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Labeling of carbon pools in Bradyrhizobium japonicum bacteroids in intact nodules following feedings of sup 14 CO sub 2 for periods of <= six minutes

Conference · · Plant Physiology, Supplement; (United States)
OSTI ID:5562492
;  [1]
  1. Ohio State Univ., Wooster (United States)

A system was established for isolation of bacteroids from soybean nodules in 5 min by centrifugation of crude brei through silicone oil in a microfuge. Using marker enzymes {Beta}-hydroxybutyrate dehydrogenase (DH) and cytochrome C oxidase, it was found that bacteroids were not ruptured during isolation, that mitochondrial contamination was {<=} 2%, and that bacteroid recovery was {approx} 93%. About 40 {mu}Ci {sup 14}CO{sub 2} were supplied to 300 mg fresh wt of intact nodules to label the 4-carbon acid pools in the host cytoplasm of infected cells. Following incubations of 1 to 6 min label in the cytosol was largely in malate (MAL), as expected. Label in aspartate and citrate was about 20% that in MAL. {sup 14}C in succinate (SUC) was only {approx} 3% of that in MAL, and fumarate was essentially unlabeled indicating limited equilibration of label via fumarase and SUC DH. In bacteroids MAL was most rapidly labeled, but glutamate {sup 14}C was about 50% that in MAL; this is consistent with other results. Aspartate and alanine were labeled at a rate approx 10% that of MAL, and SUC, citrate and fumarate were slowly labeled. Results are consistent with 4-carbon acids (mainly malate) as the main source of reduced carbon for bacteroids.

OSTI ID:
5562492
Report Number(s):
CONF-9107184--
Journal Information:
Plant Physiology, Supplement; (United States), Journal Name: Plant Physiology, Supplement; (United States) Vol. 96:1; ISSN PPYSA; ISSN 0079-2241
Country of Publication:
United States
Language:
English