The Bradyrhizobium uaiense strain UFLA 03-164T enhanced yield performance of cowpea in soils with low or high phosphorus content

The objective of this study was to evaluate the agronomic efficiency of rhizobia strains already approved or in the selection stage for cowpea, in the Recôncavo da Bahia region, Brazil. Two field experiments were performed in the municipalities of Cruz das Almas and Maragogipe, Bahia, Brazil. A randomized block design was used with seven nitrogen sources and four replicates. Nitrogen sources consisted of the strains UFRB FA51B1, UFRB BA72C2-1, UFLA 03-164T, UFLA 03-84, INPA 03-11B, and two controls without inoculation, one with mineral nitrogen and another without. All the strains nodulated cowpea. The efficiency of the strains was determined by the number of nodules, nodule dry matter, total dry matter, grain yield, nitrogen accumulation in shoots and grain, and relative efficiency. The strain UFLA 03-164T can be recommended for biomass production, green manure, and promotion of grain yields in both soils. In Maragogipe, the UFLA 03-84 and INPA 03-11B strains can be recommended for biomass production, green manure, and promotion of grain yields. The UFLA 03-164T strain showed great potential to promote and grain yield in the two municipalities studied. The strain INPA 03-11B can be recommended for Maragogipe soil. UFLA 03-84 can also be recommended to increase grain yield in Cruz das Almas.

Optimization of soybean yield in ultisols through adaptive varieties screening and plant growth promoting Rhizobacter

This research was aimed at obtaining varieties of soybean adaptive to acid soils and to obtain Plant Growth-Promoting Rhizobacteria (PGPR) isolates that can improve the agronomic characteristics of soybean and increase the ultisols fertility. This research was conducted in two-stages research on Sampali Village, Percut Sei Tuan sub-District, Deli Serdang District, North Sumatra Province, Indonesia from August 2019 until March 2020. The first stage (adaptive varieties screening) using the non-factorial Randomized Block Design (RBD) with the varieties of Argomulyo, Wilis, Kaba, Dena-1, Devon-1, Dega-1, Demas-1, Burangrang, Detam-1, and Kipas Merah. The second stage (application of PGPR isolates singly and in combinations) using the factorial RBD, the first factor of applicative single and the combination of PGPR isolates, the second factor of adaptive varieties including Detam-1 and Wilis. Data were analyzed with ANOVA and followed by DMRT at P<0.05. The results showed that the Detam-1 and Wilis varieties had significantly higher yield per plant of 14.73 g and 14.54 g, respectively, than other varieties. The applications of a single and combination of PGPR isolates significantly increased the number of branches, stem diameter, plant height, yield per plant, soil pH, organic-C, available-P, and total-N and decreased the soil C/N. The Detam-1 variety had the higher in yield per plant compared to Wilis variety. The isolates combination of Rhizobium leguminosarum+Rhizobium sp2+Bacillus sp+Burkholderia sp for Detam-1 and Wilis varieties can be recommended to support the growth and yield of soybean on ultisols.

Tuber borchii Shapes the Ectomycorrhizosphere Microbial Communities of Corylus avellana

In this study, eight-month-old ectomycorrhizae of Tuber borchii with Corylus avellana were synthesized to explore the influence of T. borchii colonization on the soil properties and the microbial communities associated with C. avellana during the early symbiotic stage. The results showed that the bacterial richness and diversity in the ectomycorrhizae were significantly higher than those in the control roots, whereas the fungal diversity was not changed in response to T. borchii colonization. Tuber was the dominant taxon (82.97%) in ectomycorrhizae. Some pathogenic fungi, including Ilyonectria and Podospora, and other competitive mycorrhizal fungi, such as Hymenochaete, had significantly lower abundance in the T. borchii inoculation treatment. It was found that the ectomycorrhizae of C. avellana contained some more abundant bacterial genera (e.g., Rhizobium, Pedomicrobium, Ilumatobacter, Streptomyces, and Geobacillus) and fungal genera (e.g., Trechispora and Humicola) than the control roots. The properties of rhizosphere soils were also changed by T. borchii colonization, like available nitrogen, available phosphorus and exchangeable magnesium, which indicated a feedback effect of mycorrhizal synthesis on soil properties. Overall, this work highlighted the interactions between the symbionts and the microbes present in the host, which shed light on our understanding of the ecological functions of T. borchii and facilitate its commercial cultivation.

