RESUMEN
Patulin is one of the mycotoxins frequently detected in apples and derivatives, representing a major food safety risk. This study aimed to validate a high-performance liquid chromatography (HPLC) method with an ultraviolet (UV) detector for patulin quantification and assess its occurrence in apple beverages marketed in Morocco. The validation parameters showed satisfactory results with adequate linearity (R > 0.997), a relative standard deviation below 2.5%, repeatability between 3.6 and 7.1%, reproducibility between 3.9 and 11.5%, a limit of quantification (LOQ) of 4 µg/L, and recoveries close to 100% for three levels. Analysis of 30 samples revealed patulin levels ranging from 0 to 16.36 µg/L, with 50% of samples showing negative levels. All positive results remained below the regulatory maximum limit of 50 µg/L. These findings affirm the efficacy of the HPLC proposed method in ensuring compliance with patulin regulations in apple beverages, underlining its importance in safeguarding food safety.
Asunto(s)
Contaminación de Alimentos , Malus , Patulina , Patulina/análisis , Malus/química , Cromatografía Líquida de Alta Presión , Marruecos , Contaminación de Alimentos/análisis , Bebidas/análisis , Frutas/química , Jugos de Frutas y Vegetales/análisisRESUMEN
Dittrichia viscosa belongs to the Dittrichia genus, it grows abundantly in the east and northeast of Morocco, and traditionally its fresh leaves are crushed and given for topical application after burns, wounds, and infections. In this study, we examine the wound-healing activity of Dittrichia viscosa lipidic extract in vivo, assess its anti-microbial effect, and explore the specific compounds that contribute to these effects. To assess the effectiveness of wound healing, a burn-induced wound model was employed in Wistar rats, and the levels of hydroxyproline as well as histopathological changes in the skin tissues were evaluated. Furthermore, the antimicrobial potential against Bacillus subtilis, Staphylococcus aureus, Escherichia coli, Pseudomonas aeruginosa, Candida albicans, Candida glabrata, and Malassezia furfur was investigated using the agar disc diffusion method. Gas Chromatography-mass spectrometry (GC-MS) and high-performance liquid chromatography (HPLC) techniques were employed to analyze the composition of fatty acids, phytosterols, and tocopherols. Topical application of Dittrichia viscosa lipidic fraction ointment exhibited significant improvements in wound contraction, achieving an impressive rate of 82% within 21 days. Additionally, the lipidic extract of Dittrichia viscosa displayed notable efficacy against various microbial strains, including Candida albicans (25.07 ± 0.2), Candida glabrata (24 ± 0.6), and Malassezia furfur (22 ± 0.7). The primary fatty acids identified in the sample were linolenic acid (58.95% ± 0), oleic acid (16.75% ±0.04), and linoleic acid (11.97% ± 0.1). Notably, the sample contained significant amounts of γ-Tocopherols (732.08 ± 21mg/kg), while the sterol fraction primarily consisted of 7-Campesterol (1937 ± 0 mg/kg), 7-ß-Sitosterol (1621 ± 0 mg/kg), and Stigmasterol (1439 ± 26 mg/kg). By its richness in active compound content, Dittrichia viscosa effectively accelerates wound healing while safeguarding against microbial infections.
Asunto(s)
Antiinfecciosos , Extractos Vegetales , Ratas , Animales , Extractos Vegetales/farmacología , Extractos Vegetales/química , Ratas Wistar , Antioxidantes/farmacología , Estructura Molecular , Antiinfecciosos/farmacología , Antiinfecciosos/química , Cicatrización de Heridas , Candida albicans , Tocoferoles/farmacología , Ácidos GrasosRESUMEN
The Moroccan endemic Vachellia gummifera grows wild under extreme desert conditions. This plant could be used as an alternative fodder for goats, and camels, in order to protect the Argan forests against overgrazing in Central and Southwestern Moroccan semiarid areas. With the aim to improve the V. gummifera population's density in semiarid areas, we proposed its inoculation with performing plant growth-promoting bacteria. Hence, 500 bacteria were isolated from the plant rhizosphere. From these, 291 isolates were retained for plant growth-promoting (PGP) activities assessment. A total of 44 isolates showed the best phosphates solubilization potential, as well as siderophore and auxin production. The combination of REP-PCR (repetitive extragenic palindromic-polymerase chain reaction) fingerprinting, PGP activities, and phenotypic properties, allowed the selection of three strains for the inoculation experiments. The three selected strains' 16S rRNA sequencing showed that they are members of the Enterobacter and Pseudomonas genera. The inoculation with three strains had diverse effects on V. gummifera growth parameters. All single and combined inoculations improved the plant shoot weight by more than 200%, and the root length by up to 139%, while some combinations further improved protein and chlorophyll content, thereby improving the plant's forage value. The three selected strains constitute an effective inoculum for use in the arid and semiarid zones of southern Morocco.
