Rootstock and Crop Load Effects on 'Honeycrisp' Photosynthetic Performance and Carbohydrate Accumulation.

Rootstock selection and crop load adjustment are key practices in apple orchard management; nevertheless, the effects of rootstocks and crop load levels on important physiological processes of the scions, such as photosynthetic performance and carbohydrate accumulation, are still unclear. To investigate the impact of different rootstocks and crop load levels on scion photosynthesis and carbohydrate buildup, in 2020, 'Honeycrisp' trees grafted on rootstocks 'G.41', 'G.935', and 'M.9-T337' were thinned to low and high crop load levels, and photosynthetic performance and carbohydrate accumulation in leaves and fruit were evaluated. Leaves from 'G.935' showed the highest net photosynthesis and electron use efficiency of photosynthesis and the lowest activity for non-net carboxylative processes, all together indicative of enhanced photosynthetic performance. High crop load determined an increase in gas exchange, suggesting a positive feedback of high fruit competition on carbon assimilation. While rootstock 'M.9-T337' showed a higher accumulation of starch in leaves, no pattern regarding the composition of leaf-soluble sugars among rootstocks could be identified. Conversely, by the end of the harvest season, leaves from low-cropping trees had higher fructose, glucose, and sorbitol than those from high-cropping trees, but differences in starch content were not significant. Fructose and sorbitol concentrations were affected by rootstock and crop load, respectively. Overall, this study showed that high cropping enhanced photosynthesis in 'Honeycrisp' apple and determined lower accumulation of some soluble carbohydrates (fructose, glucose, sorbitol) in leaves. This study also provided insights into how rootstocks affect photosynthetic performance of 'Honeycrisp', highlighting 'G.935' as the rootstock conferring the highest photosynthetic capacity under the present experimental conditions.

Effective Pollination Period and Parentage Effect on Pollen Tube Growth in Apple.

Flower receptivity is a limiting factor for the fertilization of several tree fruit. The effective pollination period (EPP) can be used to determine flower longevity and identify limiting factors by assessing stigmatic receptivity, pollen tube growth rate, and ovule longevity. EPPs were determined for three apple cultivars under natural field conditions in Washington State in 2019 and 2020. In addition, a greenhouse study, performed under semi-controlled conditions, evaluated the influence of six maternal parents on the pollen tube growth performance of six pollen sources. The duration of the stigmatic receptivity ranged from 6.3 to 8.1 days, depending on the cultivar and year-pollen tubes required between 5.5 and 7.0 days from the stigma to reach the ovules. Ovule longevity of non-pollinated flowers varied between 8.2 and 11.3 days. Combinations of these factors resulted in EPPs ranging from 3.0 days for 'Rubinstar' to 5.6 days for 'Olsentwo Gala' in the present experimental conditions. The greenhouse study revealed that parentage affected pollen tube growth performance. Importantly, a significant interaction between maternal and paternal factors indicated that the performance of different pollen sources depended on the maternal parent and that general recommendations on pollination need to account for the maternal parent.

Novel Metrics to Characterize In Vitro Pollen Tube Growth Performance of Apple Cultivars.

In vitro germination assays are frequently used in screening trials to evaluate the pollen viability of pollinizers. To be effective, screening trials must have defined threshold criteria, from which individuals can then be assessed. However, despite decades of research on pollen viability, no established threshold is available to categorize apple cultivars based on their in vitro pollen tube lengths. This study aimed to identify and characterize the subgroups of cultivars based on their pollen tube growth performance. In vitro pollen tube lengths of 41 individuals were determined by incubating samples on artificial germination media at 15 and 25 °C. A six-number summary statistic was calculated, and hierarchical clustering on principal component (HCPC) analysis was used to determine and characterize subgroups. Furthermore, a decision tree model was used to predict class membership for future datasets. HCPC analysis partitioned the 41 individuals into three subgroups with different performances. The decision tree quickly predicted the cluster membership based on the second quartile at 15 °C and the third quartile at 25 °C. The thresholds from the decision tree can be used to characterize new observations. The use of the methods will be demonstrated using a case study with 29 apple accessions.

