Alcohol Dehydrogenase of Apple

The alcohol dehydrogenase prepared from apple (Malus domesticaBorkh.) possesses both NADH and NADPH-linked activities, whenassayed with acetaldehyde as substrate. The pyridine nucleotidesbind to the same catalytic site on the enzyme. The alcohol dehydrogenasecan also catalyse the reduction of C₃–C₆ aldehydes witheither NADH or NADPH as cofactor.     

Carboxylic Ester Hydrolases of Apple

p-Nitrophenyl esters of carboxylic acids (C₂–C₈) wereused to characterize the carboxylic ester hydrolases of peeltissue of apple (Malus domestica Borkh.). The paper presentsevidence for the presence of five ester hydrolases in the peeltissue which show differences in substrate specificity and molecularweight.     

RNA, proteins and polyamines during tube growth in germinating apple pollen

Variations of RNA, protein, and free- and trichloroacetic acid-soluble bound polyamine levels were determined during tube growth in germinating Malus domestica Borkh. cv. Starkrimson pollen.     

Initiation of Rapid Ethylene Synthesis by Apple and Pear Fruits in Relation to Storage Temperature

The influence of storage temperature on the onset of rapid ethyleneproduction was investigated for fruits of Conference pear (Pyruscommunis L.) and five cultivars of apple (Malus domestica Borkh.).The time taken from harvest to rapid ethylene production wasshorter and more uniform at 3 ?C than at 18–20 ?C forConference pears and Golden Delicious apples. Increases in internalethylene concentration, 1-amino cyclopropane-1-carboxylic acidconcentration and ethylene production were simultaneous in GoldenDelicious apples at 3 ?C. When Golden Delicious apples wereheld at 3 ?C for 48 h and then kept at 20 ?C the mean time ofonset of ethylene production was similar to that for applesheld continuously at 20 ?C. However, two periods of 48 h at3 ?C caused earlier ethylene production. Conversely, ethyleneproduction at 3 ?C was delayed by transfer to 20 ?C for twoperiods of 48 h. Cox's Orange Pippin and other apple cultivarstended to show more synchronous ethylene production at 3 ?Cthan at higher temperatures but the mean time of onset was eitherunaffected by temperature or slighdy delayed at lower temperature.Acceleration of the onset of ethylene production by low temperaturewas never observed in Cox's Orange Pippin apples harvested atweekly intervals from 10 August to 17 September. Key words: Ethylene, Storage temperature, Pyrus communis, Malus domestica     

Estimation of Provisional Seed Viability Constants for Apple (Malus domestica Borkh. cv. Greensleeves)

Estimates of viability constants have been calculated for apple(cv. Greensleeves) seeds, stored hermetically in a range ofenvironments. The constants allow reasonable prediction of seedstorage life (orthodox), at least in the short and medium term. Apple, Malus domestica Borkh. cv. Greensleeves, seed storage, viability constants, influence of seed moisture content and temperature     

A Further Study of {beta}-Galactosidase Activity in Apples Ripening in Store

Apples (Malus domestica Borkh.) were ripened in 2% O₂ at 3·3°C. The loss of galactose residues from the cortical cellwalls was initiated after 27 d although an increase in the ß-galactosidaseactivity of the tissue did not occur until after 50 d. ß-Galactosidase activity associated with the cellwall was determined using discs of cortical tissue. A substantiallevel of activity was observed prior to picking and subsequentlyduring ripening of the apples in air.     

Response of Developing Apple Fruits to Ethylene Treatment

Knee, M., Hatfield, S. G. S. and Bramlage, W. B. 1987. Responseof developing fruits to ethylene treatment.—J. exp. Bot.38: 972–979. Fruits of apple (Malus domestica Borkh. cv. Cox's Orange Pippin)were treated with various concentrations of ethylene usuallyfor 48 h to determine their response in relation to stage ofdevelopment. The main response recorded was the reduction byethylene of the delay in onset of rapid ethylene production(DEP) in individual fruits. Early in development low concentrationsof ethylene had little effect but DEP was progressively reducedby concentrations up to 10⁷ mm³ m^–3. As the fruit approachedthe natural onset of rapid ethylene synthesis concentrationsof 10² and 10³ mm³ m^–3 became increasingly effective.Increasing the duration of treatments with a fixed concentrationreduced DEP proportionately. Delay after harvest in applyinga 48 h treatment had little effect on the relation between DEPand concentration of ethylene applied. Although resistance todiffusion of gas in fruits increased during fruit developmentthis resistance was never large enough to affect the relationof concentration and response. Key words: ethylene, fruit ripening, Malus domestica     

