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The book explains background knowledge about rhizobia and follows this up with a broad perspective on rhizobial diversity, information on characteristics specific to each group of rhizobia, the relationship among rhizobial groups as well as genetic factors contributed to rhizobial diversity.

Rhizobiaceae. --- Molecular microbiology. --- Microorganisms --- Molecular bacteriology --- Microbiology --- Molecular biology --- Aerobic bacteria --- Gram-negative bacteria --- Nitrifying bacteria --- Molecular aspects

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Agrobacterium Protocols offers beginning and experienced researchers the most comprehensive collection of step-by-step protocols for the genetic manipulation of plants using Agrobacterium. The topics range from the maintenance of bacterial culture collections to aspects of the metabolism and physiology of transformed tissues and transgenic plants. Drawing on the work of leading scientists from laboratories around the world, Agrobacterium Protocols provides a wealth of techniques for introducing specific DNA sequences into target plant species and discusses the environmental implications of genetically engineered plants. Its detailed procedures will facilitate rapid transfer of advanced techniques to other laboratories and their exploitation in fundamental and applied plant biology.

Agricultural biotechnology --- Agrobacterium --- Agricultural biotechnology. --- Agrobacterium. --- Agro-biotechnology --- Biotechnology --- Polymonas --- Rhizobiaceae --- Cytology. --- Cell Biology. --- Cell biology --- Cellular biology --- Biology --- Cells --- Cytologists

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This volume reviews various facets of Agrobacterium biology, from modern aspects of taxonomy and bacterial ecology to pathogenesis, bacterial cell biology, plant and fungal transformation, natural transgenics, and biotechnology. Agrobacterium-mediated transformation is the most extensively utilized platform for generating transgenic plants, but modern biotechnology applications derive from more than 40 years of intensive basic scientific research. Many of the biological principles established by this research have served as models for other bacteria, including human and animal pathogens. Written by leading experts and highlighting recent advances, this volume serves both as an introduction to Agrobacterium biology for students as well as a more comprehensive text for research scientists.

Agrobacterium. --- Polymonas --- Rhizobiaceae --- Microbiology. --- Microbial genetics. --- Microbial genomics. --- Agriculture. --- Medical Microbiology. --- Microbial Genetics and Genomics. --- Farming --- Husbandry --- Industrial arts --- Life sciences --- Food supply --- Land use, Rural --- Genomics --- Microbial genetics --- Microorganisms --- Genetics --- Microbiology --- Microbial biology --- Biology --- Medical microbiology.

