Ethylene Co-ordinates Petal Abscission in Red Raspberry (Rubus idaeus L.) Flowers

The time-course of flower development of Rubus idaeus L. cv.Glen Clova was studied on detached buds opened in the laboratory.After sepal and petal opening petal abscission occurred withthe petals from an individual flower being shed over 3-4 h.Abscission was accompanied by a peak in ethylene production.Treatment of flowers with aminoethoxyvinylglycine eliminatedthe peak in ethylene production but did not prevent petal abscission.However, petal loss was much slower, taking place over a periodof days rather than hours. Abscission was more effectively retardedby silver thiosulphate. Exogenous ethylene accelerated the rateof petal abscission and senescence. The increase in ethyleneproduction coincident with petal abscission appears to accelerateand co-ordinate the shedding of the separate petals on an individualflower. If ethylene is important in the induction of abscissionit would appear that the low rate of production sustained inthe presence of aminoethoxyvinylglycine must be sufficient.Copyright1993, 1999 Academic Press Rubus idaeus L., raspberry, flower, petal, abscission, ethylene     

Elimination of Apple Mosaic Virus and Raspberry Bushy Dwarf Virus from Infected Red Raspberry (Rubus idaeus L.) by Tissue Culture

Experiments with ApMV infected ‘Malling Landmark’ and RBDV infected ‘Schamp;önemannamp;’ and ‘Trentamp;’ plants were carried out to evaluate a) the dependence of virus eradication on explant size and mass propagation. b) the reliability of results of ELISA tests on in vitro plantlets. With ApMV a correlation between virus elimination and explant size was observed, whereas with RBDV even plantlets from the smallest established explants were still infected. With ApMV, in vitro multiplication for three subsequent subcultures did not lead to further virus elimination, with RBDV this was observed in two cases. ELISA test results for both viruses, ApMV and RBDV, were identical when small in vitro plantlets, long-term stored plants, or potted plants from the same origin were tested, indicating that virus tests are possible with very young plant material and can be used to select virus-free plants in vitro. Tissue culture permits long-term storage of plant viruses. It is also suitable tor plant virus propagation and could be a useful aid in plant virus purification. For commercial multiplication only virus-indexed plant material should be used for establishment and further propagation in vitro.     

Interaction of endodormancy and paradormancy in raspberry (Rubus idaeus L.)

Bud break in protected Northern European raspberry crops is often poor and uneven with many of the sub-apical buds remaining in a dormant state. In order to improve bud break and therefore yields, the mechanism controlling bud dormancy must be determined. Canes of the biennial cultivar ‘Glen Moy’ were forced as isolated single nodes, trisections, or as intact canes after different lengths of cold storage chill unit (CU) accumulation in order to determine whether the buds were in an endodormant or paradormant state. The results showed that buds on the lower parts of the intact canes remained in a dormant state long after buds from higher up the intact cane and also the single nodes from all parts of the cane had emerged from the deepest phase of endodormancy. This would imply that these buds were being held in a paradormant state until large amounts of chilling units (>1000 CU) had been accumulated. The trisected cane portions revealed almost no significant differences in bud break levels throughout the experiment when compared with the single nodes. This suggests that removal of the apical part of the cane would be effective in improving bud break by reducing the paradormant condition. A period of secondary dormancy was also observed in the intact canes which may also exacerbate the poor bud break observed in protected crops. This was not seen in the single nodes or the trisected canes which indicates that treatments which reduce paradormancy may also minimise the risk of secondary dormancy. By identifying the phase of bud dormancy which causes poor bud break, attention can now be focused on methods which overcome paradormancy in protected crops. Such methods might include tipping (removal of the cane apex), horizontal training methods, more efficient chilling methods, and chemical treatments.     

The Histology of Seed Development in Diploid and Tetraploid Raspberries (Rubus idaeus L.)

Ovules of the raspberry, Rubus idaeus L., resembled those ofother Rubus species in being unitegmic, crassinucellate, andanatropous. In one cultivar, ‘M. 69/139’, both eggcell and embryo were inserted slightly laterally to the micropyle.Of the tetraploid forms, 4n M. 69/139 showed a proportion ofembryo sacs with delayed differentiation, and 4n ‘M. Jewel’some with four antipodal cells. Embryo development showed littleuniformity in timing, but followed the Asterad type. Occasional delayed endosperm development seemed to be a varietalpeculiarity of M. 69/139. At maturity, the ergastic materialin healthy endosperms consisted of oil globules and aleurongrains. The hypostase formed a projecting pedestal below theendosperm at 8 days. The inner epidermal layer of the testaappeared specialized, both in the early stages of seed growth,when it is suggested that it functioned as an endodermis, andat maturity, when it appeared to form an osmotic barrier.     