Phenotypic, genetic and symbiotic characterization of Erythrina velutina rhizobia from Caatinga dry forest

Abstract Erythrina velutina ("mulungu") is a legume tree from Caatinga that associates with rhizobia but the diversity and symbiotic ability of "mulungu" rhizobia are poorly understood. The aim of this study was to characterize "mulungu" rhizobia from Caatinga. Bacteria were obteined from Serra Talhada and Caruaru in Caatinga under natural regeneration. The bacteria were evaluated to the amplification of nifH and nodC and to metabolic characteristics. Ten selected bacteria identified by 16S rRNA sequences. They were tested in vitro to NaCl and temperature tolerance, auxin production and calcium phosphate solubilization. The symbiotic ability were assessed in an greenhouse experiment. A total of 32 bacteria were obtained and 17 amplified both symbiotic genes. The bacteria showed a high variable metabolic profile. Bradyrhizobium (6), Rhizobium (3) and Paraburkholderia (1) were identified, differing from their geographic origin. The isolates grew up to 45 °C to 0.51 mol L-1 of NaCl. Bacteria which produced more auxin in the medium with l-tryptophan and two Rhizobium and one Bradyrhizobium were phosphate solubilizers. All bacteria nodulated and ESA 90 (Rhizobium sp.) plus ESA 96 (Paraburkholderia sp.) were more efficient symbiotically. Diverse and efficient rhizobia inhabit the soils of Caatinga dry forests, with the bacterial differentiation by the sampling sites.

Genomic comparisons of Rhizobium species using in silico AFLP-PCR, endonuclease restriction, and AMPylating enzymes

Background: The whole-genome sequences of nine Rhizobium species were evaluated using different in silico molecular techniques such as AFLP-PCR, restriction digest, and AMPylating enzymes. The entire genome sequences were aligned with progressiveMauve and visualized by reconstructing phylogenetic tree using NTSYS pc 2.11X. The "insilico.ehu.es" was used to carry out in silico AFLP-PCR and in silico restriction digest of the selected genomes. Post-translational modification (PTM) and AMPylating enzyme diversity between the proteome of Rhizobium species were determined by novPTMenzy. Results: Slight variations were observed in the phylogeny based on AFLP-PCR and PFGE and the tree based on whole genome. Results clearly demonstrated the presence of PTMs, i.e., AMPylation with the GS-ATasE (GlnE), Hydroxylation, Sulfation with their domain, and Deamidation with their specific domains (AMPylating enzymes) GS-ATasE (GlnE), Fic, and Doc (Phosphorylation); Asparagine_hydroxylase and Collagen_prolyl_lysyl_hydroxylase; Sulfotransferase; and CNF (Cytotoxic Necrotizing Factors), respectively. The results pertaining to PTMs are discussed with regard to functional diversities reported in these species. Conclusions: The phylogenetic tree based on AFLP-PCR was slightly different from restriction endonuclease- and PFGE-based trees. Different PTMs were observed in the Rhizobium species, and the most prevailing type of PTM was AMPylation with the domain GS-ATasE (GlnE). Another type of PTM was also observed, i.e., Hydroxylation and Sulfation, with the domains Asparagine_hydroxylase and Collagen_prolyl_lysyl_hydroxylase and Sulfotransferase, respectively. The deamidation type of PTM was present only in Rhizobium sp. NGR234. How to cite: Qureshi MA, Pervez MT, Babar ME, et al. Genomic comparisons of Rhizobium species using in silico AFLP-PCR, endonuclease restrictions and ampylating enzymes.

