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1.
WEIMBERG RALPH; LERNER H. R.; POLJAKOFF-MAYBER A. 《Journal of experimental botany》1983,34(10):1333-1346
For determination of the effects of polymyxin B, polymyxin E,or ethylenediamine tetra-acetic acid (EDTA) on plant cell membranes,the rates at which three solutes, K+, P1, and sugar, leakedfrom treated tissue culture cell suspensions of Nicotiana tabacumwere measured. The kinetics of leakage from cells treated witheither of the polymyxins was biphasic, whereas kinetics forcells treated with EDTA was monophasic. Only K+ leaked frompolymyxin-treated cells during the first phase, and all threesolutes leaked during the second phase. The slower first phaseis interpreted as leakage of K+ from the Donnan free space andcytoplasm, and the faster second phase as the leakage of solutesfrom the vacuole. The monophasic kinetics of EDTA treatmentindicated that solutes were leaking simultaneously from cytoplasmand vacuole. Of the divalent cations tested, only Ca++ and Mn++counteracted the effects of polymyxin and EDTA. Ca++ even restoredP1 and sugar uptake. Addition of Mg++ or Sr++ to polymyxin-treatedcells did not stop solute leakage but actually enhanced theleakage rates. A model is presented that suggests that polymyxinor EDTA induces solute leakage by forming pores in plant cellmembranes. The effects of divalent cations on membranes oncethe pores are formed are also discussed. Key words: Polymyxin, EDTA, Nicotiana tabacum, Solute leakage 相似文献
2.
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 相似文献
3.
Hayashida Nobuaki; Izuchi Shin-ichi; Sugiura Masahiro; Obokata Junichi 《Plant & cell physiology》1992,33(7):1031-1034
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. 相似文献
4.
VAN DEN ENDE G.; BARENDSE G. W. M.; KEMP A.; CROES A. F. 《Journal of experimental botany》1984,35(12):1853-1859
The effect of glucose on flower bud formation was studied inthin-layer tissue cultures of epidermal strips from flower stalksof Nicotiana tabacum L. cv. Samsun. A minimum concentration of 30 mol m3 glucose in the MS-mediumcontaining 1.0 mmol m3 of both NAA and BA was necessaryfor flower bud formation. With 150 mol m3 glucose a minimumstay of 10 d was required for optimal flower bud formation. Withholding glucose for a limited period at different time intervalsafter the onset of culture caused a delay in flower bud formationand did not affect previous development on glucose. The resultsindicated that competence for flower bud initiation is not restrictedto the early stage of culture. The process may start at anytime later at the appropriate glucose concentration. However,for both optimal initiation and further development of flowerbuds the presence of a metabolizable sugar is required. Incubationof the tissue on glucose is associated with higher respirationrate. Key words: Flower formation, Glucose, mannitol, Nicotiana tabacum, Respiration, tissue culture 相似文献
5.
Molecular Cloning and Characterization of a cDNA Clone That Encodes a Cdc2 Homolog from Nicotiana tabacum 总被引:1,自引:0,他引:1
Setiady Yulius Yulianto; Sekine Masami; Hariguchi Norimitsu; Kouchi Hiroshi; Shinmyo Atsuhiko 《Plant & cell physiology》1996,37(3):369-376
We have isolated a cDNA clone (cdc2Nt1) that encodes a homologof p34cdc2/CDC28 kinase from tobacco (Nicotiana tabacum). Thecdc2Ntl protein showed extensive similarity to other homologsof Cdc2 from plants. Complementation studies showed that thecdc2Ntl gene was able to overcome cell cycle arrest at boththe G1/S and the G2/M transitions of cdc28ts mutants of buddingyeast, demonstrating that the cdc2Ntl protein was able to replacethe Cdc28 kinase at both the G1/S and the G2/M transitions.Analysis of gene expression demonstrated that the cdc2Ntl genewas transcribed constitutively throughout the cell cycle butthat it was preferentially expressed in actively dividing tobaccoBY-2 cells. (Received July 13, 1995; Accepted February 15, 1996) 相似文献
6.
Embryogenesis and Plantlet Formation through Direct Culture of Isolated Pollen of Nicotiana tabacum cv. Samsum and Nicotiana rustica cv. Rustica 总被引:1,自引:0,他引:1
Pollen embryos and plantlets of Nicotiana tabacum cv. Samsunand Nicotiana rustica cv. Rustica were obtained through directpollen culture without prior treatment or prior culture of anthersor buds. Isolated pollen was cultured first in a medium withoutsucrose, then transferred into Nitsch's H medium containing2% sucrose and 5 mM glutamine. The optimum medium for the initialculture was water and the optimum period of culture was ca.6 days when binucleate pollen was used.
