A specific immunoabsorbent for the isolation of fraction I protein

Antibodies produced against pure Fraction I protein (ribulose 1,5-bis-phosphate carboxylase) from Nicotiana tabacum have been covalently linked to a Sepharose 4B matrix. Fraction I protein present in crude extracts of green leaves of higher plants can be absorbed on a column of the immobilised antibody. The Fraction I protein is eluted from the column with 8 M urea and is in the form of dissociated subunits uncontaminated with other proteins. This material is ideal for an examination of the subunit polypeptide composition of the Fraction I protein by isoelectric focusing. By this procedure, Fraction I protein from sugar beet, Beta vulgaris, has been shown to consist of four different types of polypeptides, three large subunit polypeptides and a single small subunit polypeptide.     

The evolution of fraction I protein during the origin of a new species ofNicotiana

The polypeptide composition of Fraction I protein from Nicotiana digluta, a synthetic species which arose by chromosome doubling following the interspecific hybridization of N. glutinosa and N. tabacum, has been examined by isoelectric focusing. The composition of the protein from N. digluta, which was identical to the protein from the infertile F1 hybrid N. glutinosa x N. tabacum, showed 3 polypeptides in the large subunit and 4 polypeptides in the small subunit. The large subunit polypeptides were identical to those from N. glutinosa, the maternal parent in the original hybridization, whereas the small subunit polypeptides were a composite of the small subunit polypeptides from both N. glutinosa and N. tabacum. This analysis demonstrates how the polypeptide composition of Fraction I protein evolves during the origin of new species of Nicotiana.     

Gibberellin A12 Is Converted to Gibberellin A53 in Cultured Cells of Nicotiana tabacum and Catharanthus roseus

The metabolism of GA₁₂ and its precursors was investigated incultured cells of seven cell lines of Nicotiana tabacum andthree cell lines of Catharanthus roseus using ^l4C-labeled substrates.The presence of a metabolic pathway from ent-7-hydroxykaurenoicacid to GA₅₃ via GA₁₂-aldehyde and GA₁₂ was demonstrated inthe cultured cells. GA₁₂ was effectively converted to GA₅₃ incells of BY-2, 2b-4, 2b-13 and CG from N. tabacum. By contrast,GA₅₃ was not converted to any other GAs in all of the linesof cells examined. The metabolism of C₁₉-GAs was also examinedusing ³H-labeled substrates. The conversion of GA₂₀ to GA₂₉and GA, and of GA₄ to GA₃₄ occurred more efficiently in cellsfrom C. roseus than in cells from N. tabacum. However, 13-hydroxylationof GA₄ and GA₉ was not observed in any of the cell culturesexamined. Among the various metabolites, GA₅₃, GA₂₉ and GA₃₄were identified by full-scan GC/MS. (Received December 20, 1990; Accepted May 27, 1991)     

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.     

The Composition of Phloem Exudate and Xylem Sap from Tree Tobacco (Nicotiana glauca Grah.)

The composition of xylem sap and exudate from stem incisionsof Nicotiana glauca Grah. was compared in detail. Exudationfrom stem incisions occurred over a 5 min period in certainplants, enabling collection of 5–30 µl of sap. Therate of exudation showed an exponential decline. Exudate hada high dry matter content (170–196 mg ml^–1) andhigh sugar (sucrose) levels. Xylem sap had a low pH (5.8) andexudate a pH of 7.9. Glutamine dominated the amino compoundsin xylem sap and exudate, and K⁺ was the major cation. Totalamino compounds in stem exudate reached 10.8 mg ml^–1 whereasxylem sap contained much lower levels (0.28 mg ml^–1).All mineral elements and amino compounds with the exceptionof calcium were more concentrated in stem exudate than in xylemsap. Sucrose was labelled heavily in stem exudate following pulsingof an adjacent leaf with ¹⁴CO₂. A concentration gradient ofsugar (2.1 bar m^–1) was recorded for stems. Levels ofsucrose, amino compounds and K⁺ ions in stem exudate showeda diurnal periodicity. Each commodity reached maximum concentrationat or near noon and minimum concentration about dawn. The evidencesuggests that exudate from stem incisions of N. glauca is arepresentative sample of solutes translocated in the phloem. Nicotiana glauca Grah., phloem sap, xylem sap, sucrose, amino compounds, mineral ions     

Effect of kinetin on auxin uptake and distribution in normal and tumor-prone Nicotiana shoots

