Malus ×domestica Borkh. cv. Honeycrisp has been widely planted in North America during the past two decades. However, it is susceptible to many disorders that result in high postharvest losses. Excessive vegetative vigor in apple trees can reduce fruit calcium (Ca) concentrations and increase bitter pit incidence in apple fruit. Plant growth regulators are used routinely in tree fruit orchards to control vegetative growth to increase light penetration into the canopy. The objective of this study was to determine whether shoot growth inhibition using the application of prohexadione-calcium (P-Ca; Apogee®) or stimulation via application of gibberellic acid (GA3; ProGibb®) affected bitter pit incidence in ‘Honeycrisp’ apple. In 2016 and 2017, the experiment was conducted in a commercial ‘Honeycrisp’ orchard with five treatments [untreated control, 62.5 mg·L–1 P-Ca (low P-Ca); 125 mg·L–1 P-Ca (high P-Ca); 16 mg·L–1 GA3 (low GA3); and 32 mg·L–1 GA3 (high GA3)]. Treatments were applied twice during the growing season. Shoot length and the number of internodes for new growth were measured 4 weeks apart after treatment. Overall yield and fruit quality were assessed at harvest, and bitter pit incidence was assessed after 4 months of storage. Low and high P-Ca rates limited shoot growth extension; high GA3 increased shoot extension compared with the untreated control. However, the number of internodes did not change substantially for each shoot. The number of internodes is one of the primary factors affecting leaf area and, consequently, the transpiration balance between fruit and leaves. In both years, treatments with either GA3 or P-Ca did not affect fruit elemental concentration or bitter pit incidence. These results indicate that growth-inhibiting plant growth regulators that reduce shoot extension may not be useful for managing bitter pit incidence in ‘Honeycrisp’ apple.
Corina Serban and Lee Kalcsits
Raquel Gomez and Lee Kalcsits
‘Honeycrisp’ is among the most widely grown apple cultivars in the United States and ‘WA 38’ is a new apple cultivar released in Washington State. ‘Honeycrisp’ is highly susceptible to bitter pit and other physiological disorders; however, ‘WA 38’ is not susceptible to bitter pit but little is known about its susceptibility to other disorders. Bitter pit is a calcium-related disorder that has been associated with localized calcium deficiencies in fruit in addition to the proportions of calcium relative to the presence of other nutrients like potassium and magnesium. The objective of this study was to compare physiological differences and fruit quality between ‘Honeycrisp’ and ‘WA 38’ to determine how these differences might correspond to differences in mineral nutrient composition and bitter pit susceptibility. Here, ‘Honeycrisp’ and ‘WA 38’ elemental composition in leaves, fruit, and xylem sap was measured every 20 days starting 30 days after full bloom and compared with leaf gas exchange and stem water potential. ‘Honeycrisp’ had greater foliar transpiration rates that corresponded with greater calcium in the leaves and lower leaf K+Mg/Ca ratio, when compared with ‘WA 38’. In contrast, fruit calcium concentrations were higher for ‘WA 38’ with lower K+Mg/Ca ratios. Xylem conductance was higher during late summer in ‘WA 38’ compared with ‘Honeycrisp’. ‘WA 38’ fruit was denser than ‘Honeycrisp’ and more research is needed to determine whether differences in fruit structure may affect susceptibility to bitter pit in apple.
Nadia A. Valverdi and Lee Kalcsits
‘Honeycrisp’ apple is susceptible to bitter pit, which is associated with fruit mineral nutrient composition. Rootstock genotypes can affect nutrient acquisition, distribution, and fruit yields, which all affect fruit nutrient composition and bitter pit susceptibility. However, the changes of these traits among different rootstock genotypes in response to abiotic stress under semiarid conditions are relatively unknown. The objective of this study was to evaluate the influence of different rootstocks and irrigation on nutrient uptake and partitioning with ‘Honeycrisp’ apple grown in an irrigated, semiarid environment. ‘Honeycrisp’ apple trees were grafted on four different rootstocks, Geneva 41 (‘G.41’), Geneva 890 (‘G.890’), M.9-T337 (‘M.9’), and Budagovsky 9 (‘B.9’), and these were planted at high density (3000 trees/ha). Irrigation was applied as either a water-limited treatment where volumetric soil water content was maintained near 50% field capacity (FC) and a well-watered control where soil water content was maintained near 100% FC. ‘G.890’, the most vigorous rootstock, had lower nitrogen and higher potassium content in leaves, while ‘B.9’, the least vigorous rootstock, had lower potassium and higher nitrogen content. Rootstock genotype did not affect calcium uptake. Interestingly, water-limited conditions increased the nutrient content in root and stems but not in leaves. Water-limited trees partitioned more nitrogen and calcium to roots, while well-watered trees in the control partitioned more nutrients to the stems. Fruit size was the largest for ‘G.890’ and smallest for ‘B.9’. Both ‘G.41’ and ‘G.890’ had higher bitter pit incidence, which was associated with higher potassium content in leaves and fruit. These results suggest that rootstock-induced vigor and irrigation can both contribute to nutrient imbalances in leaves and fruit that could affect the development of physiological disorders in ‘Honeycrisp’ apple.
