RESUMEN

Utilizing coastal land for agriculture presents challenges such as low water content, high soil salinity, and low organic compound content. To support plant growth under these conditions, biofertilizers composed of plant growth promoting Rhizobacteria (PGPR), especially those inhabiting coastal areas, are needed. The Parangkusumo sand dunes on the southern coast of Java, Indonesia, is a unique coastal ecosystem characterized by arid conditions, high temperatures, and high soil salinity. To date, no studies have reported the isolation of PGPR from this ecosystem. This study is the first to isolate and identify PGPR associated with Spinifex littoreus, a dominant plant species in the Parangkusumo sand dunes, which are adapted to the harsh condition of Parangkusumo sand dunes. Ten rhizobacterial isolates were obtained, with five identified as members of the Bacillaceae family. All isolates demonstrated phosphate solubilization activity, while seven exhibited cellulolytic activity. One isolate, Priestia aryabhattai strain 2, notably showed phosphate solubilization and nitrogen fixation activities. The findings of this PGPR activity screening offer valuable insights for developing biofertilizers tailored for coastal agricultural applications.

Asunto(s)

Microbiología del Suelo , Indonesia , Arena/microbiología , Fijación del Nitrógeno

RESUMEN

Peanut production could be increased through plant growth-promoting rhizobacteria (PGPR). In this regard, the present field research aimed at elucidating the impact of PGPR on peanut yield, soil enzyme activity, microbial diversity, and structure. Three PGPR strains (Bacillus velezensis, RI3; Bacillus velezensis, SC6; Pseudomonas psychrophila, P10) were evaluated, along with Bradyrhizobium japonicum (BJ), taken as a control. PGPR increased seed yield by 8%, improving the radiation use efficiency (4-14%). PGPR modified soil enzymes (fluorescein diacetate activity by 17% and dehydrogenase activity by 28%) and microbial abundance (12%). However, PGPR did not significantly alter microbial diversity; nonetheless, it modified the relative abundance of key phyla (Actinobacteria > Proteobacteria > Firmicutes) and genera (Bacillus > Arthrobacter > Pseudomonas). PGPRs modified the relative abundance of genes associated with N-fixation and nitrification while increasing genes related to N-assimilation and N-availability. PGPR improved agronomic traits without altering rhizosphere diversity.

Asunto(s)

Arachis , Bacillus , Bradyrhizobium , Metagenómica , Pseudomonas , Rizosfera , Microbiología del Suelo , Suelo , Arachis/microbiología , Arachis/crecimiento & desarrollo , Arachis/metabolismo , Arachis/genética , Bacillus/genética , Bacillus/metabolismo , Bradyrhizobium/genética , Bradyrhizobium/metabolismo , Bradyrhizobium/crecimiento & desarrollo , Bradyrhizobium/fisiología , Pseudomonas/genética , Pseudomonas/fisiología , Pseudomonas/crecimiento & desarrollo , Suelo/química , Producción de Cultivos/métodos , Bacterias/genética , Bacterias/clasificación , Bacterias/metabolismo , Bacterias/enzimología , Bacterias/aislamiento & purificación , Biodiversidad , Fijación del Nitrógeno , Raíces de Plantas/microbiología , Raíces de Plantas/crecimiento & desarrollo , Raíces de Plantas/metabolismo

RESUMEN

We present new genomes from the bacterial symbiont Candidatus Dactylopiibacterium carminicum obtained from non-domesticated carmine cochineals belonging to the scale insect Dactylopius (Hemiptera: Coccoidea: Dactylopiidae). As Dactylopiibacterium has not yet been cultured in the laboratory, metagenomes and metatranscriptomics have been key in revealing putative symbiont functions. Dactylopiibacterium is a nitrogen-fixing beta-proteobacterium that may be vertically transmitted and shows differential gene expression inside the cochineal depending on the tissue colonized. Here we found that all cochineal species tested had Dactylopiibacterium carminicum which has a highly conserved genome. All Dactylopiibacterium genomes analyzed had genes involved in nitrogen fixation and plant polymer degradation. Dactylopiibacterium genomes resemble those from free-living plant bacteria, some found as endophytes. Notably, we found here a new putative novel function where the bacteria may protect the insect from viruses, since all Dactylopiibacterium genomes contain CRISPRs with a spacer matching nucleopolyhedrovirus that affects insects.

