Effect of benzimidazole on the senescence of wheat chloroplasts and their boat shape transformation

Benzimidazole efficiently retards the senescence of detachedwheat leaves. Electron microscopic study has revealed that nodamage was visible on the fine structure of chloroplasts inleaves which retained their green color by benzimidazole treatment,whereas chloroplasts of water floated leaves showed strikingalterations in their color, shape and fine structure as senescenceproceeded. Moreover, the chloroplast lamellar system in benzimidazoletreated leaves even seemed to be organized over those in immediatelydetached leaves. The bending tendency for lamellar array within chloroplastswas more marked in benzimidazole treated leaves than in thecontrol. Chloroplasts showed, more frequently and intensely,transformation into the "boat shape" the extreme curvature ofthe granafretwork skeleton of the chloroplasts forming the keelof the boat. ¹This work was supported by Grants to Prof. E.R. WAYGOOD fromthe Canada Department of Agriculture (EMR-14) and the NationalResearch Council of Canada (PRBT-10) ²This work was undertaken during the author's stay at the Departmentof Botany, University of Manitoba, Winnipeg, Manitoba, Canada,with Fellowship granted to the author by the National ResearchCouncil of Canada, 1963–1965 (Received July 29, 1969; )     

Effect of malformin on root growth

Malformin induces curvatures, stimulates root hair and lateralroot formation, promotes radial expansion, inhibits elongation,wet and dry weight, cell division and cell wall synthesis inroots of Zea mays, but has no effect on protein synthesis. Thegrowth curves (elongation, wet and dry weight) of Z. mays rootstreated with malformin are cubic. Processes which are involvedin inhibition of elongation are considered the primary causeof root curvatures by malformin. ¹This research was supported by grant GB-7158 from the NationalScience Foundation and grant E-146-F from the American CancerSociety. Journal Paper No. 3536 of the Purdue Agricultural ExperimentStation. ²Present address: Volcani Institute of Agricultural Research,P.O.B. 6, Bet-Dagan, Israel. (Received December 24, 1969; )     

Possible role of the cytosol pathway of acetyl-CoA supply in terpene biosynthesis in sweet potato infected with Ceratocystis fimbriata

Pyruvate was converted into acetyl-CoA by the cytosol enzymefraction prepared from sweet potato root tissue infected withCeratocystis fimbriata. The conversion was dependent on thiaminepyrophosphate, NAD⁺, ATP and CoA. Activities of pyruvate decarboxylase,aldehyde dehydrogenase and acetyl CoA synthetase increased indiseased tissue. These results indicate the operation of thecytosol pathway of acetyl-CoA supply for terpene biosynthesisin C. fimbriata-infected tissue which consists of the abovethree enzymes. ¹This paper constitutes Part 135 of the Phytopathological Chemistryof Sweet Potato with Black Rot and Injury. This work was supportedin part by a Grant-in-Aid from the Ministry of Education, Scienceand Culture. ²Present address: Biochemistry Department, University of Queensland,St. Lucia 4067, Queensland, Australia. (Received April 21, 1980; )     

Isolation of microbodies from, sweet potato root tissue and their development during aging of slices

Microbodies were isolated from, sweet potato root tissue bydifferential and linear sucrose density gradient centrifugation.When the tissue was homogenized in the presence of PolyclarAT, the microbodies sedimented together with the mitochondriathrough the sucrose gradients. The microbodies had a densityof 1.25 g/cm³, and contained catalase and urate oxidase, butnot malate dehydrogenase, isocitrate lyase, glycolate oxidase,hydroxypyruvate reductase and the cyanide-insensitive palmitoylCoA-oxidation system. A small amount of o-diphenol oxidase alsoseemed to be present. Catalase, but not urate oxidase, activity in the crude extractincreased during aging of the sliced tissue. A similar resultwas obtained with the microbody fraction after linear sucrosedensity gradient centrifugation. We propose that microbodiescontaining only catalase develop during aging of sliced sweetpotato root tissue. ¹ This work was supported in part by a Grant-in-Aid (No. 311908)for Scientific Research from the Ministry of Education, Scienceand Culture, Japan. (Received June 20, 1979; )     

