylation pattern. The core region of rhizobia mainly consists of hexoses, uronic acids, N-acetylquinohozamine, and Kdo, but has no heptose region typical for enterobacteria. The O-PSs may have a different structure even among strains of the same species. They are built of various monosaccharides and are often hydrophobic. An appropriate structure of LPS domains is required for establishment of an effective symbiosis between bacteria and their plant host. Changes in the structure of LPS (most often caused by mutations) resulted in a decrease in efficiency or failure of atmospheric nitrogen fixation. Complete LPS protects symbiotic bacteria penetrating plant cells and determines the proper organization and maturation of symbiosomes.


Assuntos
Lipopolissacarídeos/química , Lipopolissacarídeos/metabolismo , Plantas/metabolismo , Plantas/microbiologia , Rhizobium/química , Rhizobium/metabolismo , Simbiose , Fixação de Nitrogênio
15.
Proc Biol Sci ; 287(1934): 20201493, 2020 09 09.
Artigo em Inglês | MEDLINE | ID: mdl-32873201

RESUMO

Legumes can meet their nitrogen requirements through root nodule symbiosis, which could also trigger plant systemic resistance against pests. The pea aphid Acyrthosiphon pisum, a legume pest, can harbour different facultative symbionts (FS) influencing various traits of their hosts. It is therefore worth determining if and how the symbionts of the plant and the aphid modulate their interaction. We used different pea aphid lines without FS or with a single one (Hamiltonella defensa, Regiella insecticola, Serratia symbiotica) to infest Medicago truncatula plants inoculated with Sinorhizobium meliloti (symbiotic nitrogen fixation, SNF) or supplemented with nitrate (non-inoculated, NI). The growth of SNF and NI plants was reduced by aphid infestation, while aphid weight (but not survival) was lowered on SNF compared to NI plants. Aphids strongly affected the plant nitrogen fixation depending on their symbiotic status, suggesting indirect relationships between aphid- and plant-associated microbes. Finally, all aphid lines triggered expression of Pathogenesis-Related Protein 1 (PR1) and Proteinase Inhibitor (PI), respective markers for salicylic and jasmonic pathways, in SNF plants, compared to only PR1 in NI plants. We demonstrate that the plant symbiotic status influences plant-aphid interactions while that of the aphid can modulate the amplitude of the plant's defence response.


Assuntos
Afídeos/fisiologia , Medicago truncatula/fisiologia , Fixação de Nitrogênio/fisiologia , Animais , Nitratos , Nitrogênio/metabolismo , Ácido Salicílico , Serratia , Simbiose
16.
Int J Syst Evol Microbiol ; 70(10): 5453-5459, 2020 Oct.
Artigo em Inglês | MEDLINE | ID: mdl-32910750

RESUMO

The members of the genus Frankia are, with a few exceptions, a group of nitrogen-fixing symbiotic actinobacteria that nodulate mostly woody dicotyledonous plants belonging to three orders, eight families and 23 genera of pioneer dicots. These bacteria have been characterized phylogenetically and grouped into four molecular clusters. One of the clusters, cluster 1 contains strains that induce nodules on Alnus spp. (Betulaceae), Myrica spp., Morella spp. and Comptonia spp. (Myricaceae) that have global distributions. Some of these strains produce not only hyphae and vesicles, as other cluster 1 strains do, but also numerous sporangia in their host symbiotic tissues, hence their phenotype being described as spore-positive (Sp+). While Sp+ strains have resisted repeated attempts at cultivation, their genomes have recently been characterized and found to be different from those of all described species, being markedly smaller than their phylogenetic neighbours. We thus hereby propose to create a 'Candidatus Frankia alpina' species for some strains present in nodules of Alnus alnobetula and A. incana that grow in alpine environments at high altitudes or in subarctic environments at high latitudes.