Asunto(s)
Fabaceae , Rizosfera , Pseudomonas/metabolismo , Marruecos , ARN Ribosómico 16S/genética , Enterobacter , Fabaceae/microbiología , Raíces de Plantas/microbiología , Microbiología del SueloRESUMEN
Objectives: Due to the limited data available within the Moroccan context, the aim of the study was therefore to estimate the prevalence of Chlamydia trachomatis (CT), Neisseria gonorrhoeae (NG) and Trichomonas vaginalis (TV) infection and co-infection among men who have sex with men (MSM) as well as to update the behavioral indicators for this population. Methods: During the period of November 2020 to January 2021, 275 and 303 MSM in Agadir and Fes respectively, were recruited by using respondent-driven sampling protocol (RDS). Eligibility criteria for participants included men identified as having anal sex with another man in the last 6 months, aged 18 years or older and residing in either Agadir or Fes, regardless of their nationality, for the past 6 months.Anal swabs were collected from 445 respondents for molecular investigation of CT, NG, and TV. GeneXpert (Cepheid, USA) was used to test all samples. A survey on the socio-demographic, and risk behavior was then administered to participants. Results: Most MSM subjects were identified as being young, and homosexual. CT prevalence was 11.3% (95%CI, 7.2 to 15.4) and 12.5% (95%CI, 7.5 to 17.5) in Agadir and Fes respectively; NG was 13.3% (95%CI, 8.5 to 18.1) in Agadir and 5.5% (95%CI, 1.9 to 9.2) in Fes. Meanwhile, TV prevalence was 0.4% (95%CI, 0 to 1.1) in Agadir and 0.2% (95%CI, -0.2 to 0.6) in Fes. A CT/NG co-infection was found in 4.5% (95%CI, 3.5 to 5.9) of cases in Agadir and 2.7% (95%CI, 1.9 to 3.9), in Fes. Conclusion: It follows that a regular risk assessment and Sexually Transmitted Infectious (STIs) screening should be administered in these two cities as part of a global strategy to enhance the sexual health of the key populations in question.
RESUMEN
Retama dasycarpa is an endemic Retama species native to the cold semi-arid bioclimates of the High Atlas Mountains in Morocco. In this work, we analyzed the diversity of the microsymbionts nodulating this plant and their different phenotypic and symbiotic characteristics. Phylogenetic analysis of the 16S rRNA gene revealed that the tested isolates clustered in the Bradyrhizobium genus. Multilocus sequence analyses of four housekeeping genes (recA, gyrB, glnII and atpD) for 12 selected strains grouped them into four clusters close to B. lupini USDA 3051T, B. frederickii CNPSo 3446T, B. valentinum LmjM3T and B. retamae Ro19T. The individual phylogenies of these core genes and the symbiotic genes nodC, nodA and nifH were congruent. These isolates showed a broad host range, being able to nodulate different legume hosts, such as R. sphaerocarpa, R. monosperma, Lupinus luteus, Cytisus grandiflorus andChamaecytisus albidus, but not Phaseolus vulgaris or Glycine max. They all had a similar metabolic capacity, using the majority of the carbohydrates and amino acids tested as sole sources of carbon and nitrogen. Furthermore, out of the 12 selected strains, some displayed plant growth-promoting features, with six of them solubilizing phosphate and three of them producing siderophores. The present work provides, for the first time, a detailed description about the microsymbionts associated with the endemic legume R. dasycarpa.