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De symbiose tussen planten van de familie Leguminosae en sommige bacteriën wordt gekenmerkt door de vorming van nieuwe plantorganen, de zogenoemde nodules of wortelknolletjes, op de wortels of stengel. De cellen van deze nieuwe organen worden geïnfecteerd door de specifieke microsymbionten. De best gekende symbiotische bacteriën behoren tot de groep van de Rhizobiaceae met de genera Rhizobium, Bradyrhizobium, Sinorhizobium (Ensifer), Mesorhizobium, Azorhizobium , en Allorhizobium , collectief rhizobia genoemd. Vlinderbloemige planten zijn een belangrijke schakel in duurzame landbouw. Symbiotische stikstoffixatie (SNF), door de interactie van vlinderbloemige planten en rhizobia, draagt in belangrijke mate bij aan de stikstofvoeding van deze planten en als dusdanig tot de stikstofhuishouding in plantaardige productiesystemen. De gewone boon ( Phaseolus vulgaris L.) is wereldwijd het belangrijkste vlinderbloemige gewas voor humane consumptie en traditioneel van bijzonder belang in grote delen van Centraal- en Zuid-Amerika en Afrika. Symbiotische stikstoffixatie bij bonenteelt is echter weinig efficiënt in vergelijking met andere vlinderbloemige gewassen. Het opzet van deze studie was het identificeren en kwantificeren van gunstige interacties tussen boongenotypes, rhizobia en plantengroeibevorderende bacteriën (PGPR). Hiertoe werden verschillende rhizobia-PGPR combinaties geëvalueerd via inoculatie van twee boongenotypes die courant gebruikt worden door de boeren in Cuba. Nodulatie- en plantgroeiparameters worden gunstig beïnvloed door co-inoculatie van gewone boon met Rhizobium-Azospirillum of Rhizobium-Azotobacter , en dit zowel in potexperimenten als onder veldcondites. Onder veldcondities werd echter duidelijk een effect van het plantgenotype waargenomen wat betreft groei en opbrengst. De combinatie Rhizobium-Azospirillum en de bemestingscontrole leverden de beste resultaten op voor de ICA Pijao variëteit, terwijl voor de BAT-304 variëteit de beste resultaten bekomen werden met enkelvoudige Rhizobium inoculatie. Morfologische en genetische karakterisatie van bacteriën geïsoleerd uit boonvelden op Cuba lieten toe een, weliswaar beperkt, beeld te geven van de bacteriële diversiteit. Sommige van deze geïsoleerde bacteriën werden getest voor hun interactie met de gewone boon. Geïsoleerde Rhizobium stammen bleken een aantal interessante eigenschappen, zoals vroege nodulatie, te vertonen. Bovendien werden in co-inoculatietesten gunstige effecten waargenomen, wat wijst op een compatibiliteit tussen rhizobia en sommige PGPR in de interactie met de gewone boon. De genetische karakterisatie van de bacteriële isolaten uit de Cubaanse bodems leidde via 16SrDNA sequenering tot de identificatie van 8 genera: Agr obacterium, Rhizobium, Ochrobactrum, Sphingomonas, Stenotrophomonas, Bacillus, Brevibacillus en Paenibacillus . In stalen genomen van nodules vertoonden 37,5% van de isolaten 100% gelijkenis met hetzij Agrobacterium tumefaciens of Rhizobium species. Twee Rhizobium species werden geïdentificeerd, met name Rhizobium et li en Rhizobium tropici . In een laatste deel werd een bijdrage geleverd in het zoeken naar plantengenen die in de interactie van de gewone boon met respectievelijk Rhizobium , een pathogene schimmel ( Fusarium solani f. sp. phaseoli ), en een controlebehandeling, differentiëel tot expressie komen. Hiertoe werd de techniek van “cDNA-Amplified Fragment Length Polymorphism (cDNA-AFLP)” gebruikt. In silico analyse van een aantal geïdentificeerde DNA fragmenten leverde een aantal interessante “transcript derived fragments (TDFs)” op. DNA sequentie-analyse van een aantal van deze TDFs liet toe verwantschap op te sporen met reeds bekende genen. Deze verwante genen bleken betrokken te zijn in stressresponses en koolhydraatmetabolisme. The symbiosis between plants of the Leguminosae family and prokaryotic partners is typically characterized by the formation of specialized organs, called nodules, on plant roots or stems that are invaded by the specific microsymbionts. These include the well-known alpha-proteobacterial group of Rhizobiaceae containing the genera Rhizobium, Bradyrhizobium, Sinorhizobium ( Ensifer ), Mesorhizobium, Azorhizobium, and Allorhizobium , collectively referred as rhizobia. Legumes play a crucial role in sustainable agriculture. Symbiotic nitrogen fixation (SNF) through interaction between legumes and rhizobia, contributes to nitrogen (N) nutrition of most legumes and legume cropping systems. Common bean ( Phaseolus vulgaris L.) is the most important legume for direct human consumption worldwide and particularly in many parts of Latin America and Africa. However, the application of SNF in common bean in the field is often low compared to the nitrogen fixing capacity of beans under optimal conditions and as compared to the amounts of nitrogen fixed by other legumes. The aim of our study is to identify, quantify and enhance the phytostimulatory effect of the interplay between Rhizobium , bean genotypes and plant growth promoting rhizobacteria (PGPR) under different growth conditions and to contribute to the understanding of the molecular mechanisms involved in the Rhizobium -bean interaction. To reach this objective, combinations of Rhizobium -PGPR were evaluated under different growth conditions in Cuba using two local bean genotypes. The nodulation and plant growth parameters were significantly stimulated with the combination of Rhizobium-Azospirillum and Rhizobium-Azotobacter under pot experiment condition, as well as in a field trial. Variations