Heidi Hargarten, Sumyya Waliullah, Lee Kalcsits, and Loren A. Honaas
Complex changes in gene expression occur during postharvest storage of apple (Malus ×domestica) and often precede or accompany changes in ripening and disorder development. Targeted gene expression analysis fundamentally relies on previous knowledge of the targeted gene. Minimally, a substantial fragment of the gene sequence must be known with high accuracy so that primers and probes, which bind to their targets in a complimentary fashion, are highly specific. Here, we describe a workflow that leverages publicly available transcriptome data to discover apple cultivar–specific gene sequences to guide primer design for quantitative real-time polymerase chain reaction (qPCR). We find that problematic polymorphisms occur frequently in ‘Granny Smith’ and ‘Honeycrisp’ apple when candidate primer binding sites were selected using the ‘Golden Delicious’ genome. We attempted to validate qPCR-based gene expression measurements with RNA sequencing (RNA-Seq) analysis of the same RNA samples. However, we found that agreement between the two technologies was highly variable and positively correlated with the similarity between cultivar-specific genes and RNA-Seq reference genes. Thus, we offer insight that 1) improves the accuracy and efficiency of qPCR primer design in cultivars that lack sufficient sequence resources and 2) better guides the essential step of validation of RNA-Seq data with a subset of genes of interest examined via qPCR.
Lee Kalcsits, Gregory van der Heijden, Michelle Reid, and Katie Mullin
Calcium (Ca) sprays are commonly used to control Ca-related disorders such as bitter pit in apple. Increases in the frequency and the amount of Ca applied directly to the fruit have increased fruit Ca levels and are associated with a reduction in bitter pit incidence. However, the absorption efficiency at different fruit developmental stages is poorly understood. Here, the absorption efficiency was measured using 44Ca stable isotope applied to 30 individual fruit at five different times every 2 weeks between June drop and 2 weeks before harvest in a medium-density ‘Honeycrisp’ orchard. Fruit size, spray adhesion, and Ca and potassium (K) content were monitored weekly for 12 weeks between 26 May and 13 Aug. 2015. At harvest, the 44Ca-labeled fruit was picked and separated into peel and inner fruit for mass balance analysis of 44Ca absorption to regions of the fruit that are important to prevent Ca-related disorders. As expected, δ44Ca was greater in the peel than the interior of the fruit. However, there was a significant amount of 44Ca present in the inner fruit at harvest for all five applications applied during the growing season. Using a stable isotope tracer approach, we present evidence that Ca is absorbed throughout fruit development. These findings support current recommendations for frequent Ca applications in low concentrations throughout fruit development to increase fruit Ca levels and reduce the incidence of bitter pit in ‘Honeycrisp’ apple.
Corina Serban, Lee Kalcsits, Jennifer DeEll, and James P. Mattheis
‘Honeycrisp’ apples are susceptible to bitter pit, a physiological disorder that impacts peel and adjacent cortex tissue. ‘Honeycrisp’ is also susceptible to chilling injury (CI) that can be prevented by holding fruit at 10 to 20 °C after harvest for up to 7 days. This temperature conditioning period reduces CI risk but can enhance bitter pit development. Previous research demonstrated a controlled atmosphere (CA) established during conditioning can reduce ‘Honeycrisp’ bitter pit development without inducing other physiological disorders. The objective of this research was to evaluate the duration of CA needed to reduce bitter pit development. Experiments were conducted in 2014, 2016, and 2017 with fruit obtained from commercial orchards in Washington State and, in 2017 only, Ontario, Canada. Half the fruit were treated with 42 µmol·L−1 1-methycyclopropene (1-MCP) for 24 hours at 10 °C immediately following harvest. The untreated fruit were held at the same temperature (10 °C) in a different cold room. Following 1-MCP treatment, all fruit were conditioned at 10 °C for an additional 6 days, then fruit was cooled to 2.8 °C. During conditioning, fruit were held in air or CA (2.5 kPa O2, 0.5 kPa CO2) established 1 day after harvest, for 1 to 8 weeks, then in air. All fruit were removed from cold storage after 4 months and then held 7 days at 20 °C. Fruit from most orchards/years stored in CA developed less bitter pit compared with fruit stored continuously in air. CA during conditioning also reduced poststorage peel greasiness but CA for 2 weeks or longer enhanced cortex cavity development in some orchard lots. Treatment with 1-MCP did not reduce bitter pit but enhanced development of peel leather blotch and core browning for some orchards/years. 1-MCP–treated fruit slowed the loss of soluble solids content, titratable acidity, and reduced internal ethylene concentration. Results suggest the potential for postharvest management of bitter pit development in ‘Honeycrisp’ apples by CA established during conditioning with minimal development of other postharvest disorders.