Asunto(s)
Bradyrhizobium , Fabaceae , Lupinus , Nódulos de las Raíces de las Plantas , Filogenia , ARN Ribosómico 16S/genética , ADN Bacteriano/genética , ADN Bacteriano/química , Análisis de Secuencia de ADN , Simbiosis/genéticaRESUMEN
Out of 70 bacterial strains isolated from root nodules of Lupinus albus and L. angustifolius grown in the soils from the Maamora forest in Morocco, 56 isolates possessed the nodC symbiotic gene, as determined by nodC-PCR, and they were able to renodulate their original hosts. The phenotypic analysis showed that many strains had great potential for using different carbon compounds and amino acids as sole carbon and nitrogen sources. The majority of strains grew in media with pH values between 6 and 8. Only one strain isolated from L. angustifolius was able to grow at low pH values, whereas fourteen strains nodulating L. albus grew at pH 5. No strain developed at 40 °C, and eighteen strains grew at NaCl concentrations as high as 855 mM. A total of 17 strains solubilized phosphates, whereas 20 produced siderophores and seven produced IAA. Only three strains, Lalb41, Lang10 and Lang16, possessed all three plant growth promoting activities. The strains were grouped into eight genetic groups by rep-PCR. Analysis of the 16S rRNA sequences of eight strains representing the different groups showed that they were members of the genus Bradyrhizobium. The sequencing of the five housekeeping genes atpD, glnII, dnaK, gyrB and recA, from the eight representative strains, and the phylogenetic analysis of their concatenated sequences, showed that both plants were nodulated by different Bradyrhizobium species. Accordingly, two strains, Lalb41 and Lalb5.2, belonged to B. lupini, whereas two strains, Lalb2 and Lang17.2, were affiliated to B. cytisi, and one strain, Lang2, was close to B. canariense. The fourth group of strains, Lalb25, Lang14.3 and Lang8.3, which had similarity values of less than 96% with their closest named species, B. cytisi, may belong to two new genospecies in the genus Bradyrhizobium. All the strains nodulated Lupinus cosentinii, L. luteus, Retama sphaerocarpa, R. monosperma, Chamaecytisus albus, but not Vachellia gummifera, Phaseolus vulgaris or Glycine max. The nodA, nodC and nifH sequence analyses and their phylogeny confirmed that the strains isolated from the two lupines were members of the symbiovar genistearum.
Asunto(s)
Bradyrhizobium , Lupinus , Carbono , ADN Bacteriano/genética , Bosques , Lupinus/microbiología , Marruecos , Filogenia , ARN Ribosómico 16S/genética , Nódulos de las Raíces de las Plantas/microbiología , Análisis de Secuencia de ADN , Simbiosis/genéticaRESUMEN
Acacia saligna is an invasive alien species that has the ability to establish symbiotic relationships with rhizobia. In the present study, genotypic and symbiotic diversity of native rhizobia associated with A. saligna in Tunisia were studied. A total of 100 bacterial strains were selected and three different ribotypes were identified based on rrs PCR-RFLP analysis. Sequence analyses of rrs and four housekeeping genes (recA, atpD, gyrB and glnII) assigned 30 isolates to four putative new lineages and a single strain to Sinorhizobium meliloti. Thirteen slow-growing isolates representing the most dominant IGS (intergenic spacer) profile clustered distinctly from known rhizobia species within Bradyrhizobium with the closest related species being Bradyrhizobium shewense and Bradyrhizobium niftali, which had 95.17% and 95.1% sequence identity, respectively. Two slow-growing isolates, 1AS28L and 5AS6L, had B. frederekii as their closest species with a sequence identity of 95.2%, an indication that these strains could constitute a new lineage. Strains 1AS14I, 1AS12I and 6AS6 clustered distinctly from known rhizobia species but within the Rhizobium leguminosarum complex (Rlc) with the most closely related species being Rhizobium indicum with 96.3% sequence identity. Similarly, the remaining 11 strains showed 96.9 % and 97.2% similarity values with R. changzhiense and R. indicum, respectively. Based on nodC and nodA phylogenies and cross inoculation tests, these 14 strains of Rlc species clearly diverged from strains of Sinorhizobium and Rlc symbiovars, and formed a new symbiovar for which the name sv. "salignae" is proposed. Bacterial strains isolated in this study that were taxonomically assigned to Bradyrhizobium harbored different symbiotic genes and the data suggested a new symbiovar, for which sv. "cyanophyllae" is proposed. Isolates formed effective nodules on A. saligna.
Asunto(s)
Acacia , Bradyrhizobium , Rhizobium leguminosarum , Rhizobium , ADN Bacteriano/genética , Filogenia , ARN Ribosómico 16S/genética , Rhizobium leguminosarum/genética , Nódulos de las Raíces de las Plantas/microbiología , Análisis de Secuencia de ADN , Simbiosis/genética , TúnezRESUMEN
BACKGROUND: Mining activity in the Touissit district of Eastern Morocco has led to an unprecedented accumulation of heavy metals, mainly lead and zinc, in the tailing ponds of the open-air mines. This poses a real danger to both the environment and local population. OBJECTIVES: The goal of this work was to characterize the Plant Growth Promoting Rhizobacteria (PGPR) isolated from the rhizosphere soil of R. pseudoacacia plants grown wild in the abandoned Pb- and Zn-contaminated tailing ponds in the mining district of Touissit, in Eastern Morocco. MAIN RESULTS: One hundred bacterial strains were isolated from the rhizosphere of black locust (Robinia pseudoacacia L.) plants growing naturally in the Touissit mine tailings. Quantitative determination of indole-acetic and siderophores production, inorganic phosphate solubilization, hydrolysis of 1-aminocyclopropane-1-carboxylic acid (ACC deaminase activity), and ability to act as a biocontrol agent allowed selection of the 3 strains, 7MBT, 17MBT and 84MBT with improved PGP properties. The three strains grew well in the presence of high concentration of Pb-acetate and ZnCl2; and the addition of Pb or Zn to the culture medium differently affected the PGP properties analyzed. NOVELTY STATEMENT: Inoculation of black locust grown with the 3 selected strains, in the presence 1000 µg ml-1 of Pb-acetate, produced varying effects on the plant dry weight. The strain 84MBT alone or in combination with strains 7MBT and 17MBT increased significantly the dry weight of the plants by 91, 62, and 85% respectively. The 16S rRNA gene sequence analysis of each strain showed that the strains 7MBT 17MBT and 84MBT had 99.34, 100, and had 99.72% similarity with Priestia endophytica (formerly B. endophyticus), B. pumilus NBRC 12092T, and B. halotolerans NBRC 15718T, respectively.
