MILLS. 1971. Degradation of rutin by Aspergillr~s pavrts. Purification of the dioxygenase, quercetinase. Can. J. Microbiol. 17: 11 1-1 18. Evidence is presented that a single enzyme, quercetinase, is responsible for the degradation of quercetin by Aspergillus flavlts to yield carbon monoxide and a depside, 2-protocatechuoylphloroglucinol carboxylic acid. A procedure for the isolation of the dioxygenase as a homogeneous protein is described. The most purified preparation degraded 10 800 umoles of quercetin/h mg protein and was homogeneous as judged by ultracentrifugation and by electrophoresis. The molecular weight was determined as 111 000 + 4000. K,,, values for quercetin and oxygen as substrates were 5.2 X 1 0 -W and 1.2 X 10-4 M respectively. The enzyme is a glycoprotein containing 27.5% carbohydrate and the amino acid composition is presented.
A survey of 108 isolates of Rhizobium leguminosarum was conducted to determine the variation in H2 uptake and relative efficiency of N2 fixation in Pisum sativum L. root nodules and the relation of relative efficiency to plant dry weight and N content. Only 14 of the isolates exhibited significant uptake hydrogenase activity and none of these had sufficient hydrogenase activity to recycle all of the H2 produced by nitrogenase. In 74 of the isolates tested relative efficiencies of N2 fixation were less than 0.60.Twenty-nine of the isolates were ineffective, since total plant N at harvest did not differ significantly from uninoculated controls. The remaining 79 effective isolates could be divided into low-and high-efficiency groups. Plants which were inoculated with isolates from the two groups and harvested after 4 weeks did not differ significantly in plant dry weight or N content. Among the isolates with high relative efficiency of N2 fixation, two groups could be recognized: one possessing significant uptake hydrogenase activity, the other lacking hydrogenase activity but in which H2 evolution was low. Although the two groups did not differ with respect to plant dry weight or N content, the identification of this latter group may be of some significance for optimizing the efficiency of N2 fixation.
Pea plants (Pisum sativum L. cv. Trapper) were inoculated and grown in controlled-environment chambers at two irradiance levels. Shoot and root dry weights and nitrogen contents, total leaf and stipule areas, and rates of C2H2 reduction were determined during growth in different treatments of NH4NO3 addition. Although overall growth increased with irradiance, the growth responses to combined nitrogen addition were similar at both light levels. Two phases of early vegetative growth were identified by their different responses to combined nitrogen. During the first phase, low levels of NH4NO3 greatly increased the relative growth rate, the growth per unit leaf area, and the percentage of nitrogen in the tissues. This indicated a period of nitrogen stress which lasted only until the 3rd week. Over the next 2 weeks, combined nitrogen increased the relative growth rates to a lesser extent, primarily through an increased partitioning of assimilates to shoot development. This distribution effect was rapidly reversible on changing nutrient conditions. The early stimulation of leaf area development by addition of combined nitrogen during these two growth phases resulted in greater capacity for symbiotic fixation after NH4NO3 was removed.
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