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Sustainable practices are key to the improvement of soil fertility and quality in apple (Malus x domestica Borkh.) orchards. Rootstock genotype and fertilizer inputs can alter soil biology, as well as aboveground traits including nutrient acquisition. In this study, a factorial design was used to assess the interaction between two apple rootstocks, 'Geneva 41' ('G.41') and 'Malling 9' ('M.9') with four fertilizer treatments [chicken-litter compost, yardwaste compost, fertigation using Ca(NO3)(2), and an unamended control]. The bacterial community in the rhizosphere was assessed for its impact on both plant and soil properties for each rootstock x fertilizer treatment combination. The bacterial community was dominated by Acidobacteria, Proteobacteria, and Planctomycetes, but Verrucomicrobia and Chloroflexi were the most responsive to the fertilizer treatments. The chicken litter and yardwaste treatments had a greater effect on bacterial community structure than the control. Yardwaste, in particular, was associated with increased relative abundance of Chloroflexi, which was correlated with soil nutrient concentrations. Malling 9 had a greater bacterial diversity than G.41, but the rootstock treatment had no independent effect on the rhizosphere community structure. There was, however, a strong interaction between the rootstock and fertilizer treatments. Carbon cycling was the most prominent functional change associated with the soil bacterial community. These results suggest that compost amendments have a more positive effect on soil bacterial activity and nutrient availability than Ca(NO3)(2). Our work shows that waste-stream amendments can lead to multiple positive responses, such as increasing aboveground tree biomass, thus potentially improving orchard productivity.

Sustainable practices are key to the improvement of soil fertility and quality in apple (Malus x domestica Borkh.) orchards. Rootstock genotype and fertilizer inputs can alter soil biology, as well as aboveground traits including nutrient acquisition. In this study, a factorial design was used to assess the interaction between two apple rootstocks, 'Geneva 41' ('G.41') and 'Malling 9' ('M.9') with four fertilizer treatments [chicken-litter compost, yardwaste compost, fertigation using Ca(NO3)(2), and an unamended control]. The bacterial community in the rhizosphere was assessed for its impact on both plant and soil properties for each rootstock x fertilizer treatment combination. The bacterial community was dominated by Acidobacteria, Proteobacteria, and Planctomycetes, but Verrucomicrobia and Chloroflexi were the most responsive to the fertilizer treatments. The chicken litter and yardwaste treatments had a greater effect on bacterial community structure than the control. Yardwaste, in particular, was associated with increased relative abundance of Chloroflexi, which was correlated with soil nutrient concentrations. Malling 9 had a greater bacterial diversity than G.41, but the rootstock treatment had no independent effect on the rhizosphere community structure. There was, however, a strong interaction between the rootstock and fertilizer treatments. Carbon cycling was the most prominent functional change associated with the soil bacterial community. These results suggest that compost amendments have a more positive effect on soil bacterial activity and nutrient availability than Ca(NO3)(2). Our work shows that waste-stream amendments can lead to multiple positive responses, such as increasing aboveground tree biomass, thus potentially improving orchard productivity.

The recent growth in the U.S. hard-cider industry has increased the demand for cider apples (Malus xdomestica Borkh.), but little is known about how to manage orchard soil fertility best to optimize horticultural performance and juice characteristics for these cultivars. To assess whether nitrogen fertilizer applied to the soil can improve apple juice and cider quality, calcium nitrate (CaNO3) fertilizer was applied at different rates to the soil beneath 'Golden Russet' and 'Medaille d'Or' trees over the course of three growing seasons. The experiment started when the trees were in their second leaf. The trees were cropped in their third and fourth leaf. At the end of the first growing season of the experiment, the greatest fertilizer rate increased tree trunk cross-sectional area (TCSA) by 82% relative to the control, but this difference did not persist through to the end of the study. Yield and crop load were unaffected by the nitrogen fertilization treatments. Increasing the nitrogen fertilizer rate correlated positively with more advanced harvest maturity in 'Golden Russet' fruit, which resulted in greater soluble solid concentration (SSC). Fruit from the greatest fertilizer rate treatment had an average starch pattern index (SP1) that was 1 U greater than in the control, and an SSC that was 3% greater than the control. The fertilizer treatments did not affect juice pH, titratable acidity (TA), or total polyphenol concentrations. Yeast assimilable nitrogen (YAN) concentrations were increased by nitrogen fertilization for both cultivars in both harvest years. The greatest fertilizer treatment increased juice primary amino nitrogen by 103% relative to the control. Greater nitrogen fertilization rates correlated positively with less hydrogen sulfide production during the fermentation of 'Golden Russet' juice from the first, but not the second, harvest. During the first year, cumulative hydrogen sulfide production for the 'Golden Russet' control treatment was 29.6 m

