agriculture

Article
Foliar Calcium Corrects a Deficiency Causing Green
Fruit Drop in ‘Draper’ Highbush Blueberry
(Vaccinium corymbosum L.)
Eric Martin Gerbrandt 1, *, Charles Mouritzen 2 and Mark Sweeney 3
1 British Columbia Blueberry Council, #275-32160 South Fraser Way, Abbotsford, BC V2T 1W5, Canada
2 Southwest Crop Consulting, Chilliwack, BC V2R 5G2, Canada; macmour@shaw.ca
3 British Columbia Ministry of Agriculture, Abbotsford, BC V3G 2M3, Canada; markesweeney55@gmail.com
* Correspondence: emg690@mail.usask.ca; Tel.: +1-604-308-0748




Received: 5 March 2019; Accepted: 21 March 2019; Published: 24 March 2019


Abstract: ‘Draper’ northern highbush blueberry (Vaccinium corymbosum L.) is a widely-planted

mid-season cultivar with excellent fruit quality. Under the climatic conditions of Southwestern
British Columbia, Canada, and Northwestern Washington, USA, it expresses a physiological disorder
causing spontaneous green fruit drop (GFD) of up to half of the developing crop just prior to onset of
the fruit coloring phenophase. Reduction of economic losses due to GFD required identification of
the cause of this disorder and development of an agronomic solution that would reduce fruit drop.
In 2014, two initial experiments were conducted to compare three foliar Ca products under a range of
N fertilization rates. In 2015 and 2016, three locations were used in a first step to optimizing rates
and timings of foliar Ca application. Initial experiments determined that higher N fertilization rates
exacerbate GFD but that foliar Ca corrects the condition. Multi-site, multi-year trials identified key
rates and timings for foliar Ca application to provide an agronomic solution for commercial growers.
These trials identified an acute fruit Ca deficiency as the cause of GFD, and that foliar calcium applied
frequently at high concentration from mid-bloom onward can be effective in reducing GFD, often
to negligible levels. This condition has now been reported in several production regions around
the world.

Keywords: application rate; application timing; calcium chloride; calcium deficiency; efficacy trial

1. Introduction
‘Draper’ highbush blueberry (Vaccinium corymbosum L.) was developed by the Michigan State
University breeding program released in 2003 [1]. Though its genetic background is primarily
comprised of the northern highbush blueberry species, V. corymbosum, its pedigree also has
contributions from southern highbush blueberry species, V. tenellum, V. ashei and V. darrowi [1].
While ‘Draper’ is recommended for northern production regions based on a sufficiently high chilling
requirement, producers in the Fraser Valley, British Columbia (BC), Canada have found that it requires
a higher level of horticultural management (e.g., preventative sprays for fungal canker) in comparison
to other standard cultivars [2].
‘Draper’ has large, firm fruit with a strong waxy cuticle and ripens just prior to ‘Bluecrop’, the
primary mid-season cultivar in BC and many other production regions [3]. With superior flavor to
the primary early-season cultivar (‘Duke’), relatively loose clusters, excellent firmness, and a long
shelf-life, the fruit have a concentrated ripening period and can be hand- or machine-harvested [1,3].
Contrasting with these outstanding fruit quality characteristics, a 2012 survey of field performance
for ‘Draper’ and other new cultivars identified several production challenges, the most economically

Agriculture 2019, 9, 63; doi:10.3390/agriculture9030063 www.mdpi.com/journal/agriculture