Asunto(s)

Sistemas CRISPR-Cas , Genoma Bacteriano , Hemípteros , Simbiosis , Hemípteros/microbiología , Hemípteros/virología , Animales , Genoma Bacteriano/genética , Genómica , Filogenia , Fijación del Nitrógeno

RESUMEN

Brazil stands out in research, industrial development, and farmers' use of microbial inoculants, with an emphasis on getting benefits from the biological nitrogen fixation process with the soybean crop. Nowadays, about 140 million doses of inoculants are commercialized annually for the soybean in the country, and strain identification is achieved by rep-PCR, an effective but time-consuming method. Aiming to develop an easy, low-cost, and low-time-consuming method, we used a complete genome-based approach based on the unequivocal identification of unique genes present in the genomes of each of the four Bradyrhizobium strains used in commercial inoculants: Bradyrhizobium elkanii strains SEMIA 587 and SEMIA 5019, Bradyrhizobium japonicum SEMIA 5079, and Bradyrhizobium diazoefficiens SEMIA 5080. The unique pairs of primers able to amplify genomic regions of different sizes allowed the identification of the four strains in a simple multiplex polymerase chain reaction (PCR). Validation was confirmed by using single colonies, multiple cultures, and commercial inoculants. The number of labor hours of a technician was 3.08 times higher, and the final cost was 3.25 times higher in the rep-PCR than in the multiplex PCR. Most importantly, the results for multiplex PCR were obtained on the same day, in contrast with 15 days in the traditional methodology. The genomic approach developed can be easily applied to a variety of microbial inoculants worldwide, in addition to studies of ecology and evaluation of the competitiveness of the strains.

Asunto(s)

Bradyrhizobium , Glycine max , Reacción en Cadena de la Polimerasa Multiplex , Bradyrhizobium/genética , Bradyrhizobium/clasificación , Bradyrhizobium/aislamiento & purificación , Glycine max/microbiología , Reacción en Cadena de la Polimerasa Multiplex/métodos , Genoma Bacteriano , Inoculantes Agrícolas/genética , Inoculantes Agrícolas/clasificación , Genómica/métodos , Brasil , ADN Bacteriano/genética , Fijación del Nitrógeno

RESUMEN

Non-symbiotic N2-fixation would greatly increase the versatility of N-biofertilizers for sustainable agriculture. Genetic modification of diazotrophic bacteria has successfully enhanced NH4+ release. In this study, we compared the competitive fitness of A. vinelandii mutant strains, which allowed us to analyze the burden of NH4+ release under a broad dynamic range. Long-term competition assays under regular culture conditions confirmed a large burden for NH4+ release, exclusion by the wt strain, phenotypic instability, and loss of the ability to release NH4+. In contrast, co-inoculation in mild autoclaved soil showed a much longer co-existence with the wt strain and a stable NH4+ release phenotype. All genetically modified strains increased the N content and changed its chemical speciation in the soil. This study contributes one step forward towards bridging a knowledge gap between molecular biology laboratory research and the incorporation of N from the air into the soil in a molecular species suitable for plant nutrition, a crucial requirement for developing improved bacterial inoculants for economic and environmentally sustainable agriculture. KEY POINTS: • Genetic engineering for NH4+ excretion imposes a fitness burden on the culture medium • Large phenotypic instability for NH4+-excreting bacteria in culture medium • Lower fitness burden and phenotypic instability for NH4+-excreting bacteria in soil.

Asunto(s)

Compuestos de Amonio , Azotobacter vinelandii , Microbiología del Suelo , Azotobacter vinelandii/genética , Azotobacter vinelandii/metabolismo , Compuestos de Amonio/metabolismo , Fijación del Nitrógeno , Nitrógeno/metabolismo , Aptitud Genética , Fenotipo , Suelo/química , Medios de Cultivo/química , Ingeniería Genética

