Effects of fruit thinning agents on apple tree canopy photosynthesis and dark respiration

Effects on photosynthesis of the fruit thinning agents naphthaleneaceticacid (NAA) and three commercial plant growth regulator formulations,naphthaleneacetic acid ('Rhodofix') and naphthaleneacetamide('Amidthin') and 2-chloroethylphosphonic acid('Ethrel')were evaluated with respect to the stress they impose on the fruit tree, usingthe alternate-bearing sensitive apple cv. 'Elstar'. This work wasbased on the hypothesis that plant stress in the form of large reductions inleaf photosynthesis are a pre-requisite for successful fruit thinning. A newtechnology was employed for continuous recording of tree canopyphotosynthesis, dark respiration and carbon balance of apple trees. This wasbased on six canopy chambers, which enclosed apple trees under naturalconditions in the field, with on-line measurements and continuous analysis ofCO₂ exchange and automated data acquisition. All employed thinningagents reduced whole tree canopy photosynthesis consistently by3–34% on the five days following their application, withphotosynthesis still declining thereafter in the case of the NAA and'Amid-thin' application. The reduction after application of either'Rhodofix' or 'Ethrel', declined within five days, suchthat most of the original photosynthetic potential was restored, indicatingacceptable phytotoxicity of these three plant growth regulators at theconcentrations used. The effects on dark respiration differed markedly. NAA and'Ethrel' increased dark respirationover-proportionally by up to 106%, whereas 'Amid-thin' and'Rhodofix' decreased it by up to 46%inthe first night after application, thereby drastically affecting the carbonbalance of the tree in opposite ways. These results are integrated into ahypothesis linking basipetal auxin transport, phloem loading, translocation anddeficiency of photoassimilates.     

Plant regeneration from leaf protoplasts of apple

Protoplasts were isolated from young leaves or etiolated shoot apices. For initiation of divisions the protoplasts were embedded in sodium alginate and cultivated in MS or MI medium supplemented with 2.2 M BA, 2.6 M NAA and 2.2 M 2,4-dichlorophenoxyacetic acid. The protoplasts of all seven lines tested developed to protocalluses at high frequencies. No genotypic differences were observed. When BA was used in combination with NAA in the regeneration experiments, only a few protocalluses (highest frequency 3%) exhibited shoot organogenesis. When BA was replaced with thidiazuron, the percentage of protocalluses that developed shoots increased in two of three tested lines to 7% and 56%, respectively. Shoot development was achieved under light conditions. The shoots were then rooted and transferred into soil.Abbreviations ABA abscisic acid - BA 6-benzyladenine - 2,4-d 2,4-dichlorophenoxyacetic acid - FW fresh weight - GA₃ gibberellic acid - IBA indole-3-butyric acid - MES 2-N-morpholinoethane sulphonic acid - NAA -naphthaleneacetic acid     

The influence of cation and gelling agent concentrations on vitrification of apple cultivars in vitro

Shoot tips of York and Vermont Spur Delicious apples (Malus domestica Borkh.) were cultured in vitro to test the influence of K⁺, Mg⁺⁺ and gelling agent concentrations on vitrification. These concentrations were 20.05, 14.05 and 8.05 mM K⁺, 1.5 and 3.0 mM Mg⁺⁺, 7.0 g/l Difco Bacto agar and 1.0, 1.5 and 2.0 g/l Gelrite. The lowest K⁺ level produced a higher percentage of vitrified shoots, affected tissue appearance, reduced shoot number and shoot elongation and apparently altered shoot metabolic activity. Gelrite consistently produced vitrified leaves and stems, even though media gelled with 1.5 g/l Gelrite presented the same apparent gel firmness as using 7 g/l Difco Bacto agar, which did not induce vitrification. Less shoot elongation, fewer total shoots, and more usable shoots of York were obtained on Bacto-agar, while similar but less noticeable effects were obtained with Vermont Spur Delicious. The results presented here show that vitrification can be studied in a standardized system in which the only change is substitution of one gelling agent for another.     

