Significance of skin flavonoids for UV-B-protection in apple fruits

An attempt has been made to assess the UV-B-protective capacity of phenolic compounds accumulated in superficial structures of plants using apple fruit as a model. Two apple (Malus domestica Borkh.) cultivars (Braeburn and Granny Smith) differing in response to high fluxes of solar radiation were selected and exposed to increasing doses of UV-B radiation. The extent of UV-B-induced damage to photosystem II of apple skin correlated with its quercetin glycoside (but not anthocyanin) content. Granny Smith apples did not demonstrate a pronounced response to high sunlight in terms of the accumulation of phenolic substances in the skin and exhibited similar patterns of Fo, Fm, and Fv/Fm changes in the course of UV-B irradiation both on sun-exposed and shaded surfaces of a fruit. Unlike Granny Smith, Braeburn fruits were characterized by a significant accumulation of quercetin glycosides in sun-exposed skin, however, shaded skin contained these compounds in similar amounts to those in Granny Smith. Accordingly, photosystem II in sun-exposed skin of Braeburn apples was resistant to high doses of UV-B radiation (up to 97 kJ m-2), whereas the susceptibility of the photosynthetic apparatus in shaded skin of Braeburn to UV-B-induced damage was much higher and similar to that of both sun-exposed and shaded skin of Granny Smith fruits. Anthocyanins, at least in the amounts found in Braeburn, did not show an additional effect in UV-B protection.     

A study of gene expression changes at the <Emphasis Type="Italic">Bp-2</Emphasis> locus associated with bitter pit symptom expression in apple (<Emphasis Type="Italic">Malus pumila</Emphasis>)

Bitter pit is a physiological disorder of apples that develops in the latter stages of fruit development and during storage. It is characterized by localized necrotic cells that collapse and form pits in the epidermis and outer cortex of fruit. The disorder has been associated with low calcium concentrations, and poor calcium distribution within the fruit. To date, the mechanism that leads to individual cell necrosis, while surrounding cells remain healthy, is not fully understood. In order to ascertain the underlying process of bitter pit incidence in apple fruit, a large mapping population of “Braeburn” (susceptible to bitter pit)?×?“Cameo” (resistant to bitter pit) was used to map the trait over two growing seasons. A subset of 94 genotypes from the mapping population representing the full range of phenotypes in the same ratio as the full population were selected for genotyping and functional characterization. RNA-Seq analysis on fruit samples of three resistant and three susceptible lines at seven developmental stages (21, 42, 63, 84, 105, 126, and 147 days post fertilization) identified a number of candidate genes displaying differential gene expression. A subset of candidate genes selected based on their position within the identified QTL interval on linkage group 16 were validated by RT-qPCR, and two candidate genes displaying differential gene expression were highlighted as strong candidates for the control of bitter pit symptom expression at the Bp-2 locus.     

Up-regulation of two cDNA clones encoding metallothionein-like proteins in apple fruit during cool storage

We are investigating the molecular basis of low temperature responses in apples, by identifying and characterising fruit genes which show altered expression in response to cool-storage, Two independent cold-regulated clones (pAMTI and pAMT2) were isolated from a cDNA library derived from cool-stored apple ( Malus domestics Borkh. cv. Granny Smith) fruit. These clones share only 27% amino acid identity with each other, but both show high similarity to plant metallothionein (MT)-like proteins. The polypeptide encoded by pAMTI shares similarity with type 2 MT-like sequences, while that encoded by pAMT2 is similar to others which share a different distribution of cysteine residues. We suggest, these form a 'type 3' group of MT-like clones. Genomic Southern analysis confirmed that there is a family of MT-like genes in apple. There are differing patterns of pAMTI and pAMT2 expression during apple fruit development, amt 1 RNA was abundant in flowers and during the early stages of development, and decreased as the fruit approached maturity, while amt 2 RNA was barely detectable in flowers and young fruit and accumulated with fruit development. In ripe fruit. amt 1. expression was up-regulated, while amt 2 expression was down-regulated. In leaves, both genes showed increased expression with leaf age. In Granny Smith, Cox's Orange Pippin and Braeburn apple cultivars. both genes were up-regulated in cool-stored fruit. In Granny Smith contical tissue, amt RNA levels were elevated within the first 45 min at both 0.5°C and 4°C, but not at 12.5°C. The different patterns of amt 1 and amt 2 expression during fruit development and in different tissues suggest that the respective genes have distinct controlling elements and may be functionally different. The in vivo roles of the encoded polypeptides, particularly in relation to chilling tolerance or acclimation, are as yet unknown.     