Agronomic efficiency of Rhizobium strains from the Amazon region in common bean / Eficiência agronômica de estirpes de Rhizobium isoladas da Amazônia sobre o feijoeiro-comum

To maximize the contribution of biological nitrogen fixation in common bean, it is necessary to use bacterial strains that are more adapted, competitive, and efficient in the symbiotic process. In this regard, the aim of this study was to evaluate the agronomic efficiency (AE) of three bacterial strains isolated from acid soils with high Al content from the Amazon region in an Argissolo Vermelho Distrófico típico soil (Typic Rhodustults - USDA Classification) from the municipality of Formiga, MG, Brazil. We compared their AE to that of the reference strain CIAT 899T and of two controls without inoculation (one without and another with 80 kg ha-1 of N-urea). The results indicated that inoculation with the strains UFLA 02-100 and UFLA 02-127 provides grain yield equivalent to inoculation with the reference strain and to the control with mineral N. Thus, both have potential for recommendation as inoculants for common bean.

Para maximizar a contribuição da fixação biológica de nitrogênio no feijoeiro-comum é necessária a utilização de estirpes de bactérias mais adaptadas, competitivas e eficientes no processo simbiótico. Nesse sentido, objetivou-se avaliar, em um Argissolo Vermelho Distrófico típico do município de Formiga-MG, a eficiência agronômica (EA) de três estirpes isoladas de solos ácidos e com alto teor de Al da Amazônia e comparar suas EA à da estirpe referência CIAT 899T e à de dois controles sem inoculação (um sem e outro com 80 kg ha-1 of N-ureia). Os resultados indicaram que a inoculação com as estirpes UFLA 02-100 e UFLA 02-127 propicia rendimento de grãos equivalente ao da estirpe referência e ao do controle com N mineral e que por isso, ambas têm potencial para ser recomendadas como inoculantes para o feijoeiro-comum.

Compatibility of Azospirillum brasilense and Pseudomonas fluorescens in growth promotion of groundnut ( Arachis hypogea L)

ABSTRACT We attempted to study the compatibility among plant beneficial bacteria in the culture level by growing them near in the nutrient agar plates. Among all the bacteria tested, Rhizobium was found to inhibit the growth of other bacteria. From the compatible group of PGPR, we have selected one biofertilizer (Azospirillum brasilense strain TNAU) and one biocontrol agent (Pseudomonas fluorescens strain PF1) for further studies in the pot culture. We have also developed a bioformulation which is talc powder based, for individual bacteria and mixed culture. This formulation was used as seed treatment, soil application, seedling root dip and foliar spray in groundnut crop in vitro germination conditions. A. brasilense was found to enhance the tap root growth and P. fluorescens, the lateral root growth. The other growth parameters like shoot growth, number of leaves were enhanced by the combination of both of the bacteria than their individual formulations. Among the method of application tested in our study, soil application was found to be the best in yielding better results of plant growth promotion.

Displacers improve the selectivity of phosphopeptide enrichment by metal oxide affinity chromatography / Uso de desplazadores para mejorar la selectividad de los fosfopéptidos enriquecidos por cromatografía de afinidad con óxidos de metales

Abstract: Background: A key process in cell regulation is protein phosphorylation, which is catalyzed by protein kinases and phosphatases. However, phosphoproteomics studies are difficult because of the complexity of protein phosphorylation and the number of phosphorylation sites. Methods: We describe an efficient approach analyzing phosphopeptides in single, separated protein by two-dimensional gel electrophoresis. In this method, a titanium oxide (TiO2)-packed NuTip is used as a phosphopeptide trap, together with displacers as lactic acid in the loading buffer to increase the efficiency of the interaction between TiO2 and phosphorylated peptides. Results: The results were obtained from the comparison of mass spectra of proteolytic peptides of proteins with a matrix-assisted laser desorption-ionization-time of flight (MALDI-TOF) instrument. Conclusions: This method has been applied to identifying phosphoproteins involved in the symbiosis Rhizobium etli-Phaseolus vulgaris.

