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1.
Welbaum G.E.; Bian D.; Hill D.R.; Grayson R.L.; Gunatilaka M.K. 《Journal of experimental botany》1997,48(3):643-654
The freezing tolerance of many plants, such as pea (Pisum sativum),is increased by exposure to low temperature or abscisic acidtreatment, although the physiological basis of this phenomenonis poorly understood. The freezing tolerance of pea shoot tips,root tips, and epicotyl tissue was tested after cold acclimationat 2C, dehydration/rehydration, applications of 104M abscisic acid (ABA), and deacclimation at 25C. Tests wereconducted using the cultivar Alaska, an ABA-deficientmutant wil, and its wildtype. Freezinginjury was determined graphically as the temperature that caused50% injury (T50) from electrical conductivity. Endogenous ABAwas measured using an indirect enzyme-linked immunosorbant assay,and novel proteins were detected using 2-dimensional polyacrylamidegel electrophoresis. The maximum decrease in T50 for root tissuewas 1C for all genotypes, regardless of treatment. For Alaskashoot tips and epicotyl tissue, exogenous ABA increased thefreezing tolerance by 1.5 to 4.0C, while coldtreatment increased the freezing tolerance by 7.5 to14.8C. Cold treatment increased the freezing toleranceof shoot tips by 9 and 15C for wiland wild-type, respectively. Cold acclimationincreased endogenous ABA concentrations in Alaskashoot tips and epicotyls 3- to 4-fold. Immunogold labeling increasednoticeably in the nucleus and cytoplasm of the epicotyl after7 d at 2C and was greatest after 30 d at the time of maximumfreezing tolerance and soluble ABA concentration. Cold treatmentinduced the production of seven, three, and two proteins inshoot, epicotyl, and root tissue of Alaska, respectively.In Alaska shoot tissue, five out of seven novelproteins accumulated in response to both ABA and cold treatment.However, only a 24 kDa protein was produced in wiland wild-type shoot and epicotyl tissues aftercold treatment. Abscisic acid and cold treatment additivelyincreased the freezing tolerance of pea epicotyl and shoot tissuesthrough apparently independent mechanisms that both resultedin the production of a 24 kDa protein. Key words: Pisum sativum, cold acclimation, immuno-localization 相似文献
2.
SINGH N; SINGH S N; SRIVASTAVA K; YUNUS M; AHMAD K. J; SHARMA S C.; SHARGA A N 《Annals of botany》1990,65(1):41-44
Five Gladiolus cultivars, namely Aldebaran, BrightEye, Illusion, Manisha andManmohan, were exposed to 1 and 2 µg l1sulphur dioxide to test their relative-sensitivity toleranceto the pollutant Plants were fumigated experimentally for 2h daily Foliar injury symptoms were observed first in Manishafollowed by Aldebaran and Illusionat the higher dose Photosynthetic pigments and leaf extractpH were significantly decreased, particularly in Manishaand Illusion Overall disturbances in the plantmetabolism due to SO2 treatment led to retarded growth of plants,as evident from decreased shoot length and phytomass valuesThe order of sensitivity of the five Gladiolus cultivars toSO2 was as follows, with the greatest first Manisha, Illusion,Aldebaran, Bright Eye, Manmohan Cultivars, Gladiolus, sensitivity, sulphur dioxide, tolerance 相似文献
3.
Cytological examination showed that day 3 pea embryo cells wereundifferentiated in terms of morphological or gross cytologicalappearance. Histogenesis had commenced by day 4 and was accompaniedby cytological differentiation with a gradient in vacuolationapparent along the root/shoot axis. Day 3 embryonic cells werecytologically different from meristematic (day 4 and 5) cellsof the shoot apex. Cells of the embryo base appeared to havean intimate association with the middle suspensor cells. Pisum sativum L. cv. Alaska, pea, morphology, cytology, histogenesis, development 相似文献
4.