Transcriptomics of Differential Ripening in 'd'Anjou' Pear (Pyrus communis L.).

Estimating maturity in pome fruits is a critical task that directs virtually all postharvest supply chain decisions. This is especially important for European pear (Pyrus communis) cultivars because losses due to spoilage and senescence must be minimized while ensuring proper ripening capacity is achieved (in part by satisfying a fruit chilling requirement). Reliable methods are lacking for accurate estimation of pear fruit maturity, and because ripening is maturity dependent it makes predicting ripening capacity a challenge. In this study of the European pear cultivar 'd'Anjou', we sorted fruit at harvest based upon on-tree fruit position to build contrasts of maturity. Our sorting scheme showed clear contrasts of maturity between canopy positions, yet there was substantial overlap in the distribution of values for the index of absorbance difference (I AD ), a non-destructive spectroscopic measurement that has been used as a proxy for pome fruit maturity. This presented an opportunity to explore a contrast of maturity that was more subtle than I AD could differentiate, and thus guided our subsequent transcriptome analysis of tissue samples taken at harvest and during storage. Using a novel approach that tests for condition-specific differences of co-expressed genes, we discovered genes with a phased character that mirrored our sorting scheme. The expression patterns of these genes are associated with fruit quality and ripening differences across the experiment. Functional profiles of these co-expressed genes are concordant with previous findings, and also offer new clues, and thus hypotheses, about genes involved in pear fruit quality, maturity, and ripening. This work may lead to new tools for enhanced postharvest management based on activity of gene co-expression modules, rather than individual genes. Further, our results indicate that modules may have utility within specific windows of time during postharvest management of 'd'Anjou' pear.

Determination of Post-Harvest Biochemical Composition, Enzymatic Activities, and Oxidative Browning in 14 Apple Cultivars.

Phenolic compounds in fruit provide human health benefits, and they contribute to color, taste, and the preservation of post-harvest fruit quality. Phenolic compounds also serve as modifiers of enzymatic activity, whether inhibition or stimulation. Polyphenol oxidases (PPO) and peroxidases (POD) use phenolic compounds as substrates in oxidative browning. Apple browning leads to flesh color, taste, texture, and flavor degradation, representing a drawback for the variety and its' market appraisal. This study was conducted to investigate the process of browning in 14 apple cultivars throughout post-harvest at three-time points: immediately (T0), one hour (T1), and 24 h (T2) after apples were cut in half. Color parameters L* (lightness), a* (red/green), b* (yellow/blue) were measured, and chroma (ΔC*) and color (ΔE) were calculated to quantify differences between T0₋T1 and T1₋T2 on the fruit surface. Enzymatic activity (PPO, POD) and phenolic composition were also quantified for each cultivar. 'Granny Smith' and 'Cripps Pink' browned minimally. In contrast, 'Fiesta' and 'Mondial Gala' browned severely, reporting high enzymatic activity and quantified phenolic concentration (QPC). Phenolic compound polymerization appears to play a significant role in enzymatic inhibition. 'Topaz' does not fit the high QPC, PPO, and browning formula, suggesting alternative pathways that contribute to apple browning.

Photoselective Protective Netting Improves "Honeycrisp" Fruit Quality.

