Peroxidase Isoenzyme Patterns in Cell Cultures derived from Cotyledon, Stem, Leaf and Fruit from Grapevine (Vitis vinifera cv. Monastrell)

The expression of peroxidase isoenzymes capable of oxidizing4-hydroxystilbenes was studied during the establishment of cellcultures derived from different tissues (cotyledon, stem, leafand fruit) of Vitis vinifera cv. Monastrell vines. This wascarried out in order to elucidate whether different tissuesof the same plant maintain persistent tissue-specific patternsof gene expression during in vitro culture or whether in vitrocultures are characterized by identical patterns of gene expressionirrespective of the tissue's origin. The results illustratedthat both the isozyme patterns and the substrate specificityof the peroxidase activity secreted to the medium are analogousfor the profile of acidic (Prx A) and basic (Prx B) peroxidaseisoenzymes, only quantitative differences being shown in theneutral peroxidase isoenzyme (Prx N) pattern. These resultssuggest that in vitro cultures of grapevines are characterizedby similar patterns of gene expression, no matter what theirtissue of origin.Copyright 1995, 1999 Academic Press Grapevine, vitis vinifera, cell cultures, 4-hydroxystilbene oxidizing peroxidase isoenzyme, substrate specificity     

Development and Evaluation of an In Vitro System to Study Strawberry Fruit Development

Growth and ripening of strawberry (Fragariaananassa Duch.)fruit harvested at immature stages of development was accomplishedby placing the peduncles of individual fruit in solutions composedof hydroxyquinoline hemisulphate (HQS) and sucrose. Fruit cultivarand developmental stage at harvest were the major determinantsof in vitro performance. ‘Pajaro’ fruit harvestedat 50 to 60% maturity exhibited the greatest and most uniformweight gain when placed in solutions containing 200 mol m^–3HQS and 88 mol m^–3 sucrose. Although the final fruit weightof in vitro-ripened fruit was less than that of field-ripenedfruit, colour development in vitro occurred at the same rateand to the same extent as field-grown fruit. Key words: Ripening, non-climacteric fruit     

A Comparison of Stem Peroxidases in Bush and Vine Forms of Squash (Cucurbita maxima Duch.and C. pepo L.)

Total peroxidase activities and peroxidase isozymes were comparedamong 10 bush- and vine-squash cultivars of Cucurbita maximaDuch. and C. pepo (L.) Several qualitative variations of isozymeswere detected between C. maxima and C. pepo, and some differenceswere evident among varieties of C. pepo. There were no qualitativedifferences between genetically similar bush and vine strains.Quantitative differences were difficult to resolve, althoughit did appear that in varieties of C. maxima exhibiting lowlevels of peroxidase activity, isozymes C₃, C₄, and C₆ wereless intense. Depending upon the cultivars compared, total peroxidase activityin bush forms was either higher or lower than, or the same astheir vine counterparts. Since the major bush genes in the varietiesstudied are believed to be allelic, the results indicate thatthe over-all genotype of squash varieties can influence therelative expression of peroxidase activity between bush andvine forms.     

Cell Number and Cell Size in Parthenocarpicvs. Pollinated Blueberry (Vaccinium ashei) Fruits

Gibberellic acid (GA₃) promotes parthenocarpic fruit developmentand is used commercially to increase fruit set in many crops.However, fruit size is usually smaller than that of pollinatedfruit. The purpose of this work was to determine the anatomicalbasis for differences in fruit size between pollinated and GA₃-inducedparthenocarpic blueberry (Vaccinium asheiReade) fruits. Freshweights at ripening averaged 1.6 and 2.5 g for GA₃-treatedvs.pollinated fruits, respectively. In both pollinated and GA₃-treatedfruits, mesocarp cell number comprised about 75% of the totalpericarp cell number, and increased from     

Protein Dysfunction and Inhibition of Amino Acid Incorporation in Root Segments from Wheat and Mung Bean Seedlings caused by some Amino Acid Analogues