Yeast assimilable nitrogen (YAN) can be a limiting nutritional factor for Saccharomyces cerevisiae yeast when fermenting apple (Maims Xdomestka Borkh.) juice into hard cider. Endogenous YAN concentrations in apples are often below the recommended thresholds to completely use all of the fermentable sugar and minimize the production of off-flavors, such as hydrogen sulfide. Cider producers supplement apple juice with exogenous nitrogen to increase YAN. Urea, commonly applied to apple orchards to increase fruit size and yields, was tested for its ability to increase endogenous apple juice YAN. Starting 6 weeks before harvest in 2017 and 2018, a 1% urea solution was applied to 'Red Spy' apple trees one, three, or five times to create low-, medium-, and high-rate treatments, respectively. Relative to the control, the high treatment increased YAN by 229% in 2017 and by 408% in 2018. More than 90% of the YAN in all juice samples was composed of primary amino nitrogen (PAN). Among all treatments, PAN mostly comprised asparagine, and as urea applications increased, the relative concentration of asparagine also increased. Aspartic acid and then glutamic acid were the second and third most abundant amino acids in all treatments, respectively, but comprised less of the total PAN as the number of urea applications increased. Soluble solid concentration, pH, titratable acidity, and total polyphenol concentration were not different among treatments. There was a positive correlation between increased urea application rate and the maximum fermentation rate, which resulted in a shorter fermentation duration. Increasing the number of urea applications was also correlated with greater hydrogen sulfide (H2S) production in juice fermented from fruit harvested in 2017 but not for fruit harvested in 2018. No residual H2S was found in the finished cider from any treatment. Increasing the number of urea applications was estimated to be less expensive than supplementing the juice with Fermaid

The recent growth in the U.S. hard-cider industry has increased the demand for cider apples (Malus xdomestica Borkh.), but little is known about how to manage orchard soil fertility best to optimize horticultural performance and juice characteristics for these cultivars. To assess whether nitrogen fertilizer applied to the soil can improve apple juice and cider quality, calcium nitrate (CaNO3) fertilizer was applied at different rates to the soil beneath 'Golden Russet' and 'Medaille d'Or' trees over the course of three growing seasons. The experiment started when the trees were in their second leaf. The trees were cropped in their third and fourth leaf. At the end of the first growing season of the experiment, the greatest fertilizer rate increased tree trunk cross-sectional area (TCSA) by 82% relative to the control, but this difference did not persist through to the end of the study. Yield and crop load were unaffected by the nitrogen fertilization treatments. Increasing the nitrogen fertilizer rate correlated positively with more advanced harvest maturity in 'Golden Russet' fruit, which resulted in greater soluble solid concentration (SSC). Fruit from the greatest fertilizer rate treatment had an average starch pattern index (SP1) that was 1 U greater than in the control, and an SSC that was 3% greater than the control. The fertilizer treatments did not affect juice pH, titratable acidity (TA), or total polyphenol concentrations. Yeast assimilable nitrogen (YAN) concentrations were increased by nitrogen fertilization for both cultivars in both harvest years. The greatest fertilizer treatment increased juice primary amino nitrogen by 103% relative to the control. Greater nitrogen fertilization rates correlated positively with less hydrogen sulfide production during the fermentation of 'Golden Russet' juice from the first, but not the second, harvest. During the first year, cumulative hydrogen sulfide production for the 'Golden Russet' control treatment was 29.6 m

