Isolation and partial characterization of endophytic diazotrophs associated with Japanese sugarcane cultivar (original) (raw)
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The numbers of culturable diazotrophic endophytic bacteria (CDEB) from roots, stems and leaves of sugarcane submitted to organic, inorganic or no fertilization were compared. In order to determine the size of the N2 fixing populations, the Most Probable Number technique (MPN) was used. The quantification of diazotrophic bacteria by using the acetylene reduction assay (ARA) was more accurate than observing the bacterial growth in the vials; to confirm N2 fixing capability, the detection of gene nifH was performed on a sample of 105 isolated bacteria. The production of extracellular enzymes involved in the penetration of the plants by the bacteria was also studied. The results showed that organic fertilization enhances the number of CDEB when compared with conventional fertilization used throughout the growing season. The maximum number of bacteria was detected in the roots. Roots and stems presented the greatest number of CDEB in the middle of the cropping season and in leaves numbers varied according to the treatment. Using two pairs of primers and two different methods, the nifH gene was found in 104 of the 105 tested isolates. Larger amounts of pectinase were released by isolates from sugarcane treated with conventional fertilizers (66%), whereas larger amounts of cellulase were released by strains isolated from sugarcane treated with organic fertilizers (80%).
Journal of Experimental Agriculture International, 2018
The bioprospection of microorganisms with the potential to promote sugarcane growth has been increasingly sought, mainly to minimize the insertion of inputs that increase the costs of the production chain. The objective of this work was to isolate, identify and select endophytic bacteria from sugarcane with potential ability for plant growth promotion. Also to evaluate the influence of different levels of salinity and pH values on plant growth-promotion and to analyze the genetic diversity of the isolates by the partial sequencing of the 16S rRNA gene. Samples of stems and Original Research Article
Applied and environmental microbiology, 1995
Nitrogen-fixing bacteria, isolated from the sugar solution in intercellular spaces of sugarcane stems, were compared with the type strain of Acetobacter diazotrophicus (PAL-5) and found to be congruent with it in all characters studied. These characters were 37 morphological and biochemical tests, cellular fatty acid composition, and nitrogenase activity. The nitrogenase activity was measured by acetylene reduction and H(inf2) evolution and found to be unusual in that the H(inf2) evolution was suppressed much less than expected by high concentrations of acetylene.
Brazilian Archives of Biology and Technology, 2022
Sugarcane is an important Brazilian commodity, being usually cultivated in soils with low natural fertility. This study aimed to isolate diazotrophic endophytes from sugarcane tissues and evaluate the morphological and physiological characteristics of their colonies as well as their plant growth-promoting (PGP) traits in select diazotrophic endophytic bacteria. Fifty-six bacterial isolates were identified in the sugarcane tissues, and these isolates presented distinct morphological and physiological traits. A total of thirty-five bacterial isolates were biochemically evaluated. Overall, Bacillus was the dominant genus. Isolates of Methylobacterium spp. and Brevibacillus agri were present only in leaves, while Herbaspirillum HIGHLIGHTS • The diazotrophic endophytic community found in stems were different those found in leaves of sugarcane. • Endophytes with valuable biochemical traits have been identified in the sugarcane tissues. • A unique symbiotic consortium between endophytes were identified in the sugarcane tissues.
Plant and Soil
To identify active diazotrophs in sugarcane, 16S rRNA and nifH transcript analyses were applied. This should help to better understand the basis of the biological nitrogen fixation (BNF) activity of a high nitrogen fixing sugarcane variety. A field experiment using the sugarcane variety RB 867515 was conducted in Seropédica, RJ, Brazil, receiving the following treatments: unfertilised and fertilised controls without inoculation, unfertilised with inoculation. The five-strain mixture developed by EMBRAPA-CNPAB was used as inoculum. Root and leaf sheath samples were harvested in the third year of cultivation to analyse the 16S rRNA and nifH transcript diversity. In addition to nifH expression from Gluconacetobacter spp. and Burkholderia spp., a wide diversity of nifH sequences from previously uncharacterised Ideonella/Herbaspirillum related phylotypes in sugarcane shoots as well as Bradyrhizobium sp. and Rhizobium sp. in roots was found. These results were confirmed using 16S cDNA analysis. From the inoculated bacteria, only nifH transcripts from G. diazotrophicus and B. tropica were detected in leaf sheaths and roots. Known as well as yet uncultivated diazotrophs were found active in sugarcane roots and stems using molecular analyses. Two strains of the inoculum mix were identified at the late summer harvest.