The effects of para-fluorophenylalanine (p-FPA) and azetidine-2-carboxylicacid (AZ) on acid phosphatase and peroxidase activity, and onL-leucine incorporation, in root segments of Triticum aestivumL. cv. Fenman and Vigna radial a (L.) Wilczek, were studied.Incubation with 50 mmol^–3 AZ significantly reduced phosphataseand peroxidase activities in wheat roots, but with 20 mol m^–3p-FPA, only the peroxidase activity was reduced. In mung beanroots, phosphatase activity was inhibited by both AZ and p-FPA.Effects of the ortho- and meta- isomers of FPA on wheat rootphosphatase and peroxidase, and on mung bean phosphatase, werealso examined. Leucine uptake and incorporation were not inhibitedby 5 h pre-incubation with either p-FPA or AZ, but were inhibitedafter 24 h of pre-incubation. The results support the view that,in the shorter term, the analogues inhibit enzyme activity bybecoming incorporated to produce non-functional protein and,in the longer term, metabolism is further affected by inhibitionof protein synthesis. Key words: Fluorophenylalanin, Azetidine-2-carboxylic acid, Triticum aestivum, Vigna radiata     

Computerized Optimization of the Relative Growth of Callus Cultures of Orange Fruit Tissues7

The induction and growth of callus cultures from different partsof the fruit of Coorg mandarin orange (Citrus reticulata, Blanco)and the hormonal combinations that can maximize the growth witha basal MS medium, have been explored using Response SurfaceMethodology. The relative growth, protein, peroxidase pattern,and GC fingerprinting of volatiles have been studied. Calluscultures from juice vesicles were found to be more similar totheir explant counterpart than those from albedo, for all propertiesstudied. Key words: Callus culture, Citrus, albedo, juice vesicles, response surface methodology, growth optimization     

Water Import Rate in Tomato Fruit: A Resistance Model

A model of the water import rate in tomato fruit is proposed.It compares the fruit to a hollow sphere (P) with external radiusR and internal radius R_G, corresponding to pericarp, and containingan internal spherical part (G). The pathway limiting water inputrate at any point I at a distance r from the fruit centre wasassumed to be proportional: (a) in P, to the length of the arcwhich has a radius r and which goes from I to the pedicel extension;(b) in G, to r. The water input rate at I was modelled basedon a law similar to Darcy's law which takes into account thedifference between the water potential at entry of fruit andthe water potential at point I. This latter potential was thesum of fruit osmotic potential and pressure potential due toresistance of tissue to deformation. This potential was proportionalto R-r or R_G -r. The model was expressed at fruit level by alaw such that water mass imported per unit time per unit surfacearea of fruit (f_rw) was a linear function of R. The model wascompared to linear regressions of this rate in terms of R whichhad been found during fruit swelling from published results,and which were obtained at different values of nutrient solutionsalinity. The results suggested that water input in tomato fruitis conditioned by passive forces depending on fruit size.Copyright1994, 1999 Academic Press Fruit, growth, model, resistance, salinity, size, tomato, transfer, water     

Abnormal Stomatal Behaviour and Hormonal Imbalance in flacca, a Wilty Mutant of Tomato: EFFECT OF ABSCISIC ACID AND AUXIN ON STOMATAL BEHAVIOUR AND PEROXIDASE ACTIVITY

The wilty tomato mutant flacca and the normal variety RheinlandsRuhm were compared in terms of: (1) potassium transport intoand out of the guard cells, (2) cell wall properties which includeprotein, hydroxyproline and peroxidase activity, and (3) activityof indol-3yl-acetic acid oxidase. Also studied were the effectsof auxin on stomatal behaviour and peroxidase activity whenapplied to normal plants during development, and the short-termeffect of abscisic acid on the resistance of flacca stomatato closure under plasmolysis. Potassium transport, wall protein and hydroxyproline all seemedto be equal in mutant and normal plants. Peroxidase activitywas higher in the soluble and wall fractions of the mutant,and decreased toward normal in the mutant treated with abscisicacid. More stomata were open and peroxidase activity was higherin normal plants treated with auxin during development. Thepercentage of open stomata under plasmolysis was lower and theiraperture size was smaller in the epidermal strips taken fromabscisic-acid-treated mutant plants than from control mutantplants.     