Inhibition of the growth of apple pollen tubes by EDTA, surfactants and fungicides

The effects of EDTA (ethylene diamine tetraacetic acid), Kresoxim‐methyl, Wettable Sulphur 80 WP and the surfactants Triton X‐100, Triton X‐114, Dehydol TA 5 and Dehydol TA 29 on pollen tube growth of Malus domestica, Borkh. cvs ‘Golden Delicious’ and ‘Mondial Gala’ were determined by image analysis. The ED₅₀ ‐ and ED₉₀ ‐ values (effective doses giving 50% and 90% reduction of growth) of the chemicals, except for the surfactants, were determined. The selected test substances showed an inhibiting effect on pollen tube growth, with Kresoxim‐methyl being the most active. Surfactants are components of pesticides and they also caused inhibition of pollen tube growth but only at application rates more then 10‐fold higher than typically recommended. Pollen of both apple cultivars were almost equally sensitive to the chemicals.     

Flow Cytometric Determination of Relative Nuclear DNA Contents in Bicellulate and Tricellulate Pollen

Nuclear DNA content in mature pollen was measured with a flowcytometer Pollen of Lilium longiflorum, Dendranthema grandiflora(syn Chrysanthemum monfolium) and Zea mays was chopped and stainedwith the DNA fluorochrome DAPI DNA levels, expressed as arbitraryC values, were compared with those of nuclei isolated from leafor root material of the same plants In mature tricellulate pollen the generative cell is dividedafter second pollen mitosis into two sperm cells Tricellulatepollen from maize and chrysanthemum gave rise to one large 1Cpeak and, only in the case of chrysanthemum, a much smallerone at the 2C level These results suggest that the haploid nucleiof the vegetative as well as both sperm cells in tricellulatepollen are arrested in the G₁ stage of nuclear division Thesmall 2C peak in the case of chrysanthemum probably arose froma fraction of pollen with the sporophytic chromosome number(2n pollen) In contrast to this, mature bicellulate lily pollengave rise to two identical peaks at the 1C and the 2C levelFrom this result it was concluded that in bicellulate pollen,the 1C peak is caused by the signal of the haploid vegetativenucleus arrested in the G₁ stage of nuclear division, whereasthe 2C peak originates from the haploid generative nucleus whichhas already undergone DNA synthesis and is arrested in G₂ Lilium longiflorumThunb, lily, Dendranthema grandiflora Tzelev (syn Chrysanthemum morifolium Ramat ), chrysanthemum, Zea maysL, maize, male gametophytic cells, vegetative cells, generative cells, sperm cells, unreduced pollen, sporophytic cells, relative nuclear DNA contents, replication stage     

Effect of Applied and Endogenous Auxin on Callus and Root Formation of In Vitro Shoots of the Apple: Rootstocks M26 and A2

The influence of exogenous IBA (indol-3yl-butyric acid) on rootand callus formation was studied in shoots of the apple rootstocksA2 and M26. The shoots grown in vitro were derived originallyfrom meristems of both juvenile and adult trees. Endogenousindol-3yl-acetic acid (IAA) concentrations in leaves and stemswere correlated with the responses to applied IAA. After 30 subcultures shoots from A2 and M26 rooted easily, butA2 did so more readily and even without IBA. Treatment withIBA improved percentage rooting and number of roots in bothrootstocks. Ex-adult and ex-juvenile shoots of A2 formed rootsto the same extent. However, ex-adult shoots of A2 showed ahigher IBA optimum for root number than ex-juvenile A2 and werealso less sensitive to supra-optimal IBA concentrations. Incontrast, in M26, there were no differences between ex-adultand ex-juvenile shoots. The results imply that rooting ability is associated more withdifferences between cultivars than with the origin of the explants.The best rooting occurred in ex-adult shoots of A2 which hadthe lowest endogenous IAA concentration, while callus formationwas correlated with high endogenous auxin concentration. Ex-adultA2 produced almost no callus even after exposure to high IBAconcentrations (25µM) whereas ex-adult M26 formed muchmore callus at 1/10 of the IBA concentration. Malus sylvestris (L.) Mill. var. domestica Borkh., Malus pumila Mill., apple rootstocks A2 and M26, in vitro culture, root and callus formation, HPLC analyses of IAA     