Assuntos
Alnus/microbiologia , Frankia/classificação , Fixação de Nitrogênio , Filogenia , Nódulos Radiculares de Plantas/microbiologia , Técnicas de Tipagem Bacteriana , Magnoliopsida/microbiologia , Simbiose
17.
PLoS One ; 15(9): e0239677, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32986754

RESUMO

A geographically isolated maize landrace cultivated on nitrogen-depleted fields without synthetic fertilizer in the Sierra Mixe region of Oaxaca, Mexico utilizes nitrogen derived from the atmosphere and develops an extensive network of mucilage-secreting aerial roots that harbors a diazotrophic (N2-fixing) microbiota. Targeting these diazotrophs, we selected nearly 600 microbes of a collection obtained from mucilage and confirmed their ability to incorporate heavy nitrogen (15N2) metabolites in vitro. Sequencing their genomes and conducting comparative bioinformatic analyses showed that these genomes had substantial phylogenetic diversity. We examined each diazotroph genome for the presence of nif genes essential to nitrogen fixation (nifHDKENB) and carbohydrate utilization genes relevant to the mucilage polysaccharide digestion. These analyses identified diazotrophs that possessed the canonical nif gene operons, as well as many other operon configurations with concomitant fixation and release of >700 different 15N labeled metabolites. We further demonstrated that many diazotrophs possessed alternative nif gene operons and confirmed their genomic potential to derive chemical energy from mucilage polysaccharide to fuel nitrogen fixation. These results confirm that some diazotrophic bacteria associated with Sierra Mixe maize were capable of incorporating atmospheric nitrogen into their small molecule extracellular metabolites through multiple nif gene configurations while others were able to fix nitrogen without the canonical (nifHDKENB) genes.


Assuntos
Microbiota/genética , Fixação de Nitrogênio , Mucilagem Vegetal/metabolismo , Raízes de Plantas/microbiologia , Zea mays/microbiologia , Bactérias/genética , Bactérias/metabolismo , Genoma Bacteriano , México , Nitrogênio/metabolismo , Óperon , Filogenia , Raízes de Plantas/metabolismo , Sequenciamento Completo do Genoma
18.
PLoS One ; 15(9): e0239081, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32925972

RESUMO

Sierra Mixe maize is a geographically remote landrace variety grown on nitrogen-deficient fields in Oaxaca, Mexico that meets its nutritional requirements without synthetic fertilizer by associating with free-living diazotrophs comprising the microbiota of its aerial root mucilage. We selected nearly 500 diazotrophic (N2-fixing) bacteria isolated from Sierra Mixe maize mucilage and sequenced their genomes. Comparative genomic analysis demonstrated that isolates represented diverse genera and composed three major diazotrophic groups based on nitrogen fixation gene content. In addition to nitrogen fixation, we examined deamination of 1-amino-1-cyclopropanecarboxylic acid, biosynthesis of indole-3-acetic acid, and phosphate solubilization as alternative mechanisms of direct plant growth promotion (PGP). Genome mining showed that isolates of all diazotrophic groups possessed marker genes for multiple mechanisms of direct plant growth promotion (PGP). Implementing in vitro assays corroborated isolate genotypes by measuring each isolate's potential to confer the targeted PGP traits and revealed phenotypic variation among isolates based on diazotrophic group assignment. Investigating the ability of mucilage diazotrophs to confer PGP by direct inoculation of clonally propagated potato plants in planta led to the identification of 16 bio-stimulant candidates. Conducting nitrogen-stress greenhouse experiments demonstrated that potato inoculation with a synthetic community of bio-stimulant candidates, as well as with its individual components, resulted in PGP phenotypes. We further demonstrated that one diazotrophic isolate conferred PGP to a conventional maize variety under nitrogen-stress in the greenhouse. These results indicate that, while many diazotrophic isolates from Sierra Mixe maize possessed genotypes and in vitro phenotypes for targeted PGP traits, a subset of these organisms promoted the growth of potato and conventional maize, potentially through the use of multiple promotion mechanisms.