Asunto(s)
Robinia , Contaminantes del Suelo , Bacterias/genética , Biodegradación Ambiental , Marruecos , ARN Ribosómico 16S/genética , Rizosfera , Suelo , Microbiología del Suelo , Contaminantes del Suelo/análisisRESUMEN
In this work, we analyzed the symbiotic performance and diversity of rhizobial strains isolated from the endemic shrubby legume Chamaecytisus albidus grown in soils of three different agroforestry ecosystems representing arid and semi-arid forest areas in Morocco. The analysis of the rrs gene sequences from twenty-four representative strains selected after REP-PCR fingerprinting showed that all the strains belong to the genus Bradyrhizobium. Following multi-locus sequence analysis (MLSA) using the rrs, gyrB, recA, glnII, and rpoB housekeeping genes, five representative strains, CA20, CA61, CJ2, CB10, and CB61 were selected for further molecular studies. Phylogenetic analysis of the concatenated glnII, gyrB, recA, and rpoB genes showed that the strain CJ2 isolated from Sahel Doukkala soil is close to Bradyrhizobium canariense BTA-1 T (96.95%); that strains CA20 and CA61 isolated from the Amhach site are more related to Bradyrhizobium valentinum LmjM3T, with 96.40 and 94.57% similarity values; and that the strains CB10 and CB60 isolated from soil in the Bounaga site are more related to Bradyrhizobium murdochi CNPSo 4020 T and Bradyrhizobium. retamae Ro19T, with which they showed 95.45 and 97.34% similarity values, respectively. The phylogenetic analysis of the symbiotic genes showed that the strains belong to symbiovars lupini, genistearum, and retamae. All the five strains are able to nodulate Lupinus luteus, Retama monosperma, and Cytisus monspessilanus, but they do not nodulate Glycine max and Phaseolus vulgaris. The inoculation tests showed that the strains isolated from the 3 regions improve significantly the plant yield as compared to uninoculated plants. However, the strains of Bradyrhizobium sp. sv. retamae isolated from the site of Amhach were the most performing. The phenotypic analysis showed that the strains are able to use a wide range of carbohydrates and amino acids as sole carbon and nitrogen source. The strains isolated from the arid areas of Bounaga and Amhach were more tolerant to salinity and drought stress than strains isolated in the semi-arid area of Sahel Doukkala.
Asunto(s)
Bradyrhizobium , Fabaceae , Lupinus , Simbiosis/genética , Nódulos de las Raíces de las Plantas , Filogenia , Ecosistema , Marruecos , ARN Ribosómico 16S/genética , ADN Bacteriano/genética , ADN Bacteriano/química , Bradyrhizobium/genética , Lupinus/genética , Alimentación Animal , Suelo , Análisis de Secuencia de ADNRESUMEN
In the Moroccan Middle Atlas, the tailings rich in lead and other metal residues, in the abandoned Zaida mining district, represent a real threat to environment and the neighboring villages' inhabitants' health. In this semi-arid to arid area, phytostabilisation would be the best choice to limit the transfer of heavy metals to populations and groundwater. The aim of this work was to characterize the bacteria that nodulate Retama sphaerocarpa, spontaneous nitrogen fixing shrubby legume, native to the Zaida mining area, with great potential to develop for phytostabilisation. Forty-three bacteria isolated from root nodules of the plant were characterized. Based on REP-PCR and ARDRA, four strains were selected for further molecular analyzes. The 16S rRNA gene sequences analysis revealed that the isolated strains are members of the genus Bradyrhizobium, and the phylogenetic analysis of the housekeeping genes glnII, atpD, gyrB, rpoB, recA and dnaK individual sequences and their concatenation showed that the strains are close to B. algeriense RST89T and B. valentinum LmjM3T with similarity percentages of 89.07% to 95.66% which suggest that the newly isolated strains from this mining site may belong to a potential novel species. The phylogeny of the nodA and nodC genes showed that the strains belong to the symbiovar retamae of the genus Bradyrhizobium. These strains nodulate also R. monosperma, R. dasycarpa and Lupinus luteus.