Sara Serra, Rachel Leisso, Luca Giordani, Lee Kalcsits, and Stefano Musacchi
The apple variety, ‘Honeycrisp’ has been extensively planted in North America during the last two decades. However, it suffers from several agronomic problems that limit productivity and postharvest quality. To reduce losses, new information is needed to better describe the impact of crop load on productivity and postharvest fruit quality in a desert environment and the major region where ‘Honeycrisp’ expansion is occurring. Here, 7-year-old ‘Honeycrisp’ trees on the M9-Nic29 rootstock (2.5 × 0.9 m) were hand thinned to five different crop loads [from 4.7 to 16.0 fruit/cm2 of trunk cross-sectional area (TCSA)] to compare fruit quality, maturity, fruit size, elemental concentration, and return bloom. Fruit size distribution was affected by crop load. Trees with the highest crop load (16 fruit/cm2) produced smaller fruit. Index of absorbance difference (I AD) measurements (absorption difference between 670 and 720 nm), a proxy indicator of the chlorophyll content below the skin of fruit measured by a DA-meter, were made shortly after harvest (T0) and after 6 months of storage (T1). Fruit from the trees with the lowest crop load had lower I AD values indicating advanced fruit ripeness. The comparison between the I AD classes at T0 and T1 showed that fruit belonging to the lowest I AD class had significantly higher red-blushed overcolor percentage, firmness, dry matter, and soluble solid content than those in the “most unripe” class (highest I AD readings) regardless of crop load. The percentage of blushed color, firmness, titratable acidity (TA), soluble solids content, and dry matter were all higher in the lowest crop loads at both T0 and T1. Fruit calcium (Ca) concentration was lowest at the lowest crop load. The (K + Mg + N):Ca ratio decreased as crop load increased until a crop load of 11.3 fruit/cm2, which was not significantly different from higher crop loads. For return bloom, the highest number of flower clusters per tree was reported for 4.7 fruit/cm2 crop load, and generally it decreased as crop load increased. Here, we highlight the corresponding changes in fruit quality, storability, and elemental balance with tree crop load. To maintain high fruit quality and consistency in yield, careful crop load management is required to minimize bienniality and improve fruit quality and storability.
Giverson Mupambi, Stefano Musacchi, Sara Serra, Lee A. Kalcsits, Desmond R. Layne, and Tory Schmidt
Globally, apple production often occurs in semiarid climates characterized by high summer temperatures and solar radiation. Heat stress events occur regularly during the growing season in these regions. For example, in the semiarid eastern half of Washington State, historic weather data show that, on average, 33% of the days during the growing season exceed 30 °C. To mediate some of the effects of heat stress, protective netting (PN) can be used to reduce the occurrence of fruit sunburn. However, the impacts of reduced solar radiation in a high light environment on light-use efficiency and photosynthesis are poorly understood. We sought to understand the ecophysiological response of apple (Malus domestica Borkh. cv. Honeycrisp) under blue photoselective PN during days with low (26.6 °C), moderate (33.7 °C), or high (38.1 °C) ambient temperatures. Two treatments were evaluated; an uncovered control and blue photoselective PN. Maximum photochemical efficiency of PSII, or photosystem II (Fv/Fm) was significantly greater at all measurement times under blue photoselective PN compared with the control on days with high ambient temperatures. Fv/Fm dropped below 0.79, which is considered the threshold for stress, at 1000 hr in the control and at 1200 hr under blue photoselective PN on a day with high ambient temperature. On days with low or moderate ambient temperatures, Fv/Fm was significantly greater under blue photoselective PN at 1400 hr, which coincided with the peak in solar radiation. ‘Honeycrisp’ apple exhibited dynamic photoinhibition as shown by the diurnal decline in Fv/Fm. Quantum photosynthetic yield of PSII (ΦPSII) was also generally greater under blue photoselective PN compared with the control for days with moderate or high ambient temperatures. Photochemical reflectance index (ΔPRI), the difference in reflectance between a stress-responsive and nonstress-responsive wavelength, was greater under PN compared with the control on the day with high ambient temperatures, with no differences observed under low or moderate ambient temperatures. Leaf gas exchange did not show noticeable improvement under blue photoselective netting when compared with the control despite the improvement in leaf-level photosynthetic light use efficiency. In conclusion, PN reduced incoming solar radiation, improved leaf-level photosynthetic light use efficiency, and reduced the symptoms of photoinhibition in a high-light, arid environment.