ADAPTIVE FORMATION OF NITRATE REDUCING SYSTEM IN ANABAENA CYLINDRICA

1. Changes in capacities for reducing nitrate, nitrite and hydroxylaminecaused by provision or depletion of various nitrogen sourceswere investigated with a nitrogen fixing blue-green alga, Anabaenacylindrica, and adaptive nature of these reducing system wasdemonstrated.
2. It was found that, under light-aerobic conditions,nitrate-and nitrite- reducing systems were induced by nitrateor nitritebut not by N₂ ammonia and glutamate. On the otherhand, theactivity of enzymes pertaining to hydroxylamine reductionwasstimulated equally by nitrate, nitrite and N₂. The latteractivitywas suppressed markedly in the presence of ammoniaor glutamate.
3. Adaptive formation of nitrite reducing systemis completelyinhibited by chloramphenicol, a potent inhibitorof proteinsynthesis. No formation was also observed under theanaerobiccondition or in the dark.
4. On the basis of thesefindings, a tentative scheme for pathwaysof nitrate reductionand nitrogen fixation in Analaena cylindricawas proposed.

(Received August 22, 1962; )     

Induction of nitrate and nitrite reductases in Anabaena cylindrica

Induction of nitrate and nitrite reductases in Anabaena cylindrica(FOGG strain) was investigated. At various stages of algal growthin the presence of nitrate or nitrite, the levels of these enzymeswere determined using cell-free preparations. Nitrate and nitritereductases were induced by the respective substrates. Nitratedid not act either as an inducer or as a repressor of nitritereductase. ¹This work was supported by grant No. 8814 from the Ministryof Education ²Department of Biology, Faculty of Science, Tokyo MetropolitanUniversitySetagaya-ku, Tokyo, Japan (Received June 18, 1970; )     

Purification and properties of nitrite reductase from the blue-green alga Anabaena cylindrical

Nitrite reductase was purified about 40-fold from the blue-greenalga Anabaena cylindrica by acetone precipitation and chromatographyon DEAE-cellulose columns. The nitrite reductase had its pHoptima at about 7.6 with Tris-HCl and at about 7.4 with phosphatewhen reduced methyl viologen was used as an electron donor.The Km's for nitrite, methyl viologen and ferredoxin were 510^–55,210^–4 and 510^–6M, respectively. A stoichiometryof one molecule of ammonia formation per one molecule of nitritedisappearance was confirmed. Ferredoxin which had been reducedeither chemically with dithionite or enzymatically with NADPHin the presence of diaphorase was active as an electron donor.Dithionite-reduced FAD and FMN were inactive. NADPH could notgive electrons directly to nitrite reductase. Hydroxylaminereductase was segregated from nitrite reductase by DEAE-cellulosecolumn chromatography. Purified nitrite reductase showed noactivity for sulfite reduction. A molecular weight of 68,000was estimated for nitrite reductase using a calibrated SephadexG-200 column. ¹This work was supported by grants 4090 and 955008 from theMinistry of Education. ²This work was supported by grants 4090 and 955008 from theMinistry of Education. 2 Present address: Department of Botany,Faculty of Science, University of Tokyo, Tokyo.     

HYDROGENASE ACTIVITY OF CELL-FREE PREPARATION OBTAINED FROM THE BLUE-GREEN ALGA, ANABAENA CYLINDRICA

A cell-free preparation of hydrogenase was obtained from acetone-driedcells of Anabaena cylindrica. This preparation was capable ofcatalyzing the reduction of various redox dyes by molecularhydrogen and evolution of hydrogen from reduced methyl viologen.PMS, methylene blue, DPIP and toluidine blue were effective,in this order, as hydrogen acceptors. Ferricyanide, p-quinone,phenosafranine and neutral red were not reduced by the presentpreparation. PPNR obtained from this alga was effective on neitherreduction of NAD and NADP by hydrogen nor stimulation of hydrogenuptake with methylene blue and PMS. Coupled with hydrogenasereaction, there occurred reduction of nitrate to ammonia whenmethylene blue was added to the reaction mixture. ¹Present address: Department of Zoology, University of Texas,Austin, Texas, U.S.A.     

Solubilization of nitrate reductase from the blue-green alga Anabaena cylindrica

Nitrate reductase was solubilized and purified from Anabaenacylindrica by Triton X-100 treatment of particulate preparationsfollowed by adsorption on calcium phosphate gel. Reduced methylviologen, FAD or FMN, but not ferredoxin, served as an effectiveelectron donor for the nitrate reduction by solubilized nitratereductase. ¹This work was supported by a grant (4061) from the Ministryof Education (Received June 25, 1970; )     

STUDIES ON THE METABOLISM OF UREA AND OTHER NITROGENOUS COMPOUNDS IN CHLORELLA ELLIPSOIDEA: III ASSIMILATION OF UREA

Assimilation of urea by Chlorella ellipsoidea was investigatedby using ¹⁴C-urea. It was revealed that urea carbon metabolizedby the N-starved cell was almost quantitatively recovered incarbon dioxide under dark-aerobic conditions. The rate of ureaconsumption was considerably influenced by the N-content ofalgal cells, practically no metabolism of urea being observedwith the cells containing more than 7 per cent N on a dry weightbasis. Similar results were also obtained when urea was replacedwith ammonia as nitrogen source. Based on the results obtained,a tentative scheme for the assimilation process of urea in Chlorellawas proposed. (Received February 7, 1960; )     

LATERAL DISPERSAL OF ADULT AQUATIC INSECTS FOLLOWING EMERGENCE FROM THE MIDDLE REACHES OF THE CHIKUMA RIVER(HONSHU ISLAND,JAPAN)IN RELATION TO WATERFRONT VEGETATION

Adults of aquatic insects emerge fromstreamslive inthe nearby riparian zone where they can select streamsidevegetations as preferred sites in order to(a)completemetamorphosis,(b)rest while awaiting proper swarmingtime,(c)feedin order to produce eggs,or(d)mate toreproduce[1—3].Most adult aquatic insect dispersal studies have fo-cused on quantifying the degree of upstream move-ment[4,5].Some researchers have shown distinct patternsof upstreammovement related to post-mating ovipositionalbehavior[6],while there...     

Differences in response to simulated herbivory between <Emphasis Type="Italic">Quercus crispula</Emphasis> and <Emphasis Type="Italic">Quercus dentata</Emphasis>

To evaluate the responses of Quercus crispula and Quercus dentata to herbivory, their leaves were subjected to simulated herbivory in early spring and examined for the subsequent changes in leaf traits and attacks by chewing herbivores in mid summer. In Quercus crispula, nitrogen content per area was higher in artificially damaged leaves than in control leaves. This species is assumed to increase the photosynthetic rate per area by increasing nitrogen content per area to compensate leaf area loss. In Quercus dentata, nitrogen content per area did not differ between artificially damaged and control leaves, while nitrogen content per mass was slightly lower in artificially damaged leaves. The difference in their responses can be attributable to the difference in the architecture of their leaves and/or the severeness of herbivory. The development of leaf area from early spring to mid summer was larger in artificially damaged leaves than in control leaves in both species, suggesting the compensatory response to leaf area loss. Leaf dry mass per unit area was also larger in artificially damaged leaves in both species, but the adaptive significance of this change is not clear. In spite of such changes in leaf traits, no difference was detected in the degree of damage by chewing herbivores between artificially damaged and controlled leaves in both species.