Purification and Some Properties of Cytochrome b-560 from Nitrosomonas europaea

Cytochrome b-560 was purified to an electrophoretically homogeneousstate from Nitrosomonas europaea. It showed absorption peaksat 427, 530 and 560 nm in the reduced form. Its molecular weightwas estimated to be 44,000 by SDS-polyacrylamide gel electrophoresisand the same value was obtained on the basis of the contentsof haem and protein. The cytochrome was not autoxidizable anddid not react with CO. ¹Present address: Tokyo Research Center, TOSOH Corporation,Hayakawa, Ayase-shi, Kanagawa 252, Japan ²Present address: Faculty of Integrated Arts and Sciences, HiroshimaUniversity, Higashisenda-machi, Hiroshima 730, Japan (Received March 23, 1988; Accepted June 2, 1988)     

Studies on the metabolism of a sulfur-oxidizing bacterium VII. Oxidation of sulfite by a cell-free extract of Thiobacillus thiooxidans

Properties of the cell-free extract, prepared from a strainof Thiobacillus thiooxidans by sonic disruption followed byfractionation with centrifugatiori, were investigated with referenceto its sulfite-oxidizing activity. Without the addition of cofactors the particulate fraction(F-P)catalyzed oxidation of sulfite with oxygen or bacterial cytochromec-552 obtained from Pseudomonas stutzeri as electron acceptor.TMPD reduced by ascorbic acid was also oxidized by F-P. Thesoluble fraction(F-S) showed no activity in oxidizing sulfiteand TMPD, but stimulated TMPD oxidation by F-P. Oxygen uptake with either sulfite or TMPD as substrate was inhibitedby KCN, NaN₃, CO and c-phenanthroline. CO-Inhibition was reversedby light. Reduction of cytochrome c-552 by sulfite was insensitiveto these agents. Antimycin A markedly inhibited sulfite oxidation with eitheroxygen or cytochrome c-552 as electron acceptor, but was withouteffect on TMPD oxidation. DDC and SAO, both strong inhibitors of sulfur oxidation, didnot affect sulfite and TMPD oxidations. Cytochromes of the a, b and c types were contained in F-P. Thesecytochromes were rapidly reduced when F-P was incubated withsulfite. Cytochrome(s) of the c type was present in F-S, too. ¹VI.=References (3) ²Partly supported by a grant from the Ministry of Education ³Present address: Sanyo Women's College, Hatsukaichi, Hiroshima738, Japan ⁴Present address: Department of Biochemistry, Hiroshima UniversitySchool of Dentistry, Hiroshima 734, Japan (Received May 15, 1970; )     

Studies on anaerobic biphasic growth of a denitrifying bacterium, Pseudomonas stutzeri

Growth of Pseudomonas stutzeri(VAN NIEL strain) in the presenceof a limiting amount of nitrate under anaerobic conditions ischaracterized by 2 logarithmic phases separated distinctly byan intermediate phase where the growth rate is very low. Inthe first logarithmic phase nitrate is reduced stoichiometricallyto nitrite stage, and in the second phase nitrite is reducedto nitrogen gas. The nitrite reducing activity of cells in the second growthphase is 3–4 times higher than that of cells in the firstphase. The rise in nitrite reducing activity is correlated witha remarkable increase in the content of cytochromes a₂ and c-552. ¹Present address: Department of Biochemistry, Hiroshima UniversitySchool of Dentistry, Hiroshima, Japan. ²Present address: Institute of Molecular Biology, Faculty ofScience, Nagoya University, Nagoya, Japan. (Received June 16, 1969; )     

Increase of cytokinin activity in detached etiolated cotyledons of squash after illumination

Cytokinins nullified the lag period of the formation of chlorophyllin detached etiolated cotyledons of squash (Cucurbita moschataDuch. var. melonaeformis Makino cv. Tokyo). One hour after illumination,cytokinin activity in detached cotyledons rapidly increasedand maintained a certain level for another hour. ¹ Present address: Department of Enviromental Studies, Collegeof Integrated Arts and Sciences, Hiroshima University, Hiroshima730, Japan. (Received November 25, 1976; )     

N-Terminal Amino Acid Sequence and Processing Site of Sweet Potato Cytochrome c Oxidase Subunit II

The N-terminal amino acid sequence of sweet potato cytochromec oxidase subunit II polypeptide was determined. Comparisonsbetween the sequence and amino acid sequences deduced from thenucleotide sequences of other higher plant subunit II genesindicate a post-translational clevage of N-terminal extensionpart. ¹Present address: Institute of Low Temperature Science, HokkaidoUniversity, Sapporo, 060 Japan. (Received June 13, 1989; Accepted September 8, 1989)     

Effects of Manipulations of Source and Sink on the Carbon Exchange Rate and Some Enzymes of Sucrose Metabolism in Leaves of Soybean [Glycine max (L.) Merr.]