The Histology of Seed Development following Crosses between Diploid and Autotetraploid Raspberries (Rubus idaeus L.)

Seed development was studied in crosses made between the diploidforms and autotetraploid mutants of two raspberry (Rubus idaeusL.) cultivars. Drupelet abortion did not appear to be causedby failure of fertilization; it occurred mainly between thefourth and eighth day after pollination, and manifested itselfas simultaneous cessation of growth in all tissues. In healthydrupelets at eight days after pollination, growth appeared tobe proceeding at similar rates in all the tissues except thatthe embryo seemed to be growing faster than the endosperm inthe most advanced crosses. Both diploid and tetraploid ovulesappeared to be more stimulated by pollen from tetraploid plants.Embryo differentiation differed according to the direction ofcross, especially in stunted embryos. Radicles tended to beabnormally large when diploid pollen was used and smaller thanusual with haploid pollen.     

Co-ordinated gene expression during phases of dormancy release in raspberry (Rubus idaeus L.) buds

Bud break in raspberry (Rubus idaeus L.) is often poor and uneven, with many of the subapical buds remaining in a dormant state. In order to determine the dormancy status of raspberry buds, an empirical measure of bud burst in a growth-permissive environment following exposure to chilling (4 degrees C cold storage) was developed. For cv. Glen Ample, percentage bud burst in intact canes and isolated nodes was recorded after 14 d. Isolated nodes (a measure of endodormancy) achieved 100% bud burst after approximately 1500 h chilling whereas buds on intact plants (combined endo- and paradormancy) required an additional 1000 h chilling. A microarray approach was used to follow changes in gene expression that occurred during dormancy transition. The probes for the microarrays were obtained from endodormant and paradormant raspberry bud cDNA libraries. The expression profiles of 5300 clones from these libraries were subjected to principal component analysis to determine the most significant expression patterns. Sequence analysis of these clones, in many cases, enabled their functional categorization and the development of hypotheses concerning the mechanisms of bud dormancy release. Thus a set of novel candidates for key dormancy-related genes from raspberry buds have been identified. Bud dormancy is fundamental to the study of plant developmental processes and, in addition, its regulation is of significant economic importance to fruit and horticultural industries.     

Distribution of 14C Assimilates in the Cowpea (Vigna unguiculata L.) in Relation to Fruit Abscission and Treatment with Benzyladenine

The distribution of ¹⁴C assimilates from ¹⁴C-sucrose was studiedin relation to premature fruit abscission in two cowpea cultivars,Adzuki and Mala. In both cultivars most of the radioactivitywas recovered in the fruits, constituting 63–85 per centof the total ¹⁴C imported from the fed leaflet. This was followedby the root, leaves and stem in descending order, except thatin Mala, import by the stem was greater than that by the leaves.Adzuki imported 56 per cent more ¹⁴C than Mala, from the fedleaflet. In Adzuki, which exhibits a relatively low degree ofabscission of young fruits, the ratio of ¹⁴C accumulated bypeduncle 1 (oldest) fruits to that of peduncle 3 (youngest)fruits was 0·31; while in Mala it was 0·61. Ratiosof the combined accumulation by peduncles 1 and 2 fruits topeduncle 3 fruits were 0·81 for Adzuki, and 1·88for Mala. The more mature fruits of Mala thus constituted amore potent sink for ¹⁴C assimilates than those of Adzuki. In Adzuki, benzyladenine treatment of young fruits at each pedunclewas not significantly effective in reversing or modifying thenormal gradient of assimilates in fruits of different ontogeny.However, in Mala, BA treatment of the youngest fruits caused43 per cent increase in ¹⁴C import, when compared with correspondingfruits of control plants. In Adzuki, BA had no significant effecton total fruit weight, whereas in Mala the weight was increasedby about 36 per cent.     