Differential gene expression analysis of ‘Granny Smith’ apple (Malus domestica Borkh.) during fruit skin coloration

‘Granny Smith’ apples growing under normal sunlight develop green skin, whereas the peel turns red due to anthocyanin accumulation after the removal of a bagging treatment. Two anthocyanins, Cyanidin 3-O-galactoside (cy3-gal) and Cyanidin 3-O-arabinoside (cy3-ara), were detected in the red ‘Granny Smith’ apple peels, and cy3-gal was determined to be chiefly responsible for the red color. The content of cy3-gal was more than 98% of the total anthocyanin in the red ‘Granny Smith’ peels. To better understand the molecular basis of anthocyanin biosynthesis in ‘Granny Smith’ apples, we performed a quantitative real-time PCR (qRT-PCR) analysis of anthocyanin biosynthetic genes (MdCHS, MdF3H, MdDFR, MdANS, MdUFGT, and MdMYB1). Our results indicate that the expression of these genes (except MdCHS) was associated with increased anthocyanin accumulation in the skin of ‘Granny Smith’ apples. Four selected genes obtained from the ‘Granny Smith’ skin cDNA library, phytoene synthase (PSY), WD40 repeat protein, polygalacturonase (PG), and galactosidase (GAL), were also confirmed by qRT-PCR. We found that these genes were differently expressed during ‘Granny Smith’ apple skin coloration, suggesting that they are directly or indirectly involved in pigment accumulation. In conclusion, anthocyanin biosynthesis in ‘Granny Smith’ apples is the result of interactions between multiple enzymes in the anthocyanin biosynthesis pathway, and the coloring mechanism of ‘Granny Smith’ apples may be similar to that of red-skinned cultivars.     

Susceptibility of selected apple cultivars to the Mediterranean fruit fly

Ceratitis capitata (Wiedemann), the Mediterranean fruit fly, is one of the key pest species affecting deciduous fruit orchards along the Mediterranean coasts. Because of global warming, C. capitata is gradually spreading north and is becoming a major pest of apples. Determining the susceptibility of the main apple varieties grown in the region will serve as a cornerstone to the management of this pest. In this study, we show the results of a field and laboratory no‐choice test conducted to determine the Medfly preferences on different apple cultivars. The seven main varieties of apples (Gala, Red Delicious, Golden Delicious, Granny Smith, Kanzi, Morgen Dallago and Fuji) were tested. The results demonstrate that C. capitata lays eggs on all apple cultivars in both field and laboratory conditions. The Granny Smith, Red Delicious and Morgen Dallago varieties showed the lowest susceptibility in laboratory conditions, (0.75, 1.55, 2 oviposition punctures/fruit, respectively), with significant differences in oviposition compared to the Golden Delicious, Kanzi and Fuji (3.27, 3.31, 3.1 oviposition punctures/fruit, respectively) varieties, which were shown to be the most susceptible to Medfly attack in laboratory conditions. On the other hand, only slight and not statistically significant differences emerged from the field trials. In relation to the physico‐chemical characteristics, the apple cultivars showing the lowest susceptibility (Granny Smith, Red Delicious and Morgen Dallago) had harder peels and pulps and lower sugar contents than the most susceptible cultivars (Golden Delicious, Fuji and Kanzi). These results were also confirmed through evaluation of larval development on different varieties. In fact, Granny Smith, Red Delicious and Morgen Dallago were the three varieties that did not allow adequate larval and adult development and reduced the possibility of the emergence of a new generation.     