among genotypes were observed for growth parameters and yield in a second field trial. The combination Rhizobium-Azospirillum and the fertilizer treatments showed the best result in yield for ICA Pijao beans, while for BAT-304 beans the best result was obtained with the single Rhizobium inoculation. Secondly, the morphological and genetic characterization of bacterial isolates from Cuban bean fields, as well as the phenotypic characterization of Cuban Rhizobium isolates under controlled and field conditions, demonstrate the biodiversity of beneficial microbes in the common bean rhizosphere and the stimulatory effect of compatible interactions between common bean genotypes and Rhizobium strains. The genetic characterization of isolated bacterial strains form Cuban soils using 16S rDNA sequencing revealed 8 groups of bacteria belonging to the genera: Agrobacterium, Rhizobium, Ochrobactrum, Sphingomonas, Stenotrophomonas, Bacillus, Brevibacillus and Paenibacillus . In nodule samples, 37.5% of isolates were 100% similar to Agrobacterium tumefaciens or Rhizobium species. This study allowed the identification of two species of Rhizobium isolates ( Rhizhobium etli and Rhizobium tropici ) in nodule samples. In nodulation tests Agrobacterium isolates were unable to nodulate the original host. The phenotypic characterization showed the stimulation of nodulation parameters and the N fixation through the native Rhizobium isolates at early stage of common bean plants. Under field trial conditions, the nodulation, growth parameters and yield were stimulated significantly for ICA Pijao as compared with BAT-304 upon inoculation with the isolated Rhizobium strains. Furthermore, genes differentially expressed during the bean root interaction with Rhizobium etli CNPAF512, infection with Fusarium solani f. sp. phaseoli and a control respectively , were identified using the cDNA-Amplified Fragment Length Polymorphism (cDNA-AFLP) technique. In silico analysis was used to determine the differential expression profiles of transcript derived fragments (TDFs). Several TDFs were isolated, cloned, sequenced and the obtained DNA sequences were compared with sequences in the GenBank database. The sequences retrieved revealed homology with genes encoding stress/defense and cell metabolism functions for Rhizobium treatments, as well as stress/defense functions for the Fusarium condition. The results outlined in this study demonstrate the potential of selection for efficient associations among bean genotypes, rhizobia and plant growth promoting rhizobacteria in order to achieve the increase of SNF in common bean under local agro-ecosystems, as well as increase our insight of the molecular dialogue in common bean-rhizobia interaction. However, these studies should be expanded using more bean genotypes and bacterial combinations in different environmental conditions, in order to provide recommendations to farmers. Biologische stikstoffixatie (BNF) is bij uitstek een proces dat bijdraagt tot een meer duurzame landbouw. Biologische stikstoffixatie in symbiose (SNF) met vlinderbloemige planten, zoals bonen, wordt daarom reeds decennia toegepast, voornamelijk in landen die om economische redenen weinig toegang hebben tot chemische meststoffen. Symbiose verwijst naar een interactie tussen de wortels, en in sommige gevallen de stengels, van vlinderbloemige planten enerzijds en bacteriën die collectief worden aangeduid als rhizobia anderzijds. In een aantal gevallen voorziet dit proces in de volledige stikstofbehoefte van de plant. De gewone boon, Phaseolus vulgaris L., is wereldwijd de belangrijkste vlinderbloemige plant voor humane consumptie, en van groot belang in grote delen van Zuid- en Centraal-Amerika en Afrika. De symbiotische stikstoffixatie bij boon is echter weinig efficiënt onder veldcondities, in vergelijking met andere vlinderbloemige gewassen. Daarentegen, onder goed gecontroleerde omstandigheden kan SNF bij boon tot 80% van de stikstofbehoefte van de plant voorzien. Dit wijst enerzijds op het genetisch potentiëel voor efficiënte stikstoffixatie en anderzijds op het voorkomen van ongunstige omgevingsfactoren om dit potentiëel effectief te realiseren. Het doctoraatsonderzoek beoogde de interactie tussen boongenotypes, rhizobia en plantengroeibevorderende rhizobacteriën (PGPR) te bestuderen en te kwantificeren, en dit met het oog op het verbeteren van de symbiotische stikstoffixatie. Er werd hierbij uitgegaan van actuele landbouwsystemen in Cuba. Hiertoe werden verschillende rhizobia-PGPR combinaties geëvalueerd via inoculatie van twee boongenotypes die courant gebruikt worden door de boeren in Cuba. Nodulatie- en plantgroeiparameters worden gunstig beïnvloed door co-inoculatie van gewone boon met Rhizobium - Azospirillum of Rhizobium-Azotobacter , en dit zowel in potexperimenten als onder veldcondites. Onder veldcondities werd echter duidelijk een effect van het plantgenotype waargenomen wat betreft groei en opbrengst. De combinatie Rhizobium-Azospirillum en de bemestingscontrole leverden de beste resultaten op voor de ICA Pijao variëteit, terwijl voor de BAT-304 variëteit de beste resultaten bekomen werden met enkelvoudige Rhizobium inoculatie. Morfologische en genetische karakterisatie van bacteriën geïsoleerd uit boonvelden op Cuba lieten toe een, weliswaar beperkt, beeld te geven van de bacteriële diversiteit. Sommige van deze geïsoleerde bacteriën werden getest voor hun interactie met de gewone boon. Geïsoleerde Rhizobium stammen bleken een aantal interessante eigenschappen, zoals vroege nodulatie, te vertonen. Bovendien werden