ASHS 2024 Conference

Apple’s (Malus × domestica Borkh.) tendency towards biennial bearing necessitates management strategies such as fruit thinning and vigor control. Applications of plant growth regulators to manage crop load and vigor overlap in the spring. A precursor to ethylene, 1-aminocyclopropane-1-carboxylic acid (ACC; Accede®) was recently registered as a chemical fruit thinner in the USA. Prohexadione-calcium (P-Ca) is a gibberellin biosynthesis inhibitor widely used to manage vigor. It is suspected that P-Ca may interfere with ACC precursors, decreasing thinning efficacy of ACC. We evaluated the effects and interactions of ACC and P-Ca on shoot growth, ethylene evolution, leaf area, and yield responses of mature ‘Golden Delicious’. The experiment was conducted in 2023, in Mills River, NC, USA. Treatments included ACC (400 mg∙L-1), P-Ca (250 mg∙L-1), ACC P-Ca, and an untreated control. Application of P-Ca occurred on three dates. ACC was applied when average fruit diameter was ~20 mm. Across all measured responses, there were no interactions between ACC and P-Ca. P-Ca had a negative main effect on shoot growth rate (48.7–75.4% reduction). ACC reduced leaf number of spurs 4 and 9 days after treatment (DAT). Ethylene evolution in fruit and spurs was increased by ACC 1, 4, and 9 DAT (spur: 1.1e4%, 5300%, and 515%, respectively; fruit: 9200%, 725%, and 222%, respectively). ACC had a negative effect on fruit number, yield, and crop density (60.1%, 56.5%, and 60.3% reduction, respectively). These results indicate that ACC and P-Ca can be used simultaneously to manage crop load and vigor, respectively

Crop load management of apple (Malus x domestica Borkh.) is one of the most important orchard management practices to ensure adequate fruit size and quality. Fruitlets are most sensitive to thinner applications early in the season, between 6mm and 20mm fruitlet diameter. During this time, abscission is associated with environmental and biological factors that influence fruitlet development, growth and sink strength. The carbohydrate economy of the fruitlet and its position within the cluster (king flower vs. lateral flowers) are primary factors contributing to the sink strength of individual fruitlets. We evaluated the relationship between the non-structural carbohydrate status of apple fruitlets in three different size classes (i.e., 90th, 50th, and 10th percentile of the population) and their respective likelihood to persist throughout the season. Persistence probabilities of these size classes were generated by an empirical fruit set prediction model based on the distribution of fruitlet mass and confirmed by actual measures of fruit set. Soluble carbohydrates and starch were quantified with an enzymatic assay procedure. Following chemical thinner application, the concentration of soluble sugars (glucose, fructose, sucrose, and sorbitol) was significantly reduced on a gram / gram fresh weight basis. Fruitlets in the 10th percentile generally had the lowest concentration of soluble sugars and starch and the slowest recovery rate following chemical thinner application. Fruitlets in the 90th percentile generally had the highest concentrations of soluble sugars and starch. Our data support that differences in the carbohydrate concentration of fruitlets contribute to their sink strength and abscission fate as predicted by a fruit set prediction model.

Post-bloom apple chemical thinning relies on the application of plant bioregulators to induce fruitlet abscission, reducing crop load to improve fruit quality and promote return bloom. Abscission rates following chemical thinner application are influenced by carbohydrate status of the tree and thus widely variable. Multiple applications are often needed for growers to reach a desired crop load. However, trees are only perceptive to chemical thinners for a 3 to 4 week period shortly after bloom. Rapid prediction of chemical thinner efficacy is critical to allow growers to make necessary applications in this narrow period of time without the risk of over-thinning. In 2021 and 2022 a predictive model using reflectance spectroscopy measured with a handheld spectrometer (Felix F-750; Felix Instruments; Camas, WA, USA) was developed to predict chemical thinner efficacy for ‘Honeycrisp’ in North Carolina, USA. This model was found to predict chemical thinner efficacy with greater than 85% accuracy as early 3 days following a chemical thinner application. In 2023 and 2024 validation of this model occurred on multiple cultivars: ‘Red Delicious’, ‘Granny Smith’, ‘Gala’, and ‘Fuji’; and in multiple growing regions: Michigan, New York, and North Carolina, USA. Performance of the developed model to predict chemical thinning efficacy in this wide range of situations will be discussed to determine commercial applicability of the model.

