Nitrogen fixation (acetylene reduction) by Klebsiella pneumoniae in association with 'Park' Kentucky bluegrass (Poa pratensis L.)

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Abstract
Turfs of 'Park' Kentucky bluegrass reestablished in the greenhouse and inoculated with Klebsiella pneumoniae (W6) showed significantly increased nitrogen fixation (acetylene reduction) compared with control turfs. Mean ethylene production rates per pot were 368 nmol h−1 for K. pneumoniae treated turfs, 55 nmol h−1 for heat-killed K. pneumoniae treated turfs, and 44 nmol h−1 for untreated turfs. Calculated lag periods before activity was observed were generally very short (less than 1 h).When 'Park' Kentucky bluegrass was grown from seed on soil-less medium of Turface, a fired aggregate clay, inoculation with K. pneumoniae (W6) resulted in 9 of 11 turfs showing nitrogenase activity (mean ethylene producion rate per pot was 195 nmol h−1). Only 3 of 11 turfs treated with heat-killed K. pneumoniae showed any activity and their mean rate of ethylene production (40 nmol h−1 per pot) was significantly lower than that for turfs treated with K. pneumoniae.Using the 'Park'–Turface soil-less model system it was shown that acetylene reducing activity was (i) root associated, (ii) generally highest at a depth of 1–4 cm below the surface, (iii) enhanced by washing excised roots, and (iv) inhibited by surface sterilization of excised roots. Klebsiella pneumoniae was recovered from Turface and roots showing acetylene reducing activity.

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