Resumen: Introducción: Un proceso clave en la regulación celular es la fosforilación de proteínas, que se lleva a cabo por cinasas y fosfatasas. Sin embargo, los estudios de fosfoproteómica son difíciles debido a la complejidad de la fosforilación proteica y el número de sitios de fosforilación. Métodos: En el presente trabajo se describe una eficiente estrategia metodológica para analizar fosfopéptidos de proteínas separadas mediante electroforesis bidimensional. En este método, una columna con microesferas de dióxido de titanio (TiO2/NuTip) se utilizó para atrapar los fosfopéptidos en la superficie del TiO2 previamente empacado en una punta. El uso de desplazadores en el buffer de carga, como el ácido láctico, mejoró significativamente la selectividad. Resultados: Los resultados se obtuvieron mediante la comparación de los espectros de masas de péptidos proteolíticos de proteínas analizados utilizando un instrumento de desorción/ionización láser asistida por matriz-tiempo de vuelo (MALDI-TOF). Conclusiones: Este método se ha aplicado para la identificación de fosfoproteínas involucradas en la simbiosis del Rhizobium etli con Phaseolus vulgaris.

Characterization of rhizobia isolates obtained from nodules of wild genotypes of common bean

Abstract This study aimed to evaluate the tolerance to salinity and temperature, the genetic diversity and the symbiotic efficiency of rhizobia isolates obtained from wild genotypes of common bean cultivated in soil samples from the States of Goiás, Minas Gerais and Paraná. The isolates were subjected to different NaCl concentrations (0%, 1%, 2%, 4% and 6%) at different temperatures (28 °C, 33 °C, 38 °C, 43 °C and 48 °C). Genotypic characterization was performed based on BOX-PCR, REP-PCR markers and 16S rRNA sequencing. An evaluation of symbiotic efficiency was carried out under greenhouse conditions in autoclaved Leonard jars. Among 98 isolates about 45% of them and Rhizobium freirei PRF81 showed a high tolerance to temperature, while 24 isolates and Rhizobium tropici CIAT899 were able to use all of the carbon sources studied. Clustering analysis based on the ability to use carbon sources and on the tolerance to salinity and temperature grouped 49 isolates, R. tropici CIAT899 and R. tropici H12 with a similarity level of 76%. Based on genotypic characterization, 65% of the isolates showed an approximately 66% similarity with R. tropici CIAT899 and R. tropici H12. About 20% of the isolates showed symbiotic efficiency similar to or better than the best Rhizobium reference strain (R. tropici CIAT899). Phylogenetic analysis of the 16S rRNA revealed that two efficient isolates (ALSG5A1 and JPrG6A8) belong to the group of strains used as commercial inoculant for common bean in Brazil and must be assayed in field experiments.

Symbiotic potential and survival of native rhizobia kept on different carriers

Native rhizobia are ideal for use as commercial legume inoculants. The characteristics of the carrier used to store the inoculants are important for the survival and symbiotic potential of the rhizobia. The objective of this study was to investigate the effects of peat (PEAT), perlite sugarcane bagasse (PSB), carboxymethyl cellulose plus starch (CMCS), and yeast extract mannitol supplemented with mannitol (YEMM) on the survival, nodulation potential and N₂ fixation capacity of the native strains Sinorhizobium mexicanum ITTG R7^T and Rhizobium calliandrae LBP2-1^T and of the reference strain Rhizobium etli CFN42^T. A factorial design (4 × 3) with four repetitions was used to determine the symbiotic potential of the rhizobial strains. The survival of the strains was higher for PEAT (46% for strain LBP2-1^T, 167% for strain CFN42^T and 219% for strain ITTG R7^T) than for the other carriers after 240 days, except for CFN42^T kept on CMCS (225%). All the strains kept on the different carriers effectively nodulated common bean, with the lowest number of nodules found (5 nodules) when CFN42^T was kept on CMCS and with the highest number of nodules found (28 nodules) when ITTG R7^T was kept on PSB. The nitrogenase activity was the highest for ITTG R7^T kept on PEAT (4911 μmol C₂H₄ per fresh weight nodule h⁻¹); however, no activity was found when the strains were kept on YEMM. Thus, the survival and symbiotic potential of the rhizobia depended on the carrier used to store them.

Developing novel bacterial based bioformulation having PGPR properties for enhanced production of agricultural crops.