Yeast assimilable nitrogen (YAN) can be a limiting nutritional factor for Saccharomyces cerevisiae yeast when fermenting apple (Maims Xdomestka Borkh.) juice into hard cider. Endogenous YAN concentrations in apples are often below the recommended thresholds to completely use all of the fermentable sugar and minimize the production of off-flavors, such as hydrogen sulfide. Cider producers supplement apple juice with exogenous nitrogen to increase YAN. Urea, commonly applied to apple orchards to increase fruit size and yields, was tested for its ability to increase endogenous apple juice YAN. Starting 6 weeks before harvest in 2017 and 2018, a 1% urea solution was applied to 'Red Spy' apple trees one, three, or five times to create low-, medium-, and high-rate treatments, respectively. Relative to the control, the high treatment increased YAN by 229% in 2017 and by 408% in 2018. More than 90% of the YAN in all juice samples was composed of primary amino nitrogen (PAN). Among all treatments, PAN mostly comprised asparagine, and as urea applications increased, the relative concentration of asparagine also increased. Aspartic acid and then glutamic acid were the second and third most abundant amino acids in all treatments, respectively, but comprised less of the total PAN as the number of urea applications increased. Soluble solid concentration, pH, titratable acidity, and total polyphenol concentration were not different among treatments. There was a positive correlation between increased urea application rate and the maximum fermentation rate, which resulted in a shorter fermentation duration. Increasing the number of urea applications was also correlated with greater hydrogen sulfide (H2S) production in juice fermented from fruit harvested in 2017 but not for fruit harvested in 2018. No residual H2S was found in the finished cider from any treatment. Increasing the number of urea applications was estimated to be less expensive than supplementing the juice with Fermaid

Multiple analytical methods are used for quantification of total polyphenols and total flavanols in fruit juices and beverages. Four methods were evaluated in this study: Folin-Ciocalteu (F-C), Lowenthal permanganate (L-P), 4-dimethylaminocinnamaldehyde (DMAC), and the bovine serum albumin (BSA) precipitation method. Method validation parameters, including working range, limit of detection, limit of quantitation, precision (repeatability), accuracy, and specificity, were assessed and compared. The F-C method was not specific to polyphenols, and the L-P method had the widest working range but lacked accuracy. The DMAC method was the most specific to flavanols, and the BSA method was not suitable for quantification of smaller flavanols, such as catechin and epicatechin. Quantitative performance was evaluated using commercial fruit juice samples (n = 14), apple juice samples of different cultivars (n = 22), and commercial ciders (n = 17). In general, the L-P titration method and DMAC method resulted in higher quantitative values than the F-C method and BSA precipitation method, respectively. However, ratios of results obtained by the L-P and F-C method ranged from 1 to 28, and ratios of results obtained by the DMAC and BSA precipitation method ranged from <1 to 280. This tremendous variation is likely due to variation in polyphenol composition and sample matrix. This information provides perspective for comparison of results obtained through these different methods, and a basis for choosing the most appropriate analytical method for quantification of polyphenols to address a specific research question when working with commercial fruit juice, apple juice from different apple cultivars, and commercial ciders. Practical Application This study compared results obtained when four common polyphenol quantification methods were applied to a diverse selection of fruit juices and beverages with distinct polyphenol composition and sample matrix. The matrix and polyphenol composi

Multiple analytical methods are used for quantification of total polyphenols and total flavanols in fruit juices and beverages. Four methods were evaluated in this study: Folin-Ciocalteu (F-C), Lowenthal permanganate (L-P), 4-dimethylaminocinnamaldehyde (DMAC), and the bovine serum albumin (BSA) precipitation method. Method validation parameters, including working range, limit of detection, limit of quantitation, precision (repeatability), accuracy, and specificity, were assessed and compared. The F-C method was not specific to polyphenols, and the L-P method had the widest working range but lacked accuracy. The DMAC method was the most specific to flavanols, and the BSA method was not suitable for quantification of smaller flavanols, such as catechin and epicatechin. Quantitative performance was evaluated using commercial fruit juice samples (n = 14), apple juice samples of different cultivars (n = 22), and commercial ciders (n = 17). In general, the L-P titration method and DMAC method resulted in higher quantitative values than the F-C method and BSA precipitation method, respectively. However, ratios of results obtained by the L-P and F-C method ranged from 1 to 28, and ratios of results obtained by the DMAC and BSA precipitation method ranged from <1 to 280. This tremendous variation is likely due to variation in polyphenol composition and sample matrix. This information provides perspective for comparison of results obtained through these different methods, and a basis for choosing the most appropriate analytical method for quantification of polyphenols to address a specific research question when working with commercial fruit juice, apple juice from different apple cultivars, and commercial ciders. Practical Application This study compared results obtained when four common polyphenol quantification methods were applied to a diverse selection of fruit juices and beverages with distinct polyphenol composition and sample matrix. The matrix and polyphenol composi