Integrated control of apple pests in New Zealand 9. Development of European red mite populations under minimal control strategies

Abstract

Development of European red mite (Panonychus ulmi) populations was studied from 1975 to 1977 in an orchard released from regular application of acaricides and supporting insignificant numbers of natural enemies of the mite. Populations grew rapidly from mid January, and reached higher levels on Hawke’s Bay Red Delicious than on Cox’s Orange Pippin or Granny Smith. Extensive early defoliation occurred in all cultivars in response to heavy feeding pressure. Application of cyhexatin in February 1976 failed to control populations in Granny Smith and Cox’s Orange Pippin, and only temporarily suppressed populations in Hawke’s Bay Red Delicious. Removal of all acaricide applications in 1976–77 on Hawke’s Bay Red Delicious showed that high population levels (365 mites per leaf) could develop. Stratification of sampling in Granny Smith showed that population densities decreased from the tree centre, through outer fruiting clusters, to current season’s shoots. Winter egg deposition on apples was studied in relation to various P. ulmi densities. Early harvest of Cox’s Orange Pippin prevented large numbers of winter eggs being laid on the fruits in spite of high P. ulmi densities; late harvest of Granny Smith exposed fruit to greater winter egg deposition. The intermediate harvest date of Hawke’s Bay Red Delicious resulted in fewer winter eggs on apples than for Granny Smith, in spite of much higher P. ulmi densities. On Hawke’s Bay Red Delicious, oviposition preference was for calyces of fruits not exposed to direct sunlight. A relationship between maximum P. ulmi densities and numbers of winter eggs on fruits is demonstrated.     

Vessel development and the importance of lateral flow in water transport within developing bundles of current-year shoots of grapevine (<Emphasis Type="Italic">Vitis vinifera</Emphasis> L.)

In the developing xylem bundles of young stems, the presence of immature living vessel elements can strongly restrict or even block axial hydraulic conductance, especially in newly matured vessels. Lateral connections between vessels may provide an alternative pathway for water movement to bypass these closed, living elements. Using the grapevine as a model system, the present study aimed to demonstrate the effects of living vessel elements on water movement patterns, and the importance of lateral flow for effective water conductivity in the developing bundles. Living vessel elements were detected using dye staining and the pattern of vessel development and maturation was then monitored. The importance of lateral flow was confirmed using several approaches: (1) capacity for lateral flow, (2) effect of increasing the distance of water transport, and (3) effect of ion concentrations. Living vessel elements were found along the developing bundles, they occupied a significant proportion of the distal and peripheral parts of the flow path, forming a substantial barrier to apoplastic water flow. Water in the developing xylem bundles could move easily from vessel to vessel and between secondary and primary xylem. Furthermore, data from increasing the transport length and altering the ion concentrations supported the critical contribution of the lateral flow to the total hydraulic conductance within the developing bundles. The hydraulic architecture of the developing xylem bundles is described. The results are discussed in terms of reliability and efficiency of water transport during shoot growth and development.     

A test of fruit varieties on entry rate and development by neonate larvae of the codling moth,Cydia pomonella

The rate of entry by neonate larvae of the frugivorous codling moth, Cydia pomonella (L.) (Lepidoptera: Tortricidae), into fruit material was investigated. We used no‐choice bioassays in climate‐controlled rooms to assay larval entry across four host plant species (apple, pear, quince, walnut) and three varieties within a single fruit species (apple). Larvae successfully entering apples were reared to adulthood, and we collected tissue samples from apples which were successfully colonized in order to determine sucrose concentrations. This information was used to evaluate differences in adult moth size, development time, and pulp sucrose concentration due to apple variety. Four important findings emerged: (1) neonate larvae had the highest frequency of entry (86% of larvae) into apple fruits, compared with pear (78%), quince (56%), and walnut (32%); (2) the frequency of larval entry into immature apples differed across apple varieties, and larval entry rate was highest in variety Golden Delicious (72%), compared with Granny Smith (46%) and Red Delicious (64%); (3) on average, adult moths were larger and development times were shorter on the variety with the highest entry frequency (Golden Delicious); and (4) apple pulp sucrose concentrations were higher for Golden Delicious (17.5 μg mg^?1) than for either Granny Smith (15.9 μg mg^?1) or Red Delicious (15.1 μg mg^?1) varieties, which correlates positively with entry and development data. We conclude that host fruit species and varietals within a species affect the entry rate of neonate codling moth larvae in no‐choice assays. We hypothesize that larval development is influenced by mean sucrose concentrations or other phytochemical differences associated with host fruit varieties.     