ABA and Low Temperature Induce Freezing Tolerance via Distinct Regulatory Pathways in Wheat 总被引:1,自引:0,他引:1
Dallaire Sylvain; Houde Mario; Gagne Yves; Saini Hargurdeep S.; Boileau Serge; Chevrier Normand; Sarhan Fathey 《Plant & cell physiology》1994,35(1):1-9
The role of ABA in the induction of freezing tolerance was investigatedin two wheat (T. aestivum L.) cultivars, Glenlea (spring var)and Fredrick (winter var). Exogenous application of ABA (5x105M for 5 days at 24°C) increased the freezing tolerance ofintact plants by only 3°C (LT50) in both cultivars. Maximalfreezing tolerance (LT50 of 9°C for Glenlea and 17°Cfor Fredrick) could only be obtained with a low temperaturetreatment (6/2°C; day/night) for 40 days. These resultsshow that exogenously applied ABA cannot substitute for lowtemperature requirementto induce freezing tolerance in intactwheat plants. Furthermore, there was no increase in the endogenousABA level of wheat plants during low temperature acclimation,suggesting the absence of an essential role for ABA in the developmentof freezing tolerance in intact plants. On the other hand, ABAapplication (5x105 M for 5 days at 24°C) to embryogenicwheat calli resulted in an increase of freezing tolerance similarto that achieved by low temperature. However, as in intact plants,there was no increase in the endogenous ABA level during lowtemperature acclimation of calli. These results indicate thatthe induction of freezing tolerance by low temperature is notassociated with an increase in ABA content. Using an antibodyspecific to a protein family associated with the developmentof freezing tolerance, we demonstrated that the induction offreezing tolerance by ABA in embryogenic wheat calli was correlatedwith the accumulation of a new 32 kDa protein. This proteinis specifically induced by ABA but shares a common antigenicitywith those induced by low temperature. These results suggestthat ABA induces freezing tolerance in wheat calli via a regulatorymechanism different from that of low temperature. (Received June 15, 1993; Accepted September 16, 1993) 相似文献
5.
The auxins contained in 5-mm. tips of horizontal Vicia fabaroots have been compared with those in tips of vertical rootsafter cold ethanol extraction, paper-chromatographic separation,and Avena mesocotyl bioassay. At about the time curvature commencesin horizontal roots there is a marked increase in the contentof an auxin corresponding to AP(ii) of pea roots(Rf 0.350.65 in isobutanol/methanol/water). There areindications that this is not due to its release from an inactivebound state but that it is either synthesized de novo or maybe converted from another auxin corresponding to AP(iii)of pea roots (Rf 0.751.0). The literature dealing with the auxins of geotropically stimulatedorgans is reassessed and it is concluded that, with the exceptionof the Avena coleoptile, there is very little evidence favouringa simple transport redistribution of auxin under gravity; themajority of the data favour an effect of gravity on auxin metabolism. 相似文献
6.
Extracrts of the shoot tips of normal and frenchedtobacco plants were chemically separated into acidic, neutral,and basic ethersoluble fractions. On chromatograms ofthese, some plant growth regulators were assayed using the Avenacoleoptile section extension test. The acidic auxins and an acids and a neutral growth inhibitorwere found. One auxin, with the samew RF value as indole-3-aceticacid, was four times more concentrated on normal as in frenchedplants. No differences could be established between the twotypes of plants in regard to other growth regulators detected. It is argued that the symptoms of the physiological diseasefrenching could be explained in terms of a auxindeficiency. 相似文献
7.