Stem segments of non-tumorous Nicotiana glauca and N. langsdorffiiplants and of their tumor-producing amphidiploid F₁ hybrid weretreated with 6-furfurylaminopurine (kinetin) prior to transporttests with applied labeled indoleacetic acid (IAA-2-¹⁴C). Kinetin-treatmentsincreased the uptake of IAA in non-tumorous shoots; the IAAuptake by N. langsdorffii segments was increased up to 3-fold.The auxin uptake in stem-segments of the tumor-forming hybrid,however, could not be increased significantly by kinetin. Theeven distribution of IAA-¹⁴C in segments of normal and tumorproneNicotiana shoots is stimulated by kinetin. Data are discussedin conjunction with previous results on auxin transport andtumorformation in Nicotiana. (Received August 8, 1972; )     

Effects of sulfite and pH on abscisic acid-dependent transpiration and on stomatal opening

In rice, alday, wheat and tobacco (Nicotiana tabacum L. Samsunand Samsun NN) plants which contained large amounts of ABA,the transpiration rate decreased rapidly with 2 ppm SO₂ fumigationand reached 20 to 65% of the initial level after 5- to 30-minexposure depending on their ABA contents. In the cases of broadbean and tobacco (N. glutinosa L.) with low ABA contents, therate slightly increased for 20 and 40 min, respectively, afterthe start of the fumigation and then decreased gradually. Thetranspiration rates of corn and sorghum, in spite of their extremelylow ABA contents, pronouncedly decreased with SO₂ fumigationand reached 65 and 50%, respectively, of the initial levelsafter 40-min exposure. Foliar application of 0.04 N HC1 to N.tabacum L. Samsun NN leaves remarkably depressed the transpirationrate, while the application of 0.04 M Na₂SO₃ decreased the rateonly to the same level as water treatment. Foliar applicationof either HCl or Na₂SO₃ to N. glutinosa L. leaves exerted littlechange in the transpiration rate. When 10^–4M ABA was appliedto broad bean leaves prior to HCl and Na₂SO₃ treatment, theirtranspiration rate was decreased by HCl, but not by Na₂SO₃ application.In sonicated epidermal strips peeled from broad bean leaves,Na₂SO₃ produced a slight increase in the stomatal aperture sizein the absence of ABA, but showed no effect in the presenceof ABA. The aperture size was identical in the pH range of 3.0to 7.0 in the incubation medium. In the presence of ABA in themedium, the aperture size was small in the acidic region ofpH with a minimal value at pH 4.0. ABA decreased the aperturesize at concentrations above 10^–9 M at pH 4.0 and 10^–6M at pH 7.0 in the medium. [2_–14C] ABA uptake by epidermalstrips was large in the acidic region, especially at pH 4.0. (Received February 28, 1980; )     

The involvement of ethylene in in vitro maturation of mid-binucleate pollen of Nicotiana tabacum

The role of ethylene during in vitro maturation of Nicotianatabacum pollen from the mld-binucleate (MB) stage was analysedby the addition of aminooxyacetic acid (AOA), aminoethoxyvinylglycine(AVG), CoCl₂ and AgNO₃ to the maturation medium (AMGLu). Anincrease in ethylene production was obtained in both isolatedpollen and pollen surrounded by sporophytic tissue during insitu maturation. in vitro maturation of pollen was inhibitedby AOA and AVG; ACC and ethrel were able to overcome this inhibitoryeffect. Cyclohexylamine (CHA) reverted the inhibition provokedby both Ag⁺ and Co²⁺ The results reported in this paper indicatethat ethylene is one of the factors implicated in in vitro maturationof MB pollen of Nicotiana tabacum. Key words: Nicotiana tabacum, maturation, germination, pollen, ethylene     

Constitution of Mitochondrial and Chloroplast Genomes in Male Sterile Tobacco Obtained by Protoplast Fusion of Nicotiana tabacum and N. debneyi