1 Present address: Friedrich Miescher Inst., P.O.B. 273, CH-4002Basel, Switzerland. (Received January 18, 1982; Accepted March 19, 1982) 相似文献
7.
Regulation of Chloroplast Gene Expression is Affected in ali, a Novel Tobacco Albino Mutant 总被引:7,自引:0,他引:7
Bae Chang-Hyu; Abe Tomoko; Matsuyama Tomoki; Fukunishi Nobuhisa; Nagata Noriko; Nakano Takeshi; Kaneko Yasuko; Miyoshi Kazumitsu; Matsushima Hisashi; Yoshida Shigeo 《Annals of botany》2001,88(4):545-553
8.
Video images of the distributional pattern of membrane-associatedcalcium (Ca2+) and calmodulin (CaM) have been documented andanalysed during pollen hydration, germination and tip growthin Nicotiana tabacum. Digitization of fluorescence microscopeimages of chlorotetracycline (CTC) and fluphenazine (FPZ)-fluorescenceemissions reveal that there is a maximum concentration of membrane-associatedCa2+ and also CaM in the vicinity of germination apertures ofhydrated pollen. With the onset of germination relatively higheramounts of Ca2+ and CaM were found to regionalize towards theaperture through which the pollen tube would emerge Both shortand long growing pollen tubes manifest tip-to-base Ca2+ andCaM gradients which are disturbed in non-growing tubes. Tubegrowth and the Ca2+-gradient were significantly affected byvanadate and verapamil suggesting that both a vanadate-sensitiveCa2+-transport system and verapamil-sensitive Ca2+ channelsare involved in maintaining Ca2+ homeostasis during pollen germinationand tube growth. The possible interactions of Ca2+ and CaM withdifferent cytoskeletal proteins modulating organelle movementare also briefly discussed. Image analysis, calcium, calmodulin, Nicotiana tabacum L., pollen germination, pollen tube, tip growth, Ca2+-channels, Ca2+ transport ATPase 相似文献
9.
Matoh Toru; Ishigaki Ken-ichi; Mizutani Masaharu; Matsunaga Waki; Takabe Keiji 《Plant & cell physiology》1992,33(8):1135-1141
Cultured cells of tobacco (Nicotiana tabacum L. cv. Bright Yellow2) grown under the standard culture conditions (1 mg boron liter1medium as boric acid) contained boron at a concentration of2.26 mg boron kg1 oven-dried cells and the protoplastcontained 1.26% of the boron in the cells. The cells requiredboron for growth and the half-maximum growth rate was obtainedwith 0.056 mg of boron liter1 medium. Subculturing thecells in media with lower concentrations of boron allowed selectionof cells that can grow even in the presence of 1 µg boronliter1 medium. Cell walls of the selected cells seemedto be thicker than those of the control cells and Golgi bodieswere accompanied by more secretory vesicles than those in thecontrol cells. (Received May 25, 1992; Accepted September 10, 1992) 相似文献
10.
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; ) 相似文献
11.
Ethylene and Ethane Production in 2,4-D Treated and Salt Treated Tobacco Tissue Cultures 总被引:1,自引:0,他引:1
Gas chromatography was used to measure ethylene (ethene) andethane production by tobacco (Nicotiana tabacum cv. Wisconsinno. 38) callus tissues grown on media containing inorganic saltsaccording to Murashige and Skoog (1962), sucrose, myo-inositol,thiamine-HCl kinetic according to Linsmaier and Skoog (1965),and either 2,4-dichiorophenoxyacetic acid (2,4-D) in the range0100 mgl1 or 2 mgl1 indoi-3-ylacetic acidplus NaCl in the range 0200 Meq l1. Ethylene productionrates were high (> 500 nl h1 g1 fresh weight)initially in all treatments. Subsequently, ethylene productiondeclined in rapidly growing cultures but remained high in moderatelyand severely 2,4-D (> 0·5 mgl1) stressed andin severely NaCl (150 Meql1) stressed cultures. Highinitial rates of ethane production (> 200 nl h1 g1fresh weight) were obtained under conditions of severe stresscaused by 2,4-D or NaCl but not in control or moderately inhibitedcultures. With further incubation ethane production declinedin the severely stressed cultures. It is concluded that ethyleneproduction can be used as an index of moderate 2,4-D stressand severe NaCl stress by virtue of the high persisting ratesof ethylene production in stressed cultures. Ethane productioncan be used as an early index of severe stress caused by either2,4-D or NaCl in vitro. Nicotiana tabacum L., tobacco, ethylene, ethenen, ethane, 2,4-dichlorophenoxyacetic acid, auxin, stress, callus tissue 相似文献
12.