High temperatures, wind, and excessive sunlight can negatively impact yield and fruit quality in semi-arid apple production regions. Netting was originally designed for hail protection, but it can modify the light spectrum and affect fruit quality. Here, pearl, blue, and red photoselective netting (≈20% shading factor) was installed in 2015 over a commercial "Cameron Select® Honeycrisp" orchard. Our research objectives were to (1) describe the light quantity and quality under the colored nets compared to an uncovered control and (2) investigate the effect of Photoselective nets on "Honeycrisp" apple quality for two growing seasons. Light transmittance and scattering for each treatment were measured with a spectroradiometer, and samples for fruit quality analyses were collected at harvest. PAR (photosynthetic active radiation), UV, blue, red, and far-red light were lower underneath all netting treatments compared to an uncovered control. The scattered light was higher under the pearl net compared to other colors, while red and far-red light were lower under the blue net. For two consecutive years, trees grown under the photoselective nets intercepted more incoming light than the uncovered trees with no differences among the three colors. In both years, trees under red and blue nets had more sunburn-free (clean) apples than pearl and control. Red color development for fruit was lower when nets were used. Interestingly, bitter pit incidence was lower underneath red nets for both years. Other than red color development, "Honeycrisp" fruit quality was not appreciably affected by the use of netting. These results highlight the beneficial effect of nets in improving light quality in orchards and mitigating physiological disorders such as bitter pit in "Honeycrisp" apple.

Determination of Biochemical Composition in Peach (Prunus persica L. Batsch) Accessions Characterized by Different Flesh Color and Textural Typologies.

The rising interest in beneficial health properties of polyphenol compounds in fruit initiated this investigation about biochemical composition in peach mesocarp/exocarp. Biochemical evaluation of phenolic compounds and ascorbic acid were quantified through high-performance liquid chromatography (HPLC) in relation to three flesh colors (white, yellow and red) and four flesh typologies (melting, non-melting, slow softening and stony hard) within six commercial cultivars and eight breeding selections of peach/nectarine in 2007. While in 2008, quality and sensorial analyses were conducted on only three commercial cultivars ('Big Top', 'Springcrest' and 'Ghiaccio 1'). The red flesh selection demonstrated the highest levels of phenolic compounds (in mesocarp/exocarp) and ascorbic acid. Total phenolic concentration was approximately three-fold higher in the exocarp than the mesocarp across all accessions. Breeding selections generally reported higher levels of phenolics than commercial cultivars. Flesh textural typologies justified firmness differences at harvest, but minimally addressed variations in quality and phenolic compounds. Flesh pigmentation explained variation in the biochemical composition, with the red flesh accession characterized by an abundancy of phenolic compounds and a high potential for elevated antioxidant activity. Sensorial analyses ranked the cultivar with high soluble solids concentration:titratable acidity (SSC:TA) and reduced firmness the highest overall. Red flesh is a highly desirable trait for breeding programs aiming to improve consumption of peaches selected for nutraceutical properties.

Canopy attachment position influences metabolism and peel constituency of European pear fruit.

BACKGROUND: Inconsistent pear fruit ripening resulting from variable harvest maturity within tree canopies can contribute to postharvest losses through senescence and spoilage that would otherwise be effectively managed using crop protectant and storage regimes. Because those inconsistencies are likely based on metabolic differences, non-targeted metabolic profiling peel of 'd'Anjou' pears harvested from the external or internal canopy was used to determine the breadth of difference and link metabolites with canopy position during long-term controlled atmosphere storage. RESULTS: Differences were widespread, encompassing everything from expected distinctions in flavonol glycoside levels between peel of fruit from external and internal canopy positions to increased aroma volatile production and sucrose hydrolysis with ripening. Some of the most substantial differences were in levels of triterpene and phenolic peel cuticle components among which acyl esters of ursolic acid and fatty acyl esters of p-coumaryl alcohol were higher in the cuticle of fruit from external tree positions, and acyl esters of α-amyrin were elevated in peel of fruit from internal positions. Possibly the most substantial dissimilarities were those that were directly related to fruit quality. Phytosterol conjugates and sesquiterpenes related to elevated superficial scald risk were higher in pears from external positions which were to be potentially rendered unmarketable by superficial scald. Other metabolites associated with fruit aroma and flavor became more prevalent in external fruit peel as ripening progressed and, likewise, with differential soluble solids and ethylene levels, suggesting the final product not only ripens differentially but the final fruit quality following ripening is actually different based on the tree position. CONCLUSIONS: Given the impact tree position appears to have on the most intrinsic aspects of ripening and quality, every supply chain management strategy would likely lead to diverse storage outcomes among fruit from most orchards, especially those with large canopies. Metabolites consistently associated with peel of fruit from a particular canopy position may provide targets for non-destructive pre-storage sorting used to reduce losses contributed by this inconsistency.