Assuntos
Produtos Agrícolas/crescimento & desenvolvimento , Produtos Agrícolas/microbiologia , Fixação de Nitrogênio , Zea mays/crescimento & desenvolvimento , Zea mays/microbiologia , Bactérias/genética , Bactérias/isolamento & purificação , Fenômenos Fisiológicos Bacterianos , Ácidos Indolacéticos/metabolismo , Fosfatos/metabolismo , Solanum tuberosum/crescimento & desenvolvimento , Solanum tuberosum/microbiologia
19.
Proc Natl Acad Sci U S A ; 117(38): 23823-23834, 2020 09 22.
Artigo em Inglês | MEDLINE | ID: mdl-32900931

RESUMO

By analyzing successive lifestyle stages of a model Rhizobium-legume symbiosis using mariner-based transposon insertion sequencing (INSeq), we have defined the genes required for rhizosphere growth, root colonization, bacterial infection, N2-fixing bacteroids, and release from legume (pea) nodules. While only 27 genes are annotated as nif and fix in Rhizobium leguminosarum, we show 603 genetic regions (593 genes, 5 transfer RNAs, and 5 RNA features) are required for the competitive ability to nodulate pea and fix N2 Of these, 146 are common to rhizosphere growth through to bacteroids. This large number of genes, defined as rhizosphere-progressive, highlights how critical successful competition in the rhizosphere is to subsequent infection and nodulation. As expected, there is also a large group (211) specific for nodule bacteria and bacteroid function. Nodule infection and bacteroid formation require genes for motility, cell envelope restructuring, nodulation signaling, N2 fixation, and metabolic adaptation. Metabolic adaptation includes urea, erythritol and aldehyde metabolism, glycogen synthesis, dicarboxylate metabolism, and glutamine synthesis (GlnII). There are 17 separate lifestyle adaptations specific to rhizosphere growth and 23 to root colonization, distinct from infection and nodule formation. These results dramatically highlight the importance of competition at multiple stages of a Rhizobium-legume symbiosis.


Assuntos
Rhizobium leguminosarum , Rizosfera , Simbiose/genética , Fabaceae/microbiologia , Genes Bacterianos/genética , Fixação de Nitrogênio/genética , Rhizobium leguminosarum/genética , Rhizobium leguminosarum/fisiologia , Nódulos Radiculares de Plantas/genética , Nódulos Radiculares de Plantas/microbiologia
20.
PLoS One ; 15(9): e0238004, 2020.
Artigo em Inglês | MEDLINE | ID: mdl-32877417

RESUMO

Fire severity affects both ecosystem N-loss and post-fire N-balance. Climate change is altering the fire regime of interior Alaska, although the effects on Siberian alder (Alnus viridis ssp. fruticosa) annual N-fixation input (kg N ha-1 yr-1) and ecosystem N-balance are largely unknown. We established 263 study plots across two burn scars within the Yukon-Tanana Uplands ecoregion of interior Alaska. Siberian alder N-input was quantified by post-fire age, fire severity, and stand type. We modeled the components of Siberian alder N-input using environmental variables and fire severity within and across burn scars and estimated post-fire N-balance using N-loss (volatilized N) and N-gain [biological N-fixation and atmospheric deposition]. Mean nodule-level N-fixation rate was 70% higher 11-years post-fire (12.88 ± 1.18 µmol N g-1 hr-1) than 40-years post-fire (7.58 ± 0.59 µmol N g-1 hr-1). Structural equation modeling indicated that fire severity had a negative effect on Siberian alder density, but a positive effect on live nodule biomass (g nodule m-2 plant-1). Post-fire Siberian alder N-input was highest in 11-year old moderately burned deciduous stands (11.53 ± 0.22 kg N ha-1 yr-1), and lowest in 11-year old stands that converted from black spruce to deciduous dominance after severe fire (0.06 ± 0.003 kg N ha-1 yr-1). Over a 138-year fire return interval, N-gains in converted black spruce stands are estimated to offset 15% of volatilized N, whereas N-gains in burned deciduous stands likely exceed volatilized N by an order of magnitude. High Siberian alder density and nodule biomass drives N-input in burned deciduous stands, while low N-fixer density (including Siberian alder) limits N-input in high severity black spruce stands not underlain by permafrost. A severe fire regime that converts black spruce stands to deciduous dominance without alder recruitment may induce progressive N-losses which alter boreal forest ecosystem patterns and processes.


Assuntos
Alnus/crescimento & desenvolvimento , Ecossistema , Fogo , Fixação de Nitrogênio , Nitrogênio/análise , Nitrogênio/metabolismo , Árvores/crescimento & desenvolvimento , Alaska , Alnus/metabolismo , Taiga , Árvores/metabolismo
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