Soybean plants [Glycine max (L.) Merr. cv. AGS129], two andthree weeks after depodding and defoliation, respectively, wereused to examine the possibility of end-product regulation onthe carbon exchange rate and activities of enzymes involvedin sucrose metabolism in leaves. Removal of one and two lateralleaflets per trifoliate leaf reduced the total leaf area by20% and 47%, respectively. Removal of one pod per node reducedthe total pod number by 23% per plant. Dry weights of roots,stems and petioles decreased with reductions in leaf area. Bycontrast, removal of pods resulted in an increase in these parameters.The carbon exchange rate and transpiration rate of leaves increasedwith defoliation and decreased with depodding. The intercellularconcentration of CO₂ in leaves was reduced by defoliation andincreased by depodding. Furthermore, defoliation increased thelevel of leaf chlorophyll in leaves while depodding decreasedit. Removal of pods decreased the activities of sucrose-phosphatesynthase and -amylase but increased that of sucrose synthase.A significant positive correlation was found between the activityof leaf sucrose-phosphate synthase and both the carbon exchangerate and the sucrose content of leaves. Thus, manipulation ofthe sink and source in soybean plants influenced the relationshipbetween sucrose metabolism and the carbon exchange rate in intactleaves. ³Faculty of Agriculture, Okayama University, Tsusimanaka Okayama,700 Japan ⁴Faculty of Agriculture, Saga University, Honjo-machi, Saga,840 Japan ¹Present address: Faculty of Agriculture, Sriwijaya University,J1 Raya Indralaya, OK1 30662, Indonesia ²Present address: Faculty of Agriculture, Saga University, Honjo-machi,Saga, 840 Japan     

The Deduced Amino Acid Sequence of Phytochrome from Adiantum Includes Consensus Motifs Present in Phytochrome B from Seed Plants

A cDNA for the phytochrome of the fern Adiantum capillus-venerisL. was cloned and sequenced. The deduced phytochrome is 50{smalltilde}55% identical to phytochromes of seed plants, and 68%identical to Selaginella phytochrome. Regions resemble thosein previously characterized phytochromes from ferns, lower plantsand seed plants. ³Present address: Yamanouchi Pharmaceutical Co., Ltd., 21 Miyukigaoka,Tsukuba-shi, Ibaraki, 305 Japan ⁴Present address: Plant Growth Regulation Laboratory, The Instituteof Physical and Chemical Research (RIKEN), Hirosawa 2-1, Wako-shi,Saitama, 351-01 Japan ⁵Present address: Advanced Research Laboratory, Hitachi, Ltd.,Hatoyama, Saitama, 350-03 Japan     

The growth promoting effect of phytic acid on callus tissues of rice seed

The effects of phytic acid, myo-inositol hexaphosphoric acid,in the induction and growth of rice seed callus were studied.Phytic acid was substituted for myo-inositol in the inductionof the callus and preferable effects were shown on the growthof the callus. ¹Present address: Hyogo Agricultural Experiment Station, Akashi,Hyogo, Japan (Received June 30, 1970; )     

Suppression of Acid Invertase Activity by Antisense RNA Modifies the Sugar Composition of Tomato Fruit