Agriculture 2019, 9, 63 2 of 23

damaging being the pervasive incidence of green fruit drop (GFD) [2]. Affecting 5–50% of the crop,
depending on the field, GFD is characterized by spontaneous drop of green fruit between the end of
June and the beginning of July as the first fruit begin to turn blue [4]. The fruit drop over the course
of one or two weeks, and they appear normal on the exterior, contain white developing seeds within
brown decaying internal tissues [4].
In 2014, this cultivar-specific phenomenon had not been observed in other production regions,
and it did not present the typical etiology of a new pest or pathogen. It occurred to varying degrees
within and across fields and years with increased severity under high plant vigor. These observations,
combined with the low transpiration conditions preceding onset of drop, indicated similarities to other
Ca deficiencies such as blossom end rot in tomato (Solanum lycopersicum L.) and bitter pit in apples
(Malus pumila Miller) [5–7]. In Oregon, relative to other cultivars, ‘Draper’ showed low fruit Ca and
high leaf Ca [8].
Ca is an essential plant macronutrient required for maintaining cell wall and membrane integrity,
facilitating intracellular communication and controlling vacuolar osmotic balance [9]. Low soil pH
environments have low Ca availability [10], but blueberry is a calcifuge, being adapted to an optimal
pH range between 4.5–5.5 where Ca availability is limited [11]. Though some symplastic uptake of Ca
occurs, most uptake is apoplastic, occurring mostly through the newest roots [12], requiring active
root extension to access relatively immobile soil Ca and the rate of Ca uptake depending on the rate of
transpiration [10,13]. In strawberry (Fragaria ˆ ananassa Duch.) and tomato, Ca transport depends on
positive pressure in the xylem [14,15]. In general, there is very limited re-mobilization of Ca from leaves
to fruit by the phloem in most crops, meaning that adequate supply of Ca via the xylem to the fruit at
key loading stages is critical for normal fruit development [16]. Compared to blueberry leaves, the fruit
accumulate (i.e., load) Ca in limited concentrations but do so primarily during early developmental
stages, which is also observed in other crops [17,18]. Consequently, conditions of low transpiration
that reduce fruit Ca loading can result in localized Ca deficiencies when initial Ca concentrations
are diluted during fruit sizing [19]. This is related to competition between low-transpiring fruit and
high-transpiring leaves, but the role of cation balance for cation exchange sites is also important and
the underlying mechanisms of Ca deficiencies are not fully understood [20].
Foliar application of Ca can be effective in correcting acute deficiencies in susceptible cultivars of
other crops, while uptake, allocation, and movement of Ca can be modulated by cultural practices such
as reduced N management or application of exogenous plant growth regulators [7,21–23]. Previous
work on application of foliar Ca has shown mixed results in blueberry, resulting in increased fruit Ca
concentration and influencing fruit quality under some conditions [24–26] and having no significant
effects in others [27,28]. Correction of GFD in ‘Draper’ using foliar applications is not reported in the
literature. The objective of this study was to identify the role of Ca deficiency in GFD and determine
key foliar Ca application rates and timings to reduce yield losses to negligible levels.

2. Materials and Methods

In 2014, two preliminary trials were conducted in a 2008 commercial planting of ‘Draper’ in
Abbotsford, BC to evaluate the effect of foliar Ca products under high vigor conditions (Experiment 1)
and under low vigor conditions with variable rates of granular N fertilization (Experiment 2). In
2015 and 2016, comparison of foliar Ca application rates and timings was conducted across three
sites (Experiment 3), using separate sections of the same field used in the previous year (Site 1),
another 2008 planting in Abbotsford (Site 2) and a 2009 planting in Langley (Site 3). Soil types were
a Lynden gravelly silt loam, Lynden silt loam and Everett gravelly sandy loam for Sites 1, 2, and 3,
respectively [29].
For Experiment 1, a section of the field at Site 1 was top-dressed with non-composted broiler
chicken manure on 20-May, which is routine practice for the collaborating grower but not a common
production practice in the region. The rate of manure was calculated to deliver approximately
75 kg N¨Ha´1 . Manure application was withheld from the section of field used for Experiment 2
Agriculture 2019, 9, 63 3 of 23