Tryptophan Oxidizing Enzyme and Basic Peroxidase Isoenzymes in Arabidopsis thaliana (L.) Heynh.: Are They Identical?

The in vitro conversion of [³H]tryptophan by a plasma membraneenriched fraction from Arabidopsis thaliana (L.) Heynh. seedlings,grown in liquid culture, revealed indole-3-acetaldoxime (IAOX)as the only detectable reaction product. The pH optimum of thereaction was at pH 8, the K_m value for tryptophan 12 µM.The formation of IAOX was stimulated about 10-fold by H₂O₂ Incubationexperiments with solubilized proteins and membrane vesiclesshowed that the investigated enzyme(s) were bound covalent tothe plasma membrane. Tryptophan oxidizing enzyme (TrpOxE) andperoxidase activity were not only found in the plasma membrane,but also in the culture medium. Specific IAOX forming activitywas 74-fold and 6-fold higher compared to the crude extractand the plasma membrane fraction, respectively. After isoelectricfocusing of solubilized plasma membrane and precipitated mediumproteins, TrpOxE activity co-migrated with two prominent highpI peroxidase bands stained with benzidine-guaiacol. The zonesof the IEF gel with peroxidase and TrpOxE activity were analyzedby SDS PAGE and revealed in all fractions a main protein bandof ca. 55 kDa. TrpOxE activity and peroxidase activity wereboth inhibited by antisera directed against tobacco and horseradishperoxidase. TrpOxE activity and peroxidase activity were determinedduring plant development. TrpOxE activity peaked after 8 and42 days, whereas peroxidase activity was consistently presentduring the whole life cycle. The inhibitory effects of indolederivatives, especially indole-3-glyoxylic acid, on (i) seedlingdevelopment and (ii) on TrpOxE and peroxidase activity werealso compared. (Received November 1, 1991; Accepted September 2, 1992)     

Separate Assays Specific for Ascorbate Peroxidase and Guaiacol Peroxidase and for the Chloroplastic and Cytosolic Isozymes of Ascorbate Peroxidase in Plants

Ascorbate (AsA) peroxidase can be inactivated both by p-chloromercuribenzoateand by the depletion of AsA but guaiacol peroxidases, such ashorseradish peroxidase, cannot. The cytosolic isozymes of AsAperoxidase are less sensitive to depletion of AsA than the chloroplasticisozymes, which include stromal [Chen and Asada (1989) PlantCell Physiol. 30: 987] and thyla-koid-bound [Miyake and Asada(1992) Plant Cell Physiol. 33: 541] enzymes. Exploring theseproperties, we established simple methods for separate assaysof AsA peroxidase and guaiacol peroxidase and of the three isozymesof AsA peroxidase in plant extracts. These methods were usedto characterize the guaiacol peroxidases and isozymes of AsAperoxidase in plants and algae. (Received October 20, 1993; Accepted February 7, 1994)     

Fruit Development and Structure in Some Indian Bamboos

Fruit structure and development of seven species belonging tofive genera of Indian bamboos are described. The fruit in fourspecies is a caryopsis typical of the family Poaceae. The ovuleis bitegmic; the outer surface of the cells of nucellar epidermisbecomes cutinized and forms the seed coat. Three species beara fleshy fruit with a unitegmic ovule. In a mature fruit theendosperm is either completely absorbed by the embryo or ispresent only in small quantity. The developing embryo comesin direct contact with the fruit wall due to the disintegrationof the nucellus and integument. The embryo is covered by a thickbrown mat from the disorganized cells of the inner layers ofthe fruit wall. Poaceae, Bambusoideae, Bambusa, Dendrocalamus, Melocalamus, Ochlandra, Pseudostachyum (fleshy fruits), fruit wall     

Plant Growth Regulators in Cucumis melo L. var. flexuosus Naud Fruit during Rapid Growth