cDNA for an acid invertase (EC 3.2.1.26 [EC] ) of tomato (Lycopersiconesculentum Mill.) fruit was introduced into tomato plants underthe control of the cauliflower mosaic virus 35S promoter inthe antisense orientation. The antisense gene effectively suppressedthe invertase activity in soluble and cell wall fractions fromripening fruits. The sucrose content of fruits of the transformantswas markedly increased, while the hexose content was reduced.These results indicate that acid invertase is one of main determinantsof the sugar composition of tomato fruit. The invertase activityin the cell wall fraction of the leaf tissues of the transformantswas not suppressed to the same extent as that in the solublefraction. Wounding of the control leaf tissues induced invertaseactivity in both soluble and cell wall fractions. The inductionof activity in the soluble fraction was suppressed by the antisensegene, while that in the cell wall fraction was unaffected. Thesefindings suggest that mRNA for some other invertase, in particular,the mRNA for a cell wall-bound invertase, was present in leaves. ¹Present address: Plant Breeding and Genetics Research Laboratory,Japan Tobacco Inc., 700 Higashibara, Toyoda, Iwata, Shizuoka,438 Japan. ²Present address: National Institute of Agrobiological Resources,Kannondai, Tsukuba, Ibaraki, 305 Japan.     

Turgor Regulation in a Brackish Water Charophyte, Lamprothamnium succinctum III. Changes in Cytoplasmic Free Amino Acids and Sucrose Contents during Turgor Regulation

Cytoplasm and cell sap of Lamprothamnium succinctum were analyzedseparately for the contents of free amino acids and sucroseto find whether they contribute to turgor regulation. In thevacuole, both amino acids and sucrose were found to be minorcomponents contributing to the generation of osmotic pressure.Their amounts were almost insensitive to changes in externalosmotic pressure. In the cytoplasm, both amino acids and sucrosein the cytoplasm contributed about 20% to the osmotic pressure.Hypotonic treatment did not affect the contents of either, buthypertonic treatment, while not affecting the amino acid contents,caused a significant increase in sucrose content. The cytoplasmicsucrose content increased linearly with an increase in externalosmotic pressure, accounting for 40% of the increased osmoticpressure. ¹ Present address: Department of Biology, Osaka Medical College,Sawaragi-cho, Takatsuki, Osaka 569, Japan ² Present address: Department of Applied Physiology, NationalInstitute of Agrobiological Resources, Yatabe, Tsukuda, Ibaragi305, Japan (Received November 25, 1986; Accepted March 18, 1987)     

Nucleotide Sequence of cDNA Clones Encoding the PS I-H Subunit of Photosystem I in Tobacco

cDNA clones encoding the PS I-H subunit of photosystem I wereisolated from Nicotiana tabacum and Nicotiana sylvestris. Thenucleotide sequences of three clones showed that, in both species,the mature PS I-H protein consists of 95 amino acid residuesand has a calculated molecular mass of 10.3 kDa. ³ Present address: The Institute of Physical and Chemical Research,Tsukuba, 305 Japan.     

Studies on Microbody Development in Wounded Sweet Potato Root Tissue: Proposal for the Post-Translational Transport of Catalase into Preexisting Microbodies

Pure microbody fractions could be prepared in considerable yieldsfrom sweet potato root tissue slices incubated for 16 hr and3 days. The ratio of catalase activity to phospholipid contentin the fraction from slices incubated for 3 days was about 3times that from slices incubated for 16 hr. Total catalase activityin the former slices was about twice that in the latter. Thissuggests that catalase synthesized during incubation of theslices is transported into microbodies preexisting in intacttissue. ¹ Present address: Laboratory of Food Technology, Faculty ofApplied Biological Science, Hiroshima University, Fukuyama,Hiroshima 720, Japan. ² Present address: Terumo Co., Ltd., Omiya, Fujinomiya, Shizuoka418, Japan. (Received July 1, 1982; Accepted September 24, 1982)     

Molecular Cloning and Characterization of S-Adenosyl-L-Methionine:Scoulerine-9-O-Methyltransferase from Cultured Cells of Coptis japonica