Impedance spectroscopy was used in studying frost hardinessof leaves of two diploid rhododendron cultivars, RhododendronL. PJM and R. Cunningham's White,and their tetraploid derivatives, R. Northern Starburst(NSB) and CW4. After the growing season and initial hardeningin a greenhouse, plants were subjected to an acclimation regimein a phytotron: 3 consecutive weeks at +5, +1 and -2°C each.Hardiness was studied with controlled freezing tests beforeeach decrease in temperature and at the end of the experiment,based on data of extracellular resistance reand relaxation time of the frost-exposed leaves. The correlation of the two estimateswas 0.92. Generally, the diploid clones had better frost hardinessthan the tetraploid clones. At the end of the experiment, frosthardiness of the diploid PJM was -28.7°C andthat of the tetraploid NSB -20.6°C. Leaves of the diploidCunningham's White and of the tetraploid CW4 hardenedto -32.0°C and -20.9°C, respectively. Frost hardinessestimated by impedance spectroscopy correlated well with earlierresults based on visual scoring (r = 0.810.86) and electrolyteleakage tests (r = 0.840.90), but results from impedancespectroscopy indicated weaker hardiness than the other tests.The difference between the results from impedance spectroscopyand the other tests was smaller and more coherent within theCunningham's White clones than within PJMand NSB. Changes in extracellular and intracellular resistanceof non-frozen leaves during the acclimation correlated withthe changes in frost hardiness of Cunningham's Whiteclones, but not with those of PJM and NSB, whichbelong to another subspecies.Copyright 2000 Annals of BotanyCompany Cold resistance, evergreen, frost hardiness, impedance spectroscopy, polyploid, Rhododendron, tetraploid 相似文献
8.
Increased Synthesis of ABA in Partially Dehydrated Root Tips and ABA Transport from Roots to Leaves 总被引:27,自引:3,他引:24
Zhang, J. and Davies, W. J. 1987. Increased synthesis of ABAin partially dehydrated root tips and ABA transport from rootsto leaves.J. exp. Bot. 38: 20152023. Isolated root tips of pea (Pisum sativum L. cv. Feltham First)and Commelina communis L. were air-dried until they lost between10% and 40% of their fresh weight, followed by a period of incubationat these reduced water contents. These treatments resulted inincreased ABA production, suggesting that root tips of bothspecies have the capacity to synthesize ABA in increased amountswhen water deficits develop in the root. The ABA concentrationin pea roots increased linearly as turgors fell below about0·15 M Pa and relative water contents (R WC) fell below90%. Commelina roots produced more ABA when RWC fell below asimilar value but the threshold turgor for increased ABA productionin Commelina roots was around 0·30 MPa. Roots of intact plants loaded with ABA as a result of incubationin solutions of varying concentrations provided ABA to leaveswhich resulted in increased ABA concentrations in the leaveswhen these were assayed several hours later. This occurred whenthese roots were not contributing substantially to transpirationalflux. Leaves on shoots that were enclosed and darkened and thereforenot transpiring, did not accumulate ABA from loadedroots. A role for root-sourced ABA in root-to-shoot communication ofthe effects of soil drying is discussed. Key words: ABA, roots, water relations 相似文献
9.
Effect of Inorganic Phosphate on the Biosynthesis of Purine and Pyrimidine Nucleotides in Suspension-Cultured Cells of Catharanthus roseus 总被引:1,自引:0,他引:1
The levels of purine and pyrimidine nucleotides in suspensioncultures of Catharanthus roseus were determined 24 h after stationary-phasecells were transferred to fresh complete (+Pi)or phosphate-deficient (Pi) Murashige-Skoogmedium. The levels of ATP, GTP, UTP and CTP were from approx.3 to 5-fold greater in the cells grown in +Pimedium than in the cells grown in Pi medium.The levels of almost all other nucleotides were slightly higherin the cells in +Pi medium. The rates of de novoand salvage biosynthesis of purine and pyrimidine nucleotideswere estimated from the rates of incorporation of radioactivityfrom [14C]formate, [214C]glycine, NaH14CO3, [614C]orotate,[814C]adenine, [814C]adenosine, [214C]uraciland [214C]uridine. The results indicated that the activityof both the de novo and the salvage pathway was higher in thecells in +Pi medium than in the cells in Pimedium. The rate of degradation estimated from the rate of releaseof 14CO2 from labelled purines and pyrimidines indicated thatdegradation of uridine was significantly reduced in the cellsin +Pi medium, but no significant difference wasfound in the degradation of adenine, adenosine and uracil. Thepossible role of Pi in the control of the biosynthesis of nucleotidesand in the degradation of uridine is discussed. Catharanthus roseus, Madagascar periwinkle, suspension culture, inorganic phosphate, nucleotides, purines, pyrimidines, biosynthesis, degradation 相似文献
10.