Plant growth promoting rhizobacteria (PGPR) are beneficial rhizobacteria which enhance plant growth as well as the productivity by a variety of mechanisms. PGPR were isolated from the rhizosphere region of som plants (Machilus bombycina King) maintained at the Central Muga Eri Research and Training Institute, Lahdoigarh, Jorhat. A bacterial based bioformulation was prepared and sprayed over the experimental crops including tomato (Solanum lycopersicum), cauliflower (Brassica oleracea var botrytis), chili (Capsicum annuum) and brinjal (Solanum melongena). Biochemical analysis was done on these PGPR treated crops as well as the untreated crops. The bioformulations prepared from Bacillus cereus (MTCC 8297), Pseudomonas rhodesiae (MTCC 8299) and Pseudomonas rhodesiae (MTCC 8300) was found to be the most effective in increasing the shoot height, number of leaves, early fruiting and total biomass content of the plants after treatment.

Caracterización de rizobacterias promotoras de crecimiento en plántulas de Eucalyptus nitens / Characterization of growth-promoting rhizobacteria in Eucalyptus nitens seedlings

Se aislaron bacterias rizosféricas y endófitas a partir de rizósfera y tejidos de raíz de árboles de Eucalyptus nitens con el objetivo de evaluar su capacidad de promover el crecimiento en plántulas de la misma especie en condiciones de invernadero. Los aislamientos que incrementaron el crecimiento de las plántulas fueron identificados y caracterizados por su capacidad de producir ácido indolacético (AIA), solubilizar fosfato y expresar la 1-aminociclopropano-1-carboxilato (ACC) desaminasa. Los 105 aislamientos obtenidos fueron morfológicamente diferentes y solo 15 promovieron significativamente el crecimiento de plántulas de E. nitens. Los máximos incrementos observados fueron en el peso seco aéreo (142 %) y de la raíz (135 %); también aumentaron la altura de las plantas (50 %) y el largo de raíces (45 %) de las mismas. Las rizobacterias pertenecieron a los géneros Arthrobacter, Lysinibacillus, Rahnella y Bacillus. Los aislados identificados como A. phenanthrenivorans 21 y B. cereus 113 incrementaron la emergencia de E. nitens a los 12 días en un valor promedio de 3,15 veces con relación al control. R. aquatilis aislado 78 presentó la mayor producción de AIA (97,5 ± 2,87 μg/ml) en presencia de triptófano y el mayor índice de solubilización de fósforo (2,4). B. amyloliquefaciens aislado 60 fue positivo para la actividad ACC desaminasa. Los resultados obtenidos indican el potencial de las rizobacterias estudiadas como promotoras de emergencia y crecimiento de plántulas de E. nitens y su posible uso como inoculantes, ya que presentan más de un mecanismo de acción asociado a la promoción del crecimiento.

Rhizospheric and endophytic bacteria were isolated from the rizosphere and root tissue of Eucalyptus nitens. The objective of this work was to evaluate their capacity to promote growth in seedlings of the same species under greenhouse conditions. The isolates that improved seedling growth were identified and characterized by their capacity to produce indoleacetic acid (IAA), solubilize phosphates and increase 1-aminocyclopropane-1-carboxylate (ACC) deaminase activity. One hundred and five morphologically different strains were isolated, 15 of which promoted E. nitens seedling growth, significantly increasing the height (50%), root length (45%) as well as the aerial and root dry weight (142% and 135% respectively) of the plants. Bacteria belonged to the genus Arthrobacter, Lysinibacillus, Rahnella and Bacillus. Isolates A. phenanthrenivorans 21 and B. cereus 113 improved 3.15 times the emergence of E. nitens after 12 days, compared to control samples. Among isolated R. aquatilis, 78 showed the highest production of IAA (97.5±2.87 μg/ml) in the presence of tryptophan and the highest solubilizer index (2.4) for phosphorus, while B. amyloliquefaciens 60 isolate was positive for ACC deaminase activity. Our results reveal the potential of the studied rhizobacteria as promoters of emergence and seedling growth of E. nitens, and their possible use as PGPR inoculants, since they have more than one mechanism associated with plant growth promotion.