Vacuolar accumulation of malic acid, the predominant organic acid in apple (Malus domestica) and many other fleshy fruits, largely determines fruit acidity, a key trait for fruit taste and flavor. ALUMINUM-ACTIVATED MALATE TRANSPORTER 9 (ALMT9/Ma1) underlies a major genetic locus, Ma, for fruit acidity in apple. A natural mutation at base 1455 leads to a pre-mature stop codon that truncates the Ma1 protein by 84 amino acids to ma1, and this truncation significantly reduces Ma1’s malate transport activity by disrupting a conserved C-terminal domain, leading to low fruit acidity in recessive homozygous ma1ma1 genotypes. However, it remains unclear how Ma1 functions to transport malate across the tonoplast membrane. When the coding sequence of Ma1 was overexpressed in ‘Royal Gala’ apple (Ma1ma1) to determine its in planta function, to our surprise, the transgenic fruit had drastically reduced fruit acidity compared with the wild-type (WT). This led us to uncover that Ma1 undergoes alternative splicing. Two isoform proteins are generated by alternative splicing: Ma1beta being 68 amino acids shorter with much lower expression than the full-length protein Ma1alpha. Ma1beta does not transport malate itself, but interacts with the functional Ma1alpha, creating synergy with Ma1alpha for malate transport in a threshold manner when Ma1beta is equal to or exceeds 1/8 of Ma1alpha. In WT ‘Royal Gala’, Ma1 operates at this threshold. Overexpression of Ma1alpha triggers feedback inhibition on the native Ma1 expression via transcription factor MYB73, decreasing the Ma1beta level well below the threshold that leads to significant reductions in Ma1 function and malic acid accumulation in fruit. Overexpression of Ma1alpha and Ma1beta or genomic Ma1 increases both isoforms proportionally and enhances fruit malic acid accumulation. These findings reveal an essential role of alternative splicing in Ma1-mediated malate transport underpinning apple fruit acidity.

Biennial or alternate bearing is characterized by an abundance of blooming and fruiting in “on” years and a lack of blooming and fruiting in “off” years. Biennial bearing in fruit tree production has been extensively studied. However, it continues to be problematic in certain crops and cultivars. Fundamental physiological mechanisms controlling this behavior are still poorly understood. Along with carbohydrates in plant tissues, reserve nitrogen influences early-season growth in apples, with the percent contribution of reserve nitrogen being dependent on the current nitrogen supply. Because flowering and fruit set occur in spring alongside bud break, reserve nitrogen can significantly affect these processes too. We sought to understand the role carbohydrate and nitrogen storage in various plant tissues has on biennial bearing for ‘Honeycrisp’ apple. Apical buds, terminal shoots, and one-year-old spurs were collected in 2022

In apple orchards, addressing pre-harvest fruit drop in early-maturing, commercially valuable cultivars like ‘Honeycrisp’ and ‘Gala’ is crucial. This issue is largely linked to ethylene, a ripening hormone that, despite its role in enhancing fruit qualities, triggers premature fruit drop. From 2018 to 2021, extensive field trials were conducted across commercial and experimental orchards to refine the application of two ethylene-inhibiting products, ReTain (AVG-15%) and Harvista (1-MCP-1.3%), in mitigating this challenge in ‘Gala’ apples. Our findings confirmed AVG’s effectiveness in reducing fruit drop and stem-end splitting; however, AVG application consistently resulted in diminished fruit coloration. In the subsequent seasons of 2023 and 2024, our research pivoted towards identifying plant growth regulator (PGR) formulations that could balance the dual objectives of minimizing fruit drop and preserving color development in ‘Honeycrisp’ and ‘Gala’ apples. Key findings include: a) the multiple applications of Accede (ACC-10%) and Motivate (Ethephon-21.7%) starting from three weeks before harvest significantly enhanced red color formation in ‘Honeycrisp’ and ‘Gala’ apples, surpassing control and ReTain treatments, though Motivate increased the rate of fruit drop; b) ReTain, both alone and in combination with Accede, notably reduced fruit drop, with the ReTain-Accede mix also enhancing fruit coloration; c) combining ReTain and Motivate neither decreased fruit drop nor enhanced color; d) the positive impact of the ReTain-Accede mix on color was biochemically and molecularly validated; e) Accede's color-enhancing effect appears to be ethylene-independent. This research underscores the potential of specific PGR mixes in achieving a harmonious balance between reducing fruit drop and enhancing fruit coloration.