A comparison of nine species of arbuscular mycorrhizal fungi on the development and nutrition of micropropagated Rubus idaeus L. cv. Glen Prosen (Red Raspberry)

Growth, development and nutrient status of micropropagated Rubus idaeus cv. Glen Prosen in response to inoculation with nine species of arbuscular mycorrhizal (AM) fungi from three different genera was investigated. The nine species of AM fungi included, Glomus clarum, G. etunicatum, G. intraradices, Gigaspora rosea, Gi. gigantea, Gi. margarita, Scutellospora calospora, S. heterogama and S. persica. Plant responses to AM fungi varied from growth enhancement to growth depression. Depressive growth effects were specific to Gigaspora species. Furthermore, particular species of AM fungi had unique effects on the mineral status of the raspberry plants. Importance of isolate selection for inoculation of micropropagated raspberry plants is discussed. This revised version was published online in June 2006 with corrections to the Cover Date.     

The Role of Ethylene in the Shedding of Red Raspberry Fruit

The production of ethylene by red raspberry (Rubus idaeus L.cv. Glen Clova) fruit increased climacterically during development.The concentration of ethylene within green fruit was low butincreased substantially as fruit abscission and ripening commenced.The receptacle contained higher concentrations than the drupeletsat all stages measured. In the mature ripening fruit the ethyleneconcentrations were found to be physiologically significant,and would accelerate the abscission of large green non-abscisingfruit if supplied as a fumigant. The addition of ethylene toripe fruit did not accelerate abscission, probably because saturatinglevels occurred naturally within these fruit. Reduction of ethylenesynthesis rates using the inhibitor of ethylene production aminoethoxyvinylglycine(AVG) reduced the rate of abscission zone weakening which occursin detached large green fruit. The rate of ethylene productionwas found to be dependent on the supply of the precursor l-aminocyclopropane-l-carboxylicacid (ACC). This only accumulated to any extent in those ripefruit with high rates of ethylene production. Rubus idaeus, raspberry, abscission, fruit ripening, ethylene, aminocyclopropane-l-carboxylic acid     

A comparison of the composition of epicuticular wax from red raspberry (Rubus idaeus L.) and hawthorn (Crataegus monogyna Jacq.) flowers

Epicuticular waxes have been characterised from the flowers of raspberry and hawthorn, on both of which adult raspberry beetles (Byturus tomentosus) can feed. The flower wax from both species had similar alkane profiles and also contained long-chain alcohols, aldehydes and fatty acids. The range of the carbon numbers detected for these classes of compounds was broadly similar in both but the relative amounts of each differed between species. Raspberry flower wax also contained fatty acid methyl esters, a group of compounds that has rarely been detected in plant epicuticular waxes, however, these were not observed in hawthorn flower wax. Long-chain alcohol-fatty acid esters with carbon numbers ranging from C36 to C48 were also detected in both plant species. However, an examination of their constituent acids indicated that in hawthorn the esters based on the C16 fatty acid predominated, whilst in raspberry flower wax, esters based on the C20 fatty acid were most abundant. Both species also contained pentacyclic triterpenoids, which accounted for, on average, over 16 and 48% of the total wax extracted from raspberry and hawthorn flowers respectively. In the former, ursolic and oleanolic acids accounted for over 90% of the pentacyclic triterpenes, whilst hawthorn flower wax, in addition to containing these acids, also contained high relative concentrations of both free and esterified alpha- and beta-amyrins.     

Accessory Buds in the Genus Rubus with Particular Reference to R. idaeus L.

Accessory (secondary) axillary buds occurred in 21 out of 32Rubus species, including wild raspberry, R. idaeus L. The adaptivevalue of the secondary-bud character is discussed. Disbuddingexperiments on three raspberry cultivars and their inbred derivativesrevealed considerable variability in proportion of secondarylaterals developing and in numbers of flowers per secondarylateral, suggesting the possibility of selection for high secondarycrop potential.     

The Relation Between Seed and Fruit Development in the Peach (Prunus persica L.)

The ontogeny of the peach seed and its organs correlates wellwith the stages of growth of the pericarp of peach fruit. Theinitial stages of rapid f.wt (FW I) and d.wt (DW I) increasecoincide with the period of rapid f.wt increase of the wholeseed and rapid size increase of the endosperm respectively.The period of slow f.wt increase of the fruit (FW II) coincideswith the period of rapid size increase of the embryo. The seed and pericarp compete strongly for assimilates throughoutfruit development and the seed is the weaker competitor. Thereis no period however when the growth of the seed or one of itsorgans inhibits the sink strength of the pericarp to allow theseed to develop. The data presented indicate that the periodsof high nutrient demand (DW I and DW III) are created in thepericarp in response to a stimulatory signal from the seed andthe period of low nutrient demand (DW II) results from the absenceof this signal after the endosperm reaches full size.     