Crescimento de cultivares de arroz (Oryza sativa L. ) influenciado pela inoculação com bactérias promotoras de crescimento de plantas / Growth of rice cultivars (Oryza sativa L. ) as affected by inoculation with plant growth-promoting bacteria

Este trabalho objetivou avaliar o efeito da inoculação de seis estirpes de bactérias promotoras de crescimento de plantas (BPCP), Burkholderia spp. (B7 e B16), Pseudomonas spp. (P18 e P21) e Rhizobium spp. (R65 e R82) e duas misturas contendo uma estirpe de cada gênero no crescimento de seis cultivares de arroz (BRS Bonança, BRS Aimoré, BRS Talento, BRSMG Curinga, Japonês e Bolinha). As características fisiológicas das bactérias foram determinadas em ensaios in vitro. Foi conduzido um experimento sob condições de casa de vegetação onde as BPCP foram inoculadas em cultivares de arroz, cultivados em vasos contendo 1 kg de solo, em delineamento de blocos casualizados com 4 repetições. Aos 60 dias após a emergência foram avaliados parâmetros de crescimentos relacionados à parte aérea e ao sistema radicular dos cultivares de arroz. Entre as bactérias, somente duas (B7 e R65) não apresentaram capacidade de produção in vitro de auxina. Todas as estirpes apresentaram atividade de catalase e formação de película em meio NFb semi-sólido sem N ou presença de nódulo em feijoeiro, indicando capacidade de fixação biológica de nitrogênio. A inoculação das BPCP em arroz proporcionou aumentos médios de 10% em relação ao controle para a área foliar e massa seca da parte aérea e de 9% para o comprimento e massa seca de raiz. Entre os cultivares, BRS Aimoré apresentou maior interação com as estirpes estudadas e a estirpe P21 de Pseudomonas spp. proporcionou diferença significativa em relação ao controle em parâmetros relacionados à parte aérea e ao sistema radicular.

Aiming to evaluate the effect of the inoculation of six strains of plant growth-promoting bacteria (PGPBs) Burkholderia spp. (B7 and B16), Pseudomonas spp. (P18 and P21) and Rhizobium spp. (R65 and R82) and two mixtures containing one strain of each genera on the growth of six rice cultivars (BRS Bonança, BRS Aimoré, BRS Talento, BRSMG Curinga, Japonês and Bolinha). Physiological characteristics of the bacteria were determined on in vitro assays. Rice cultivars were inoculated with PGPB under greenhouse conditions and cultivated on 1 kg pots filled with soil on a randomized block design with four replicates. At 60 days after emergence growth parameters related with the shoot and root were evaluated for rice cultivars. Among the evaluated bacteria only two (B7 and R65) were not able to produce auxin under in vitro conditions. All strains showed catalase activity and film formation in semisolid medium NFb without N or nodulation in common bean, indicating the ability of biological nitrogen fixation. Inoculation of PGPBs in rice provided increases of 10% as compared to the control regarding leaf area and dry mass of shoots and 9% for length and root dry weight. Among the cultivars BRS Aimoré showed greater interaction with the strains studied and the strain P21 of Pseudomonas spp. promoted significant difference compared to the control for the parameters related with the shoot and root system.

Phenotypic and Genotypic Characteristics of Rhizobia Isolated from Meknes-tafilalet Soils and Study of Their Ability to Nodulate Bituminaria bituminosa.

Aims: The objectives were to isolate and characterize phenotypically and genotypically the rhizobial strains from the soils belonging to the Meknes-Tafilalet region in order to select strains that are able to nodulate Bituminaria bituminosa. Study Design: An experimental study. Place and Duration of Study: Department of biology (Soil & Environment Microbiology Unit) Faculty of Sciences, Moulay Ismail University and Technical Support Unit for Scientific Research, CNRST in Rabat; between January and August 2010. Methodology: Samples from 23 different sites belonging to the Meknes-Tafilalet region were collected in order to select rhizobial strains that are able to nodulate Bituminaria bituminosa. The morphological, cultural and phenotypic parameters of isolated strains were studied. The phenotypic characteristics include colony morphology, growth speed, tolerances to temperature, salt and pH. To assess the genotypic diversity among the isolates, molecular characteristics based on 16S rDNA gene sequencing were performed. Results: The majority of the isolated strains showed fast-growing capacity (75%). Most strains tolerate neutral to alkaline pH, however some strains (18%) showed weak growth capacity at pH 4. All isolates were tolerant to high salt stress ([NaCl] = 3%). The genotypic characterization based on16S rDNA gene sequencing of the twelve strains showed a high diversity between the isolates. Conclusion: Taken together, our results highlight the important biodiversity of the isolated rhizobial strains and open opportunities for the development of new bio-fertilizer.

