Nitrogen fixation by a Bangladesh deepwater rice-field calothrix

Abstract

In order to study the influence on blue-green algal nitrogenase activity of environmental variables in deepwater rice-fields (DWR), a laboratory study was planned on a DWR isolate of Calothrix (D764). The variables chosen were light, oxygen, combined nitrogen, phosphorus and iron. As availability of P is likely to play an especially important role for growth and nitrogen fixation in DWR, studies on phosphatase activity of the isolate were also included. The method used for measuring nitrogenase activity was acetylene reduction assay (ARA). In order to convert nitrogenase activity to nitrogen fixation, the conversion ratio of N(_2) : C(_2)H(_2) reduced was determined by comparing the total amount of N fixed with total C(_2)H(_2) reduced. The ratio was 1 : 4.1 and 1 : 5.2 at 85 and 10 µmol photon m(^-2) s(^-1), respectively. Changes in nitrogenase activity in batch culture were studied in relation to growth characteristics. Maximum activity (10.5 nmol C(_2)H(_4) mg d. wt(^-1) min(^-1)) was observed after two days of growth. During this period, juvenile trichomes (hence maximum heterocyst frequency) were abundant and cyanophycin granules were absent; chl a, phycobiliprotein and algal N decreased. It is suggested that the juvenile filament is the most active nitrogen-fixer during the growth of the alga. The response of nitrogenase to changes in light flux (down- or upshift) was rapid. The alga showed a marked drop in nitrogenase activity in the dark, but subsequent changes were slow, with detectable activity after 24 h. Higher nitrogenase activity was observed when the dark grown alga was re-illuminated, than the maximum activity found under continuous illumination. Nitrogen fixation and heterocyst differentiation were suppressed when 10 mg 1(^-1) NH(_4)-N was added to a batch culture. Fe-deficient cultures had lower nitrogenase activity and N content than Fe-sufficient cultures. Fe- deficiency led to the development of a series of new heterocysts apical to the basal ones. Addition of Fe to Fe-deficient cultures led to a marked increase in nitrogenase activity and loss of the degenerated basal heterocysts. The alga was capable of using a number of organic P substrates as the sole source of phosphorus and showed both cell-bound phosphomono- and phosphodiesterase activities. In batch culture, phosphatase activity was detected when cellular P content dropped to 0.98%. A brief study on the influence of the environmental factors on cell-bound phosphatase activities of the alga has been included. A brief comparison in nitrogenase activity of a UK field Rivularia population and bacterised laboratory isolate Rivularia D403 was made and probable behaviour of algae in DWR is discussed.