Abscisic acid (ABA) has been shown to increase freezing toleranceof bromegrass (Bromus in-ermis Leyss cv. Manchar) cell suspensioncultures from a LT50 (the temperature at which 50% cells werekilled) of 7 to 30?C in 5 days at 23?C. Our objectivewas to study the qualitative changes in the translatable RNApopulation during ABA induced frost tolernace. In vitro translationproducts of poly(A)+ RNA isolated from bromegrass cells withor without 75 µM ABA treatment for various periods oftime were separated by 2D-PAGE and visualized by fluorography.SDS soluble proteins from the same treatments were also separatedby 20-PAGE. After 5 days treatment, at least 22 new or increasedabundance SDS soluble polypeptides were observed. From fluorographs,29 novel or increased abundance in vitro translation productscould be detected. The pattern of changes between ABA inducedSDS-soluble proteins and translation products from the 2D gelswere similar. A time course study (07 days) showed that17 of the 29 translation products were detected after 1 dayABA treatment, and at least 14 were present after 1 h. Coldtreatment (+4?C) induced fewer changes in the pool of translatableRNA than with ABA treatment. Three translation products inducedby cold appear to be similar to 3 of the ABA induced translationproducts. The majority of the ABA inducible translatable RNAsappeared at 10 µM or higher which coincides with the inductionof freezing tolerance. Many of these ABA inducible RNAs persisted7 days after ABA was removed from the media and correspondinglythe LT50 (17?C) was still well above the control level(17?C). The results suggest that ABA alters the poolof translatable RNAs during induction of freezing tolerancein bromegrass suspension culture cells.
1Oregon Agricultural Experiment Station Technical Paper No.9256. (Received August 3, 1990; Accepted October 18, 1990) 相似文献
11.
Seeds ofTaxus maireiare known for their deep dormancy whichcan only be broken by a procedure involving warm stratificationfollowed by cold stratification. Treatments with alternatingtemperatures of 25/15 or 23/11 °C (12 h light) for 6 monthsfollowed by 5 °C for 3 months were successful in overcomingseed dormancy. After 6 months of warm stratification, cytologicalchanges observed included: enlargement of the embryo; a decreasein the number of lipid bodies; appearance of ER; and increasesin mitochondria, plastids, dictyosomes, vacuoles and microbodiesin the shoot apical meristem. Cold stratification followingthe warm treatment induced cell division, and one or two distinctnucleoli in the shoot apical meristem cells were observed. Bothwarm and cold stratification reduced endogenous ABA concentrationsfrom the original 8888 pg per freshly harvested seed to 392and 536 pg, respectively. Treatment with exogenous gibberellinsafter seeds had been warm-stratified showed that GA4and GA7wereeffective at promoting seed germination, but GA3was not. Theseresults suggest that the strong seed dormancy ofT. maireicouldbe caused by a high ABA content and underdevelopment of theembryos in freshly shed seeds. We conclude that warm stratificationwith alternating temperatures increases the growth of embryosby cell expansion and enlargement and decreases ABA content,but seeds still remain ungerminated. Cold stratification mayinduce the response to GAs and initiate cell division resultingin release from physiological dormancy and subsequent germinationofT. maireiseeds.Copyright 1998 Annals of Botany Company Taxus mairei; ultrastructure; abscisic acid; gibberellin; seed dormancy; stratification; germination. 相似文献
12.