The influence of rootstock and time of crop load adjustment on ‘Honeycrisp’ apple fruit quality at harvest and after storage was studied over four seasons. Yield per trees varied from year to year because of biennial bearing. Trees on V.1 and G.30 were higher while those on G.969 often had lower yield per tree. Fruit from trees on M.26EMLA and G.30 were larger, while those on G.202 and G.969 were smaller than those on other rootstocks in two of three years. Fruit firmness at harvest and after storge was often inversely proportional to their size, so that fruits from trees on G. 202 often had higher but those on G.30 often had lower firmness at harvest and after storage. Percentages of fruit bitter pit among rootstocks at harvest was different than those after storage. However, fruit from trees on M.26EMLA, V.1, and G.30 generally had higher bitter pit than those on G.969 and G.202 rootstocks. Fruit fresh weight continually grew during growing season. However, the increase in percentage fruit dry weight reached a plateau on August 9, after which it leveled off. Trees receiving an “Early Heavy Thinning” and “Thinned Early to 1 Fruit/bunch” had lower yield but larger fruit with more bitter pit than Un-thinned-Control trees. Trees receiving a thinning on June 11 and June 25 had similar fruit size in the same season. However, bitter pit was increased, while fruit size decreased, as the hand thinning was delayed until July 8. Late hand thinning resulted in smaller fruit size with less bitter pit. Fruit potassium/ calcium (K/Ca) ratio did not change significantly between June 20 and August 8 within the same year. This research unveils importance of rootstock, time, and level of and thinning in the fruit quality of ‘Honeycrisp’ apple.

Insufficient transport of calcium to and throughout the cortex of apple is a primary factor contributing to bitter pit, a disorder characterized by dark lesions on the fruit surface. Calcium transport, in-planta, occurs solely in the xylem. Progressive transport dysfunction of xylem limits calcium concentration in the peel and subtending tissue. Rapid fruit growth may accelerate xylem dysfunction due to the stretching, shearing, or compressing of xylem elements. The relative time course of these events and subsequent effect on calcium concentration of peel tissues is not well understood. Growth rate of fruit was manipulated by manually adjusting the crop load of 6th leaf ‘Honeycrisp' trees to either 4 (low), 8 (moderate), or 16 (high) fruit per cm2 of trunk cross-sectional area via bud extinction. Fruit mass, xylem functionality, and peel calcium content were assessed weekly throughout the season. Fruit tissues were frozen weekly to evaluate the expression of genes related to xylogenesis and calcium transport and for quantification of hormones. At maturity, 25 trees were harvested from each crop load treatment. Fruit were assessed for bitter pit, stored for three months in regular air (3⁰C), then reassessed. Crop load influenced both xylem functionality and bitter pit incidence of fruit. Low crop load fruit lost 80% of xylem functionality by 8 weeks after bloom, which was two weeks earlier than moderate crop. By seven and five weeks before harvest, total xylem dysfunction was observed in fruit of low and moderate crop load, respectively; xylem in high crop load fruit never became fully dysfunctional. Final fruit mass from low crop load trees was only 20 grams larger than fruit from moderate crop load trees, yet, these fruit had a roughly 100% increase in bitter pit incidence at harvest (27.3% vs. 12.6%). Only 4.1% of high crop load fruit were affected by bitter pit. Many previous studies have ascribed the positive relationship between fruit mass and bitter pit incidence to calcium dilution. Our data suggest, however, that the issue is one of transport limits that appear to begin very early in fruit development, exacerbated by the growth rate of fruit, resulting in less peel calcium and higher bitter pit incidence at harvest than fruit of relatively lower growth rates. Future analysis of endogenous hormone concentrations and expression of genes relating to xylogenesis and calcium allocation will facilitate a better understanding of the processes governing xylem dysfunction and its association to bitter pit disorder.