Laboratory tests carried out onMicrothrix inconspicuella [Lep.: Pyralidae], a biological control agent forEmex australis in Australia

Host specificity tests of the moth,Microthrix inconspicuella Ragonot in Australia, indicated that larvae could feed and develop on young apple leaves. Additional tests in South Africa on leaves and fruit of the 4 apple varieties, Jonathan, Starking (Red Delicious), Granny Smith and Golden Delicious, showed that apples were not a preferred food. Little feeding occurred and pupation happened infrequently. No 2^nd generation resulted whenM. inconspicuella colonies were confined on apple fruit or leaves.      

根域限制对葡萄果实不同发育期维管束水分运输的影响

本文采用质外体染料示踪法研究了葡萄果实不同发育期根域限制和非根域限制（对照）处理果实维管束水分运输变化。结果表明,在葡萄果实发育的第一次快速生长期,根域限制和对照处理的果实周缘维管束被染色数量最多,染料溶液在葡萄果实中的运输速率也最高,分别为1.72和1.63cm·h-1,根域限制处理果实中周缘维管束和中央维管束中染料溶液的运输速度低于对照处理,但胚珠维管束中染料运输速度要高于对照,为1.32cm·h-1;进入生长停滞期后,根域限制和对照处理的葡萄果实中周缘维管束被染色数目降低,染料溶液在中央维管束中的运输速度分别降低为0.72和0.70cm·h-1;在果实的第二次快速生长期,根域限制和对照处理果实周缘维管束染色数目仍然低于果实发育的第一次快速生长期,根域限制果实周缘维管束染色范围和染料运输速率都高于对照处理。     

Ionic control of the lateral exchange of water between vascular bundles in tomato

Ions can enhance water flow through the xylem via changes in the hydraulic resistance at border pit membranes. Because flow between adjacent xylem vessels occurs primarily via bordered pit fields, it is hypothesized that xylem sap ion concentrations would affect lateral movement of water more than longitudinal flow. Using tomato as a model system, evidence is presented for ion-mediated changes in xylem hydraulic resistance and the lateral transport of water. Water flow between adjacent xylem bundles increased by approximately 50% in the presence of ions while longitudinal flow only increased by approximately 20%. However, the enhancement of lateral exchange due to ions was magnified by the presence of a pressure difference between vascular bundles. These results indicate that the degree of nutrient-sharing among sectors of a plant may depend on both nutrient concentration and the availability of water in the root zone.     

The Vascular System of Maize Stems Revisited: Implications for Water Transport and Xylem Safety

The plexus of vascular bundles in the nodes of grasses is notoriouslycomplex, where long axial bundles pass through a network oftransverse bundles. The xylem pathways for water in maize stemshave been investigated anatomically and with dye and particulatetracers, revealing some of the details of this complexity. Onlyapprox. 3% of axial vessels pass through nodes without beinginterrupted by end walls. Axial bundles at nodes differ fromthose in internodes in having the metaxylem and protoxylem vesselsconnected by small tracheary elements. So it is only at nodesthat exchange of sap occurs between the large vessels withina bundle. End walls, acting as filters for particles and gasbubbles, always separate axial vessels from vessels in transversebundles. The high redundancy of bundle connections in the nodalplexus is interpreted as providing alternative water pathwaysto bypass embolisms and damaged or diseased sections of thexylem. The pores in the filters at the base of nodes and betweenaxial and transverse vessels within nodes are <20 nm in diameter.Where axial vessels connect to transverse vessels, a varietyof unusual shapes of vessel elements mediate two- and three-wayconnections within the plexus.Copyright 2000 Annals of BotanyCompany Zea mays, cryoSEM, maize, node, pits, pit membranes, vessel ends, vessels, xylem embolism, xylem pathogens     

The three-dimensional structure of vascular tissues in Agrobacterium tumefaciens-induced crown galls and in the host stems of Ricinus communis L.