Stylet penetration of Cacopsylla pyri; an electrical penetration graph (EPG) study.

Detailed information on plant penetration activities by pear psylla Cacopsylla pyri L. (Hemiptera Psyllidae) is essential to study phytoplasma transmission of "Candidatus Phytoplasma pyri" responsible of pear decline disease (PD) and to trace and evaluate resistant traits in new pear tree selections for advanced breeding programs. The electrical penetration graph technique or (full) EPG may relevantly contribute to this knowledge. C. pyri EPG waveforms were characterized on basis of amplitude, frequency, voltage level, and electrical origin. Additionally, stylet tracks and the putative location of stylet tips in the plant tissue were histologically related to EPG waveforms by light and transmission electron microscopy observations after stylectomy. More than one waveform occurred in the same tissue: PA, PB, PC1 and PC2 were all detected in the mesophyll, and PE1 and PE2 were both recorded in the phloem. Waveform PE1 was always preceded by transient waveform PD, as previously described in other psyllids. Interestingly, no brief intracellular punctures (potential drop waveforms) were observed during plant penetration, opposite of what is usually recorded in aphids and other Sternorrhyncha.

Mapping of an anthocyanin-regulating MYB transcription factor and its expression in red and green pear, Pyrus communis.

'Max Red Bartlett' is a red bud mutation of the yellow pear (Pyrus communis L.) cultivar 'Williams' (known as 'Bartlett' in North America). Anthocyanins are the most important pigments for red colour in fruits. Synthesis of anthocyanins is mediated by a number of well-characterized enzymes that include chalcone synthase (CHS), flavanone-3-hydroxylase (F3H), dihydroflavonol-4-reductase (DFR), anthocyanidin synthase (ANS), and UDP-glucose:flavonoid-3-O-glucosyltransferase (UFGT). Expression of the genes encoding these five enzymes was examined in pear fruit skin in order to elucidate the molecular mechanism for red coloration. In addition, the gene PcMYB10, encoding an R2R3 MYB transcription factor involved in anthocyanin biosynthetic pathway regulation, was isolated from both 'Williams' and 'Max Red Bartlett'. Analysis of the deduced amino acid sequence suggests that this gene is an ortholog of anthocyanin regulators known in other plant species. Its expression level was significantly higher in 'Max Red Bartlett' (red pear) compared with the original yellow variety 'Williams'. Although the map position of PcMYB10 corresponds to that of MdMYBa and MdMYB10, which control pigmentation of apple fruit skin, PcMYB10 is not directly responsible for red versus yellow colour in the two pear varieties, as the mutation underlying this difference maps to a different region of the pear genome.

Use of homologous and heterologous gene expression profiling tools to characterize transcription dynamics during apple fruit maturation and ripening.

BACKGROUND: Fruit development, maturation and ripening consists of a complex series of biochemical and physiological changes that in climacteric fruits, including apple and tomato, are coordinated by the gaseous hormone ethylene. These changes lead to final fruit quality and understanding of the functional machinery underlying these processes is of both biological and practical importance. To date many reports have been made on the analysis of gene expression in apple. In this study we focused our investigation on the role of ethylene during apple maturation, specifically comparing transcriptomics of normal ripening with changes resulting from application of the hormone receptor competitor 1-methylcyclopropene. RESULTS: To gain insight into the molecular process regulating ripening in apple, and to compare to tomato (model species for ripening studies), we utilized both homologous and heterologous (tomato) microarray to profile transcriptome dynamics of genes involved in fruit development and ripening, emphasizing those which are ethylene regulated.The use of both types of microarrays facilitated transcriptome comparison between apple and tomato (for the later using data previously published and available at the TED: tomato expression database) and highlighted genes conserved during ripening of both species, which in turn represent a foundation for further comparative genomic studies. The cross-species analysis had the secondary aim of examining the efficiency of heterologous (specifically tomato) microarray hybridization for candidate gene identification as related to the ripening process. The resulting transcriptomics data revealed coordinated gene expression during fruit ripening of a subset of ripening-related and ethylene responsive genes, further facilitating the analysis of ethylene response during fruit maturation and ripening. CONCLUSION: Our combined strategy based on microarray hybridization enabled transcriptome characterization during normal climacteric apple ripening, as well as definition of ethylene-dependent transcriptome changes. Comparison with tomato fruit maturation and ethylene responsive transcriptome activity facilitated identification of putative conserved orthologous ripening-related genes, which serve as an initial set of candidates for assessing conservation of gene activity across genomes of fruit bearing plant species.