S-Adenosyl-L-methionine : scoulerine-9-O-methyltransferase (SMT)catalyzes the transfer of the S-methyl group of S-adenosyl-L-methionineto the 9-hydroxyl group of scoulerine during the biosynthesisof berberine. We have isolated functionally active cDNA clones(pCJSMTs) from a cDNA library prepared from cultured cells ofCoptis japonica. The longest cDNA insert (pCJSMT1) had an openreading frame that encoded 351 amino acids, but the calculatedmolecular mass (38,364 Da) of the deduced product was slightlylower than the experimentally determined molecular mass of purifiedSMT. Rapid amplification of the 5' end of the cDNA indicatedthat the full-length cDNA of SMT consisted of 1,458 nucleotidesthat encoded 381 amino acids. When the full-length cDNA wasexpressed in E. coli, the molecular mass of the expressed SMTwas greater than that of native SMT in Coptis cells. This resultsuggests that SMT might be produced in a pre-mature form andprocessed post-translationally. SMT was also found to exhibitsequence homology to other O-methyltransferases from plantsand N-terminal region of the SMT polypeptide appeared to benecessary for enzymatic activity. ¹Present address: High Quality Life Research Laboratories, SumitomoMetal Industries, Ltd., 3-5 Hikaridai, Seika, Sourakugun, Kyoto,619-02 Japan ²Present address: Suntory Research Center, 1-1-1 Wakayamadai,Shimamoto, Mishima-gun, Osaka, 618 Japan ³Present address: Department of Cell Biology, The Scripps ResearchInstitute, La Jolla, CA 92037 U.S.A.     

Effects of growth conditions on formation of cytochrome system of a denitrifying bacterium, Pseudomonas stutzeri

Cytochrome systems in cells of a denitrifying bacterium, Pseudomonasstutzeri (VAN NIEL strain), grown under different atmosphericconditions were compared with reference to the effects of nitrateand nitrite on cytochrome synthesis. When a culture was sufficiently aerated (aerobic conditions),synthesis of all cytochrome components was repressed, regardlessof the presence or absence of nitrate and nitrite. When aeratedmoderately (semi-aerobic conditions), both soluble and paniculatecytochromes c-552 and cytochrome b-558 contents markedly increasedeven in the absence of nitrate and nitrite. Under anaerobic or semi-aerobic conditions, nitrite inducedcytochrome a₂–c synthesis. This inductive effect of nitritewas counteracted by nitrate. Nitrate also repressed particulatecytochrome c-552 synthesis to some extent but nitrite did not. ¹Present address: Department of Biochemistry, Hiroshima UniversitySchool of Dentistry, Hiroshima, Japan (Received June 24, 1969; )     

Properties of the Respiratory NAD(P)H Dehydrogenase Isolated from the Cyanobacterium Synechocystis PCC6803

Activity staining with NADPH-nitroblue tetrazolium after native-PAGEof membrane proteins of Synechocystis PCC6803, solubilized with3-[(3-cholamidopropyl)dimethylammonio]-1-propanesulfonate (CHAPS),revealed four NAD(P)H dehydrogenase (NDH) activities; an NDHcomplex of the respiratory chain, a ferredoxin NADP⁺ reductase(FNR), a drgA product which oxidized both NADH and NADPH, andan uncharacterized NADH-specific enzyme. The NDH complex waspurified with anion exchange and gel filtration chromatographies.The purified complex had a molecular mass of 376 kDa and wascomposed of 9 subunits. Western analysis showed that the complexcontained the NDH-H subunit, but not NDH-A or B. The enzymereduced ferricyanide much faster than plastoquinone and usedNADPH as its prefered electron donor rather than NADH. The enzymaticactivity was inhibited by diphenyleneiodonium chloride and salicylhydroxamicacid, but not by rotenone, p-chloromercuribenzoate, N-ethylmaleimide,flavon, dicumarol, or antimycin A. These results suggest thatthe purified complex is a hydrophilic subcomplex which containsan NADPH binding site and flavin, and is dissociated from ahydrophobic subcomplex, which contains quinone binding site. ¹Present address: Division of Applied Life Sciences, GraduateSchool of Agriculture, Kyoto University, Sakyo, Kyoto, 606-8502Japan ³Present address: Department of Biotechnology, Faculty of Engineering,Fukuyama University, 1 Gakuencho, Fukuyama, Hiroshima, 729-0292Japan     

Effects of Growth Regulators on the Induction of Anthocyanin Synthesis in Carrot Suspension Cultures