《Journal of experimental botany》1977,28(6):1247
Effects of coupled solute and water flow in plant roots withspecial reference to Brouwer's experiment. Edwin L. Fiscus. p. 71 Abstract: Line 3 delete interval insertinternal. p. 73 Materials and Methods: line 6: delete diversion insert division line 9 equation should read Jv=Lp PRT(C0C1). 74 Last line of figure legend: 101 should read 1011. 75 Line 11: delete seems insert seem. le 1 column heading106 should read 1011. 77 delete ...membrane in series of... insert membranein series or... Delete final paragraph. 相似文献
13.
The Sensitivity of Net Photosynthesis in Several Plant Species to Short-term Fumigation with Sulphur Dioxide 总被引:4,自引:0,他引:4
The effects of exposure of up to 2 h with sulphur dioxide ona range of plant species was observed by measuring changes inthe rate of net photosynthesis under closely controlled environmentalconditions. Ryegrass, Lolium perenne S23 was thespecies most sensitive to SO2; significant inhibition was detectedat 200 nl l1. Fumigations at 300 nl l1 also inhibitedphotosynthesis in field bean (Vicia faba cv. Three FoldWhite and Blaze) and in barley (Hordeumvulgare cv. Sonja). No effect was detected inwheat (Triticum aestivum cv. Virtue) at concentrationsup to 600 nl l1 SO2, or in oil-seed rape (Brassica napuscv. Rafal) except at 800 nl l1 SO2). Recoverycommenced immediately after the fumigation was terminated andwas complete within 2 h when inhibition had not exceeded 20%during the SO2 treatment. Key words: Sulphur dioxide, short-term fumigation, photosynthesis 相似文献
14.
Influence of Flooding on Net CO2Assimilation, Growth and Stem Anatomy of Annona Species 总被引:1,自引:0,他引:1
NUNEZ-ELISEA ROBERTO; SCHAFFER BRUCE; FISHER JACK B.; COLLS ANGEL M.; CRANE JONATHAN H. 《Annals of botany》1999,84(6):771-780
A series of experiments was conducted to assess net CO2assimilationand growth responses to waterlogging of grafted and seedlingtrees in the genus Annona. Seedlings of A. glabra, A. muricataandA. squamosa L., and scions of Gefner atemoya(A. squamosaxA. cherimola Mill.), 49-11 (GefneratemoyaxA. reticulata L.), 4-5 (PriestleyatemoyaxA. reticulata), A. reticulata grafted onto either A.glabra, A. reticulata orA. squamosa rootstocks were floodedfor up to 60 d. Soil anaerobiosis occurred on the third dayof flooding. Seedlings ofA. glabra and A. muricata, and thescions 49-11, Gefner atemoya, andA. reticulata grafted onto A. glabra rootstock were consideredflood tolerant based on their ability to survive and grow inflooded conditions. Scions of the normally flood-sensitive A.reticulata, Gefner atemoya, and 49-11tolerated root waterlogging when grafted onto the flood-tolerantspecies, A. glabra. In contrast, flooding of A. squamosa seedlingsand rootstocks, and A. reticulata rootstocks greatly reducedgrowth and net CO2assimilation rates, and resulted in 2080%tree mortality. Stem anatomical responses to long-term flooding(12 continuous months) were assessed in seedlings of A. glabraand A. muricata, and trees of 49-11 grafted ontoA. glabra. Flooded trees developed hypertrophied stem lenticels,particularly in A. glabra, and enlarged xylem cells resultingin thicker stems with reduced xylem density. Flooding did notincrease air spaces in pre-existing xylem near the pith or inxylem tissue that was formed during flooding. Thus, flood tolerancedid not involve aerenchyma formation in the stem. Copyright1999 Annals of Botany Company Flood tolerance, net CO2assimilation, photosynthesis, stem anatomy, shoot growth, anaerobiosis, Annonaceae. 相似文献
15.