Asunto(s)
Bradyrhizobium , Fabaceae , Minería , Filogenia , Bradyrhizobium/clasificación , Bradyrhizobium/aislamiento & purificación , ADN Bacteriano/genética , Fabaceae/microbiología , Genes Bacterianos , Plomo , Marruecos , ARN Ribosómico 16S/genética , Nódulos de las Raíces de las Plantas/microbiología , Análisis de Secuencia de ADN , SimbiosisRESUMEN
Out of 54 isolates from root nodules of the Moroccan-endemic Chamaecytisus albidus plants growing in soils from the Maamora cork oak forest, 44 isolates formed nodules when used to infect their original host plant. A phenotypic analysis showed the metabolic diversity of the strains that used different carbohydrates and amino acids as sole carbon and nitrogen sources. The isolates grew on media with pH values ranging from 6 to 8. However, they did not tolerate high temperatures or drought and they did not grow on media with salt concentrations higher than 85â¯mM. REP-PCR fingerprinting grouped the strains into 12 clusters, of which representative strains were selected for ARDRA and rrs analyses. The rrs gene sequence analysis indicated that all 12 strains were members of the genus Bradyrhizobium and their phylogeny showed that they were grouped into two different clusters. Two strains from each group were selected for multilocus sequence analysis (MLSA) using atpD, recA, gyrB and glnII housekeeping genes. The inferred phylogenetic trees confirmed that the strains clustered into two divergent clusters. Strains CM55 and CM57 were affiliated to the B. canariense/B. lupini group, whereas strains CM61 and CM64 were regrouped within the B. cytisi/B. rifense lineage. The analysis of the nodC symbiotic gene affiliated the strains to the symbiovar genistearum. The strains were also able to nodulate Retama monosperma, Lupinus luteus and Cytisus monspessulanus, but not Phaseolus vulgaris or Glycine max. Inoculation tests with C. albidus showed that some strains could be exploited as efficient inocula that could be used to improve plant growth in the Maamora forest.
Asunto(s)
Bradyrhizobium , Fabaceae/microbiología , Filogenia , Nódulos de las Raíces de las Plantas/microbiología , Bradyrhizobium/genética , ADN Bacteriano/genética , Bosques , Marruecos , Nodulación de la Raíz de la Planta , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , SimbiosisRESUMEN
The aim of this work was to characterize and identify some bacteria isolated from the root nodules of Retama monosperma grown in Sidi Boubker lead and zinc mine tailings. Very few root nodules were obtained on the root nodules of R. monosperma grown in these soils. The three bacteria isolated from the root nodules were tolerant in vitro to different concentrations of heavy metals, including lead and zinc. The rep-PCR experiments showed that the three isolates have different molecular fingerprints and were considered as three different strains. The analysis of their 16S rRNA gene sequences proved their affiliation to the genus Bradyrhizobium. The analysis and phylogeny of the housekeeping genes atpD, glnII, gyrB, recA, and rpoB confirmed that the closest species was B. valentinum with similarity percentages of 95.61 to 95.82%. The three isolates recovered from the root nodules were slow-growing rhizobia capable to renodulate their original host plant in the presence of Pb-acetate. They were able to nodulate R. sphaerocarpa and Lupinus luteus also but not Glycine max or Phaseolus vulgaris. The phylogeny of the nodA and nodC nodulation genes as well as the nifH gene of the three strains showed that they belong to the symbiovar retamae of the genus Bradyrhizobium. The three strains isolated could be considered for use as inoculum for Retama plants before use in phytoremediation experiments.