to facilitate evaluation of the effects of foliar Ca under a range of granular N treatments. For
both experiments, randomized complete block designs with four replications were used to compare
treatments applied to five-plant plots with one-plant buffers between plots. Experiment 1 compared
three Ca products (Terralink Horticulture Ltd., Abbotsford, BC, Canada) at various rates with a
no-spray control: DowFlake™ Xtra (83–87% CaCl2 ¨4H2 O) calcium chloride (CaCl2 ), TigerClaw
Power-Cal 4% calcium phosphite (Ca3 (PO3 )2 ), and CaTs® 6% calcium thiosulphate solution (CaO3 S2 ).
CaCl2 , Ca3 (PO3 )2 , and CaO3 S2 were applied at spray concentrations of 1263, 400, and 450 ppm Ca in
500, 494, and 935 L water¨ha´1 , respectively. The latter two product rates were based on their label
rates, but CaCl2 does not have a label rate in blueberry and so was calculated to emulate standard
applications used in treatment of bitter pit in apple. Experiment 2 used broadcast application of
granular ammonium sulphate, (NH4 )2 SO4 (21-0-0-24[S]; Terralink Horticulture Ltd., Abbotsford, BC,
Canada), at four rates: 0 (0-N), 56 (56-N), 112 (112-N), and 168 (168-N) kg N¨Ha´1 . An incomplete
factorial design was used to compare the same foliar Ca products as in Experiment 1. Specifically, the
no-spray control was evaluated under all four N rates, and the three foliar Ca products were only
applied for the low and high N treatments (0-N and 168-N, respectively). Ammonium sulphate was
applied to each plot by hand as a 50/50 split on 20-May and 03-June-2014. There was significant
rainfall the day after and four days after the first and second applications, respectively, dissolving
the fertilizer and washing it into the rootzone. Weekly foliar Ca applications were made using a 15-L
backpack sprayer (Solo USA, Newport News, VA) flat fan 8004 nozzle (TeeJet Technologies, Springfield,
Illinois) from 20-May to 17-June-2014 at approximately 500 L¨Ha´1 under slow drying conditions
from 6:00–8:30 a.m. with no precipitation for the following eight hours or more, depending on the
application date (Table 1). No other foliar or granular N or Ca were applied by the collaborating
grower in this commercial field.
In 2015 and 2016, randomized complete block designs, each with four replications, were used to
compare ten rate and timing treatment combinations across Sites 1, 2, and 3. Different sections of each
field were used for each year of the trial to eliminate carry-over of experimental treatments from the
previous year. A no-spray control was compared with a factorial design of four application timings
and two rates of CaCl2 using TETRA Flake (77% CaCl2 ¨2H2 0) calcium chloride (Terralink Horticulture
Ltd., Abbotsford, BC, Canada). The low (L-CaCl2 ) and high (H-CaCl2 ) rates were mixed to 1360 ppm
and 2720 ppm solutions of Ca, respectively. Signature® SST 8% Calcium (SST), a formulated foliar Ca
product (Evergro, Abbotsford, BC, Canada) with a relatively high Canadian label rate, was applied at
4.9 L¨Ha´1 (800 ppm) as a comparison to the factorial treatment of CaCl2 application rates and timings.
Using a backpack sprayer (as described above) and 0.1% of a non-ionic surfactant, LI700 (Evergro,
Abbotsford, BC, Canada), the tank mix concentration for each treatment was maintained as a constant
throughout the experiment. Applications were made to run-off, resulting in variation in spray volume
from 371 to 914 L water¨Ha´1 , depending on plant size and crop development stage across sites and
application timings (Table 1). L-CaCl2 and H-CaCl2 were applied three times at weekly intervals, and
there was an overlapping week between early (Early), middle (Mid), and late (Late) timing treatments.
A fourth application timing (All) overlapped the seven application timings from Early to Late timing.
SST was applied at Mid timing only. Dates and plant phenological stages for these applications varied
across locations and years, starting as early as 40% of open flowers for the Early timing to three or four
weeks into green fruit development for the Late and All timings.
Agriculture 2019, 9, 63 4 of 23

Table 1. Dates, plant phenological stages and foliar Ca application spray volumes made to ‘Draper’ highbush blueberry across three farm sites in the Fraser Valley,
British Columbia, Canada in 2015 and 2016.