in co-inoculatietesten gunstige effecten waargenomen, wat wijst op een compatibiliteit tussen rhizobia en sommige PGPR in de interactie met de gewone boon. De genetische karakterisatie van de bacteriële isolaten uit de Cubaanse bodems leidde via 16SrDNA sequenering tot de identificatie van 8 genera: Agrobacterium, Rhizobium, Ochrobactrum , Sphingomonas, Stenotrophomonas, Bacillus, Brevibacillus en Paenibacillus . Twee Rhizobium species werden geïdentificeerd, met name Rhizobium etli en Rhizobium tropici . In een laatste deel werd een bijdrage geleverd in het zoeken naar plantengenen die in de interactie van de gewone boon met respectievelijk Rhizobium , een pathogene schimmel ( Fusarium solani f. sp. phaseoli ), en een controlebehandeling, differentiëel tot expressie komen. Hiertoe werd de techniek van “cDNA-Amplified Fragment Length Polymorphism (cDNA-AFLP)” gebruikt. In silico analyse van een aantal geïdentificeerde DNA fragmenten leverde een aantal interessante “transcript derived fragments (TDFs)” op. DNA sequentie-analyse van een aantal van deze TDFs liet toe verwantschap op te sporen met reeds bekende genen. Deze verwante genen bleken betrokken te zijn in stressresponses en koolhydraatmetabolisme. International emphasis on environmentally sustainable development with the use of renewable resources is likely to focus attention on the potential role of biological nitrogen (N) fixation (BNF) in supplying N for agriculture to counteract the indiscriminate use of nitrogenous fertilizers, which has resulted in unacceptable levels of pollution to groundwater and the atmosphere, low crop yields, decrease in the protein content of food and declined soil fertility. It is largely known that legumes play a crucial role in sustainable agriculture. Symbiotic N fixation (SNF) through interaction between legumes and micro-organisms collectively named rhizobia, contributes to N nutrition of most legumes and legume cropping systems. Common bean ( Phaseolus vulgaris L.) is the most important legume for direct human consumption worldwide and particularly in many parts of Latin America and Africa. However, the application of SNF in common bean in the field is often low compared to the N fixing capacity of beans under optimal conditions and as compared to the amounts of N fixed by other legumes. This doctoral thesis aims to identify, quantify and enhance the phytostimulatory effect of the interplay between Rhizobium , bean genotypes and plant growth promoting rhizobacteria (PGPR) under different growth conditions and to contribute to the understanding of the molecular mechanisms involved in the Rhizobium -bean interaction. Combinations of Rhizobium -PGPR were evaluated under different growth conditions in Cuba using two local bean genotypes. The nodulation and plant growth parameters were significantly stimulated with the combination of Rhizobium-Azospirillum and Rhizobium-Azotobacter under pot-controlled condition, as well as in a field trial. The combination Rhizobium-Azospirillum and the fertilizer treatments showed the best result in yield for ICA Pijao beans, while for BAT-304 beans the best result was obtained with the single Rhizobium inoculation. Secondly, the morphological and genetic characterization of bacterial isolates from Cuban intercropping bean fields, demonstrate the biodiversity of beneficial microbes in the common bean-rhizosphere, revealing 8 groups of bacteria detected by 16S rDNA belonging to the genera: Agrobacterium, Rhizobium , Ochrobactrum, Sphingomonas, Stenotrophomonas, Bacillus, Brevibacillus and Paenibacillus . This study allowed the identification of two species of Rhizobium isolates ( Rhizhobium etli and Rhizobium tropici ) in nodule samples. In nodulation tests Agrobacterium isolates were unable to nodulate the original host. The phenotypic characterization of Rhizobium isolates under growth controlled and field conditions showed stimulatory effect of compatible interactions between common bean genotypes and Rhizobium strains. The nodulation parameters and the N fixation were increased through the native Rhizobium isolates at early stage of common bean plants and under field conditions the nodulation, growth parameters and yield were stimulated significantly for ICA Pijao as compared with BAT-304 upon inoculation with the isolated Rhizobium strains. However, Rhizobium tropici isolated strain RL-2 showed the best result in yield as compared with the control, reference strain CIAT899 and fertilizer treatments. Furthermore, genes differentially expressed during the root interaction with Rhizobium etli CNPAF512 in common bean (BAT-477) were identified using the cDNA-Amplified Fragment Length Polymorphism (cDNA-AFLP) technique, having as control conditions the infection with Fusarium solani f. sp. phaseoli and a treatment without rhizobia inoculation or Fusarium infection . In silico analysis was used to determine the differential expression profiles of transcript derived fragments (TDFs). The sequences retrieved revealed homology with genes encoding stress/defense and cell metabolism functions for Rhizobium treatments, as well as stress/defense functions for the Fusarium condition. However the studies should continued to verify the genes detected by qRT-PCR. The results outlined in this study demonstrate the potential of selection for efficient associations among bean genotypes, rhizobia and plant growth promoting rhizobacteria in order to achieve SNF in common bean under local agro-ecosystems, as well as to increase our insight in molecular signal transduction pathways in Rhizobium -bean interplay.