Yeast assimilable nitrogen (YAN) is essential for yeast growth and metabolism during apple (Malus x domestica Borkh.) cider fermentation. YAN concentration and composition can impact cider fermentation kinetics and the formation of volatile aroma compounds by yeast. The YAN concentration and composition of apples grown in Virginia, USA over the course of two seasons was determined through analysis of both free amino nitrogen (FAN) and ammonium ion concentration. FAN was the largest fraction of YAN, with a mean value of 51 mg N L⁻¹ FAN compared to 9 mg N L⁻¹ ammonium. Observed YAN values ranged from nine to 249 mg N L⁻¹, with a mean value of 59 mg N L⁻¹. Ninety-four percent of all samples analyzed in this study contained <140 mg N L⁻¹ YAN, a concentration generally considered the minimum level needed in grape-based wines for yeast to fully utilize all of the fermentable sugars. FAN concentration was correlated with total YAN concentration, but ammonium concentration was not. Likewise, there was no correlation between FAN and ammonium concentration.

Yeast assimilable nitrogen (YAN) concentration and composition impact hydrogen sulphide (H2S) production and fermentation kinetics during wine fermentation, but this phenomenon has not been extensively studied in cider fermentation. Our hypothesis was that H2S production during cider fermentation could be decreased through pre‐fermentation modification of concentrations of individual amino acids. Apple juice (53 mg L−1 YAN) was supplemented with asparagine, arginine, methionine or ammonium and fermented with EC1118 and UCD522 yeast strains. No difference in H2S production among fermentations was observed with addition of asparagine, arginine or ammonium. Methionine addition of 5 mg L−1 decreased H2S production by yeast strain EC1118 at 53 mg L−1 YAN. With 153 mg L−1 initial YAN, only methionine addition of 50 mg L−1 decreased H2S production, and no tested methionine rates decreased H2S production with 253 mg L−1 initial YAN. Supplementation to 153 mg L−1 YAN resulted in increased H2S production at all methionine concentrations tested. Sensory differences in aroma were detected in samples supplemented with ammonium and methionine, and these differences were correlated with observed differences in H2S production. Our results indicate that supplementing cider fermentations with methionine leads to lower H2S formation, especially in apple juice containing low YAN.

BACKGROUND Fungicide residues on fruit may adversely affect yeast during cider fermentation, leading to sluggish or stuck fermentation or the production of hydrogen sulfide (H₂S), which is an undesirable aroma compound. This phenomenon has been studied in grape fermentation but not in apple fermentation. Low nitrogen availability, which is characteristic of apples, may further exacerbate the effects of fungicides on yeast during fermentation. The present study explored the effects of three fungicides: elemental sulfur (S⁰) (known to result in increased H₂S in wine); fenbuconazole (used in orchards but not vineyards); and fludioxonil (used in post-harvest storage of apples). RESULTS Only S⁰ led to increased H2S production. Fenbuconazole (≥0.2 mg L⁻¹) resulted in a decreased fermentation rate and increased residual sugar. An interactive effect of yeast assimilable nitrogen (YAN) concentration and fenbuconazole was observed such that increasing the YAN concentration alleviated the negative effects of fenbuconazole on fermentation kinetics. CONCLUSION Cidermakers should be aware that residual fenbuconazole (as low as 0.2 mg L⁻¹) in apple juice may lead to stuck fermentation, especially when the YAN concentration is below 250 mg L⁻¹. These results indicate that fermentation problems attributed to low YAN may be caused or exacerbated by additional factors such as fungicide residues, which have a greater impact on fermentation performance under low YAN conditions.