CHANGE OF B-TYPE HAEM CONTENT IN RELATION TO PEROXIDASE BIOSYNTHESIS IN INJURED SWEET POTATO ROOTS

The methods of quantitative analysis of b-type haem in plantswere investigated. With an improved method developed was determinedthe haem content in the supernatant, mitochondrial, and microsomalfractions of sweet potato tissue. The activities of peroxidase,catalase, and cytochrome oxidase, as well as the contents ofb-type haem and acid-insoluble nitrogen in the cellular fractionswere determined at different incubation times after cuttingof sweet potato tissue. Peroxidase and catalase increased withtime in each celluler fraction, following a short lag phase.In the mitochondrial fraction, b-type haem, cytochrome oxidase,and acid insoluble nitrogen increased linearly with time. Inthe microsomal and supernatant fraction, b-type haem increasedwith time following a short lag phase. The increase in haemcontent of the supernatant fraction appeared to be associatedwith peroxidase formation. Time course analysis showed that ⁵⁹Fe-incorporation into b-typehaem of the supernatant fraction increased with time and thatincorporation was markedly inhibited by blasticidin S. The incorporationof ⁵⁹Fe into mitochondrial haem did not increase with time andwas not inhibited by blasticidin S. Blasticidin S inhibited⁵⁹Fe-incorporation into microsomal haem. Time course analysisof b-type haem content, ⁵⁹Fe-incorporation into b-type haem,and peroxidase activity suggest that in the injured tissue haemis synthesized from low molecular weight compounds and is incorporatedinto peroxidase as the haem moiety. ¹ This paper constitutes Part 57 of the Phytopathological Chemistryof Sweet Potato with Black Rot. ² Present address: Institute for Plant Virus Research, Chiba.     

COMPARISON OF EUTHECOSOMATOUS PTEROPODS IN THE PLANKTON AND SEDIMENTS OFF BARBADOS, WEST INDIES

The euthecosomatous pteropod fauna of Barbados plankton wascompared with euthecosome shells occurring in recent sediments.Twenty species of euthecosomes were collected in the same relativeabundance in both provinces. Differences in the average sizesof Spiratella inflata and Creseis virgula conica, the most commonspecies were not statistically significant although differencesin the size-frequency histograms were noted. It is concludedthat euthecosomes deposited in the upper sediment layers offBarbados accurately reflect the species composition and relativeabundance of euthecosome species in the plankton. ¹Supported by a Postgraduate Scholarship from the National ResearchCouncil of Canada and NRC Grant No. A5248 to Dr. C.M. Lalli. ²Present address: Department of Oceanography, Dalhousie University,Halifax, Nova Scotia, Canada.     

Dual pattern of DL-leucine absorption by duckweed root tips

The rates of cellular absorption of DL-leucine from solutionsof different concentrations indicate the presence of dual uptakemechanisms in root tips of Lemna minor, one in the low (0.01to 0.2 mM) and the other in the high (5 to 25 mM) concentrationrange. The absorption pattern for the lower concentrations conformsto Michaelis-Menten kinetics. ¹Supported in part by a Grant-in-Aid of Research from the Societyof Sigma Xi. (Received June 19, 1973; )     

Some properties of shikimate: NADP oxidoreductase produced in sweet potato root tissue after slicing

The activity of shikimate: NADP oxidoreductase [EC 1. 1. 1.25] in sweet potato root tissue increased soon after slicing.Enzyme preparations obtained from both sliced tissue and fromfresh tissue probably contained a single enzyme component, andthey showed identical chromatographical behaviour. K_m values of the enzyme for NADP and shikimate were 1.0x10^–4Mand 1.3 x 10^–3M, respectively. Enzyme activity was potentlyinhibited by SH-inhibitors such as p-chloromercuribenzoate andoxidized glutathione. Enzyme activity was affected neither by mononucleotides suchas ATP, ADP and AMP, divalent cations, Mg⁺⁺, Ca⁺⁺ and Mn⁺⁺,nor by metabolites such as tryptophan, phenylalanine, tyrosineand t-cinnamic acid which are involved in aromatic compoundsyntheses. The enzyme rapidly lost its activity. This inactivation reactionshowed a time course consisting of two steps of the first-orderreaction. The inactivated enzyme preparation was not reactivatedby thiol compounds such as cysteine, 2-mercaptoethanol and glutathione,although these reagents, to a certain extent, protected theenzyme from inactivation. The results suggest that denaturationof the enzyme protein was involved in inactivation of the enzyme. ¹Part 74 of the phytopathological chemistry of sweet potatowith black rot and injury. ²Present address: Department of Biology, Faculty of Science,Tokyo Metropolitan University, Setagaya-ku, Tokyo. (Received August 5, 1968; )     