RESUMEN

AIM: Bacteria that promote plant growth, such as diazotrophs, are valuable tools for achieving a more sustainable production of important non-legume crops like rice. Different strategies have been used to discover new bacteria capable of promoting plant growth. This work evaluated the contribution of soil diazotrophs to the endophytic communities established in the roots of rice seedlings cultivated on seven representative soils from Uruguay. METHODS AND RESULTS: The soils were classified into two groups according to the C and clay content. qPCR, terminal restriction fragment length polymorphism (T-RFLP), and 454-pyrosequencing of the nifH gene were used for analyzing diazotrophs in soil and plantlets' roots grown from seeds of the same genotype for 25 days under controlled conditions. A similar nifH abundance was found among the seven soils, roots, or leaves. The distribution of diazotrophs was more uneven in roots than in soils, with dominance indices significantly higher than in soils (nifH T-RFLP). Dominant soils' diazotrophs were mainly affiliated to Alphaproteobacteria and Planctomycetota. Conversely, Alpha, Beta, Gammaproteobacteria, and Bacillota were predominant in different roots, though undetectable in soils. Almost no nifH sequences were shared between soils and roots. CONCLUSIONS: Root endophytic diazotrophs comprised a broader taxonomic range of microorganisms than diazotrophs found in soils from which the plantlets were grown and showed strong colonization patterns.

Asunto(s)

Endófitos , Oryza , Raíces de Plantas , Microbiología del Suelo , Suelo , Oryza/microbiología , Oryza/crecimiento & desarrollo , Raíces de Plantas/microbiología , Endófitos/genética , Endófitos/aislamiento & purificación , Endófitos/clasificación , Suelo/química , Polimorfismo de Longitud del Fragmento de Restricción , Bacterias/genética , Bacterias/aislamiento & purificación , Bacterias/clasificación , Fijación del Nitrógeno , Oxidorreductasas/genética

RESUMEN

Bradyrhizobium is known for fixing atmospheric nitrogen in symbiosis with agronomically important crops. This study focused on two groups of strains, each containing eight natural variants of the parental strains, Bradyrhizobium japonicum SEMIA 586 (=CNPSo 17) or Bradyrhizobium diazoefficiens SEMIA 566 (=CNPSo 10). CNPSo 17 and CNPSo 10 were used as commercial inoculants for soybean crops in Brazil at the beginning of the crop expansion in the southern region in the 1960s-1970s. Variants derived from these parental strains were obtained in the late 1980s through a strain selection program aimed at identifying elite strains adapted to a new cropping frontier in the central-western Cerrado region, with a higher capacity of biological nitrogen fixation (BNF) and competitiveness. Here, we aimed to detect genetic variations possibly related to BNF, competitiveness for nodule occupancy, and adaptation to the stressful conditions of the Brazilian Cerrado soils. High-quality genome assemblies were produced for all strains. The core genome phylogeny revealed that strains of each group are closely related, as confirmed by high average nucleotide identity values. However, variants accumulated divergences resulting from horizontal gene transfer, genomic rearrangements, and nucleotide polymorphisms. The B. japonicum group presented a larger pangenome and a higher number of nucleotide polymorphisms than the B. diazoefficiens group, possibly due to its longer adaptation time to the Cerrado soil. Interestingly, five strains of the B. japonicum group carry two plasmids. The genetic variability found in both groups is discussed considering the observed differences in their BNF capacity, competitiveness for nodule occupancy, and environmental adaptation.IMPORTANCEToday, Brazil is a global leader in the study and use of biological nitrogen fixation with soybean crops. As Brazilian soils are naturally void of soybean-compatible bradyrhizobia, strain selection programs were established, starting with foreign isolates. Selection searched for adaptation to the local edaphoclimatic conditions, higher efficiency of nitrogen fixation, and strong competitiveness for nodule occupancy. We analyzed the genomes of two parental strains of Bradyrhizobium japonicum and Bradyrhizobium diazoefficiens and eight variant strains derived from each parental strain. We detected two plasmids in five strains and several genetic differences that might be related to adaptation to the stressful conditions of the soils of the Brazilian Cerrado biome. We also detected genetic variations in specific regions that may impact symbiotic nitrogen fixation. Our analysis contributes to new insights into the evolution of Bradyrhizobium, and some of the identified differences may be applied as genetic markers to assist strain selection programs.