Chloroplast DNA (cpDNA) and mitochondrial DNA (mtDNA) of malesterile tobacco plants obtained by fusion of Nicotiana tabacumprotoplasts and X-irradiated N. debneyi protoplasts were analyzed.Digestion of cpDNA isolated from ten male sterile lines withfour restriction endonucleases (EcoRI, XhoI, SmaI and HindIII)indicated that these lines possessed either one or the otherparental chloroplast genome. Neither mixture of two types ofcpDNA nor unique restriction fragments were detected in anyof the cases examined. The genetic constitution of chloroplastgenomes identified by restriction analysis of cpDNA showed goodagreement with that based on isoelectric focusing of the largesubunit of the Fraction I protein. The mtDNA from five fusion-derivedmale sterile plants showed banding patterns quite differentfrom each other and from the parental plants. Each plant exhibitednew restriction fragments not found in the parental species.These findings indicate that recombinational events in the mitochondrialgenomes take place rather frequently in the mixed cytoplasmsafter protoplast fusion, whereas the mixed chloroplasts becomesegregated to homogeneity. (Received June 19, 1987; Accepted October 5, 1987)     

Use of a Disarmed Ri Plasmid Vector in Analysis of Transformed Root Induction

Nicotiana glauca, N. tabacum, Solanian dulcamara and S. nigrumwere transformed by Agrobacteriun rhizogenes strain BN1010 (TL^–TR⁺).The TR-DNA stimulated agropine-positive root induction and wastransformation competent in the absence of the TL-DNA. An unusualpattern of root induction was seen when stem explants were inoculatedwith this strain; occasionally, agropine-positive roots wereinduced at the inoculation sites, but prolific agropine-negativeroots were formed in profusion down the stems. The utility ofBN1010 as an efficient co-integrating vector was demonstratedby the separate transfer of a fragment containing rol ABC (BN1010::pEM15) and of a chimeric nopaline synthase-kanamycin resistancegene (BN1010:: Neo) into plants. Root cultures of S. dulcamaratransformed with BN1010:: Neo had an unusual, positively geotropicphenotype. Strain BN1010:: pEM15 (rol ABC⁺D^–TR⁺) incitedmore roots down stem explants than strain A4T. This indicatesthat rol D may act to suppress agropine-negative root productionin N. glauca and N. tabacum. Key words: Agrobacterium rhizogenes, TL-DNA, TR-DNA, disarmed Ri vector, transformed roots, Nicotiana glauca, N. tabacun, Solatium dulcamara, S. nigrum     

Experimentally Synthesized Plant Chimeras 2. A Comparison of In vitro and In vivo Techniques for the Production of Interspecific Nicotiana Chimeras

In vitro and in vivo techniques were compared for synthesizingchimeras between Nicotiana glauca Grahm and N tabacum L Interspecificchimeral callus, produced from mixed callus cultures in vitro,was placed on media which favoured only N tabacum shoot formationNone of the 474 regenerated N tabacum shoots incorporated Nglauca cells into their meristems When chimeral callus was regeneratedunder hormonal conditions favouring simultaneous organogenesis,of 397 shoots, only non-chimeral shoots of both species aroseIn vivo, reciprocal splice grafts between species were decapitatedjust above the graft union and treated with or without auxin—lanolinpastes Auxin increased callus formation but inhibited adventitiousshoot formation Three of 209 adventitious shoots arising fromthe graft union were interspecific mericlinal chimeras whichwere later stabilized as periclinal chimeras All three chimerasformed when N glauca was the understock Two of the chimerasarose on untreated shoots which produced no visible callus,indicating that excessive callus formation may be unnecessaryfor multiple cell origin of adventitious shoots to occur Chimeras, tobacco, Nicotiana glauca, Nicotiana tabacum, tissue culture, graft chimeras, callus cultures     

Identification of several chloroplast DNA genes which code for the large subunit of Nicotiana Fraction I proteins

Summary Electrofocusing of carboxymethylated, crystalline Fraction I proteins in polyacrylamide gels containing 8 M urea resolves the large subunit into three major peptides and the small subunit into one or more peptides. Electrofocusing of proteins isolated from leaves of the reciprocal, F₁, hybrids: N. glutinosa x N. tabacum, N. glauca x N. tabacum, N. glauca x N. langsdorfii and the parental species confirms that coding information for the large subunit is inherited only by the maternal line whereas both parents contribute coding information for the small subunit. The analysis shows that one or more of the three peptides of the large subunit of Fraction I proteins from different Nicotiana species differ in isoelectric point and therefore serve as phenotypic markers for chloroplast DNA genes.     