Reciprocal transfers of Nicotiana tabacum cv. Xanthi nc. leafexplants were made daily between root inducing medium (RIM)and shoot inducing medium (SIM), SIM and a basal medium containingno growth regulators (BM), and RIM and BM. It was found thatthe explants became determined for shoot production after 6d, while roots were produced after only 1 d on RIM before transferto BM. The competence of the explant to produce roots was greatlyreduced by culture on BM prior to culture on RIM. There wasfar less reduction in shoot numbers with preculture on BM. Explantswere found to be only weakly canalized for both caulogenesisand rhizogenesis for the first 2 d after determination. Thereafterthey became strongly canalized. Transfers were also made fromBM to SIM and back to BM, which revealed that the explants becamecompetent for caulogenesis in the absence of cytokinins priorto determination. The period for which SIM is required can bereduced to only 1 d. Key words: Nicotiana tabacum, in vitro, organogenesis, competence, determination 相似文献
13.
In synchronously cultured tobacco cells (Nicotiana tabacum cv.Xanthi), the incorporation of U-14C-adenosine into butanol-solublecytokinins in vivo was studied. The radioactivity was incorporatedinto zeatin, ribosylzeatin, isopentenyladenosine and glucosylzeatinafter 20 min. The radioactive cytokinins were identified bythin-layer chromatography and high performance liquid chromatography.From the short time course of the incorporation of 14C-adenosineinto butanol-soluble cytokinins, the presence of the followingbiosynthetic pathway in vivo was suggested: adenosine is converedinto isopentenyladenosine and then into zeatin via ribosylzeatin.The biosynthetic pathway of free cytokinins in vivo is comparedwith that in vitro. (Received June 20, 1980; ) 相似文献
14.
In synchronously cultured tobacco cells (Nicotiana tabacum cv.Xanthi), the incorporation of U-14C-adenosine into butanol-solublecytokinins in vivo was studied. The radioactivity was incorporatedinto zeatin, ribosylzeatin, isopentenyladenosine and glucosylzeatinafter 20 min. The radioactive cytokinins were identified bythin-layer chromatography and high performance liquid chromatography.From the short time course of the incorporation of 14C-adenosineinto butanol-soluble cytokinins, the presence of the followingbiosynthetic pathway in vivo was suggested: adenosine is converedinto isopentenyladenosine and then into zeatin via ribosylzeatin.The biosynthetic pathway of free cytokinins in vivo is comparedwith that in vitro. (Received June 20, 1980; ) 相似文献
15.
Kost Benedikt; Galli Alessandro; Potrykus Ingo; Neuhaus Gunther 《Journal of experimental botany》1995,46(9):1157-1167
An efficient system has been established that allows well controlledDNA microinjection into tobacco (Nicotiana tabacum) mesophyllprotoplasts with partially regenerated cell walls and subsequentanalysis of transient as well as stable expression of injectedreporter genes in particular targeted cells or derived clones.The system represents an effective tool to study parametersimportant for the successful transformation of plant cells bymicroinjection and other techniques. Protoplasts were immobilizedin a very thin layer of medium solidified with agarose or alginate.DNA microinjection was routinely monitored by coinjecting FITC-dextranand aimed at the cytoplasm of target cells. The injection procedurewas optimized for efficient delivery of injection solution intothis compartment. Cells were found to be at the optimal stagefor microinjection about 24 h after immobilization in solidmedium. Embedded cells could be kept at this stage for up to4 d by incubating them at 4 C in the dark. Within 1 h successfuldelivery of injection, solution was routinely possible into2040 cells. Following cytoplasmic coinjection of FITC-dextran and pSHI913,a plasmid containing the neo (neomycin phosphotransferase II)gene, stably transformed, paromomycin-resistant clones couldbe recovered through selection. Transgenic tobacco lines havebeen established from such clones. Injection solutions containingpSHI913 at a concentration of either 50 µg ml1or 1 mg ml1 have been tested. With 1 mg ml1 plasmidDNA the percentage of resistant clones per successfully injectedcell was determined to be about 3.5 times higher. Incubationof embedded protoplasts at 4C before microinjection was foundto reduce the percentage of resistant clones obtained per injectedcell Protoplasts were immobilized above a grid pattern and the locationof injected cells was recorded by Polaroid photography. Thefate of particular targeted cells could be observed. Isolationand individual culture of clones derived from injected cellswas possible. Following cytoplasmic coinjection of FITC-dextranand 1 mg ml1 plasmid DNA on average about 20% of thetargeted cells developed into microcalli and roughly 50% ofthese calli were stably transformed. Transient expression ofthe firefly luciferase gene (Luc) was nondestructively analysed24 h after injection of pAMLuc. Approximately 50% of the injectedcells that were alive at this time point expressed the Luc genetransiently. Apparently, stable integration of the injectedgenes occurred in essentially all transiently expressing cellsthat developed into clones. Key words: DNA microinjection, firefly luciferase, FITCdextran, Nicotiana tabacum, protoplast transformation 相似文献
16.