Seasonal patterns of growth, dehydrins and water-soluble carbohydrates in genotypes of Dactylis glomerata varying in summer dormancy 总被引:1,自引:0,他引:1
Background and Aims Summer dormancy in perennial grasseshas been studied inadequately, despite its potential to enhanceplant survival and persistence in Mediterranean areas. The aimof the present work was to characterize summer dormancy anddehydration tolerance in two cultivars of Dactylis glomerata(dormant Kasbah, non-dormant Oasis)and their hybrid using physiological indicators associated withthese traits. Methods Dehydration tolerance was assessed in a glasshouseexperiment, while seasonal metabolic changes which produce putativeprotectants for drought, such as carbohydrates and dehydrinsthat might be associated with summer dormancy, were analysedin the field. Key Results The genotypes differed in their ability tosurvive increasing soil water deficit: lethal soil water potential(s) was 3·4 MPa for Kasbah (althoughnon-dormant), 1·3 MPa for Oasis,and 1·6 MPa for their hybrid. In contrast, lethalwater content of apices was similar for all genotypes (approx.0·45 g H2O g d. wt1), and hence the greater survivalof Kasbah can be ascribed to better drought avoidancerather than dehydration tolerance. In autumn-sown plants, Kasbahhad greatest dormancy, the hybrid was intermediate and Oasishad none. The more dormant the genotype, the lower the metabolicactivity during summer, and the earlier the activity declinedin spring. Decreased monosaccharide content was an early indicatorof dormancy induction. Accumulation of dehydrins did not correlatewith stress tolerance, but dehydrin content was a function ofthe water status of the tissues, irrespective of the soil moisture.A protein of approx. 55 kDa occurred in leaf bases of the mostdormant cultivar even in winter. Conclusions Drought avoidance and summer dormancy arecorrelated but can be independently expressed. These traitsare heritable, allowing selection in breeding programmes. 相似文献
16.
Ludwig-Mller Jutta; Schubert Birgit; Pieper Kerstin 《Journal of experimental botany》1995,46(4):423-432
The rate of indole-3-butyric acid (IBA) synthesis in maize seedlingsis dependent on the culture conditions of the plants. When theseedlings were grown on filter paper soaked with different amountsof water, the activity of IBA synthetase differed strongly.High amounts of water (150 and 200 ml per bowl) inhibited IBAsynthesis completely in vitro, whereas 30 and 50 ml water perbowl increased the activity dramatically. Under conditions whereIBA synthetase was inhibited (150 ml H2O), an increase of enzymeactivity was observed when abscisic acid (ABA) was exogenouslyadded in concentrations between 5104 to 5107M. Under drought conditions (50 ml H2O per bowl)the same ABA concentrations were inhibitory. Jasmonic acid andsalicylic acid also enhanced IBA synthetase activity to someextent, whereas indole-3-acetic acid (IAA) and kinetin had noeffect. Activity could also be enhanced by osmotic stress (NaCIand sorbitol), but not under temperature stress. In accompanyinginvestigations the endogenous contents of IAA, IBA, and ABAunder the different culture conditions have been determinedas well as the energy charge of the seedlings. Similar observationshave been made with Amaranthus, wheat and pea seedlings Key words: Abscisic acid, Amaranthus paniculatus, drought stress, inole-3-butyric acid biosynthesis, Pisum sativum, Triticum aestivum, Zea mays 相似文献
17.
《Journal of experimental botany》1960,11(2):150
On page 235, Table I: Equation (1) for Node 4 should read A/Ac=0·840+0·0006Ac;Equation (2) for Node 4 should read A=0·89Acand Equation (2) for Node 510 should read A=0·813Ac. 相似文献
18.