Anais da Academia Brasileira de Ciências
The growing need for the use of clean energy has led society to seek alternatives to fossil fuels in order to less pollute the environment. Sugarcane has been known to produce enough biomass and to have associative microorganisms, such as plant growth-promoting bacteria (PgPB), which have the potential to improve the yield of crops. The objective of this study is to evaluate the efficiency and the isotopic dilution of 15 N of microbial inoculants containing endophytic diazotrophic bacteria, individually inoculated and in mixture, on productivity, technological quality and biological nitrogen fixation (BNF) of sugarcane variety RB92579. The experiment was conducted for three consecutive years, comprising different crop cycles, in a completely randomized block design with four replications. The evaluations were at 11 months after planting for plant-cane, and at 12 months after cutting for ratoon cane and second ratoon cane. The contribution of BNF by 15 N natural abundance technique to inoculated treatments ranged from 18% to 57.31%. This study showed that no strain promoted the improvement on yield and biological nitrogen fixation during the three cycles of sugarcane (plant-cane, ratoon cane and second ratoon cane). However, industrial characteristics of sugar cane can be affectd by PGPB inoculation.
2001
Twenty-two endophytic bacterial isolates from the roots of sugarcane were compared morphologically, biochemically and genetically. Gram staining, colony pigment, texture and other cultural characteristics were taken for morphological characterization. Oxidation-fermentation tests for D -glucose and D -sucrose, production of acid and hydrogen from different carbon source, oxidase activity, antibiotic and drug resistance patterns were chosen as the biochemical and physiological criteria. Twelve random decamer primers were used to analyze and compare these isolates through RAPD among themselves as well as with known standard diazotrophic strains. The isolates were compared through dendrograms constructed on the basis of similarity patterns obtained from biochemical and RAPD analysis. The estimated diversity through RAPD analysis was more evident than the diversity on the basis of morphological and biochemical characters. Within Acetobacter group, the isolates showed substantial genetic diversity for future exploitation as PGPRs and diazotrophic associative endophytes.
Evaluation of Performance of Sugarcane Genotypes Inoculated with Endophytic Diazotrophic Bacteria
ainfo.cnptia.embrapa.br
The present experiment was aimed to evaluate the response of sugarcane varieties inoculated with endophytic diazotrophic bacteria. Sixteen sugarcane varieties were used for three kind of treatrnents (no inoculation and inoculation with 120 kg N/hectare). The inoculant was a mixture of five strains of bacterial species isolated from sugarcane. There were significant differences between genotypes (G) and genotype x treatment interaction (GxT) for ali the traits. The variety SP 71-6949,SP 78-4764and RB 867515were found responsive to biological nitrogen fixation and can be used in breeding programs which are directed to study and selection of promising genotypes responsible for the biological nitrogen fixation.
International Journal of Systematic Bacteriology, 1989
Results of deoxyribonucleic acid (DNA)-ribosomal ribonucleic acid and DNA-DNA hybridizations, together with a phenotypic and chemotaxonomic analysis, revealed that nitrogen-fixing bacteria isolated from roots and stems of sugarcane belong to a new species in the genus Acetobacter, for which the name Acetobacter diazotrophicus sp. nov. is proposed. Strain LMG 7603 (= Dobereiner PA1 5 = ATCC 49037) is the type strain. New microaerobic, gram-negative, N,-fixing bacteria were isolated from roots and stems of sugarcane in Brazil (1). Because of their ability to grow at low pH values and their ability to form acetic acid from ethanol, these organisms could belong to the acetic acid bacteria (the genera