Inhibition of Adventitious Root Growth in Tradescantia by Calmodulin Antagonists and Calcium Inhibitors

When cuttings of Tradescantia fluminensis stem were incubatedin distilled water, the buds located at the node grew into adventitiousroots. The root growth could be inhibited by calmodulin antagonists,trifluoperazine, chlorpromazine, compound 48/80 and calmidazolium,in a concentration-dependent manner. The divalent cation chelatorethyleneglycol-bis-(ß-aminoethyl ether)-N, N, N, N-tetraaceticacid had no effect, however, the intracellular chelator TMB-8completely inhibited root growth. The growth was also inhibitedby calcium ionophore A23187 [GenBank] , lanthanum, a competitive inhibitorof Ca²⁺ uptake and verapamil, a calcium channel inhibitor. AWestern blot of the adventitious root extract followed by immunostainingwith an anti-spinach calmodulin antibody clearly showed thepresence of calmodulin in this tissue. These results stronglysuggest the involvement of calmodulin and calcium in the growthof Tradescantia advenitious roots. ¹A part of this work has been published in abstract form in"Molecular and Cellular Aspects of Calcium in Plant Development"(Editedby A.J.Trewavas, Plenum Publishing Co. 1986. ³Present address: Plant Laboratory , Kirin Brewerry Co. Ltd.,Kitsuregawa-cho, Tochigi-ken 329-14 ,Japan (Received May 2, 1987; Accepted September 17, 1987)     

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     

Physiological gradients and shoot initiation in tobacco callus cultures

Support for the idea that physiological gradients of substancesinto the tissue may be the operative factors promoting organinitiation in vitro is presented. Evidence for this conceptwas obtained through measurement of the starch content and respiratoryactivity of upper and lower segments of tobacco (Nicotiana tabacumL. cv. Wisconsin 38) callus and inversion of the tissue duringculture. ¹This investigation was supported by NRC of Canada grant A-6467to T.A.T. (Received October 16, 1972; )     

Trifluralin and root growth

Trifluralin inhibited root elongation and induced root tip swellingof Zea mays and Triticum aeslivum. Time-course experiments showedthese effects occurred within 6 hr of treatment. As the rootstreated with trifluralin enlarged, there was a concomitant increasein root growth inhibition. Bioassays were devised to quantitativelymeasure the radial enlargement of trifluralin-treated roots.Histological observations indicated that swollen root tissuewas growing in a non-polar manner. The root swelling effectof trifluralin was inhibited 70% by the reducing agent, 2,3-dimercaptopropanol.Trifluralin had no effect on SH content of root tip proteinof Z.mays. ¹ Published with the approval of the Director of die West VirginiaAgricultural Experiment Station as Scientific paper No. 1177. ² Present address: Department of Biology, Mercyhurst College,Erie, Pennsylvania 16501, U.S.A. (Received March 19, 1971; )     

ATP-Dependent Ca2+ Transport in Tonoplast Vesicles from Apple Fruit

Protoplasts and vacuoles were isolated from immature apple fruit(Malus pumila Mill. cv. Golden Delicious). ATP-stimulated Ca²⁺uptake was identified in both protoplast vesicles and tonoplastvesicles. The apparent K_m for Ca²⁺ of the tonoplast transportsystem was 43.4 µM. The pH optima were 7.2 and 6.7 forCa²⁺ transport by protoplast and tonoplast vesicles, respectively.Ca²⁺ transport in tonoplast vesicles was strongly inhibitedby the calmodulin antagonists fluphenazine and N-(6-aminohexyl)-5-chloro-l-naphthalensulfonamidehydrochloride (W-7), while N-aminohexyl)-l-naphthalensulfonamidehydrochloride (W-5) was relatively ineffective. Addition ofexogenous calmodulin stimulated transport by 35%. Ca²⁺ uptakewas inhibited by vanadate, but not by the ionophores carbonylcyanidem-chlorophenyl hydrazone (CCCP) or valinomycin. The resultsindicate that apple tonoplasts have a Ca²⁺ transport systemthat is driven by the direct hydrolysis of ATP, and may be calmodulindependent. ¹Present address: Morioka Branch, Fruit Tree Research Station,Ministry of Agriculture, Forestry and Fisheries, Shimokuriyagawa,Morioka 020-01, Japan. To whom reprint requests should be addressed. (Received October 18, 1985; Accepted January 29, 1986)     