The effects of plant growth regulators were investigated onanthocyanin synthesis induced by removing auxin from carrotsuspension cultures. Of the auxins tested, 2,4-D showed thestrongest inhibiting effect on anthocyanin synthesis and hadthe strongest promoting effect on undifferentiated growth. When2,4-D was added to anthocyanin synthesizing cells, in whichcell division had ceased, anthocyanin synthesis was repressedimmediately, accumulated anthocyanin disappeared and cell divisionresumed. All cytokinins examined promoted anthocyanin synthesisin the absence of auxin. Both gibberellic acid (GA₃) and abscisicacid inhibited anthocyanin synthesis in media lacking 2,4-D,though GA³ showed no effect on cell division. These effectsof growth regulators on anthocyanin synthesis are similar tothose reported for their effects on embryogenesis [Fujimuraand Komamine (1975) Plant Sci. Lett. 5: 359, (1979) Z. Pflanzenphysiol.95: 13, (1980) Z. PJlanzenphysiol. 99: 1]. The relationshipbetween the induction of anthocyanin synthesis, metabolic differentiation,and embryogenesis are discussed. ¹ Present address: Department of Biology, College of Arts andSciences, The University of Tokyo, Komaba, Meguro-ku, Tokyo153, Japan. ² Present address: Biological Institute, Faculty of Science,Tohoku University, Sendai, Miyagi 980, Japan. (Received November 28, 1985; Accepted July 23, 1986)     

Synergistic Function of rolB, rolC, ORF13 and ORF14 of TL-DNA of Agrobacterium rhizogenes in Hairy Root Induction in Nicotiana tabacum

Comparison of the frequency of rooting in the tobacco leaf segmentsinoculated with Agrobacterium tumefaciens harboring variouscombinations of rolB, rolC, ORF13 and ORF14 of TL-DNA of Riplasmid (pRiHRI) revealed that the genes differ in their functionto stimulate adventitious root induction. A single gene rolBinduced roots, while rolC, ORF13 and ORF14 independently promotedthe root induction by the rolB gene. The effects of these geneson the rolB-mediated rooting were in the order of ORF13>rolCORF14. Present address: Laboratory of Phylogenetic Botany, Departmentof Biology, Chiba University, 1-33 Yayoi-cho, Inage-ku, Chiba,263-8522 Japan. ² Present address: Department of Chemical and Biological Sciences,Faculty of Science, Japan Women's University, 2-8-1 Mejirodai,Bunkyo-ku, Tokyo, 112-8681 Japan.     

Localization and Binding Characteristics of a High-Affinity Binding Site for N--Acetylchitooligosaccharide Elicitor in the Plasma Membrane from Suspension-Cultured Rice Cells Suggest a Role as a Receptor for the Elicitor Signal at the Cell Surface

A high-affinity binding site for N-acetylchitooligosac-chlarideelicitor was found to localize in the plasma membrane from suspension-culturedrice cells. Binding kinetics as well as the specificity of thisbinding site corresponded well with the behavior of the ricecells to the editor. These characteristics suggest that thebinding site represents a functional receptor for N-acetylchitooligosaccharideelicitor in rice. ²Present address: Okinawa Prefectural Livestock ExperimentalStation, 2009-5 Shoshi, Nakijin-son, Okinawa, 905-04 Japan. ³Present address: School of Hygiene and Public Health, The JohnsHopkins University, 615 North Wolfe Street, Baltimore, Maryland,21205 U.S.A. ⁴Present address: University of Tenessee, Microbiology, knoxville,Tennessee, 37996 U.S.A.     

Novel Light-Dark Change of Proline Levels in Halophyte (Mesembryanthemum crystallinum L.) and Glycophytes (Hordeum vulgare L. and Triticum aestivum L.) Leaves and Roots under Salt Stress

Proline accumulation was determined in a facultative halophyte,Mesembryanthemum crystallinum and glycophytes, barley (Hordeumvulgare L.) and wheat (Triticum aestivum L.) Proline accumulationpreceded the shift of CAM in M. crystallinum and did not occurin the continuous darkness. The novel light-dark change of prolinelevel (high in the light and low in the dark) was observed inleaves of all three plants. Proline levels of shoots in barleyand wheat also showed the same light-dark change, suggestingthat proline accumulated in the leaves in the light was nottranslocated to other tissues in the dark period. These resultssuggest that proline has a bifunctional role in the acclimationto high salt stress; an osmoregulant role in the light, anda substrate for dark respiration to supply energy to compartmentationof ions into vacuole in the dark. ¹Present address: Kyoto Biological Res. Lab., Bio-Chiba Inc.Watsuka,Soraku, Kyoto, 619-12 Japan ²Present address: Kobayashi Pharmaceutical Co., Ltd. Doshomachi,Chuo-ku, Osaka, 541 Japan