This study was undertaken to characterize the nod and sdh operons of Bradyrhizobium japonicum and Sinorhizobium meliloti. Ten putative B. japonicum mutants with altered nod gene induction characteristics were isolated by screening mutants for genistein-independent nod gene expression. The mutants were found to have higher nodY expression than the wild-type in the presence of genistein. The increased sensitivity of all mutants to genistein was more apparent under suboptimal inducer concentration (0.1muM) and/or temperature (15°C). The expression of nodY gene induction was determined for five strains (Bj30050, 53, 56, 57, 58) under different temperature and inducer conditions. These five strains were also found to produce more lipochitooligosaccharide than the wild-type, at both 25°C and 15°C. Three of the ten mutant strains (including Bj30056 and 57) were unable to fix nitrogen with soybeans grown at optimal temperatures. Based on nodY gene expression and symbiotic phenotype the B. japonicum mutants were classified into three groups. / A molecular genetic approach was taken to investigate the regulation of expression of succinate dehydrogenase (SDH) in S. meliloti. The sdhCDAB genes encoding SDH were shown by RT-PCR to be co-transcribed and thus constitute an operon. The transcriptional start site and putative promoter region of the first gene in the operon, sdhC , were identified by 5'-RACE and DNA sequence analysis. Transcriptional lacZ fusions to sdhC indicated that expression of the operon is regulated by carbon source in the growth medium but not by growth phase. The highest expression of the sdh operon was observed in cells grown with acetate, arabinose and glutamate, as sole carbon sources, and the lowest expression was observed in cells grown with glucose and pyruvate as sole carbon sources. / Also presented is the isolation and characterization of the first defined sdh mutant in a rhizobial species. The mutants helped demonstrate that the total lack of SDH activity would be lethal to S. meliloti cells. Symbiotic phenotype of the mutants indicated that SDH is required for N2-fixation.

No description available.

Twelve F(,1) families of alfalfa (Medicago sativa L.) plants having different potential for nitrogenase activity, and the two parental populations were tested for response to mycorrhizal inoculation in a low-phosphate soil mixture in the greenhouse. The purpose of this study was to: (a) determine the effects of vesicular-arbuscular mycorrhizae on growth, nutrition and nodulation of these 14 populations, (b) determine if differences existed between the populations with regard to several morphological characteristics, and (c) determine if certain characteristics can be transmitted across generations. The 14 populations were evaluated under four treatments: control no Myorrhizae, no Rhizobium; Rhizobium alone; Mycorrhizae alone; and the combination of Mycorrhizae and Rhizobium. The growth parameters measured differed significantly among the treatments and among the 14 populations studied, and no significant interaction between the populations and the treatments were found. The dual Mycorrhizae and Rhizobium treatment significantly increased plant height at 30 days and 60 days after planting, leaf area per plant, and plant top-dry-weight at two harvest dates. Mycorrhizal inoculation, however, decreased specific-leaf-weight significantly. Nitrogen fixation parameters such as nodule mass score, fibrous root score, and nitrogenase activity were increased significantly by the dual inoculation of mycorrhizae and rhizobium. The 14 populations differed significantly in nodule mass score, fibrous root score, and nitrogenase activity. Mycorrhizal inoculation increased nitrogen fixation more than plant growth. Correlation coefficients indicated that increased Nitrogenase activity is positively correlated with increased nodule mass, increased fibrous root mass, greater top-dry-weight, and leaf area. A step wise multiple regression showed that 49% of the variation in nitrogenase activity can be explained by the variation due to nodule mass, fibrous root mass, top-dry-weight, and leaf area. Several morphological characters showed a heritable response. Plants selected for high nitrogenase activity and high top dry weight transmitted these characteristics to their progenies.