Involvement of Calcium in Adventitious Bud Initiation in Torenia Stem Segments

In Torenia stem segments cultured in vitro, active meristematicdivisions are induced in the epidermis by treatment with cytokinin,resulting in the formation of adventitious buds. Applicationof the calcium ionophore A23187 [GenBank] was found to induce meristematicdivisions in the absence of cytokinin. The induction by A23187 [GenBank] was inhibited by simultaneous addition of auxin, but not byanti-cytokinin. A two hour pre-treatment with A23187 [GenBank] was alsoeffective, but only when it was applied to the explants justafter their excision from mother plants. The A23187 [GenBank] -inducedmeristematic zones developed into dome-shaped structures, butnot into complete adventitious buds. Complete elimination ofcalcium from the culture medium caused 50% inhibition of A23187 [GenBank] -and/or cytokinin-induced initiation of meristematic divisions.When the explants were preincubated with EGTA and then culturedon a Ca-free medium containing EGTA, cytokinin failed to inducebud initiation. Similar inhibition was also obtained by lanthanum,a calcium antagonist, by verapamil, a calcium channel inhibitor,and by trifluoperazine and chlorpromazine, calmodulin inhibitors.These results support the idea that adventitious bud initiationinduced by cytokinin in Torenia stem segments may be mediated,at least partially, by an increase in the level of intracellularCa²⁺. ¹Bioscience Research Center, Mitsui Petrochemical IndustriesLtd., Waki-cho, Kuga-gun, Yamaguchi 740, Japan. (Received May 9, 1985; Accepted October 5, 1985)     

Studies on chlorophyll formation in Chlorella protothecoides III. Effects of chloramphenicol, cycloheximide, and ethionine on chlorophyll formation

Effects of chloramphenicol, cycloheximide, puromycin and ethionineon the light-independent and subsequent light-dependent processesof chlorophyll formation in "glucose-bleached" cells of Chlorellaprotothecoides were studied. These substances, except puromycin,strongly suppressed different phases of chlorophyll formation.Ethionine most strongly suppressed the light-independent phaseand chloramphenicol an early, relatively short process in thelight-dependent phase of chlorophyll formation. Cycloheximideseverely suppressed all phases of chlorophyll formation. Possibleimplications of these results for the biosynthesis of chlorophyllin algal cells are discussed. ¹ Present address: National Food Research Institute, Ministryof Agriculture and Forestry, Koto-ku, Tokyo 135, Japan. ² Laboratory of Entomology, Faculty of Agriculture, TamagawaUniversity, Machida-shi, Tokyo, Japan (Received October 5, 1972; )     

Separation and Partial Characterization of Membranes from Prochloron sp.

Two differently colored membrane preparations were separatedfrom the prochlorophyte, Prochloron sp., by mechanical disintegrationof the cells followed by sucrose density gradient centrifugation.An orange-colored preparation, containing zeaxanthin as themajor constituent pigment, seemed to comprise the cytoplasmicmembrane. The other green-colored membrane preparation, containingß-carotene and chlorophyll a and b as major pigmentconstituents, was identified as the thylakoid membrane. Thetwo types of membranes were compared as to their absorptionspectra and buoyant densities. ¹ This work is one of the results of the 8th International Expeditionon Prochloron organized by Dr. R. A. Lewin, University of Californiaat San Diego. ⁵ Present address: Solar Energy Research Group, The Algatron,The Institute of Physical and Chemical Research (RIKEN), Wako-shi,Saitama 351, Japan. ⁶ Present address: National Institute for Basic Biology, Okazaki444, Japan. (Received October 19, 1984; Accepted January 7, 1985)     

Zooplankton growth rates: the larvaceans Appendicularia, Fritillaria and Oikopleura in tropical waters

The growth rates of Appendicularia sicula, Fritillaria borealissargassi, Fritillaria haplostoma, Oikopleura dioica and Oikopleuralongicauda were determined from microcosms incubated in situat 23C in Jamaican waters. Experiments were conducted fromoligotrophic offshore waters, through mesotrophic Lime Cay andeutrophic Kingston Harbour in both natural and nutrient-enhancedphytoplankton communities. Length-weight relationships werecalculated for two of these species: O.longicauda log W=2.47log TL –6.10 and F.haplostoma log W=2.44 log TL –7.37,where weight (W) is in micrograms and trunk length (TL) is inmicrometres. Instantaneous growth rates averaged 1.7–2.5day^–1 for the five species and were observed as high as3.3 day^–1 These instantaneous rates are equivalent todaily specific growth rates averaging 4.6–11.4 and rangingup to 28. In larger genera, growth rates were related positivelyto picoplankton and nanoplankton concentration, and negativelyto the biomass of larvaceans, but in the smallest species growthwas unrelated to these factors. However, because the variabilityin these two factors within microcosms exceeded their naturalrange of variability, growth rates of larvaceans may normallybe unlimited by resources or population density effects. ¹Present address :Monterey Bay Aquarium Research Institute 7700Sandholdt Road, Moss Landing, CA 95039-0628, USA ²Present address :Bedford Institute of Oceanography PO Box 1006,Dartmouth, Nova Scotia B2Y 4A2, Canada     