Nontranslation of foreign genetic information for fraction 1 protein under circumstances favorable for direct transfer ofNicotiana gossei isolated chloroplasts intoN. Tabacum protoplasts

Summary The nuclei and cytoplasm ofN. gossei andN. tabacum are compatible to the extent that reciprocal, interspecific F₁ hybrids can be produced by conventional breeding techniques. Conditions were established in which manyN. gossei isolated chloroplasts could be seen by phase and fluorescence microscopy to adhere to 40% of the population of protoplasts obtained from white tissue of variegatedN. tabacum plants and to remain attached after washing the protoplasts. Chloroplasts also could be seen to enter the interior of the protoplasts. After treating albino protoplasts withN. gossei chloroplasts, the protoplasts were subjected to further conditions whereby 65 calluses containing shoots developed. TwentyN. tabacum protoplasts not treated with foreign chloroplasts also produced calluses with shoots to serve as a control. All calluses developed chlorophyll irrespective of whether or not the albino protoplasts had been treated with isolatedN. gossei chloroplasts. The Fraction 1 protein ofN. tabacum has a different electrophoretic mobility from the protein ofN. gossei or anN. gossei xN. tabacum F₁ hybrid. The Fraction 1 protein large subunit is coded by chloroplast DNA, whereas the small subunit is coded by nuclear DNA. Fraction 1 protein was isolated from the variegated shoots of the 65 calluses obtained after treating albino protoplasts with foreign chloroplasts. Immunoelectrophoresis demonstrated the protein from each callus to have a mobility identical toN. tabacum protein. Therefore, under circumstances highly favorable for the direct transfer ofN. gossei isolated chloroplasts (and possibly nuclei also) intoN. tabacum protoplasts, no evidence was obtained to suggest that genetic information contained in the isolated foreign organelles was being translated into the polypeptides of either the large or small subunits of Fraction 1 protein contained in newly differentiated leaves derived from the protoplasts. Supported by Research Grant PCM-75-07368 from the National Science Foundation.     

Effects of light intensity and quality on the relative N and P requirement (the optimum N:P ratio) of marine planktonic algae

The relative requirement of N and P (the optimum N:P ratio)by Dunaliella tertiolecta, Phaeodactylum tricornutum, Prymnesiumparvum and Thalassiosira pseudonana was studied under variouslight intensities and spectra. The ratio was determined as theratio of the minimum cell N and P concentrations (q₀N and q₀pwhen either nutrient was limiting. The ratio varied widely amongspecies; under light-saturation for growth (116 µEin ^m–2s^–1 it ranged from 11.8 in ^D. tertiolecta to 36.6 in P.tricornutum. The ratio appeared to be higher at a sub-saturatingintensity (24 µEin m^–2 s^–1 in all except P.tricornutum, mainly because of higher q_oN with little changein q_oP. In T. pseudonana Q_oP also increased, resulting in aninsignificant change in the ratio. The ratio varied little withinthe range of saturation intensity. Light quality affected q_oNand q_oP as well as the ratio, and the pattern of change variedfrom species to species. The optimum ratio of individual specieswas linearly correlated to their q_oN except in P. tricornutum.q_oN for all species showed a linear correlation with cell proteinconcentrations irrespective of light conditions. The changeof optimum N:P ratios in the three species thus appears to berelated to changes in cell protein contents. The ratio of carbohydratesto protein remained constant regardless of light intensity orquality and was higher in P-limited cultures. We conclude thatchanges in light regime can strongly influence algal nutrientrequirements and species interrelationships by altering theoptimum cellular N:P ratio.     

Diurnal Variation in Acetylene Reduction and Net Hydrogen Evolution in Five Tropical and Subtropical Nitrogen-Fixing Tree Symbioses

Diurnal variations in acetylene reduction and net hydrogen evolutionwere shown in five tropical and subtropical nitrogen-fixingtree symbioses. The symbioses studied in a growth chamber were:Acacia albida x TAL 1457, Leucaena leucocephala x TAL 1145,Prosopis chilensis x TAL 600, Casuarina glauca x HFP Cc13 andC. obesa x HFP Cc13. Acetylene reduction was highest at the end of the light periodin all symbioses studied. In the A. albida x TAL 1457 symbiosis,the diurnal variations in acetylene reduction and net hydrogenevolution showed a minor synchrony, while in the other symbiosesthe diurnal pattern of acetylene reduction and net hydrogenevolution clearly differed. Also, a diurnal variation in relativeefficiency of nitrogenase was shown in the A. albida x TAL 1457symbiosis. A hydrogen uptake enzyme was detected at a low substrate concentration(24.5 mmol m^–3 H₂) for L. leucocephala x TAL 1145, C.obesa x HFP Cc13 and has earlier been found for C. glauca xHFP CcI3. A hydrogen uptake system was also found for P. chilensisx TAL 600 and A. albida x TAL 1457 at a 17-fold higher substrateconcentration. The results show that a diurnal variation in C₂H₂ reductionand H₂ evolution occurs, and that diurnal variation in the conversionfactor between C₂H₂ reduction and N₂ fixation could occur. Thisfact raises criticisms regarding the use of a point estimateof this factor. Key words: Acetylene reduction, hydrogen evolution, uptake hydrogenase, nitrogen-fixing tree symbioses     