The three-dimensional pattern of phloem and xylem in 10-d-to two-month-old tumors induced by Agrobacterium tumefaciens (C58) and in adjacent Ricinus communis L. stem tissues was studied in thick sections by clearing with lactic acid and by staining with lacmoid. The crown galls contained two types of vascular strands: treelike branched bundles, which developed towards the tumor surface in fast-growing regions, and globular bundles in the slowly developing parts. Both types of vascular bundles contained xylem and phloem and were continuous with the vascular system of the host plant. The tumor bundles were interconnected by a dense net of phloem anastomoses, consisting of sieve tubes but no vessels. These vascular patterns reflect the apparent synthesis sites, concentration gradients and flow pathways of the plant hormones additionally produced in the tumors upon expression of the T-DNA-encoded genes. The A. tumefaciens-induced crown gall affected vascular differentiation in the host stem. In the basipetal direction, the tumor induced more xylem differentiation directly below it, where the crown-gall bundles joined the vascular system of the host. In the centripetal direction, the crown gall caused the development of pathologic xylem characterized by narrow vessels, giant rays and absence of fibers. On the other hand, most probably as a consequence of its gibberellic acid content, the host plant stimulated a local differentiation of regenerative phloem and xylem fibers with unique ramifications, only at the base of the tumor. However, fibers were absent from the main body of the crown gall. The study shows that A. tumefaciens-induced crown galls are characterized by a sophisticated network of vascular tissues in the tumor and are accompanied by a perturbated vessel system in the host. The hormonal mechanisms controlling vascular differentiation in the tumor and neighboring host tissues are discussed. In addition, the gall constriction hypothesis is proposed for explaining the mechanism which gives priority in water supply to the growing gall over the host shoot.We thank Dr. Zs. Koncz (Max-Planck-Institut für Züchtungsforschung, Köln, Germany) for Agrobacterium strains and the Deutsche Forschungsgemeinschaft (SFB 199) for financial support to C.I.U.     

Uptake and Transport of Calcium and the Possible Causes of Blossom-end Rot in Tomato

The possible causes of blossom-end rot (BER) in tomato fruitwere investigated by comparing the uptake of calcium by theroots, the distribution of ⁴⁵Ca within the fruit and the vascularbundle network in the fruit of susceptible cultivars (Calypsoand Spectra) with those of a less susceptible cultivar (Counter)grown in a range of salinities (electrical conductivity of 5,10 and 15 mS cm^–1). The daily calcium uptake rates at5 mS cm^–1 as estimated from the xylem exudation of thedecapitated stem stump in young plants of Calypso and old plantsof Spectra, were lower than that of Counter. The uptake of ⁴⁵Caby, and the transport to, the distal part of the detached fruitof susceptible cultivars, especially Calypso, were less thanin Counter at 10 mS cm^–1. The number of vascular bundlesin both proximal and distal fruit tissues was similar in allcultivars and was only slightly reduced by salinity. However,the number of bundles containing lignified xylem vessels, asdetected by safranin staining, was reduced substantially bysalinity, particularly in Calypso. The estimated area of thefruit tissue served by individual xylem bundles in the BER susceptiblefruit grown at high salinity was greater than in Counter. Theincidence of BER in all trusses was linearly related to theproduct of average daily irradiance and daily temperature throughoutthe year. Temperature appears to be the major environmentalfactor which induces BER, regardless of cultivars and salinitytreatment. The most likely causes of BER in susceptible cultivarsare the interactions of (a) light and temperature on fruit enlargement,(b) inadequate xylem tissue development in the fruit and (c)competition between leaves and fruit for the available Ca. Key words: Lycopersicon esculentum, calcium transport, susceptibility to blossom-end rot, root exudation, xylem     

Role of biodiversity in integrated fruit production in eastern North American orchards