A existência de rizóbios nativos eficientes na fixação simbiótica do nitrogênio em leguminosas introduzidas nos campos do Rio Grande do Sul, como as forrageiras do gênero Lotus, é um fator determinante para adaptação ou o fracasso destas novas espécies. Este trabalho visou estudar a compatibilidade simbiótica de rizóbios simbiontes em Desmodium incanum, uma planta nativa dos campos do Rio Grande do Sul, e em L. corniculatus, L. uliginosus, L. glaber e L. subbiflorus. Para isto 35 isolados e 17 estirpes de rizóbios foram estudadas. Os isolados foram caracterizados fenotípicamente quanto a morfologia colonial, produção de ácido indol-acético e sideróforos. Os rizóbios foram estudados quanto a compatibilidade simbiótica pela inoculação cruzada com as espécies de plantas estudadas. Observou-se existem rizóbios nativos dos solos do Rio Grande do Sul capazes de produzir auxinas e sideróforos. Os rizóbios estudados neste trabalho foram sensíveis a salinidade. Também observou-se que todos os rizóbios isolados de Lotus foram capazes de induzir nodulação em pelo menos duas das cinco espécies estudadas. Os rizóbios de L. uliginosus que apresentaram compatibilidade com as cinco espécies de plantas estudadas, já os de D. incanum mostraram alta especificidade. Em experimento em casa de vegetação as plantas de L. corniculatus inoculadas com os o isolado UFRGS Lu 2 e as estirpes EEL 698 e SEMIA 816 produziram maior massa seca da parte aérea. Este isolado também foi eficiente em plantas de D. incanum. Os resultados deste trabalho mostram que existem rizóbios autóctones dos solos do Rio Grande do Sul que apresentam baixa especificidade hospedeira sendo capazes de formar simbiose com plantas de D. incanum, L. corniculatus, L. subbiflorus, L. glaber e L. uliginosus. A compatibilidade simbiótica com diferentes hospedeiros dificultou o estabelecimento de grupos de compatibilidade. A técnica de REP-PCR com o oligonucleotídeo iniciador BOX A1 mostrou que existe variabilidade entre os rizóbios estudados, sendo estes, diferentes das estirpes fornecidas pelas coleções de cultura. / The presence of native Rhizobia which are efficient in symbiotic nitrogen fixation in legumes introduced in the fields of Rio Grande do Sul, such as the forages plants of the genus Lotus, is one of the factors determining the success or the failure of the adaptation of these new plant species. This work aimed to evaluate the symbiotic compatibility of Rhizobia in association with Desmodium incanum, a native plant from the fields of Rio Grande do Sul, as well as with L. corniculatus, L. uliginosus, L. glaber and L. subbiflorus species. For this, 35 isolates and 17 lineages of Rhizobia were analyzed. The isolates were characterized phenotypically based on the colony morphology and on the indole acetic acid and siderophore production. The Rhizobia were studied for symbiotic compatibility by cross-inoculation with the plant species studied. It was observed that there are indigenous Rhizobia in the soils of Rio Grande do Sul capable of producing siderophores and auxins. All of Lotus Rhizobia were able to induce nodulation in at least two of the five studied species. Rhizobia strains isolated from L. uliginosus have shown a broad specificity, being able to induce nodulation in all studied species. On the other hand, Rhizobia isolated from D. incanum have shown to be highly specific. In a green-house experiment the plants of L. corniculatus inoculated with the isolate UFRGS Lu 2 and strains EEL 698 and SEMIA 816 produced higher shoot dry mass. This isolate was also efficient in D. incanum plants. The results show that there are indigenous Rhizobia in the soils of Rio Grande do Sul, which have low host specificity and are capable of forming symbiosis with D. incanum, L. corniculatus, L. subbiflorus, L. glaber and L. uliginosus plants. The symbiotic compatibility with different hosts made difficult the establishment of compatibility groups. The technique of REP-PCR using primer BOX A1 showed differences among the Rhizobia studied, which are different from strains provided by culture collections.

The Rhizobium-legume symbiosis is an opportunistic association between two symbiotic partners that results in the formation of the root nodule. The process depends on the expression of a number of plant and bacterial genes that are considered critical for the establishment and maintainance of the symbiotic state. The merits of a mutational approach to the analysis of symbiosis have been recognized for several years and transposon Tn5 mutagenesis of Rhizobium has led to the identification of several symbiotic genes. This study describes the use of Tn5 mutagenesis for the isolation of symbiotically defective mutants of Bradyrhizobium japonicum. Two classes, auxotrophic and cell surface-altered mutants defective in nodule formation, have been characterized. In B. japonicum USDA 122, histidine auxotrophs that are defective in nodulation have been studied. The mutagenized DNA region has been cloned and the wild-type DNA region isolated by hybridization and complementation. In B. japonicum 61A76, Tn5-induced cell surface-altered mutants have been isolated by selecting for bacteriophage resistance. Several parameters have been used to demonstrate alterations in cell surface components. It has been shown that the Tn5 insertion is not the primary cause of the mutation in two of the characterized mutants. Complementation tests have led to the isolation of a wild-type DNA-containing cosmid, pPS23A, that overcomes the symbiotic defect in one of the mutants. Analysis of the cell surface showed a partial restoration of surface components in the complemented mutant.

Typescript. / Thesis (Ph. D.)--University of Hawaii at Manoa, 1990. / Includes bibliographical references (leaves 164-173) / Microfiche. / xi, 173 leaves, bound ill. 29 cm

Thesis (Ph.D.) -- McMaster University, 1997. / Includes bibliographical references (leaves 237-260). Also available via World Wide Web.

Thesis (M.S.)--University of Wisconsin--Madison, 1966. / eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references.

Thesis (M.S.)--University of Wisconsin--Madison, 1967. / eContent provider-neutral record in process. Description based on print version record. Includes bibliographical references.

Techn. Universiẗat, Diss., 2000--Berlin.