Inactivation of the Water-Oxidizing Complex by Exogenous Reductants in PS II Membranes Depleted of Extrinsic Proteins

We have studied the inactivation of the water-oxidizing complexby exogenous, ‘general’ reductants in various typesof PS II membrane. Extraction of the 33, 23 and 17 kDa proteinsfrom PS II membranes rendered the functional Mn susceptibleto rapid solubilization by reductants such as hydroquinone,benzidine and ascorbate, while water analogs, such as NH₂OH,inactivated the complex regardless of the presence of PS IIextrinsic proteins. The extent of the inactivation was dependenton the hydrophobicity of the reductants examined. Diphenylcarbazide,an efficient electron donor to Z⁺ and D⁺, did not inactivatethe Mn complex. As reported earlier [Ghanotakis et al. (1984)Biochim. Biophys. Acta 767: 524], weak illumination deceleratedthe inactivation of the complex by the various reductants. Kineticanalyses of the flash-induced protection provided evidence aboutthe nature of the light state that was not susceptible to thereductants. This state was generated and decayed with half timesof approximately 0.5 and 9 s, respectively. However, such lightprotection was diminished under Cl^–-depleted conditions,at slightly alkaline pH, or when ascorbate was employed as areductant. Furthermore, we observed that the oxidation of N,N,N',N'-tetramethyl-p-phenylenediamine,which reacts with the Mn complex, was accomplished as a biphasicreaction. The amount of the fast phase, which was almost eliminatedafter the reconstitution of the 33 kDa protein and Ca²⁺, wasapproximately 7 electron equivalents per 200 Chl. From theseresults, it is likely that the bulky, ‘general’reductants reduce the functional Mn directly by solubilizingMn from the complex in the same way as do the water analogs.The effectiveness of these reductants in the photoactivationof the apo-water-oxidizing complex is also discussed. (Received September 13, 1989; Accepted March 12, 1990)     

Two types of voltage-dependent potassium channels in outer hair cells from the guinea pig cochlea

Cell-attached and cell-free configurations of the patch-clamptechnique were used to investigate the conductive properties andregulation of the major K⁺channels in the basolateral membrane of outer hair cells freshly isolated from the guinea pig cochlea. There were two majorvoltage-dependent K⁺ channels. ACa²⁺-activatedK⁺ channel with a high conductance(220 pS,P_K/P_Na = 8) was found in almost 20% of the patches. The inside-out activityof the channel was increased by depolarizations above 0 mV andincreasing the intracellular Ca²⁺concentration. External ATP or adenosine did not alter thecell-attached activity of the channel. The open probability of theexcised channel remained stable for several minutes without rundown andwas not altered by the catalytic subunit of protein kinase A (PKA)applied internally. The most frequentK⁺ channel had a low conductanceand a small outward rectification in symmetricalK⁺ conditions (10 pS for inwardcurrents and 20 pS for outward currents, P_K/P_Na = 28). It was found significantly more frequently in cell-attached andinside-out patches when the pipette contained 100 µM acetylcholine. It was not sensitive to internalCa²⁺, was inhibited by4-aminopyridine, was activated by depolarization above 30 mV,and exhibited a rundown after excision. It also had a slow inactivationon ensemble-averaged sweeps in response to depolarizing pulses. Thecell-attached activity of the channel was increased when adenosine wassuperfused outside the pipette. This effect also occurred with permeantanalogs of cAMP and internally applied catalytic subunit of PKA. Bothchannels could control the cell membrane voltage of outer hair cells.