Asunto(s)

Bradyrhizobium , Genoma Bacteriano , Glycine max , Fijación del Nitrógeno , Filogenia , Simbiosis , Bradyrhizobium/genética , Bradyrhizobium/clasificación , Bradyrhizobium/aislamiento & purificación , Fijación del Nitrógeno/genética , Brasil , Glycine max/microbiología , Simbiosis/genética , Variación Genética , Adaptación Fisiológica/genética , Nódulos de las Raíces de las Plantas/microbiología , Microbiología del Suelo , Genómica

RESUMEN

Tropical montane cloud forests are high altitude ecosystems characterized by very high ambient humidity, which favors organisms that depend on the environment for their water status, such as bryophytes and their nitrogen-fixing symbionts. Bryophyte-associated N2 fixation is a major source of new N in several northern environments, but their contributions to the N cycle in other ecosystems is still poorly understood. In this work, we evaluated N2 fixation rates associated with epiphytic bryophytes growing along the stems of pumpwood trees (Cecropia sp.) as well as in surrounding litter and soil from a primary and a secondary cloud forests in the Talamanca Mountain Range, Costa Rica. Nitrogen fixation was significantly higher in substrates from the secondary forest compared to those from the primary forest. Overall, N2 fixation rates associated with epiphytic bryophytes were 57 times those of litter and 270 times what was measured in soil. Further, light intensity was the major factor influencing N2 fixation rates in all substrates. Increased access to light in disturbed cloud forests may therefore favor bryophyte-associated N2 fixation, potentially contributing to the recovery of these ecosystems.

Asunto(s)

Bosques , Fijación del Nitrógeno , Costa Rica , Briófitas , Clima Tropical , Suelo/química , Luz , Monitoreo del Ambiente , Ecosistema

RESUMEN

Fertilization with nickel (Ni) can positively affect plant development due to the role of this micronutrient in nitrogen (N) metabolism, namely, through urease and NiFe-hydrogenase. Although the application of Ni is an emerging practice in modern agriculture, its effectiveness strongly depends on the chosen application method, making further research in this area essential. The individual and combined effects of different Ni application methods-seed treatment, leaf spraying and/or soil fertilization-were investigated in soybean plants under different edaphoclimatic conditions (field and greenhouse). Beneficial effects of the Soil, Soil + Leaf and Seed + Leaf treatments were observed, with gains of 7 to 20% in biological nitrogen fixation, 1.5-fold in ureides, 14% in shoot dry weight and yield increases of up to 1161 kg ha-1. All the Ni application methods resulted in a 1.1-fold increase in the SPAD index, a 1.2-fold increase in photosynthesis, a 1.4-fold increase in nitrogenase, and a 3.9-fold increase in urease activity. Edaphoclimatic conditions exerted a significant influence on the treatments. The integrated approaches, namely, leaf application in conjunction with soil or seed fertilization, were more effective for enhancing yield in soybean cultivation systems. The determination of the ideal method is crucial for ensuring optimal absorption and utilization of this micronutrient and thus a feasible and sustainable management technology. Further research is warranted to establish official guidelines for the application of Ni in agricultural practices.

Asunto(s)

Fertilizantes , Glycine max , Níquel , Suelo , Glycine max/crecimiento & desarrollo , Glycine max/efectos de los fármacos , Glycine max/metabolismo , Fertilizantes/análisis , Suelo/química , Ureasa/metabolismo , Hojas de la Planta/crecimiento & desarrollo , Hojas de la Planta/metabolismo , Hojas de la Planta/efectos de los fármacos , Fijación del Nitrógeno/efectos de los fármacos , Nitrógeno/metabolismo , Fotosíntesis/efectos de los fármacos , Semillas/crecimiento & desarrollo , Semillas/efectos de los fármacos , Semillas/metabolismo , Agricultura/métodos

RESUMEN

The actinobacterium Arthrobacter sp. UMCV2 promotes plant growth through the emission of N,N-dimethylhexadecilamine (DMHDA). The Medicago-Sinorhizobium nodulation has been employed to study symbiotic nitrogen fixation by rhizobia in nodulating Fabaceae. Herein, we isolated three Sinorhizobium medicae strains that were used to induce nodules in Medicago truncatula. The co-inoculation of M. truncatula with Arthrobacter sp. strain UMCV2 produced a higher number of effective nodules than inoculation with only Sinorhizobium strains. Similarly, the exposure of inoculated M. truncatula to DMHDA produced a greater number of effective nodules compared to non-exposed plants. Thus, we conclude that Arthrobacter sp. UMCV2 promotes nodulation, and propose that this effect is produced, at least partly, via DMHDA emission.