Date Phenology Spray Volume (L¨¨ha´ 1 )

Timings 2015 2016 Site 2015 2016 2015 2016
1 70% bloom; 10% petal fall 80% bloom; 5% petal fall 494 549
Early, All 22-April 26-April 2 50% bloom; 0% petal fall 60% bloom; 0% petal fall 371 504
3 30% bloom; 0% petal fall 40% bloom; 0% petal fall 371 413
1 100% bloom; 40% petal fall 100% bloom; 40% petal fall 494 596
Early, All 2-May 3-May 2 95% bloom; 25% petal fall 100% bloom; 15% petal fall 371 549
3 60% bloom; 5% petal fall 90% bloom; 5% petal fall 371 504
1 100% bloom; 80% petal fall 100% petal fall 556 731
Early, Mid, All 9-May 10-May 2 100% bloom; 60% petal fall 95% petal fall 494 670
3 95% bloom; 25% petal fall 90% petal fall 432 640
1 100% petal fall Green fruit 618 731
Mid, All 16-May 17-May 2 100% petal fall Green fruit 556 640
3 100% bloom; 70% petal fall Green fruit 556 640
1 Green fruit Green fruit 618 853
Mid, Late, All 23-May 24-May 2 Green fruit Green fruit 556 793
3 100% petal fall Green fruit 556 731
1 Green fruit Green fruit 803 914
Late, All 30-May 31-May 2 Green fruit Green fruit 741 823
3 Green fruit Green fruit 618 731
1 Green fruit Green fruit 803 914
Late, All 06-Jun 06-Jun 2 Green fruit Green fruit 741 823
3 Green fruit Green fruit 618 731
Agriculture 2019, 9, 63 5 of 23

In each experiment, the central three plants from each plot were used for data collection. For
Experiments 1 and Experiment 2, 50-fruit samples were taken from each plot at three times during
the ripe fruit stage on 9-, 21-, and 28-July to calculate a weighted average fruit weight (1:2:1 between
first, second and third samples, respectively). Three subsamples of ripe fruit were taken for each
of the four replicates for the no-spray control and CaCl2 treatment under both the lowest (0-N) and
highest (168-N) vigor conditions in Experiment 2, and these were washed with distilled water and
subjected to chemical nutrient analysis by a commercial laboratory (Brookside Laboratories Inc., New
Bremen, OH, USA). For Experiment 3, fruit samples were taken at the green fruit stage on 19/20-June
in 2015 and 16/17-June in 2016 and at the ripe fruit stage on 15-July in 2015 and 9-July in 2016. Soil
samples were taken as a composite of nine cores from each plot. Cores were taken at 0–30 cm soil
depth at 20–30 cm from the center of the row. Leaf samples (40-leaves per plot) were taken at the time
of green fruit sampling and later in the season, on 29/30-July in 2015 and 30/31-July in 2016, as per the
recommended leaf tissue sampling protocol developed in Oregon [30]. Fruit and leaf samples were
weighed, washed with distilled water, dried in an oven, and then weighed again before being sent for
chemical nutrient analysis (Brookside Laboratories Inc., New Bremen, OH, USA). For Experiment 3,
green fruit and early leaf samples were analyzed for all treatment combinations in 2015 but only for
the control in 2016, whereas ripe fruit and late leaf tissue samples were analyzed for all biological
replicates of each treatment combination in both years. Chemical nutrient analysis results, reported as
dry weight (DW) values, were converted to fresh weight (FW) values using individual sample weights
before and after drying.
At the same time as ripe fruit sampling, evaluation of yield components was performed for
each of the three technical replicate plants of each plot by counting: total number of clusters¨plant´1 ,
ten counts of fruit¨cluster´1 , and calculation of average fruit weight based on fruit samples. For
Experiment 1 and Experiment 2, yield estimate after GFD was compared with estimates made prior
to GFD on 16-June in 2014. While there was negligible GFD across the region in 2015, severity of
fruit drop was determined in 2014 and 2016 by counting all fruit on the ground beneath the three
central plants of each plot. Percent fruit drop was calculated from yield components and the number
of dropped fruit.
All data were analyzed in the ‘R’ statistical environment [31] via linear mixed model regression
using the ‘lme4’ package [32]. Treatments for Experiments 1 and Experiment 2 were compared
via stepwise model simplification to determine the minimally adequate model with comparisons
between successive models via the Akaike Information Criterion (AIC). All parameters remaining
in the minimally adequate model were statistically different at p < 0.05. For Experiment 3, multiple
comparisons between treatments were made using linear contrasts extracted from regression models
and pairwise comparisons using the ‘multcompView’ package [33] to determine statistically significant
differences (p < 0.05).