Simulated swards of each of two selection lines of Lolium perennecv. S23 with fast and slow ratesof mature tissue respiration were establishedin growth rooms at 20/15 °C day/night temperatures and studiedover four successive regrowth periods of 46, 30, 26 and 53 daysduration. The slow line outyielded the fast,both in harvestable shoot (above a 5 cm cut) and in root andstubble. Its advantage increased over successive regrowth periodsto 23 per cent (total biomass). Gas analysis measurements onthe entire communities (including roots), during the final regrowthperiod, showed that the slow line had a 2234per cent lower rate of dark respiration per unit dry weight.This enabled it to maintain its greater mass of tissue for thesame cost in terms of CO2 efflux per unit ground area. Halfthe extra dry weight produced by the slow line,relative to the fast, could be attributed to itsmore economic use of carbon. The rest could be traced to a 25per cent greater tiller number which enabled the slowline to expand leaf area faster (though not at a greater rateper tiller), intercept more light and fix more carbon, earlyin the regrowth period. Lolium perenne L., ryegrass, respiration, maintenance respiration, tiller production, simulated swards, canopy photosynthesis, carbon economy 相似文献
19.
Nitrate provision has been found to regulate the capacity forChara corallina cells to take up nitrate. When nitrate was suppliedto N sufficient cells maximum nitrate uptake was reached after8 h. Prolonged treatment of the cells in the absence of N alsoresulted in the apparent ability of these cells to take up nitrate.Chlorate was found to substitute partially for nitrate in theinduction step. The effects on nitrate reductionwere separated from those on nitrate uptake by experiments usingtungstate. Tungstate pretreatment had no effect on NO3uptake induced by N starvation, but inhibitedNO3 uptake associated with NO3 pretreatment. Chloridepretreatment similarly had no effect on NO3 uptake inducedby N deprivation, but inhibited NO3 uptake followingNO3 pretreatment. The data suggest that there are atleast two mechanisms responsible for the inductionof nitrate uptake by Chara cells, one associated with NO3reduction and induced by CIO3 or NO3and one associated with N deprivation. Key words: Nitrate, Chlorate, Chara corallina, Induction 相似文献
20.
A Cytochrome P450 Mediated Naringenin 3'-Hydroxylase from Sweet Orange Cell Cultures 总被引:1,自引:0,他引:1
Doostdar Hamed; Shapiro Jeffrey P.; Niedz Randall; Burke M.D.; McCollum T. Gregory; McDonald Roy E.; Mayer Richard T. 《Plant & cell physiology》1995,36(1):69-77
A microsomal flavonoid 3'-hydroxylase (F3'H) catalyzing themetabolism of naringenin to eriodictyol in Citrus sinensis (L.)Osbeck cv. Hamlin cell suspension cultures wasshown to be a cytochrome P450 enzyme. This reaction requiredO2 and NADPH and was inhibited by CO, with partial reversalof CO-inhibition by light at 450 nm. Cytochrome P450 contentranged from 1020 pmol (mg microsomal protein)1.The F3'H reaction was shown to be linear in regard to proteinconcentration between 2.5 and 25 µg of microsomal protein.The optimum pH for the reaction was 7.47.6 and the temperatureoptimum was between 30 and 37°C. The apparent Km and Vmaxfor naringenin were 24 µM±3.2 and 81.4±7.9pmol eriodictyol min1 (mg protein)1, respectively.The microsomal F3'H was also capable of forming dihydroquercetinfrom dihydrokaempferol (40 pmol min1 (mg protein)1)and of quercetin from kaempferol (3.25 pmol min1 (mgprotein1). Cytochrome c and ketoconazole were the bestinhibitors of WH activity followed by piperonyl butoxide anda-naphthoflavone. Light was shown to be an inducer of the F3'Halmost doubling the specific activity and increasing the microsomalcytochrome P450 content by 30% over that of dark grown cells.F3'H activity was also confirmed in microsomal preparationsof young (new flush) leaves from Hamlin treesand flavedo of Hamlin oranges, Marshgrapefruit, and Lisbon lemon. No activity wasobserved in older, hardened leaves and albedo of all the fruittested. Initiation of embryogenesis in the Hamlincell suspension cultures by switching from a sucrose mediumto a glycerol-based medium resulted in the down-regulation ofF3'H.
1Mention of a trademark, warranty, proprietary product, or vendordoes not constitute a guarantee by the U.S. Department of Agricultureand does not imply its approval to the exclusion of other productsor vendors that may also be suitable. 相似文献