Vascular Development and Sap Flow in Apple Pedicels

Xylem and phloem tissues of the pedicel of apple fruit increasein cross-sectional area throughout development. The increasein phloem is similar in the two cultivars examined (Cox's OrangePippin and Royal Gala) and reflects a steadily increasing phloemsap flow to the fruit. The increase in xylem tissue is due toa proliferation of non-conducting, structural, components sinceclose examination reveals no increase in the number of vesselelements from just after flowering onwards. The greater number,and the larger diameter, of the vessels in Cox's explains theinitially higher xylem conductance found in this cultivar. In vitro measurements of xylem exudation reveal a decline duringthe growing season in the xylem conductance of both cultivarsand an increasing proportion of fruit (particularly in Cox's)in which the xylem comes to be totally non-conducting. Thisobservation is in line with previously reported measurementsof xylem sap flow in vivo. The straightforward techniques used in this study offer a feasiblealternative to more arduous methods of assessing xylem and phloemsap flows and their balance during growth.Copyright 1994, 1999Academic Press Apple, xylem, phloem, vascular development, sap flow, Malus domestica Borkh     

Interconversions of 14C-labelled Sugars in Apple Tree Tissues12

Tissue pieces excised from orchard-grown apple trees duringa growing season exhibited different and changing capabilitiesof transferring ¹⁴C-label from sucrose, fructose and sorbitolto other soluble carbohydrates. All tissues incorporated fructose¹⁴C into sucrose but only leaves incorporated significant amountsof label from sucrose into sorbitol. As seeds developed andmatured, their ability to incorporate ¹⁴C from sorbitol andfrom fructose into sucrose increased. Sorbitol and sucrose arethe major translocated photosynthetic products of apple leavesbut whereas sorbitol appears to be an end-product of synthesis,sucrose may be considered as a substrate involved more directlyin carbohydrate utilization. Key words: Malus domestica, Apple, Carbohydrate interconversions     

Visualisation of transglutaminase-mediated cross-linking activity in germinating pollen by laser confocal microscopy

Abstract

Transglutaminases (TGs) are a multigenic family of calcium-dependent protein cross-linking enzymes, which are present in animal and plant cells. We have previously reported the presence of TGs in the cytosol and, more recently, in the cell wall of Malus domestica pollen, where it may be involved in pollen germination and pollen–stylar interactions. In this report we describe a simple method for the in situ visualisation of TG activity in germinating pollen. The method is based on the incorporation, mediated by pollen TG, of a fluorescently labelled exogenous diamine substrate of TG (fluorescein-cadaverine) into endogenous pollen substrates. Following the in situ TG activity reaction, the presence of cross-linked pollen proteins was visualised in fixed specimens of germinated pollen by laser confocal microscopy. Our data indicate the presence of TG cross-linking activity mainly at the apical part of the pollen tube, in the region proximal to the grain, and in the pollen grain itself. In planta, the products of this activity may provide strength to the pollen tube migrating through the style.     

ATP-Promoted Sorbitol Transport into Vacuoles Isolated from Apple Fruit

Sorbitol was transported actively into vacuoles isolated fromapple (Malus pumilla Mill, var domestica Schneid.) fruit flesh.The uptake was stimulated up to twofold by the addition of ATP,and the ATP dependent uptake showed a saturation curve as tothe substrate concentration. The optimum uptake of sorbitolwas pursued in the acidic range of pH 5 to 6. The K_m value forthe ATP dependent sorbitol uptake was about 5 mM. Sorbitol uptake was clearly inhibited by PCMB and uncouplers(CCCP and DCCD), and to a lesser extent by orthovanadate, butonly slightly by oligomycin. K⁺ stimulated sorbitol uptake.Sorbitol was converted to other sugars (glucose) only very slowlywhen transported across the tonoplast. This suggests that sorbitolis transported into vacuoles by a carrier mediated transportsystem coupled with H⁺- ATPase, localized on the tonoplast.Sucrose uptake into the vacuoles was also enhanced by ATP. (Received May 31, 1986; Accepted March 2, 1987)     

In vitro test of self-incompatibility in Malus domestica

Summary An in vitro assay in which self-incompatible pollen of Malus domestica is selectively inhibited is described. This assay involves heat-labile substances diffusing from the stylar tissues — in particular, glycoproteins found in the protein extract of styles. In the presence of the self-style extract, a dramatic decrease in total protein concentration in the culture medium was revealed at 30-min germination. Pretreatment of the self-pollen with 100 mM glucose prevented this drop in protein level; moreover, tube growth was entirely restored. A possible explanation in terms of protein-carbohydrate complementation is suggested.     