Asunto(s)
Bradyrhizobium/metabolismo , Fabaceae/microbiología , Plomo/metabolismo , Nódulos de las Raíces de las Plantas/microbiología , Zinc/metabolismo , Bradyrhizobium/clasificación , Bradyrhizobium/genética , Bradyrhizobium/aislamiento & purificación , Minería , Marruecos , Filogenia , Nodulación de la Raíz de la Planta , Glycine max/microbiologíaRESUMEN
In this work, we analyzed the diversity of seventy-six bacteria isolated from Pea and faba bean nodules in two regions of Morocco. The molecular diversity was realized using the analysis of the sequences of 16S rRNA and six housekeeping genes (recA, glnII, atpD, dnaK, rpoB and gyrB) and two symbiotic genes (nodA and nodC). The phylogeny of the 16S rRNA gene sequences revealed that all strains belong to the genus Rhizobium, being related to the type strains of R. leguminosarum, R. laguerreae, R. indigoferae, R. anhuiense and R. acidisoli. The housekeeping genes phylogenies showed that some strains formed a subclade distinct from the rhizobial species that usually nodulate Vicia faba and Pisum sativum which are closely related to R. acidisoli FH23 with sequence similarity of 98.3%. Analysis of the PGPR activities of the different isolates showed that the strains related to R. laguerreae were able to solubilize phosphates and to produce siderophores and auxin phytohormone. However, R. acidisoli strain F40D2 was unable to solubilize phosphates although they produce siderophores and IAA. The phylogenetic analysis of the nodA and nodC sequences showed that all isolated strains were closely related with the strains of symbiovar viciae. The nodulation tests confirmed the ability to nodulate V. faba and P. sativum but not Cicer arietinum or Phaseolus vulgaris. Hence, in Morocco P. sativum is nodulated by R. laguerreae; whereas V. faba is nodulated by R. laguerreae and the symbiovar viciae of R. acidisoli which has been not previously described in this species.
Asunto(s)
Rhizobium , Nódulos de las Raíces de las Plantas/microbiología , Simbiosis , Vicia faba/microbiología , Genes Bacterianos , Genes Esenciales , Genómica/métodos , Marruecos , Fenotipo , Filogenia , ARN Ribosómico 16S/genética , Microbiología del SueloRESUMEN
Millions tons of lead and zinc wastes from the abandoned Touissit mine are stored in the open air as dikes in the vicinity of the villages in Eastern Morocco and pose a real danger to both the environment and local populations. To prevent the movement of minerals to the nearby villages and limit the damages to the environment and health, we proposed the nitrogen-fixing leguminous shrub Retama monosperma, as a model plant to use for phytostabilization experimentations. This plant species is known by its ability to grow in hard climatic conditions and in heavy metals contaminated soils. The isolation of bacterial strains nodulating R. monosperma in the abandoned mine soils will permit the selection of rhizobia to inoculate young plant seedlings before their use for the phytostabilization of the mine tailings. In this work, 44 bacteria were isolated from the root nodules of R. Monosperma grown in the Touissit abandoned mine. Twenty-four isolates were considered as true rhizobia as they possess a copy of the nodC symbiotic gene and were able to renodulate their original host. The phenotypic characterization showed that all the strains are tolerant in vitro to different concentrations of heavy metals. The analysis of the 16S rRNA sequences of two selected representative strains showed they were related to different strains of Ensifer aridi isolated from different legumes in three continents deserts. The glnII, recA, and gyrB housekeeping genes analysis confirmed the affiliation of the strains to E. aridi. Moreover, the phylogenic analysis of nodA, nodC, and nifH symbiotic genes showed that the strains are more related to E. aridi JNVUTP6 species isolated from Tephrosia purpurea root nodules in the Thar Desert in India. To our knowledge, this is the first report about the isolation of E. aridi from R. monosperma root nodules.
RESUMEN
The purpose of this work was to study the genetic diversity of the nodule-forming bacteria associated with native populations of Vachellia gummifera growing wild in Morocco. The nearly complete 16S rRNA gene sequences from three selected strains, following ARDRA and REP-PCR results, revealed they were members of the genus Ensifer and the sequencing of the housekeeping genes recA, gyrB, dnaK and rpoB, and their concatenated phylogenetic analysis, showed that the 3 strains belong to the species E. fredii. Based on the nodC and nodA phylogenies, the 3 strains clearly diverged from the type and other reference strains of E. fredii and formed a clearly separated cluster. The strains AGA1, AGA2 and AGB23 did not form nodules on Glycine max, Phaseolus vulgaris and Medicago truncatula, and effectively nodulated V. gummifera, Acacia cyanophylla, Prosopis chilensis and Leucaena leucocephala. Based on similarities of the nodC and nodA symbiotic genes and differences in the host range, the strains isolated from Moroccan endemic V. gummifera may form a different symbiovar within Ensifer species, for which the name "vachelliae" is proposed.