Asunto(s)

Arthrobacter , Medicago truncatula , Medicago truncatula/microbiología , Arthrobacter/efectos de los fármacos , Arthrobacter/fisiología , Sinorhizobium/fisiología , Sinorhizobium/efectos de los fármacos , Nodulación de la Raíz de la Planta/efectos de los fármacos , Simbiosis , Fijación del Nitrógeno/efectos de los fármacos

RESUMEN

Rhizobia are bacteria that form nitrogen-fixing nodules in legume plants. The sets of genes responsible for both nodulation and nitrogen fixation are carried in plasmids or genomic islands that are often mobile. Different strains within a species sometimes have different host specificities, while very similar symbiosis genes may be found in strains of different species. These specificity variants are known as symbiovars, and many of them have been given names, but there are no established guidelines for defining or naming them. Here, we discuss the requirements for guidelines to describe symbiovars, propose a set of guidelines, provide a list of all symbiovars for which descriptions have been published so far, and offer a mechanism to maintain a list in the future.

Asunto(s)

Rhizobium , Simbiosis , Fabaceae/microbiología , Guías como Asunto , Fijación del Nitrógeno , Rhizobium/genética , Rhizobium/clasificación , Nódulos de las Raíces de las Plantas/microbiología

RESUMEN

Bacterial symbionts, with their shorter generation times and capacity for horizontal gene transfer (HGT), play a critical role in allowing marine organisms to cope with environmental change. The closure of the Isthmus of Panama created distinct environmental conditions in the Tropical Eastern Pacific (TEP) and Caribbean, offering a "natural experiment" for studying how closely related animals evolve and adapt under environmental change. However, the role of bacterial symbionts in this process is often overlooked. We sequenced the genomes of endosymbiotic bacteria in two sets of sister species of chemosymbiotic bivalves from the genera Codakia and Ctena (family Lucinidae) collected on either side of the Isthmus, to investigate how differing environmental conditions have influenced the selection of symbionts and their metabolic capabilities. The lucinid sister species hosted different Candidatus Thiodiazotropha symbionts and only those from the Caribbean had the genetic potential for nitrogen fixation, while those from the TEP did not. Interestingly, this nitrogen-fixing ability did not correspond to symbiont phylogeny, suggesting convergent evolution of nitrogen fixation potential under nutrient-poor conditions. Reconstructing the evolutionary history of the nifHDKT operon by including other lucinid symbiont genomes from around the world further revealed that the last common ancestor (LCA) of Ca. Thiodiazotropha lacked nif genes, and populations in oligotrophic habitats later re-acquired the nif operon through HGT from the Sedimenticola symbiont lineage. Our study suggests that HGT of the nif operon has facilitated niche diversification of the globally distributed Ca. Thiodiazotropha endolucinida species clade. It highlights the importance of nitrogen availability in driving the ecological diversification of chemosynthetic symbiont species and the role that bacterial symbionts may play in the adaptation of marine organisms to changing environmental conditions.

Asunto(s)

Bivalvos , Transferencia de Gen Horizontal , Fijación del Nitrógeno , Nitrógeno , Filogenia , Simbiosis , Simbiosis/genética , Animales , Fijación del Nitrógeno/genética , Nitrógeno/metabolismo , Bivalvos/microbiología , Bivalvos/genética , Bacterias/genética , Bacterias/clasificación , Bacterias/metabolismo , Adaptación Fisiológica/genética , Genoma Bacteriano , Región del Caribe , Panamá