A simple and efficient method for DNA extraction from grapevine cultivars andVitis species

A quick, simple, and reliable method for the extraction of DNA from grapevine species, hybrids, andAmpelopsis brevipedunculata (Vitaceae) has been developed. This method, based on that of Doyle and Doyle (1990), is a CTBA-based extraction procedure modified by the use of NaCl to remove polysaccharides and PVP to eliminate polyphenols during DNA purification. The method has also been used successfully for extraction of total DNA from other fruit species such as apple (Malus domestica), apricot (Prunus armeniaca), cherry (Prunus avium), peach (Prunus persica), plum (Prunus domestica), and raspberry (Rubus idaeus). DNA yield from this procedure is high (up to 1 mg/g of leaf tissue). DNA is completely digestible with restriction endonucleases and amplifiable in the polymerase chain reaction (PCR), indicating freedom from common contaminating compounds.     

Metabolism of 1-Aminocyclopropane-l-Carboxylic Acid during Apple Fruit Development

The metabolism of [U^–14C] 1-aminocyclopropane-1-carboxylicacid (ACC) supplied to whole fruits of apple (Malus domesticaBorkh., cv. Cox's Orange Pippin) was investigated. Radioactiveethylene was recovered in mercuric acetate traps and an acidicmetabolite was formed in proportions which varied little withthe absolute amount of substrate supplied. The amount of ACCusually supplied did not cause immediate, rapid ethylene productionby mature, pre-climacteric fruit but the onset of productionwas earlier than in untreated fruit. The radioactive acidic metabolite was purified by four chromatographicprocedures and activity was coincident with authentic 1-malonylamino)cyclopropane-1-carboxylic acid (MACC). The presence of thiscompound was confirmed by gas chromatography linked to massspectrometry. MACC was a major metabolite of [¹⁴C] ACC supplied to applesthroughout fruit development. The proportion converted to ethylenewas low but increased with endogenous ethylene production inthe final samples. MACC was shown to be a natural constituent of apple fruits andto accumulate to the amol kg^–1 level. Key words: 1-Aminocyclopropane-l-carboxylic acid, Ethylene, 1 (Malonylamino) cyclopropane-1-carboxylic acid, Malus domestica     

Ovule and Embryo Sac Development in Malus pumila L. cv. Cox's Orange Pippin, from Dormancy to Blossom

Ovule and embryo sac development in the flowers of Cox's OrangePippin apple (Malus pumila L.) were studied from dormancy topetal fall using both scanning electron and light microscopy.The relative timing was established between these developmentsand the external development of the flower bud and flower. Malus pumila L. cv. Cox's Orange Pippin, apple, Cox, SEM, ovule development, anatomy, histology     

Internal Changes in Hyphae of Rhizopus sexualis (Smith) Callen and Mucor hiemalis Wehm. associated with Zygospore Formation

The distribution of nucleic acids, nuclei, mitochondria, andreserve foods in vegetative hyphae, zygophores, and developingzygospores of Rhizopus sexualis and Mucor hiemalis were examinedby differential staining. The extreme tips and growing zones of vegetative hyphae containeda high concentration of RNA and numerous mitochondria. Nucleiwere not present at the extreme tip but were numerous just behindit. In older parts of the hyphae the concentration of RNA waslow and both nuclei and mitochondria were fewer than in thezone of elongation. Glycogen and lipids were present in all parts of the livinghyphae except the extreme tips and were more highly concentratedin the older parts of the hyphae. Young zygophores showed a much lower RNA/DNA ratio than thatfound in the vegetative hyphal tips. Transfer of colonies from20? C to temperatures of less than 10? C, which is known toprevent zygospore initiation, caused some but not all recognizablezygophores of R. sexualis, but not those of M. hiemalis, torevert to the RNA/DNA ratio characteristic of vegetative hyphae.Some zygophores of Rhizopus and most of those of Mucor developedinto sporangiophores at low temperature, retaining the relativelylow RNA/DNA ratio throughout development. It is suggested thata reduction in the RNA/DNA ratio is an early step in the changefrom the vegetative state to the reproductive one. At firstthis step is reversible, but soon becomes irreversible by anadditional step, the nature of which is unknown. For some timeafter this the reproductive hyphae are capable of either producingasexual sporangia or of conjugating to produce zygospores. Onceconjugation has taken place development either ceases or continuesuntil the spore is fully mature, but it cannot under any circumstancesthen be reversed. The development and maturation of the zygospore involves a greatincrease in number of both nuclei and mitochondria and in theconcentration of glycogen and lipids.