Asunto(s)
Fabaceae/microbiología , Variación Genética , Nódulos de las Raíces de las Plantas/microbiología , Sinorhizobium/clasificación , Simbiosis , ADN Bacteriano/genética , Genes Bacterianos/genética , Genes Esenciales/genética , Especificidad del Huésped , Marruecos , Filogenia , ARN Ribosómico 16S/genética , Análisis de Secuencia de ADN , Sinorhizobium/genética , Sinorhizobium/crecimiento & desarrollo , Simbiosis/genéticaRESUMEN
Astragalus algarbiensis is a wild herbaceous legume growing in Maamora, the most important cork oak forest in northern Africa. It is a plant of great importance as fodder in silvopastoral systems, and in the restoration of poor and degraded soils. The purpose of this study was to describe the biodiversity of rhizobia nodulating this plant and determine their identity. Out of 80 bacterial isolates, 56 strains isolated from root nodules of A. algarbiensis were characterized. ERIC-PCR fingerprinting grouped the strains in two main clusters containing 29 and 27 isolates, respectively, and the amplified ribosomal DNA restriction analysis (ARDRA) generated two different ribotypes. Based on both the ERIC-PCR and ARDRA results, representative strains As21 and As36 were selected for further genetic studies. The nearly complete 16S rRNA gene sequences of As21 and As36 showed that they were closely related to Bradyrhizobium cytisi CTAW11T with similarity values of 99.84% and 99.77%, respectively. Concatenation of atpD, recA, gyrB and dnaK housekeeping gene sequences indicated that strains As21 and As36 had a 95.22% similarity but they showed values of 95.80% and 94.97% with B. cytisi CTAW11T, respectively. The sequencing of the symbiotic nodC gene of the two strains revealed 97.20% and 97.76% identities, respectively, with that of B. cytisi CTAW11T isolated from Cytisus villosus growing in the Moroccan Rif Mountains. Furthermore, the phylogenic analysis showed that the strains isolated from A. algarbiensis clustered with B. cytisi and B. rifense within the bradyrhizobia genistearum symbiovar and may constitute two novel genospecies.
Asunto(s)
Planta del Astrágalo/microbiología , Bradyrhizobium/clasificación , Filogenia , Nodulación de la Raíz de la Planta/genética , Microbiología del Suelo , Biodiversidad , Bradyrhizobium/genética , Cytisus/microbiología , ADN Bacteriano/genética , ADN Ribosómico/genética , Genes Bacterianos/genética , Genes Esenciales/genética , Marruecos , Nódulos de las Raíces de las Plantas/microbiología , Análisis de Secuencia de ADN , Simbiosis/genéticaRESUMEN
A total of 51 bacterial strains were isolated from root nodules of Scorpiurus muricatus sampled from 6 regions of western Algeria. Strain diversity was assessed by rep-PCR amplification fingerprinting, which grouped the isolates into 28 different clusters. Partial nucleotide sequencing of the 16S rRNA gene and BLAST analysis revealed that root nodules of S. muricatus were colonized by different species close to Rhizobium vignae, Rhizobium radiobacter, Rhizobium leguminosarum, Phyllobacterium ifriqiyense, Phyllobacterium endophyticum, Starkeya sp., and Pseudomonas sp. However, none of these strains was able to form nodules on its host plant; even nodC was present in a single strain (SMT8a). The inoculation test showed a great improvement in the growth of inoculated plants compared with noninoculated control plants. A significant amount of indole acetic acid was produced by some strains, but only 2 strains could solubilize phosphate. In this report we described for the first time the diversity of bacteria isolated from root nodules of S. muricatus growing in different regions in western Algeria and demonstrated their potential use in promoting plant growth.
Asunto(s)
Bacterias/aislamiento & purificación , Fabaceae/microbiología , Argelia , Alphaproteobacteria/genética , Alphaproteobacteria/aislamiento & purificación , Bacterias/clasificación , Bacterias/genética , Biodiversidad , Ácidos Indolacéticos/metabolismo , Filogenia , Desarrollo de la Planta , Raíces de Plantas/microbiología , ARN Ribosómico 16S , Rhizobium/genética , Rhizobium/aislamiento & purificaciónRESUMEN
Polyphasic characterization of 61 bacteria isolated from root nodules of Medicago arborea (Medic tree) plants growing in 4 arid soils of the arid eastern area of Morocco was studied. All the isolates characterized were fast growers. The phenotypic, symbiotic, and cultural characteristics analyzed allowed the description of a broad physiological diversity among the isolates. The results obtained suggest that the phenotype of these rhizobia might have evolved to adapt to the local conditions. The genetic characterization consisted of an analysis of the rep-PCR (repetitive extragenic palindromic polymerase chain reaction) fingerprints and a PCR-based RFLP (restriction fragment length polymorphism) of the 16S rDNA patterns. The diversity of the isolates was investigated by rep-PCR, giving a similarity of 62%, delineated into 3 clusters, 4 groups, and 6 subclusters. This wide diversity was also observed by a phenotypic approach, where the carbohydrate assimilation test was the most discriminating. The results show a relationship between rep-PCR fingerprinting and sugar assimilation, which are complementary in diversity investigation. The nearly complete 16S rRNA gene sequence from representative strains of each soil showed they are closely related to members of the genus Ensifer of the family Rhizobiaceae within the Alphaproteobacteria and shows the highest similitude values (99.93%/100%) with Ensifer meliloti LMG 6133(T) (X67222). Sequencing of the symbiotic nodC gene from 7 representative strains revealed they had 94.89% identity with the nodC sequence of the type strain E. meliloti LMG 6133(T) (EF428922). Therefore, the 61 M. arborea isolates from the 4 different soils have the same phylogenetic affiliation, which proves the restricted host specificity among M. arborea species.