Sorbitol Synthesis in Transgenic Tobacco with Apple cDNA Encoding NADP-Dependent Sorbitol-6-Phosphate Dehydrogenase 总被引:8,自引:0,他引:8
The apple (Malus domestica) cDNA encoding NADP-dependent sorbitol-6-phosphatedehydrogenase (S6PDH) was stably integrated and expressed intransgenic tobacco (Nicotiana tabacum cv. SR1). Expression ofthe cDNA in either a sense or antisense orientation was accomplishedusing cauliflower mosaic virus regulatory sequences (CaMV35S).Sorbitol synthesis was confirmed by gas-chromatography-mass-spectroscopy(GC-MS). Sorbitol concentration in the leaves of the transgenicplants expressing the sense orientation varied from 186 to 446nmol (g fr wt)-1. The concentration positively correlates withS6PDH activity in leaves. Neither sorbitol nor S6PDH activitywas detected in the extracts of nontransformed tobacco or transgenictobacco expressing the antisense orientation. These resultsprovide key genetic evidence that S6PDH expression is sufficientfor the synthesis of sorbitol in tobacco, implicating it asa key enzyme in the sorbitol biosynthetic pathway in apple andperhaps other members of the woody Rosaceae.
1Present address: Laboratory of Pomology, Faculty of Agriculture,Kyoto University, Sakyo, Kyoto, 606-01 Japan 相似文献
17.
MCINNES E.; DAVEY M. R.; MULLIGAN B. J.; DAVIES K.; SARGENT A. W.; J.MORGAN A. 《Journal of experimental botany》1989,40(10):1135-1144
Nicotiana glauca, N. tabacum, Solanian dulcamara and S. nigrumwere transformed by Agrobacteriun rhizogenes strain BN1010 (TLTR+).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+DTR+) 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 相似文献
18.
Potassium-Ammonium Uptake Interactions in Tobacco Seedlings 总被引:6,自引:0,他引:6
SCHERER HEINRICH W.; MACKOWN CHARLES T.; LEGGETT J. EVERETT 《Journal of experimental botany》1984,35(7):1060-1070
Short-term (< 12 h) uptake experiments were conducted with67-week-old tobacco (Nicotiana tabacum L. cv. Ky 14)seedlings to determine absorption interactions between K+ andNH4+. At equal solution concentrations (0.5 mol m3) netK+ uptake was inhibited 3035% by NH4+ and NH4+ uptakewas decreased 924%. Removal of NH4+ resulted in completerecovery in K+ uptake rate, but NH4+ uptake rate did not recoverwhen K+ was removed. In both cases, inhibition of the uptakerate of one cation saturated as the concentration of the othercation was increased up to 0.5 mol m3. The relative effectof K+-NH4+ interactions was not altered when Cl- was replacedwith SO42, but the magnitudes of the uptake rates wereless in the absence of Cl-. The Vmax for NH4+ uptake was reducedfrom 128 to 105 µmol g1 dry wt. h1 in thepresence of 0.5 mol m3 K+ and the Km for NH4+ doubledfrom 12 to 27 mmol m3 in the presence of K+. The resultsof these K+-NH4+ experiments are interpreted as mixed-noncompetitiveinteractions. However, an enhanced efflux of K+ coupled to NH4+influx via an antiporter cannot be ruled out as contributingto the decrease in net K+ uptake. Key words: Nicotiana tabacum, K+, NH4+, Uptake interactions 相似文献
19.
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 auxinlanolinpastes 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 相似文献
20.
Song Nam Hi; Fujioka Shozo; Kamiya Yuji; Sakurai Akira; Liu Jang Ryol 《Plant & cell physiology》1991,32(6):783-789
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) 相似文献