Dissecting apple tree architecture into genetic, ontogenetic and environmental effects: QTL mapping

The present study aimed to dissect tree architectural plasticity into genetic, ontogenetic and environmental effects over the first 4 years of growth of an apple F1 progeny by means of quantitative traits loci (QTL) mapping. Both growth and branching processes were phenotyped on the consecutive annual shoots of different axes within a tree. For each studied trait, predicted values (best linear unbiased predictors, BLUPs) of the genotypic (G) effect or its interaction with tree age (G×A) and climatic year (G×Y) were extracted from mixed linear models of repeated data. These BLUPs, which are independent from autocorrelations between repeated measurements, were used for QTL mapping. QTL detection power was improved by this two-step approach. For each architectural process, numerous QTLs were detected and some particularly interesting co-localised in common genomic regions, for internode lengthening, top diameter, and number and percentage of axillary shoots. When several QTLs were detected for a given trait, global models were estimated, which explained a maximum of 40% of the total variance for both internode length and top diameter and 28% for branching. QTLs detected for BLUPs of G×Y effects were interpreted as resulting from the interaction between genetic maximal potential of growth and climatic factors, while those for G×A effects were interpreted in relation to tree ontogeny. Most of the latter ones were found to be concomitant with key development stages during which the trait average started to decrease, but with different magnitudes depending on genotype. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

Identification and stability of QTLs for fruit quality traits in apple

Breeding for fruit quality traits is complex due to the polygenic (quantitative) nature of the genetic control of these traits. Therefore, to improve the speed and efficiency of genotype selection, attention in recent years has focused on the identification of quantitative trait loci (QTLs) and molecular markers associated with these QTLs. However, despite the huge potential of molecular markers in breeding programmes, their implementation in practice has been limited by the lack of information on the stability of QTLs across different environments and within different genetic backgrounds. Here, we present the results from a comprehensive analysis of the inheritance of fruit quality traits within a population derived from a cross between the apple cultivars ‘Telamon’ and ‘Braeburn’ over two successive seasons. A total of 74 different QTLs were identified for all the major fruit physiological traits including fruit height, diameter, weight and stiffness, flesh firmness, rate of flesh browning, acidity, the oBrix content and harvest date. Seventeen of these QTLs were ‘major’ QTLs, accounting for over 20% of the observed population variance of the trait. However, only one third (26) of the identified QTLs were stable over both harvest years, and of these year-stable QTLs only one was a major QTL. A direct comparison with published QTL results obtained using other populations (King et al., Theor Appl Genet 102:1227–1235, 2001; Liebhard et al., Plant Mol Biol 52:511–526, 2003) is difficult because the linkage maps do not share a sufficient number of common markers and due to differences in the trait evaluation protocols. Nonetheless, our results suggest that for the six fruit quality traits which were measured in all populations, nine out of a total of 45 QTLs were common or stable across all population × environments combinations. These results are discussed in the framework of the development and application of molecular markers for fruit quality trait improvement.     

CO2-Enrichment and photosynthetic photon flux affect the growth of in vitro-cultured apple plantlets

Micro-cuttings (shoots with two small leaves) of cultivar M9 apple were cultured in-vitro for 40 d under CO₂-enriched and non-enriched (i.e., ambient air) conditions, and at a PPF of 40 or 100 μmol m^-2 s^-1 Afterward, shoot length, number of leaves, leaf area, chlorophyll content, shoot and root fresh weights, and % survival were recorded. Those plant-lets grown under CO₂- and PPF-enriched treatments were healthy and vigorous, and showed higher values for their growth parameters. In contrast, those grown without supplemental CO₂ or PPF often showed hyperhydricity. We also demonstrated that CO₂ enrichment and a relatively high PPF during in-vitro culture promoted normal photosynthesis and growth after ex-vitro transplantation.     

Biosynthesis of phenolic compounds and its regulation in apple

This paper summarises the information on the occurrence of phenolic compounds in apple Malus domestica leaves and fruits with special reference to their developmental changes and regulation of biosynthesis. Besides the ontogenetic variation in biosynthesis and accumulation, the stress-induced and pathogenesis-related changes are emphasised. Aspects of commercial importance are addressed, ranging from fruit colouration, through disease resistance, to the direct use of apple tissues, as raw material for the extraction of bioactive phenolic compounds.     