Asunto(s)
Medicago/microbiología , Sinorhizobium meliloti/fisiología , Microbiología del Suelo , Simbiosis , Proteínas Bacterianas/genética , Variación Genética , Marruecos , N-Acetilglucosaminiltransferasas/genética , Fenotipo , Filogenia , Raíces de Plantas/microbiología , ARN Ribosómico 16S/genética , Sinorhizobium meliloti/genéticaRESUMEN
Astragalus gombiformis is a desert symbiotic nitrogen-fixing legume of great nutritional value as fodder for camels and goats. However, there are no data published on the rhizobial bacteria that nodulate this wild legume in northern Africa. Thirty-four rhizobial bacteria were isolated from root nodules of A. gombifomis grown in sandy soils of the South-Eastern of Morocco. Twenty-five isolates were able to renodulate their original host and possessed a nodC gene copy. The phenotypic and genotypic characterizations carried out illustrated the diversity of the isolates. Phenotypic analysis showed that isolates used a great number of carbohydrates as sole carbon source. However, although they were isolated from arid sandy soils, the isolates do not tolerate drought stress applied in vitro. The phenotypic diversity corresponded mainly to the diversity in the use of some carbohydrates. The genetic analysis as assessed by repetitive extragenic palindromic (REP)-polymerase chain reaction (PCR) showed that the isolates clustered into 3 groups at a similarity coefficient of 81 %. The nearly-complete 16S rRNA gene sequence from a representative strain of each PCR-group showed they were closely related to members of the genus Mesorhizobium of the family Phyllobactericeae within the Alphaproteobacteria. Sequencing of the housekeeping genes atpD, glnII and recA, and their concatenated phylogenetic analysis, showed they are closely related to Mesorhizobium camelthorni. Sequencing of the symbiotic nodC gene from each strain revealed they had 83.53 % identity with the nodC sequence of the type strain M. camelthorni CCNWXJ 40-4(T.)
Asunto(s)
Planta del Astrágalo/microbiología , Mesorhizobium/clasificación , Mesorhizobium/aislamiento & purificación , Nodulación de la Raíz de la Planta , Nódulos de las Raíces de las Plantas/microbiología , Proteínas Bacterianas/genética , Técnicas de Tipificación Bacteriana , Secuencia de Bases , ADN Bacteriano/genética , Variación Genética , Mesorhizobium/genética , Marruecos , Filogenia , ARN Ribosómico 16S/genética , Rec A Recombinasas/genética , Análisis de Secuencia de ADN , SimbiosisRESUMEN
In this paper we analyze through a polyphasic approach several Bradyrhizobium strains isolated in Spain and Morocco from root nodules of Retama sphaerocarpa and Retama monosperma. All the strains have identical 16S rRNA genes and their closest relative species is Bradyrhizobium lablabi CCBAU 23086(T), with 99.41% identity with respect to the strain Ro19(T). Despite the closeness of the 16S rRNA genes, the housekeeping genes recA, atpD and glnII were divergent in Ro19(T) and B. lablabi CCBAU 23086(T), with identity values of 95.71%, 93.75% and 93.11%, respectively. These differences were congruent with DNA-DNA hybridization analysis that revealed an average of 35% relatedness between the novel species and B. lablabi CCBAU 23086(T). Also, differential phenotypic characteristics of the new species were found with respect to the already described species of Bradyrhizobium. Based on the genotypic and phenotypic data obtained in this study, we propose to classify the group of strains isolated from R. sphaerocarpa and R. monosperma as a novel species named Bradyrhizobium retamae sp. nov. (type strain Ro19(T)=LMG 27393(T)=CECT 8261(T)). The analysis of symbiotic genes revealed that some of these strains constitute a new symbiovar within genus Bradyrhizobium for which we propose the name "retamae", that mainly contains nodulating strains isolated from Retama species in different continents.