Rhizobium radiobacter recovered from blood of a pediatric patient with stem cell transplantation: a case report and characterization of antimicrobial susceptibility profile

Rhizobium radiobacter is an uncommon agent of infection and has been associated with indwelling intravascular devices such as catheter in immunocompromised patients. Here, we report a case of R. radiobacter recovered from blood cultures in stem cell transplantation in a pediatric patient and present an extensive characterization of its antimicrobial susceptibility profile. The isolate presented low MICs to many antimicrobial agents, but high MICs to ceftazidime, piperacillin-tazobactam, aztreonam, and fosfomycin.

Induction and in vitro culture of hairy roots of Dianthus caryophyllus and its plant regeneration / 生物工程学报

To use Agrobacterium rhizogenes-induced hairy roots to create new germplasm of Dianthus caryophyllus, we transformed D. caryophyllus with A. rhizogenes by leaf disc for plant regeneration from hairy roots. The white hairy roots could be induced from the basal surface of leaf explants of D. caryophyllus 12 days after inoculation with A. rhizogenes ATCC15834. The percentage of the rooting leaf explants was about 90% 21 days after inoculation. The hairy roots could grow rapidly and autonomously in liquid or solid phytohormone-free MS medium. The transformation was confirmed by PCR amplification of rol gene of Ri plasmid and silica gel thin-layer chromatography of opines from D. caryophyllus hairy roots. Hairy roots could form light green callus after cultured on MS+6-BA 1.0-3.0 mg/L + NAA 0.1-0.2 mg/L for 15 days. The optimum medium for adventitious shoots formation was MS + 6-BA 2.0 mg/L + NAA 0.02 mg/L, where the rate of adventitious shoot induction was 100% after cultured for 6 weeks. The mean number of adventitious shoot per callus was 30-40. The adventitious shoots can form roots when cultured on phytohormone-free 1/2 MS or 1/2 MS +0.5 mg/L NAA for 10 days. When the rooted plantlets transplanted in the substrate mixed with perlite sand and peat (volume ratio of 1:2), the survival rate was above 95%.

Production of L(+)-tartaric acid by immobilized Rhizobium strain BK-20 / 生物工程学报

The cis-epoxysuccinate hydrolase (CESH) from Rhizobium strain BK-20 is the key enzyme for L(+)-tartaric acid production. To establish a highly efficient and stable production process, we first optimized the enzyme production from Rhizobium strain BK-20, and then developed an immobilized cell-culture process for sustained production of L(+)-tartaric acid. The enzyme activity of free cells reached (3 498.0 +/- 142.6) U/g, and increased by 643% after optimization. The enzyme activity of immobilized cells reached (2 817.2 +/- 226.7) U/g, under the optimal condition with sodium alginate as carrier, cell concentration at 10% (W/V) and gel concentration at 1.5% (W/V). The immobilized cells preserved high enzyme activity and normal structure after 10 repeated batches. The conversion rate of the substrate was more than 98%, indicating its excellent production stability.

Effectiveness of halo-tolerant, auxin producing Pseudomonas and Rhizobium strains to improve osmotic stress tolerance in mung bean (Vigna radiata L.)

Halo-tolerant, auxin producing bacteria could be used to induce salt tolerance in plants. A number of Rhizobium and auxin producing rhizobacterial strains were assessed for their ability to tolerate salt stress by conducting osmoadaptation assay. The selected strains were further screened for their ability to induce osmotic stress tolerance in mung bean seedlings under salt-stressed axenic conditions in growth pouch/jar trials. Three most effective strains of Rhizobium and Pseudomonas containing ACC-deaminase were evaluated in combination, for their ability to induce osmotic stress tolerance in mung bean at original, 4, and 6 dS m-1 under axenic conditions. Results showed that sole inoculation of Rhizobium and Pseudomonas strains improved the total dry matter up to 1.4, and 1.9 fold, respectively, while the increase in salt tolerance index was improved up to 1.3 and 2.0 fold by the Rhizobium and Pseudomonas strains, respectively. However, up to 2.2 fold increase in total dry matter and salt tolerance index was observed due to combined inoculation of Rhizobium and Pseudomonas strains. So, combined application of Rhizobium and Pseudomonas strains could be explored as an effective strategy to induce osmotic stress tolerance in mung bean.