The fruit growth of the snake melon (Cucumis melo L. var. flexuosusNaud) and the plant hormones contained in its immature fruitwere investigated. The fruit growth started 5 days after pollinationand its rapid growth continued for about 10 days. During thisperiod the growth rate (length) was 9 cm per day. The finalsize of the fruit was about 120 cm in length and 6 cm in width25 days after pollination. The cell number of the fruit increasedto more than twice that of the fruitlet before pollination.The increase started immediately after pollination and stoppedat 10 days after pollination. On the other hand, no change incell size was observed during the first 7 days after pollination.After this period, rapid growth started and continued to theend of the fruit growth. The cell size increased to more than7 times that of the fruitlet before pollination. In rapidly developing immature fruit including placenta andimmature seeds, trans-zeatin riboside (ZR) and ABA were identifiedwith gas-liquid chromatography-selected ion monitoring or gas-liquidchromatography-mass spectrometry analysis. In addition, thepresence of trans-zeatin (Z) and another very polar cytokinin,and a novel gibberellin-like substance which is probably anisomer of GA₃ was suggested. The possible significance of theseplant hormones in fruit growth is discussed. (Received June 27, 1985; Accepted April 8, 1986)     

Content and De Novo Synthesis of Cocaine in Embryos and Endosperms from Fruit of Erythroxylum coca Lam

There is controversy as to whether the mature fruit of Erythroxylumcoca var. coca Lam. contains the cocaine alkaloid (benzoylmethylecgonine).In the present study, cocaine was monitored to determine ifit was present in embryos and endosperms of mature fruit ofE. coca var. coca Lam., and if present, the time required forde novo synthesis in imbibing seed. Seeds from mature fruitof E. coca were dissected to separate the embryos from the endosperms.The separated embryos and endosperms were analysed for cocaine.Subsequently, endosperms and embryos from seed imbibed. undera light and dark treatment were separated on days 3, 6, 9, 12and 15 and analysed for cocaine. Cocaine was present in embryos(0.005% of d. wt) and endosperms (0–001% of d. wt) ofmature fruit of E. coca. De novo synthesis of cocaine occurredonly in embryos of seed imbibed under light after day 9 of imbibition. Erythroxylum coca, alkaloid, benzoylmethylecgonine, cocaine, embryo, endosperm, seed imbibition     

Effects of Growth Regulators on Polyamine Content and Peroxidase Activity in Hevea brasiliensis Callus

3, 4-dichlorophenoxyacetic acid (3,4-D) and benzylaminopurine(BAP) at 9 µM (control medium) was compared with 4.5,2.25, and 0.45 µM for ability to induce callogenesis andembryogenesis from seed explants of Hevea brasiliensis. Supplyingthese growth regulators at 4.5 µM for 20 d improved embryogenicpotential compared with the control medium (El Hadrami, Carronand d'Auzac, 1991, Annals of Botany 67, 511–515), sustainedputrescine, spermidine and spermine at a higher level throughoutof much of the culture period (40–70 d), and maintainedlow levels of peroxidase activity. In the control medium, poorcallus embryogenesis is considered a consequence of rapid ageingof tissues characterized by (i) acceleration of an early buttransient production of polyamines, which promoted embryogeniccapacity, and (ii) an early peak in peroxidase activity thatwas positively correlated with callus browning, one of the factorslimiting embryogenesis. Somatic embryogenesis, polyamines, peroxidase, Hevea brasiliensis, rubber-tree     

Spatial Distribution of Ethylene Production by Individual Needles Along a Shoot ofPinus sylvestrisL.: Relationship with Peroxidase Activity

Spatial distribution of ethylene production by individual currentyear needles and the relationship with peroxidase activity wasstudied along a shoot of Scots pine (Pinus sylvestrisL.). Muchvariation in ethylene production between and within individualseedlings was found. There was a significant difference in meanintensity of ethylene production between orthostichies withinindividual seedlings. The capacity of individual needles toconvert 1-aminocyclopropane-1-carboxylic acid (ACC) to ethylenedid not correlate with the basal rate of ethylene production.Mean ethylene production intensity by orthostichies correlatedpositively with mean peroxidase activity. A positive correlationbetween the intensity of ethylene production by individual needlesand detachment-induced peroxidase activity was found. The datasuggests that the rate of ethylene production by individualneedles may be directly related to their peroxidase activityand at least in part to their antioxidative capacity.Copyright1998 Annals of Botany Company Pinus sylvestris,pine, ethylene production, spatial distribution, individual needles, peroxidase, seasonal changes.     