Medium,container and genotype all influence in vitro cold storage of apple germplasm

The goal of this study was to evaluate the in vitro storage of apple germplasm by screening a range of genotypes followed by more comprehensive testing of multiple parameters on two genotypes of differing species, Malus domestica cultivar Grushovka Vernenskaya and wild Malus sieversii selection TM-6. Stored plants were rated on a 6 point scale (0 low to 5 high) for plant appearance at 3 month intervals after storage at 4°C. Combinations of carbon source (sucrose and/or mannitol), nitrate nitrogen content (25, 50 or 100%) and plant growth regulators (ABA, BAP, IBA) were studied in three types of containers (tissue culture bags, test tubes or jars). An initial screen of 16 genotypes stored in tissue culture bags indicated that plantlets could be stored at 4°C for 9–14 months without subculture on standard 3% sucrose Murashige and Skoog (1962) (MS) medium with no plant growth regulators (PGRs). In subsequent in-depth studies on the two genotypes, ANOVA indicated highly significant interactions of medium, container and genotype. ‘Grushovka Vernenskaya’ shoots with no PGRs and 3% sucrose remained viable (ratings of ≥1) for 21 months of storage in bags. Storage on reduced nitrogen (MS with 25% nitrogen), PGRs, and 3% sucrose kept ‘Grushovka Vernenskaya’ shoot condition rated >2 at 21 months. Addition of 0.5 or 1 mg⁻¹ abscisic acid (ABA) also improved plant ratings at 21 months. The longest storage for ‘Grushovka Vernenskaya’ was 33–39 months with PGRs and 3% sucrose in either tubes or jars. Addition of abscisic acid (ABA) to the medium did not improve storage of plantlets in jars and tubes at 15 months. TM-6 stored best in tubes on 3% sucrose with PGRs or in jars on 2% mannitol and 2% sucrose. Overall it appears that cold storage of apple shoot cultures can be successful for 21 months in tissue culture bags with 25% MS nitrate nitrogen, 3% sucrose, and no PGRs or for 33 months in jars or tubes on MS with 3% sucrose and PGRs. Preliminary RAPD analysis found no significant differences between plants stored for 39 months and non-stored controls.     

Repeat flowering in apple caused by water stress or defoliation

Summary A series of experiments involving defoliation or water stress at different dates indicated that either of these treatments can make potted apple trees flower a second time in any one year, as long as the treatment is given near the end of July. The results suggest that the reflowering after a period of water stress was primarily a result of the loss of leaves that occurred when the plants were subsequently rewatered. Reflowering normally occurred only if flower primordia had already differentiated at the time of the treatment. There was an indication that in early July water stress was more effective than defoliation at stimulating reflowering.     

Seasonal correlations of specific leaf weight to net photosynthesis and dark respiration of apple leaves

Specific leaf weight (SLW), net photosynthesis (P_n), and dark respiration (R_d) of apple leaves were monitored for an entire growing season. Leaves were sampled from the canopy interior and periphery to provide a range of SLW. Leaf P_nwas linearly correlated with SLW until mid-August, when P_nbegan to decline. During September the relationship between SLW and P_nwas a quadratic. Leaf R_dand SLW were linearly correlated throughout the season. Leaf P_nand R_dwere significantly correlated through most of the season, but the relationship was not always linear. Specific leaf weight appears to be a reliable index of the previous light environment of a leaf, but use to estimate P_nis probably limited to the first half of the season, because of increased variation after mid-August.Former Graduate Research Assistant (presently Assistant Professor, Department of Horticulture and Forestry, Rutgers University, Cook College, New Brunswick, NJ 08903, USA) and Associate Professor, respectively.     

Identification and mapping of the novel apple scab resistance gene Vd3

Apple scab, caused by the fungal pathogen Venturia inaequalis, is one of the most devastating diseases for the apple growing in temperate zones with humid springs and summers. Breeding programs around the world have been able to identify several sources of resistance, the Vf from Malus floribunda 821 being the most frequently used. The appearance of two new races of V. inaequalis (races 6 and 7) in several European countries that are able to overcome the resistance of the Vf gene put in evidence the necessity of the combination of different resistance genes in the same genotype (pyramiding). Here, we report the identification and mapping of a new apple scab resistance gene (Vd3) from the resistant selection “1980-015-25” of the apple breeding program at Plant Research International, The Netherlands. This selection contains also the Vf gene and the novel V25 gene for apple scab resistance. We mapped Vd3 on linkage group 1, 1 cM to the south of Vf in repulsion phase to it. Based on pedigree analysis and resistance tests, it could be deduced that 1980-015-25 had inherited Vd3 from the founder “D3.” This gene provides resistance to the highly virulent EU-NL-24 strain of race 7 of V. inaequalis capable of overcoming the resistance from Vf and Vg. JMS and SGJ contributed equally to this work     