Resposta de cultivares de feijoeiro-comum à inoculação das sementes com duas estirpes de rizóbio / Common bean cultivars response to seed inoculation with two rhizobium strains

Fixação simbiótica de nitrogênio pode ser uma alternativa para reduzir, ou mesmo substituir, a adubação nitrogenada do feijoeiro-comum, cultura na qual ainda há descrédito quanto aos benefícios da inoculação e que se caracteriza por aplicações de altas doses de N mineral. Com o objetivo de verificar a resposta de cultivares de feijoeiro-comum à inoculação das sementes com duas estirpes de rizóbio, foram conduzidos dois experimentos de campo na safra de inverno 2010 (Patos de Minas e Uberaba, MG). Utilizou-se o delineamento em blocos casualizados, com três repetições e esquema fatorial 8 x 3, envolvendo oito cultivares (União, Madrepérola, Supremo, Ouro Negro, Ouro Vermelho e Majestoso) e três tipos de inoculação (sementes não inoculadas e estirpes CIAT 899T e UFLA 04-173). Na floração, avaliaram-se o número e massa seca de nódulos, massa seca de parte aérea e teor e acúmulo de nitrogênio (N) na parte aérea e, na colheita, o rendimento de grãos com seus componentes primários, teor e acúmulo de N nos grãos. Constatou-se que as populações nativas promovem nodulação equivalente à das estirpes inoculadas. As estirpes CIAT 899T e UFLA 04-173 favorecem o crescimento vegetativo do feijoeiro. A estirpe CIAT 899T proporciona maior número de vagens e maior massa de grãos que a estirpe UFLA 04-173, e em Uberaba também proporciona maior rendimento de grãos. Quando a inoculação é realizada com a estirpe UFLA 04-173, as cultivares diferem quanto ao teor foliar de N. As cultivares Madrepérola, Ouro Negro e Ouro Vermelho apresentam mais nódulos e maior massa seca de parte aérea, além de maior produtividade em Patos de Minas. Ouro Negro situa-se entre as mais produtivas em Uberaba. Em Uberaba há maior acúmulo de matéria seca de nódulos e de parte aérea, mas o ambiente Patos de Minas resulta em maiores valores do rendimento de grãos e componentes.

The symbiotic nitrogen fixation can be an alternative to reduce, or even replace, the nitrogen (N) fertilization of the common bean crop, characterized by application of high doses of this nutrient. This work aimed to verify the response of common bean cultivars to the seed inoculation with two Rhizobium strains, conducting two field experiments in the winter-spring crop of 2010, at Patos de Minas and Uberaba, Minas Gerais State, Brazil. The experiment had a randomized block design, with three replications and a 8 x 3 factorial squemme, involving eight cultivars and three levels of inoculation. At flowering, it was evaluated the number of nodules, nodules dry mass, shoots dry weight, N content and accumulation in shoots and, at harvest, the grain yield and its primary components (100 grains weight, number of pods per plant, number of grains per pod) and the N content and accumulation in the grains. The Madrepérola, Ouro Negro and Ouro Vermelho cultivars present the highest number of nodules and higher dry mass; in Patos de Minas, the same cultivars show higher productivity, but in Uberaba, only Ouro Negro cultivar is among the most productive. In Uberaba there is the greater nodules dry weight and higher shoots dry matter, but the most favorable conditions in Patos de Minas results in higher values for grain yield and its components. In Uberaba, the CIAT 899T strain overcomes the UFLA 04-173 strain in relation to grain yield. In both environments, the CIAT 899T strain also shows superiority in number of pods per plant and weight of one hundred grains.