Molecular and biochemical characterization of benzalacetone synthase and chalcone synthase genes and their proteins from raspberry (Rubus idaeus L.)

Two new members of the polyketide synthase (PKS) gene family (RiPKS4 and RiPKS5) were cloned from raspberry fruits (Rubus idaeus L., cv Royalty) and expressed in Escherichia coli. Characterization of the recombinant enzyme products indicated that RiPKS4 is a bifunctional polyketide synthase producing both 4-hydroxybenzalacetone and naringenin chalcone. The recombinant RiPKS4 protein, like the native protein from raspberry fruits [W. Borejsza-Wysocki, G. Hrazdina, Plant Physiol. 1996;110: 791-799] accepted p-coumaryl-CoA and ferulyl-CoA as starter substrates and catalyzed the formation of both naringenin chalcone, 4-hydroxy-benzalacetone and 3-methoxy-4-hydroxy-benzalacetone. Although activity of RiPKS4 was higher with ferulyl-CoA than with p-coumaryl-CoA, the corresponding product, 3-methoxy-4-hydroxy phenylbutanone could not be detected in raspberries to date. Sequence analysis of the genes and proteins suggested that this feature of RiPKS4 was created by variation in the C-terminus due to DNA recombination at the 3′ region of its coding sequence. RiPKS5 is a typical chalcone synthase (CHS) that uses p-coumaryl-CoA only as starter substrate and produces naringenin chalcone exclusively as the reaction product.     

Changes of Carbohydrates in Pepper (Capsicum annuum L.) Flowers in Relation to Their Abscission Under Different Shading Regimes

Abscission of pepper flowers is enhanced under conditions oflow light and high temperature. Our study shows that pepperflowers accumulate assimilates, particularly in the ovary, duringthe day time, and accumulate starch, which is then metabolizedin the subsequent dark period. With the exception of the petals,the ovary contains the highest total amounts of sugars and starch,compared with other flower parts and contains the highest totalactivity, as well as activity calculated on fresh mass basis,of sucrose synthase, in accordance with the role of this enzymein starch biosynthesis. Low light intensity or leaf removaldecreased sugar accumulation in the flower and subsequentlycaused flower abscission. The threshold of light intensity fordaily sugar accumulation in the sink leaves was much lower thanin flowers, resulting in higher daytime accumulation of sugarsin the sink leaves than in the adjacent flower buds under anylight intensity, suggesting a competition for assimilates betweenthese organs. Flowers of bell pepper cv. ‘Maor’and ‘899’ (sensitive to abscission) accumulatedless soluble sugars and starch under shade than the flowersof bell pepper cv. ‘Mazurka’ and of paprika cv.‘Lehava’ (less sensitive). The results suggest thatthe flower capacity to accumulate sugars and starch during theday is an important factor in determining flower retention andfruit set. Pepper; Capsicum annuum L.; abscission; shading; pepper flowers; ovary; leaves; sugars; starch; acid invertase; sucrose synthase     

The isolation of RNA from raspberry (Rubus idaeus) fruit

Previous attempts to extract high-quality, total RNA from raspberry (Rubus idaeus) fruits using published protocols have proven to be unsuccessful. Even the use of protocols developed for the extraction of RNA from other fruit tissue has resulted in low yields (1) or the isolation of degraded RNA (2). Here, we report on the development of a quick and simple method of extracting total RNA from raspberry fruit. Using this method, high yields of good quality, undegraded RNA were obtained from fruit at all stages of ripening. The RNA is of sufficient quality for northern analysis and cDNA library construction.     

Clonal Propagation of Dwarf Raspberry (Rubus pubescens Raf.) through in vitro Axillary Shoot Proliferation

A protocol for the micropropagation of dwarf raspberry (Rubus pubescens) was developed by the establishment of axenic shoot cultures from greenhouse-grown plants, induction of shoot proliferation, and rooting in vitro. Cultures were initiated from shoot tip and nodal explants on 1/2 strength MS (Murashige T. and Skoog F. 1962. Physiol. Plant. 15:473) macro-salts and MS micro-salts and vitamins containing 8.9 M N ⁶-benzyladenine (BA) and 0.98 M indole-3-butyric acid (IBA). Zeatin was more effective than BA, and induced proliferation of about 1.5–2 times as many shoots as BA in combination with 0.54–1.1 M -naphthaleneacetic acid (NAA) or 0.49–0.98 M IBA. With higher zeatin, shoots did not expand and had a high mortality rate. Shoots growing for more than 10 weeks on medium that contained 9.1 M zeatin occasionally produced adventitious shoot masses, which appeared to arise from dense calluses growing at the base of the shoots in the medium. Shoots were rooted in vitro in the same medium used for shoot proliferation, but without any growth regulators. Almost all (85–90%)in vitro plantlets survived when transferred to potting medium.     