RESUMEN

The flooding pampa is one of the most important cattle-raising regions in Argentina. In this region, natural pastures are dominated by low-productivity native grass species, which are the main feed for livestock. In this context, previous studies in the region with the subtropical exotic grass Panicum coloratum highlight it as a promising species to improve pasture productivity. Cultivable phosphate solubilizing bacteria (PSB) communities associated to native (Sporobolus indicus) and exotic (Panicum coloratum) forage grasses adapted to alkaline-sodic soils of the flooding pampa were analyzed. PSB represented 2-14% of cultivable rhizobacteria and Box-PCR fingerprinting revealed a high genetic diversity in both rhizospheres. Taxonomic identification by MALDI-TOF showed that PSB populations of P. coloratum and S. indicus rhizospheres are dominated by the phylum Proteobacteria (92,51% and 96,60% respectively) and to a lesser extent (< 10%), by the phyla Actinobacteria and Firmicutes. At the genus level, both PSB populations were dominated by Enterobacter and Pseudomonas. Siderophore production, nitrogen fixation, and indoleacetic acid production were detected in a variety of PSB genera of both plant species. A higher proportion of siderophore and IAA producers were associated to P. coloratum than S. indicus, probably reflecting a greater dependence of the exotic species on rhizospheric microorganisms to satisfy its nutritional requirements in the soils of the flooding pampa. This work provides a novel knowledge about functional groups of bacteria associated to plants given that there are no previous reports dedicated to the characterization of PSB rhizosphere communities of S indicus and P coloratum. Finally, it should be noted that the collection obtained in this study can be useful for the development of bioinputs that allow reducing the use of chemical fertilizers, providing sustainability to pasture production systems for livestock.

Asunto(s)

Bacterias , Fosfatos , Poaceae , Rizosfera , Microbiología del Suelo , Suelo , Poaceae/microbiología , Bacterias/clasificación , Bacterias/genética , Bacterias/metabolismo , Bacterias/aislamiento & purificación , Suelo/química , Fosfatos/metabolismo , Argentina , Animales , Filogenia , Sideróforos/metabolismo , Fijación del Nitrógeno , Ácidos Indolacéticos/metabolismo , Inundaciones , ARN Ribosómico 16S/genética

RESUMEN

All non-Mimosoid nodulated genera in the legume subfamily Caesalpinioideae confine their rhizobial symbionts within cell wall-bound 'fixation threads' (FTs). The exception is the large genus Chamaecrista in which shrubs and subshrubs house their rhizobial bacteroids more intimately within symbiosomes, whereas large trees have FTs. This study aimed to unravel the evolutionary relationships between Chamaecrista growth habit, habitat, nodule bacteroid type, and rhizobial genotype. The growth habit, bacteroid anatomy, and rhizobial symbionts of 30 nodulated Chamaecrista species native to different biomes in the Brazilian state of Bahia, a major centre of diversity for the genus, was plotted onto an ITS-trnL-F-derived phylogeny of Chamaecrista. The bacteroids from most of the Chamaecrista species examined were enclosed in symbiosomes (SYM-type nodules), but those in arborescent species in the section Apoucouita, at the base of the genus, were enclosed in cell wall material containing homogalacturonan (HG) and cellulose (FT-type nodules). Most symbionts were Bradyrhizobium genotypes grouped according to the growth habits of their hosts, but the tree, C. eitenorum, was nodulated by Paraburkholderia. Chamaecrista has a range of growth habits that allow it to occupy several different biomes and to co-evolve with a wide range of (mainly) bradyrhizobial symbionts. FTs represent a less intimate symbiosis linked with nodulation losses, so the evolution of SYM-type nodules by most Chamaecrista species may have (i) aided the genus-wide retention of nodulation, and (ii) assisted in its rapid speciation and radiation out of the rainforest into more diverse and challenging habitats.

Asunto(s)

Chamaecrista , Filogenia , Bosque Lluvioso , Simbiosis , Chamaecrista/fisiología , Chamaecrista/genética , Chamaecrista/crecimiento & desarrollo , Brasil , Ecosistema , Rhizobium/fisiología , Nodulación de la Raíz de la Planta/fisiología , Evolución Biológica , Fijación del Nitrógeno