Phaseolus vulgaris --- Génotype --- genotypes --- Rhizobium --- phenotypes --- Contrôle de croissance --- growth control --- Substance de croissance végétale --- plant growth substances --- ADN --- DNA --- Code génétique --- genetic code --- Cuba --- Academic collection --- 575 --- 66.098 --- 579.841.3 --- 631.847 --- 635.652 --- General genetics. General cytogenetics. Immunogenetics. Evolution. Speciation. Phylogeny --- Biological processes. Biotechnology --- Rhizobiaceae --- Vegetable nitrogen fertilizers. Bacterial and mycorrhizal inoculation of soil --- French bean. Green beans. Haricot beans. Phaseolus vulgaris --- Theses --- 631.847 Vegetable nitrogen fertilizers. Bacterial and mycorrhizal inoculation of soil --- 579.841.3 Rhizobiaceae --- 66.098 Biological processes. Biotechnology --- 575 General genetics. General cytogenetics. Immunogenetics. Evolution. Speciation. Phylogeny --- 635.652 French bean. Green beans. Haricot beans. Phaseolus vulgaris --- DNA.

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Agrobacterium is the only cellular organism on Earth that is naturally capable of transferring genetic material between the kingdoms of life, from prokaryotes to eukaryotes. Studies have uncovered a wealth of information on the process of Agrobacterium-mediated genetic transformation and on the bacterial and host cell factors involved in the infection. Agrobacterium has been shown to genetically transform, under laboratory conditions a large number of plant species and numerous non-plant organisms, indicating the truly basic nature of the transformation process. It is therefore not surprising that Agrobacterium and the genetic transformation itself have also become the focus of numerous ethical and legal debates. ‘Agrobacterium’ is a comprehensive book on Agrobacterium research, including its history, application, basic biology discoveries, and effects on human society. Although the book largely focuses on providing a detailed review of virtually all molecular events of the genetic transformation process, it also provides coverage of ethical and legal issues relevant to the use of Agrobacterium as a "genetic transformation machine". The result is an all-inclusive text which readers—including scientists and students involved in plant genetic engineering—will find useful as a reference source for all major aspects of the Agrobacterium-mediated genetic transformation of plant and non-plant organisms. About the Editors: Dr. Tzvi Tzfira is an Assistant Professor in the Department of Molecular, Cellular, and Developmental Biology at the University of Michigan. Dr. Vitaly Citovsky is a Professor in the Department of Biochemistry and Cell Biology at Stony Brook University.

Agrobacterium. --- Polymonas --- Rhizobiaceae --- Botany. --- Bacteriology. --- Biotechnology. --- Plant genetics. --- Plant diseases. --- Agriculture. --- Plant Sciences. --- Plant Genetics and Genomics. --- Plant Pathology. --- Farming --- Husbandry --- Industrial arts --- Life sciences --- Food supply --- Land use, Rural --- Botany --- Communicable diseases in plants --- Crop diseases --- Crops --- Diseases of plants --- Microbial diseases in plants --- Pathological botany --- Pathology, Vegetable --- Phytopathology --- Plant pathology --- Plants --- Vegetable pathology --- Agricultural pests --- Crop losses --- Diseased plants --- Phytopathogenic microorganisms --- Plant pathologists --- Plant quarantine --- Genetics --- Chemical engineering --- Genetic engineering --- Microbiology --- Botanical science --- Phytobiology --- Phytography --- Phytology --- Plant biology --- Plant science --- Biology --- Natural history --- Pathology --- Diseases and pests --- Diseases --- Wounds and injuries --- Plant science. --- Plant pathology. --- Floristic botany