Effect of fruiting and drought or flooding on carbon balance of apple trees

The response of fruiting or deblossomed trees to water stress such as drought or flooding was investigated in six semi open-top cuvettes each containing one apple (Malus domestica Borkh. cv. Golden Delicious) tree. Xylem water potentials of leaves dropped from -1.2 to -4.1 MPa within 7 d of drought, the effect being enhanced by fruiting. Apple trees without fruits showed smaller reductions in net photosynthetic rate (P _N ) and dark respiration rate (R _D ) after 2 d of drought and hence more positive carbon balances relative to fruiting trees. Flooding for 4 d had a more pronounced effect on P _N than on transpiration, resulting in a reduced water use efficiency (WUE). This reduction in WUE was greater in the non-fruiting trees. Flooding reduced P _N of the whole apple canopies irrespective of fruiting; aple trees without fruits increased R _D resulting in a less positive carbon balance relative to fruiting trees. Fruiting increased the sensitivity to drought of apple trees (R _D and P _N ), but decreased their sensitivity to flooding (R _D and WUE), suggesting different adaptation mechanisms for the two forms of water stress. This revised version was published online in August 2006 with corrections to the Cover Date.     

Changes in photosynthetic characteristics during leaf development in apple

A comprehensive developmental survey of leaf area, chlorophyll, photosynthetic rate, leaf resistance, transpiration ratio, CO₂ compensation point and photorespiration was conducted in apple. The largest changes in each of the photosynthetic characteristics studied took place during the earliest stages of leaf development, coinciding with the period of greatest leaf expansion and chlorophyll synthesis. During early development, photosynthesis increased 5-fold, reaching a maximum rate of 40 mg CO₂ dm^-2 hr^-1 at a leaf plastochron index (LPI) of 10. During this same period, leaf resistance, transpiration ratio, CO₂ compensation point and mesophyll resistance decreased, while carboxylation efficiency increased. Two especially interesting aspects of the data discussed are simultaneous changes that occur at a LPI of 10 and 12 in all of the photosynthetic characteristics examined and an apparent decrease in photorespiration as leaves age. From our results it is clear that stage of leaf development is an important factor affecting the rate of photosynthesis and photorespiration.Scientific Paper No. 5687, College of Agriculture, Washington State University, Pullman. This work is supported by the National Science Foundation Grant 80-10958 and the Columbia River Orchards Foundation.     

Identification of co-dominant RAPD markers tightly linked to fruit skin color in apple

A simple genetic basis for the red/yellow skincolor polymorphism in apple was verified using DNA markers. Bulked segregant analysis identified one 10-base oligomer that generated different fragments in each of the bulks. After testing the primer in four populations, two fragments were found to be associated with red skin color and another two fragments associated with yellow skin color. Three of the fragments (1160, 1180, and 1230 bp) were partly sequenced and found to share high sequence homology, suggesting these were generated from the same locus. A pair of universal primers were designed to amplify the fragments. In the Rome Beauty x White Angel population, two fragments were associated with red skin color; one fragment designated as A¹ (1160 bp) was from Rome Beauty and another fragment (A², 1180 bp) was from White Angel. Progeny possessing both fragments, or either one, had red fruit. Both parents displayed an alternate fragment, a¹ (1230 bp), associated with yellowskinned fruit. In three other crosses tested, only fragment A¹ co-segregated with red skin color; two fragments, a¹ and a² (1230 bp and 1320 bp), were associated with yellow skin color. Our results are consistent with the hypothesis that the red/yellow dimorphism is controlled by a monogenic system with the presence of the red anthocyanin pigmentation being dominant. There was no indication that other modifier genes could reverse the effect of the locus (R _f ) linked to the markers. Examination of amplification products in 56 apple cultivars and advanced breeding selections demonstrated that the universal primers could be used to correctly predict fruit skin color in most cases.     

