The Rapid Serological Identification of Rhizobia in Small Nodules

Parker, C.A.; Grove, Pamela L.

doi:10.1111/j.1365-2672.1970.tb05249.x

Cited by 10 publications

(4 citation statements)

References 3 publications

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“…8e8quipedalis (L.) van Eseltine], which nodulates with representatives of both fast-and slow-growing groups of rhizobia (Trinick, 1968). In this way, the maximum genetic difference between rhizobial strains was possible 1075 (see Graham, 1964) The nodular bacterial component was identified by reaction with the specific antiserum (dilution 1: 200) after crushing the nodules in wells on Perspex plates (Parker & Grove, 1970). In no case did the bacteroid component fail to react with the homologous antiserum.…”

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confidence: 99%

Control of leghaemoglobin synthesis in snake beans

Broughton

Dilworth

1971

Biochemical Journal

855

511

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1. The finding that the plant is the genetic determinant of leghaemoglobin production in legume nodules was further tested by inoculating snake beans with two strains of Rhizobium selected to give large genetic differences. Carbohydrate requirement patterns, immunological techniques and DNA base ratio determinations were used to demonstrate genetic differences between the two rhizobial strains. 2. Partially purified preparations of the haemoglobins from the nodules produced by the two strains showed no differences when examined by electrophoresis, isoelectric focusing or ion-exchange chromatography. 3. Two different leghaemoglobins from each type of nodule were separated by chromatography on DEAE-cellulose. One of these was isolated in the Fe3+ form and accounted for two-thirds of the total leghaemoglobin. When it was examined in the analytical ultracentrifuge and by amino acid analysis, this major component did not vary with the inoculant rhizobial strain. The molecule had an s20,w of 1.88S, a diffusion coefficient of 10.7×10-7cm2·s-1 and a mol. wt. of 16700. 4. These results strongly support the hypothesis that the mRNA for leghaemoglobin is transcribed from plant DNA.

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confidence: 99%

Control of leghaemoglobin synthesis in snake beans

Broughton

Dilworth

1971

Biochemical Journal

855

511

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“…The insoluble somatic antigens on the surface of Rhizobium bacteroids in legume nodules have (IF) immunofluorescence (aggln) agglutination Mixed nodules containing both strains been shown to be highly specific towards their homologous antisera in strain identification by immunofluorescence (Trinick 1969) and agglutination (Means et al 1964;Parker & Grove 1970). These properties were shown when using nodules which were either fresh or preserved by methods which did not involve prior heat treat-…”

Section: Discussionmentioning

confidence: 99%

“…Since the bacteroids in the nodules share common antigenic properties with the bacterial genotype in culture (Zipfel 1912), bacteroids from fresh nodules have been used directly for nodule identification by tube agglutination (Means et al 1964), immunodiffusion (Skrdleta 1969), and immunofluorescence (Trinick 1969). Nodules which could not be identified immediately have been frozen (Russell & Jones 1975;Wagner et al 1978;Berger et al 1979), dried over CaCl,, or suspended in 0.2% formalin prior to storage at 4°C (Parker & Grove 1970).…”

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Suitability of oven‐dried root nodules for Rhizobium strain identification by immunofluorescence and agglutination

Somasegaran

Woolfenden

Halliday

1983

Journal of Applied Bacteriology

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Legume root‐nodules, dried at oven temperature (70°C for 48 h) were suitable for Rhizobium strain identification by immunofluorescence and agglutination. The fluorescence of bacteroids of R. japonicum, R. leguminosarum, R. meliloti, R. phaseoli, and Rhizobium spp. from oven‐dried nodules was the same as those from frozen, desiccated, or nodules dried at room temperature (28°C). Oven‐dried nodules did not require further steaming for agglutination. Bacteroid agglutinations gave 2–16 fold lower titres than those of the cultured cells. Fresh and oven‐dried soybean rhizobia from a mixed inoculation gave exactly the same results when identified by immunofluorescence or agglutination.

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“…The agglutination technique, however, lacks the analytical resolving power of immunodiffusion, especially in distinguishing between antigenically identical and closely related nonidentical strains. Because of the greater confidence that can be placed in the identification of strains made by immunodiffusion than by agglutination (12,19,21,26), it would be helpful in ecological studies if immunodiffusion could be used conveniently for the identification of R. japonicwn isolates, especially directly from the crushed nodules.…”

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Antigenic Analysis of Rhizobium japonicum by Immunodiffusion

Dudman¹

1971

Applied Microbiology

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Immunodiffusion reactions were studied with seven strains of Rhizobium japonicum and three strains of the cowpea miscellany by using antisera against eight of the strains. Most strains yielded only weak precipitin bands when untreated cell suspensions were used as antigens in the diffusions. Ultrasonic disruption or heat treatment of the cells led to stronger bands, and immersion in boiling water for 20 min was used as the standard procedure for preparing these bacteria for immunodiffusion analysis. Heat-labile antigens were detected in only a few strains; the major antigens of all of the strains appeared to be heat-stable. Many of the strains cross-reacted, sometimes in a nonreciprocal manner; unheated cell suspensions cross-reacted more widely but more weakly than the heated suspensions. Heat-treated crushed nodule preparations reacted well in immunodiffusions. The antigens of cultured cell and nodule extract (bacteroid) forms of three strains were compared. In one of these strains, an antigen present in the cultured cells was absent from the bacteroids. Unknown strains present in soybean root nodules were readily identified by immunodiffusion.

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The Rapid Serological Identification of Rhizobia in Small Nodules

Abstract: Summary. A simple rapid method for the serological identification of bacteroids in small leguminous root nodules is described. The method may be used where very large numbers of nodule samples require routine examination. Its utility and limitations are discussed.

Cited by 10 publications

References 3 publications

Control of leghaemoglobin synthesis in snake beans

Control of leghaemoglobin synthesis in snake beans

Suitability of oven‐dried root nodules for Rhizobium strain identification by immunofluorescence and agglutination

Antigenic Analysis of Rhizobium japonicum by Immunodiffusion

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