• 1 Diversifying agricultural ecosystems to enhance biological control is a promising way of promoting sustainable pest management.
• 2 In the present study, monoculture apple and peach with standard insecticide treatments were compared with three biodiverse treatments (polyculure, monoculture with companion plants and polyculture with companion plants) with reduced standard insecticide use.
• 3 Abundance of insect predators was increased by both the presence of companion plants and extrafloral nectar but parasitism of the leafroller Platynota idaeusalis was not affected.
• 4 There were no consistent effects of biodiversity treatment on either tree growth or fruit yield. Insect injury to Empire apple and peach fruit was not consistently affected by the biodiversity treatments. Granny Smith apples were harvested later than Empire and had more fruit injury in the biodiverse treatments than the standard insecticide control.
• 5 A reduction in pesticides with added biodiversity proved to be a viable alternative to standard chemical insecticide management for temperate tree fruit. Increases in the natural control of pests resulting from increased plant diversity has promise for reducing the reliance on chemical insecticides for pest suppression.

     

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     

Inhibitory effect of diazocyclopentadiene on the development of superficial scald in Granny Smith apple

In this work, diazocyclopentadiene (DACP), an ethylene action inhibitor was used to test if ethylene is involved in the development of superficial scald of apple. Apples (Malus domestica Borkh., c.v. Granny Smith) were pre-stored at 0°C for a month before DACP treatment. After treatment, fruit were stored at 0°C for a further 17 weeks before being transferred to room temperature for a week. The incidence of superficial scald, contents of -farnesene and conjugated triene in fruit skin were analysed. Ethylene production, respiration rate, flesh firmness and soluble solids content of fruit were determined. Results indicated that superficial scald is related to chilling injury. DACP delayed ripening, and dramatically inhibited the development of superficial scald in Granny Smith apple by lowering -farnesene and conjugated triene contents. Ethylene might promote -farnesene synthesis presumably by binding to the ethylene receptor(s).     

The peripheral xylem of grapevine (Vitis vinifera). 1. Structural integrity in post-veraison berries

During the development of many fleshy fruits, water flow becomes progressively more phloemic and less xylemic. In grape (Vitis vinifera L.), the current hypothesis to explain this change is that the tracheary elements of the peripheral xylem break as a result of berry growth, rendering the xylem structurally discontinuous and hence non-functional. Recent work, however, has shown via apoplastic dye movement through the xylem of post-veraison berries that the xylem should remain structurally intact throughout berry development. To corroborate this, peripheral xylem structure in developing Chardonnay berries was investigated via maceration and plastic sectioning. Macerations revealed that, contrary to current belief, the xylem was comprised mostly of vessels with few tracheids. In cross-section, the tracheary elements of the vascular bundles formed almost parallel radial files, with later formed elements toward the epidermis and earlier formed elements toward the centre of the berry. Most tracheary elements remained intact throughout berry maturation, consistent with recent reports of vascular dye movement in post-veraison berries.     

A model considering light reabsorption processes to correct in vivo chlorophyll fluorescence spectra in apples.

Chlorophyll-a contained in the peel of Granny Smith apples emits fluorescence upon excitation with blue light. The observed emission, collected by an external detector and corrected by its spectral response, is still distorted by light reabsorption processes taking place in the fruit skin and differs appreciably from the true spectral distribution of fluorescence emerging from chlorophyll molecules in the biological tissue. Reabsorption processes particularly affect the ratio of fluorescence intensities at 680 nm and at 730 nm. A model to obtain the correct spectral distribution of the emission, from the experimental fluorescence recorded at a fluorometer detector and corrected for the detector spectral sensitivity, is developed in the present work. Measurements of the whole fruit reflectance, the peel transmittance and the flesh reflectance allow the calculation of the reabsorption-corrected spectra. The model is validated by comparing the corrected emission spectra with that obtained for a thin layer of apple-peel-chloroplasts, where no reabsorption takes place. It is recommended to correct distortions in emission spectra of intact fruits due to light reabsorption effects whenever a correlation between the physiological state of the fruit and its fluorescence spectra is investigated.