The Anatomy of Fruit Abscission in Loganberries

Loganberry fruits abscise at the base of the receptacle, justdistal to the sepals. As the fruit ripens, all cells of theabscission zone expand. The central parenchyma cells, due totheir position, appear to be the driving force behind abscission.Their expansion causes early cell-separation within a superficialzone of small cells and rupture of the epidermis at the sepal/receptaclejunction without prior dissolution of cell walls. However, othercells within the abscission zone have their walls degraded,mostly in the region of the middle lamella, as ripening progresses. Xylem transfer cells are found in abundance in the vascularbundles supplying the sepals. The outward curve of these bundlesinto the sepals brings the transfer cells into close proximitywith the abscission zone. A comparison of their distributionin loganberries with that in raspberries (MacKenzie, 1979),which are closely related but abscise at a different site, suggeststhat transfer cells may be implicated in the abscission process. The inevitable structural weakness brought about by the paucityof vascular tissue in the abscission zone relative to the morerobust pedicel may also predispose this area to separation. Anatomy, abscission, loganberry, Rubus idaeus x R, ursinus, Mailing Sunberry, transfer cells structure, fruit     

The effect of resistance to Amphorophora rubi in raspberry (Rubus idaeus) on the spread of aphid-borne viruses

The effectiveness of resistance to the aphid Amphorophora rubi in restricting the spread of aphid-borne viruses was assessed in a field experiment using six genotypes of red raspberry. In one block of the experiment, the genotypes alternated with rows of virus-infected Mailing Jewel raspberry, and in the other they alternated with virus-free Mailing Jewel. During 4 years, the numbers of A. rubi and the amount of 52V virus spread in the two blocks were similar, suggesting that this virus was mostly introduced from outside the plots. Lloyd George and Mailing Jewel raspberry became heavily infested with A. rubi and were rapidly infected with raspberry leaf mottle, raspberry leaf spot and 52V viruses. Glen Clova and Norfolk Giant raspberry, which contain minor genes for resistance to A. rubi, were infested with fewer A. rubi and virus spread more slowly in these cultivars. A. rubi were rare on Mailing Orion and an East Mailing raspberry selection (888/49) which have genes A₁ and A₁₀ respectively for resistance to A.rubi, and these plants remained largely free of virus. The role of minor and major gene resistance to A. rubi in restricting virus spread is discussed. A few Macrosiphum euphorbiae and Myzus ornatus were recorded on several of the raspberry genotypes.     

Fruit abscission and ethylene production of four blackberry cultivars (Rubus spp.)

In all cultivars the force required to harvest fruit declined during ripening. The fruit retention strength (FRS) of ripe fruit varied between cultivars with ‘Ashton Cross’ and ‘Chehalem’ being easier to remove than ‘Bedford Giant’ or ‘Oregon Thornless’. The cultivars ‘Ashton Cross’ and ‘Oregon Thornless’ showed no increase in ethylene production during fruit ripening whereas ‘Bedford Giant’ and ‘Chehalem’ had increased rates of ethylene production (EPR) in the ripe fruit. Exogenous ethylene accelerated abscission, ethylene production and pigment changes in ‘Ashton Cross’ fruit at all stages of development. 1-amino-cyclopropane-1-carboxylic acid (ACC) supplied to fruit at all stages of development was converted to ethylene at levels in excess of those found naturally. The differences between cultivars are discussed with reference to the role of ethylene in both machine harvesting and post-harvest storage of blackberry fruit.     

Resistance to Botrytis cinerea in canes of Rubus idaeus and some related species

Mycelial inoculation of canes with Botrytis cinerea proved a useful method for assessing Rubus material for resistance. When canes were inoculated in summer resistant genotypes developed relatively small lesions which produced few small sclerotia in the following spring. The size of lesions in autumn generally provided the best discrimination between genotypes. Very strong resistance was found in several species, of which Rubus pileatus and R. occidentalis are the most useful for breeding; hybrids of raspberry with these species or with R. crataegifolius also had strong resistance. The resistance shown by a derivative of red raspberry cv. Chief was less strong, as was the resistance conferred by gene H, which determines cane pubescence.