Modelling fruit-temperature dynamics within apple tree crowns using virtual plants

Background and Aims

Fruit temperature results from a complex system involving the climate, the tree architecture, the fruit location within the tree crown and the fruit thermal properties. Despite much theoretical and experimental evidence for large differences (up to 10 °C in sunny conditions) between fruit temperature and air temperature, fruit temperature is never used in horticultural studies. A way of modelling fruit-temperature dynamics from climate data is addressed in this work.

Methods

The model is based upon three-dimensional virtual representation of apple trees and links three-dimensional virtual trees with a physical-based fruit-temperature dynamical model. The overall model was assessed by comparing model outputs to field measures of fruit-temperature dynamics.

Key Results

The model was able to simulate both the temperature dynamics at fruit scale, i.e. fruit-temperature gradients and departure from air temperature, and at the tree scale, i.e. the within-tree-crown variability in fruit temperature (average root mean square error value over fruits was 1·43 °C).

Conclusions

This study shows that linking virtual plants with the modelling of the physical plant environment offers a relevant framework to address the modelling of fruit-temperature dynamics within a tree canopy. The proposed model offers opportunities for modelling effects of the within-crown architecture on fruit thermal responses in horticultural studies.     

QTL analysis for aphid resistance and growth traits in apple

The rosy apple aphid (Dysaphis plantaginea), the leaf-curling aphid (Dysaphis cf. devecta) and the green apple aphid (Aphis pomi) are widespread pest insects that reduce growth of leaves, fruits and shoots in apple (Malus × domestica). Aphid control in apple orchards is generally achieved by insecticides, but alternative management options like growing resistant cultivars are needed for a more sustainable integrated pest management (IPM). A linkage map available for a segregating F₁-cross of the apple cultivars ‘Fiesta’ and ‘Discovery’ was used to investigate the genetic basis of resistance to aphids. Aphid infestation and plant growth characteristics were repeatedly assessed for the same 160 apple genotypes in three different environments and 2 consecutive years. We identified amplified fragment length polymorphism (AFLP) markers linked to quantitative trait loci (QTLs) for resistance to D. plantaginea (‘Fiesta’ linkage group 17, locus 57.7, marker E33M35–0269; heritability: 28.3%), and to D. cf. devecta (‘Fiesta’ linkage group 7, locus 4.5, marker E32M39–0195; heritability: 50.2%). Interactions between aphid species, differences in climatic conditions and the spatial distribution of aphid infestation were identified as possible factors impeding the detection of QTLs. A pedigree analysis of simple sequence repeat (SSR) marker alleles closely associated with the QTL markers revealed the presence of the alleles in other apple cultivars with reported aphid resistance (‘Wagener’, ‘Cox’s Orange Pippin’), highlighting the genetic basis and also the potential for gene pyramiding of aphid resistance in apple. Finally, significant QTLs for shoot length and stem diameter were identified, while there was no relationship between aphid resistance and plant trait QTLs. Electronic supplementary material  The online version of this article (doi:) contains supplementary material, which is available to authorized users.     

The effects of pollen diversity on plant reproduction: insights from apple

For many plants, the number of pollen genotypes deposited on a flower’s stigma is positively related to fruit maturation and seed number; however, the mechanisms underlying this effect are less understood. Here we examined whether diversity of pollen (1, 3, or 5 donors) affects reproductive success in self-incompatible apple (Malus × domestica). Using paternity analysis, we then assessed the siring rate of individual donors to test whether diversity effects are related to the presence of a superior pollen donor or to donor × donor interactions. Increasing the diversity of compatible pollen enhanced seed number and reduced seed abortion in some but not all recipient genotypes. This effect was associated with an increased number of sires per fruit and non-random siring success among pollen donors. Two donors had consistently high siring rates, regardless of recipient genotype; however, the siring success of pollen donors was not correlated with their siring success in single-donor pollinations. Rather, siring success was affected by the identity of other pollen genotypes present on the stigma. Our results therefore suggest that the effects of pollen diversity are variable but may enhance fecundity by fostering interactions between pollen donors.