Purine Alkaloids of the Fruits of Camellia sinensis L. and Coffea arabica L. during Fruit Development

The amounts of two purine alkaloids, caffeine and theobromine,in the fruit of tea (Camellia sinensis L.) increased markedlyduring the growing season until the fruit was full-ripened anddried. In the dry fruit, the pericarp contained the most alkaloids,but there were also considerable amounts in the seed coat and,to a lesser extent, the fruit stalk and the seed. The shed seedsalso contained significant amounts of the alkaloids, especiallyin the seed coats. In contrast with the dry fruit of tea, seedsand pericarp of coffee (Coffea arabica L.) fruit contained aconsiderable amount of caffeine and a small amount of theobromide.A small amount of theophylline was also present in the pericarpof the ripened fruit. Relationships between growth and purinealkaloid content in tea and coffee fruits and their roles duringseed formation are discussed. Camellia sinensis L., tea, Coffea arabica L., coffee, purine alkaloids, fruit development, seed, seed coat, caffeine, theobromine, theophylline     

Effects of Exogenous Growth Regulators on Fruit Development in Citrus unshiu

The effects of applied growth regulators on fruit developmenthave been determined in the parthenocarpic Satsuma mandarin(Citrus unshiu Marc.). The application of either gibberellicacid or benzyladenine at flower opening, caused a transientincrease in cell division in the ovary wall, but had no significanteffect on final fruit size. Late fruit growth and final fruitsize were increased by the application of the synthetic auxin2,4,5-trichlorophenoxyacetic acid, which had a specific effecton the enlargement of the juice vesicles. The three growth regulators enhanced vascularization in thepedicel, but the growth effects observed were unrelated to theirinfluence on the transport capacity of the phloem but causedby their direct effects on the fruit tissues. The sensitivityof the fruit tissues to the applied growth regulators changedmarkedly during early fruitlet development, and was characterizedculturing the fruit tissues in vitro.Copyright 1993, 1999 AcademicPress Citrus unshiu Marc., fruit growth, hormone treatment, in vitro culture, phloem formation, phloem transport, xylogenesis     

Dry Matter and Water Import Rates in the Tomato Fruit: a Model Incorporating the Changes in Sap Viscosity and Osmotic Potential with Temperature

The change in water import rate in tomato fruit was modelledby incorporating into a previously-published model the changesin sap viscosity and osmotic potential into fruit with temperature.An experimental relationship between water and dry matter importrates was used to compare the model to dry matter import ratesin fruit measured by Walker and Thornley (Annals of Botany 41:977-985, 1997) at different temperatures. The effect of temperatureon the water import rate, calculated from the model, was alsocompared with the effect of temperature on the fruit growthrate measured by Pearce, Grange and Hardwick (Journal of HorticulturalScience 68: 1-11 and 13-23, 1993). The model accounted for alarge part of these temperature effects. It was concluded that resistances in sap transfer pathways inthe tomato fruit could be due to viscosity. These results supported,on the one hand, the hypothesis that the progressive decreaseof water import rate during fruit growth could result partlyfrom the progressive increase in transfer pathway length, and,on the other hand, the hypothesis that the ratio between waterand dry matter import rates could depend on flow conditionsin transfer pathways. The equations of the model could be usedto simulate tomato fruit growth, mass and dry matter contentin relation to fruit size, to nutrient solution salinity andto fruit temperature.Copyright 1995, 1999 Academic Press Dry matter, fruit, growth, model, resistance, sap, temperature, tomato, transfer, viscosity, water     

Peroxidase isozymes and their indoleacetate oxidase activity in the Japanese morning glory, Pharbitis nil

Distributions of peroxidase isozymes in various organs of theJapanese morning glory (Pharbitis nil CHOISY) are described.Most peroxidase isozymes had indoleacetate oxidase activity. (Received December 29, 1969; )