Carbonic Anhydrase in Chlamydomonas reinhardtii II. Requirements for Carbonic Anhydrase Induction

Carbonic anhydrase (CA) activity in wild type cells of Chlamydomonasreinhardtii was low when cells were cultured under 2% CO₃ inthe light. When the gas phase was changed to air, CA activityincresaed as much as 20 fold over the next 24 hours. In contrast,CA activity did not change markedly in cells of the mutantspet 20-8 (PS II-negative), lip 10-2 (photophosphorylation-negative),and F60 (phosphoribulokinase-negative), when they were subjectedto the same induction regimen. DCMU (10^–5 M) and cydoheximide(3 µg/ml) severely inhibited the induction in wild typecells. No induction occured when CO₂ concentration was loweredin darkness. ³Present adress: Photoconversion Research Branch, Solar EnergyResearch Institute, Golden, Colorado 80401, USA. (Received June 7, 1982; Accepted December 25, 1982)     

Malate Decarboxylation by Mitochondria of the Inducible Crassulacean Acid Metabolism Plant Mesembryanthemum crystallinum

Mitochondria isolated from leaves of Mesembryanthemum crystallinumoxidized malate by both NAD malic enzyme and NAD malate dehydrogenase.Rates of malate oxidation were higher in mitochondria from plantsgrown at 400 mil NaCl in the rooting medium and performing Crassulaceanacid metabolism (CAM) than in mitochondria from plants grownat 20 mM NaCl and exhibiting C₃-photosynthetic CO₂ fixation.The mitochondria isolated from plants both in the CAM and C₃modes were tightly coupled and gave high respiratory control.At optimum pH for malate oxidation (pH 7.0), pyruvate was themajor product in mitochondria from CAM-M. crystallinum, whereasmitochondria from C₃-M. crystallinum produced predominantlyoxaloacetate. Both the extracted NAD malic enzyme in the presenceof CoA and the oxidation of malate to pyruvate by the mitochondriafrom plants in the CAM mode had a pH optimum around 7.0 withactivity declining markedly above this pH. The activity of NAD-malicenzyme, expressed on a cytochrome c oxidase activity basis,was much higher in mitochondria from the CAM mode than the C₃mode. The results indicate that mitochondria of this speciesare adapted to decarboxylate malate at high rates during CAM. ¹Current address: Lehrstuhl für Botanik II, UniversitätWurzburg, Mittlerer Dallenbergweg 64, 8700 Würzburg, WestGermany. ²Current address: KD 120, Chemical Research Division, OntarioHydro, 800 Kipling Avenue, Toronto, Ontario M8Z5S4, Canada. ³Current address: Department of Botany, Washington State University,Pullman, Washington 99164-4230, U.S.A. (Received March 13, 1986; Accepted September 18, 1986)     

The nature of the dual effect of auxin on root formation in Azukia cuttings

In disbudded Azukia stem cuttings, auxin exerted a dual effecton root formation. The first phase of auxin action is identifiedwith the acceleration of cell division, especially longitudinaldivision. In cuttings treated with auxin during the first 24hr, longitudinally divided cells were observed in all 12 rootprimordia, while in water-treated cuttings, such cells wereobserved only in 8 root primordia. The second phase is the promotionof the reaction in which root primordia unable to develop furtherwithout auxin supply develop into roots. Irrespective of thetreatment during the first 24 hr, the auxin-treatment duringthe second 24 hr increased the number of roots protruding fromthe cuttings. Portulal applied during the first 24 hr increased the numberof root primordia which contained longitudinally divided cells.Gibberellin applied during the first 24 hr inhibited both transverseand longitudinal divisions in root primordia. ¹ Supported in part by Grant No. 139011 from the Ministry ofEducation, Japan. ² Present address: Junior College of Toyo University, Hakusan,Bunkyo-ku, Tokyo 112, Japan. (Received June 13, 1978; )