A greater understanding of apple (Malus domestica) pollen tube growth rates can improve crop load management in commercial orchards. Specifically, applications of caustic bloom-thinning chemicals need to occur when enough, but not too many, flowers have been fertilized to achieve crop load densities that balance yields with marketable fruit sizes. In this study, the pollen tube growth rates of five crabapple (Malus sp.) cultivars were measured in the styles of three maternal cultivars at 12, 18, 24, and 30 °C after 24 hours in a growth chamber. Pollen tube growth rates were greatest for ‘Selkirk’ and ‘Thunderchild’ at 12 °C, and greatest for ‘Indian Summer’, ‘Selkirk’, and ‘Thunderchild’ at 24 °C. Pollen tube growth increased with increasing temperatures until 24 °C. There were minimal pollen tube growth rate increases between 24 and 30 °C. Overall, ‘Snowdrift’ had the slowest pollen tube growth rate of the five evaluated crabapple genotypes. At 24 and 30 °C, ‘Indian Summer’ and ‘Thunderchild’ pollen tubes reached the base of the style most frequently, and ‘Snowdrift’ pollen tubes the least frequently. Pollen tube growth rate was also influenced by the maternal cultivar, with Golden Delicious having relatively faster pollen tube growth than Fuji at 24 and 30 °C. Interactions among paternal and maternal genotypes as well as temperature after pollination reveal complex biological and environmental relationships that can be used to develop more precise crop load management strategies for apple orchards.

A greater understanding of apple (Malus domestica) pollen tube growth rates can improve crop load management in commercial orchards. Specifically, applications of caustic bloom-thinning chemicals need to occur when enough, but not too many, flowers have been fertilized to achieve crop load densities that balance yields with marketable fruit sizes. In this study, the pollen tube growth rates of five crabapple (Malus sp.) cultivars were measured in the styles of three maternal cultivars at 12, 18, 24, and 30 °C after 24 hours in a growth chamber. Pollen tube growth rates were greatest for ‘Selkirk’ and ‘Thunderchild’ at 12 °C, and greatest for ‘Indian Summer’, ‘Selkirk’, and ‘Thunderchild’ at 24 °C. Pollen tube growth increased with increasing temperatures until 24 °C. There were minimal pollen tube growth rate increases between 24 and 30 °C. Overall, ‘Snowdrift’ had the slowest pollen tube growth rate of the five evaluated crabapple genotypes. At 24 and 30 °C, ‘Indian Summer’ and ‘Thunderchild’ pollen tubes reached the base of the style most frequently, and ‘Snowdrift’ pollen tubes the least frequently. Pollen tube growth rate was also influenced by the maternal cultivar, with Golden Delicious having relatively faster pollen tube growth than Fuji at 24 and 30 °C. Interactions among paternal and maternal genotypes as well as temperature after pollination reveal complex biological and environmental relationships that can be used to develop more precise crop load management strategies for apple orchards.

To assess the impact crop load has on hard cider chemistry, ‘York’ apple (Malus3domestica Borkh.) trees were hand thinned to three different crop loads: low [two apples per cm2 branch cross-sectional area (BCSA)], medium (four apples per BCSA), and high (six apples per BCSA). Higher crop loads produced smaller, less acidic fruit that were slightly more mature. In juice made from fruit from these treatments, the total polyphenol content did not differ at harvest, but, after fermentation, the medium crop load had 27% and the high crop load had 37%greater total polyphenol content than the low crop load. Yeast assimilable nitrogen (YAN) concentration in juice made from fruit from the low crop load treatment had 18%and 22% greater than the medium and high crop load, respectively. YAN concentrations in juice from the medium and high crop load treatments were similar. Our results provide apple growers and hard cider producers with a better understanding of how apple crop load impacts YAN concentrations in juice and total polyphenol concentrations in juice and cider.