     

Photosynthetic parameters, pigment composition and respiration rates of the marine diatom Skeletonema costatum grown in continuous light and a 12:12 h light-dark cycle

Nutrient-sufficient cultures of a Trondheimsfjord (Norway) cloneof the marine centric diatom Skeleionema costatum (Grev.) Clevewere grown at 75 µmol m^–2 s^–1 and 15C at24 and 12 h daylength to study diurnal variations and the effectof daylength on pigment and chemical composition, photosyntheticparameters, dark respiration rates and scaled fluorescence excitationspectra (F), the latter used as estimates for the absorptionof energy available to Photosystem II. Specific growth rateswere 1.06 and 0.56 day^– in 24 and 12 h daylength, respectively,while dark respiration rates were generally 85% of the net growthrate. The Chla-normalized photosynthetic coefficients P^B_m anda^B were {small tilde}20–25% higher in continuous lightthan at 12 h daylength, while the Chla:C ratio was {small tilde}15%lower (0.051 versus 0.061 w:w). Thus, the carbon-normalizedcoefficients P^c_m and a^c were <11% lower at 24 h than at 12h daylength. The maximum quantum yield _max, the Chla:C ratioand F differed negligibly, as did the light saturation indexl_k, the N:C ratio and the ratios Chlc:Chla and Fucoxanthin:Chla. P^B_m and l_k did not exhibit diurnal variations at 24 hdaylength, and varied within 23% of the daily mean at 12 h daylength.Predictions of the daily gross photosynthetic rate based ondata for a given time of the day should thus not be >10%in error relative to an integrated value based on several datasets collected through 24 h. _max was 0.084–0.117 mol O₂(mol photons)^– for gross oxygen evolution. However, ifused in mathematical models for predicting the gross and netgrowth rates (i.e. the gross and net carbon turnover rates),‘practical’ values of 0.076 and 0.040 g-at C (molphotons)^–, respectively, should be employed. Correspondingly,values for a^B and P^B_m should be adjusted pro rata. ¹Present address: College of Marine Studies, Sjmannsveien 27,N-6008 lesund, Norway     

Function of Ngrol Genes in the Evolution of Nicotiana glauca: Conservation of the Function of NgORF13 and NgORF14 after Ancient Infection by an Agrobacterium rhizogenes-Like Ancestor

Ngrol genes are thought to have resulted from horizontal genetransfer from an Agrobacterium rhizogenes-like ancestor earlyin the evolution of the genus Nicotiana. Four Ngrol genes (NgrolB,NgrolC, NgORF13 and NgORF14) have been found in the genome ofN. glauca, but their functions are not yet known. We have investigatedthe properties of Ngrol genes and shown that some of them areable to function in tobacco plants. Transgenic analysis revealedthat NgORF13 promotes RirolB-mediated adventitious root inductionon tobacco leaf segments. NgORF14 also promoted the RirolB-mediatedroot induction, but the intensity of this promoting effect wasweak. These promoting functions of NgORF13 and NgORF14 havemuch the same efficiency as those of the corresponding genesof A. rhizogenes, RiORF13 and RiORF14, respectively. Overexpressionof NgORF13, under control of the cauliflower mosaic virus 35Spromoter (P_35s), provoked morphological abnormalities in transgenictobacco plants. Transgenic plants that harbored the P_35s-NgORF13had rounded leaves and stout flowers resulting from suppressionof the longitudinal growth of leaf and floral leaves such assepals, petals, stamens and carpels. These results suggest thatNgORF13 and NgORF14 in the genome of N. glauca have conservedfunctional sequences since their original integration eventby an A. rhizogenes-like ancestor. 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.     

Variations in the Contents and Kinetic Properties of Ribulose-1,5-bisphosphate Carboxylases among Rice Species

The K_m(CO₂) ancl V_max of ribulose 1,5-bisphosphate (RuBP) carboxylaseand its protein ratio to total soluble protein from Oryza speciesincluding cultivars (25 varieties) and wild types (11 species,21 strains) were surveyed. Their variabilities among cultivarsof O. sativa were very small. The averages of the K_m(CO₂) andV_max values and the ratio of carboxylase to soluble protein,and their standard errors were 10.2?1.0µM, 1.72?0.13units.mg^–1(pH 8.0 and 25?C) and 52?2%, respectively. However, some differencesseemed to exist based on genome constitution in the Oryza genus.RuBP carboxylases from the species with the A^gA^g genome, O.graberrima and O. breviligulate, exhibited low K_m(CO₂) values(8.0?0.8 µM). High V_max was associated with the CC genome,O. eichingeri and O. officinalis (2.08?0.15 units.mg^–1).A higher ratio of RuBP carboxylase protein to soluble proteinwas found for the AA genome, O. sativa and O. perennis. (Received September 24, 1986; Accepted April 15, 1987)