RESUMEN

Rhizobial phosphatidylcholine (PC) is thought to be a critical phospholipid for the symbiotic relationship between rhizobia and legume host plants. A PC-deficient mutant of Sinorhizobium meliloti overproduces succinoglycan, is unable to swim, and lacks the ability to form nodules on alfalfa (Medicago sativa) host roots. Suppressor mutants had been obtained which did not overproduce succinoglycan and regained the ability to swim. Previously, we showed that point mutations leading to altered ExoS proteins can reverse the succinoglycan and swimming phenotypes of a PC-deficient mutant. Here, we report that other point mutations leading to altered ExoS, ChvI, FabA, or RpoH1 proteins also revert the succinoglycan and swimming phenotypes of PC-deficient mutants. Notably, the suppressor mutants also restore the ability to form nodule organs on alfalfa roots. However, nodules generated by these suppressor mutants express only low levels of an early nodulin, do not induce leghemoglobin transcript accumulation, thus remain white, and are unable to fix nitrogen. Among these suppressor mutants, we detected a reduced function mutant of the 3-hydoxydecanoyl-acyl carrier protein dehydratase FabA that produces reduced amounts of unsaturated and increased amounts of shorter chain fatty acids. This alteration of fatty acid composition probably affects lipid packing thereby partially compensating for the previous loss of PC and contributing to the restoration of membrane homeostasis.

Asunto(s)

Ácidos Grasos , Medicago sativa , Fosfatidilcolinas , Nodulación de la Raíz de la Planta , Sinorhizobium meliloti , Simbiosis , Sinorhizobium meliloti/fisiología , Sinorhizobium meliloti/genética , Medicago sativa/microbiología , Medicago sativa/genética , Nodulación de la Raíz de la Planta/genética , Ácidos Grasos/metabolismo , Ácidos Grasos/biosíntesis , Fosfatidilcolinas/metabolismo , Fosfatidilcolinas/biosíntesis , Proteínas Bacterianas/genética , Proteínas Bacterianas/metabolismo , Nódulos de las Raíces de las Plantas/microbiología , Nódulos de las Raíces de las Plantas/genética , Nódulos de las Raíces de las Plantas/metabolismo , Mutación , Polisacáridos Bacterianos/metabolismo , Polisacáridos Bacterianos/biosíntesis , Fijación del Nitrógeno

RESUMEN

The unicellular cyanobacterium Candidatus Atelocyanobacterium thalassa (UCYN-A) is a key diazotroph in the global ocean owing to its high N2 fixation rates and wide distribution in marine environments. Nevertheless, little is known about UCYN-A in oxygen-deficient zones (ODZs), which may be optimal environments for marine diazotrophy. Therefore, the distribution and diversity of UCYN-A were studied in two consecutive years under contrasting phases (La Niña vs. El Niño) of El Niño Southern Oscillation (ENSO) along a transect in the ODZ of the Mexican Pacific upwelling system. Of the three UCYN-A sublineages found, UCYN-A1 and UCYN-A3 were barely detected, whereas UCYN-A2 was dominant in all the stations and showed a wide distribution in both ENSO phases. The presence of UCYN-A was associated with well-oxygenated waters, but it was also found for the first time under suboxic conditions (<20 µM) at the bottom of a shallow coastal station, within the oxygen-poor and nutrient-rich Subsurface Subtropical water mass. This study contributes to the understanding of UCYN-A distribution under different oceanographic conditions associated with ENSO phases in upwelling systems, especially because of the current climate change and increasing deoxygenation in many areas of the world's oceans.

Asunto(s)

Cianobacterias , El Niño Oscilación del Sur , Cianobacterias/genética , Fijación del Nitrógeno , Océanos y Mares , Oxígeno

Asunto(s)

Medicago truncatula , Rhizobium , Sinorhizobium meliloti , Rhizobium/fisiología , Nodulación de la Raíz de la Planta , Proteínas de Plantas/metabolismo , Medicago truncatula/metabolismo , Simbiosis , Nódulos de las Raíces de las Plantas/metabolismo , Regulación de la Expresión Génica de las Plantas , Sinorhizobium meliloti/fisiología , Fijación del Nitrógeno

Asunto(s)

Phaseolus , Rhizobium , Phaseolus/genética , Simbiosis , Ciclinas/metabolismo , Factor de Unión a CCAAT/metabolismo , Nodulación de la Raíz de la Planta , Fijación del Nitrógeno , Nódulos de las Raíces de las Plantas/metabolismo , Proteínas de Plantas/genética , Proteínas de Plantas/metabolismo