QTL dynamics for fruit firmness and softening around an ethylene-dependent polygalacturonase gene in apple (Malus x domestica Borkh.).

Apple fruit are well known for their storage life, although a wide range of flesh softening occurs among cultivars. Loss of firmness is genetically coordinated by the action of several cell wall enzymes, including polygalacturonase (PG) which depolymerizes cell wall pectin. By the analysis of 'Fuji' (Fj) and 'Mondial Gala' (MG), two apple cultivars characterized by a distinctive ripening behaviour, the involvement of Md-PG1 in the fruit softening process was confirmed to be ethylene dependent by its transcript being down-regulated by 1-methylcyclopropene treatment in MG and in the low ethylene-producing cultivar Fj. Comparing the PG sequence of MG and Fj, a single nucleotide polymorphism (SNP) was discovered. Segregation of the Md-PG1(SNP) marker within a full-sib population, obtained by crossing Fj and MG, positioned Md-PG1 in the linkage group 10 of MG, co-located with a quantitative trait locus (QTL) identified for fruit firmness in post-harvest ripening. Fruit firmness and softening analysed in different stages, from harvest to post-storage, determined a shift of the QTL from the top of this linkage group to the bottom, where Md-ACO1, a gene involved in ethylene biosynthesis in apple, is mapped. This PG-ethylene-related gene has beeen positioned in the apple genome on chromosome 10, which contains several QTLs controlling fruit firmness and softening, and the interplay among the allelotypes of the linked loci should be considered in the design of a marker-assisted selection breeding scheme for apple texture.

Oxidative stress and senescence-like status of pear calli co-cultured on suspensions of incompatible quince microcalli.

This work presents a simple in vitro system to study physiological, biochemical and molecular changes occurring in a pear callus (Pyrus communis L., cv. Beurré Bosc) grown in close proximity to spatially separated undifferentiated homologous (pear) or heterologous (quince; Cydonia oblonga Mill., East Malling clone C) cells in its neighboring environment. After a 7-day co-culture period, the presence of heterologous cells produced negative effects on the pear callus, whose relative weight increase and adenylate energy charge decreased by 30 and 24%, respectively. Such behavior was associated with a higher O(2) consumption rate (+125%) which did not seem to be coupled to adenosine triphosphate synthesis. Analyses of alternative oxidase and enzymatic activities involved in reactive oxygen species (ROS) detoxification strongly suggested that the higher O(2) consumption rate, measured in the pear callus grown in the heterologous combination, may probably be ascribed to extra-respiratory activities. These, in turn, might contribute to generate metabolic scenarios where ROS-induced oxidative stresses may have the upper hand. The increase in the levels of 2-thiobarbituric acid reactive metabolites, considered as diagnostic indicators of ROS-induced lipid peroxidation, seemed to confirm this hypothesis. Moreover, reverse transcription polymerase chain reaction analysis revealed that the expression levels of a few senescence-associated genes were higher in the pear callus grown in the heterologous combination than in the homologous one. Taken as a whole, physiological and molecular data strongly suggest that undifferentiated cells belonging to a pear graft-incompatible quince clone may induce an early senescence-like status in a closely co-cultured pear callus.