RESUMEN

Paenibacillus sonchi SBR5T is a Gram-positive, endospore-forming facultative aerobic diazotrophic bacterium that can fix nitrogen via an alternative Fe-only nitrogenase (AnfHDGK). In several bacteria, this alternative system is expressed under molybdenum (Mo)-limiting conditions when the conventional Mo-dependent nitrogenase (NifHDK) production is impaired. The regulatory mechanisms, metabolic processes, and cellular functions of N2 fixation by alternative and/or conventional systems are poorly understood in the Paenibacillus genus. We conducted a comparative proteomic profiling study of P. sonchi SBR5T grown under N2-fixing conditions with and without Mo supply through an LC-MS/MS and label-free quantification analysis to address this gap. Protein abundances revealed overrepresented processes related to anaerobiosis growth adaption, Fe-S cluster biosynthesis, ammonia assimilation, electron transfer, and sporulation under N2-fixing conditions compared to non-fixing control. Under Mo limitation, the Fe-only nitrogenase components were overrepresented together with the Mo-transporter system, while the dinitrogenase component (NifDK) of Mo­nitrogenase was underrepresented. The dinitrogenase reductase component (NifH) and accessory proteins encoded by the nif operon had no significant differential expression, suggesting post-transcriptional regulation of nif gene products in this strain. Overall, this was the first comprehensive proteomic analysis of a diazotrophic strain from the Paenibacillaceae family, and it provided insights related to alternative N2-fixation by Fe-only nitrogenase. SIGNIFICANCE: In this work, we try to understand how the alternative nitrogen fixation system, presented by some diazotrophic bacteria, works. For this, we used the SBR5 lineage of P. sonchi, which presents the alternative system in which the nitrogenase cofactor is composed only of iron. In addition, we tried to unravel the proteome of this strain in different situations of nitrogen fixation, since, for Gram-positive bacteria, these systems are little known. The results achieved, through LC-MS/MS and label-free quantitative analysis, showed an overrepresentation of proteins related to different processes involved with growth under stressful conditions in situations of nitrogen deficiency, in addition to suggesting that some encoded proteins by the nif operon may be regulated at post-transcriptional levels. Our findings represent important steps toward the elucidation of nitrogen fixation systems in Gram-positive diazotrophic bacteria.

Asunto(s)

Fijación del Nitrógeno , Paenibacillus , Proteoma/metabolismo , Proteómica , Cromatografía Liquida , Espectrometría de Masas en Tándem , Nitrogenasa/metabolismo , Paenibacillus/genética , Paenibacillus/metabolismo , Molibdeno/metabolismo , Hierro/metabolismo , Nitrógeno/metabolismo

RESUMEN

Studies on the role of nickel (Ni) in photosynthetic and antioxidant metabolism, as well as in flavonoid synthesis and biological fixation nitrogen in cowpea crop are scarce. The aim of this study was to elucidate the role of Ni in metabolism, photosynthesis and nodulation of cowpea plants. A completely randomized experiment was performed in greenhouse, with cowpea plants cultivated under 0, 0.5, 1, 2, or 3 mg kg-1 Ni, as Ni sulfate. In the study the following parameters were evaluated: activity of urease, nitrate reductase, superoxide dismutase, catalase and ascorbate peroxidase; concentration of urea, n-compounds, photosynthetic pigments, flavonoids, H2O2 and MDA; estimative of gas exchange, and biomass as plants, yield and weight of 100 seeds. At whole-plant level, Ni affected root biomass, number of seeds per pot, and yield, increasing it at 0.5 mg kg-1 and leading to inhibition at 2-3 mg kg-1 (e.g. number of seeds per pot and nodulation). The whole-plant level enhancement by 0.5 mg Ni kg-1 occurred along with increased photosynthetic pigments, photosynthesis, ureides, and catalase, and decreased hydrogen peroxide concentration. This study presents fundamental new insights regarding Ni effect on N metabolism, and nodulation that can be helpful to increase cowpea yield. Considering the increasing population and its demand for staple food, these results contribute to the enhancement of agricultural techniques that increase crop productivity and help to maintain human food security.

Asunto(s)

Vigna , Humanos , Catalasa/metabolismo , Vigna/metabolismo , Fijación del Nitrógeno , Níquel/farmacología , Níquel/metabolismo , Peróxido de Hidrógeno/metabolismo