3. Results

3.1. Experiment 1
In the manured section of the field, neither the percent fruit drop nor the estimated loss of yield
were significantly reduced by CaO3 S2 , but there was a significant 4.9% increase in estimated yield
(Table 2). However, Ca3 (PO3 )2 and CaCl2 reduced percent fruit drop by 22.9% and 55.9% and the
estimated amount of yield loss by 26.0% and 55.6%, respectively. This did not translate to significant
increases in the estimated yield that was measured after GFD, indicating some compensation in yield
components in response to a change in crop load under high N management through application of
manure. There were no significant differences in fruit weight at early or late sampling dates, but foliar
Ca treatments varied in their effect on fruit weight at the middle sampling date, CaCl2 increasing
and Ca3 (PO3 )2 decreasing fruit weight in comparison to the control. The average fruit weight across
sampling dates was only significantly impacted by CaCl2 , being slightly larger than the control.
Agriculture 2019, 9, 63 6 of 23

Table 2. Effects of foliar Ca treatments on mean ˘ SE estimated fruit yield, pre-mature green fruit drop, yield loss and fruit weight in ‘Draper’ highbush blueberry in
Abbotsford, British Columbia, Canada in 2014.

Experiment 1

Calcium Nitrogen Yield Estimate Yield Loss Estimate Average Fruit Weight (g/fruit)
Treatment Fruit Drop (%)
Treatment (kg¨¨ha´ 1 ) 5 (kg¨¨ha´ 1 ) Early Mid Late Average
(kg N¨¨ha´ 1 )
Control 10,004 ˘ 474a 16.1 ˘ 1.4c 1613 ˘ 159c 1.8 ˘ 0.1a 1.9 ˘ 0.0b 1.8 ˘ 0.1a 1.9 ˘ 0.0a
CaCl2 1 Poultry 9946 ˘ 176a 7.2 ˘ 0.9a 716 ˘ 81a 1.7 ˘ 0.1a 2.2 ˘ 0.1c 1.8 ˘ 0.0a 2.0 ˘ 0.1b
Ca3 (PO3 )2 2 manure 4 9579 ˘ 250a 12.4 ˘ 0.8b 1194 ˘ 91b 1.8 ˘ 0.1a 1.9 ˘ 0.1a 1.8 ˘ 0.1a 1.8 ˘ 0.1a
CaO3 S2 3 10,496 ˘ 336b 15.6 ˘ 1.8c 1635 ˘ 185c 1.8 ˘ 0.1a 2.0 ˘ 0.1b 1.8 ˘ 0.0a 1.9 ˘ 0.1a
Experiment 2
0 12,167 ˘ 1334b 13.1 ˘ 1.6c 1530 ˘ 53d 1.9 ˘ 0.1b 2.2 ˘ 0.1c 1.8 ˘ 0.0b 2.0 ˘ 0.0d
56 11,190 ˘ 1233a 13.1 ˘ 1.7c 1408 ˘ 99d 1.9 ˘ 0.1b 2.2 ˘ 0.0b 1.8 ˘ 0.1a 2.0 ˘ 0.1c
Control
112 10,998 ˘ 570a 12.6 ˘ 1.0c 1372 ˘ 73d 1.8 ˘ 0.1b 2.1 ˘ 0.0a 1.7 ˘ 0.0a 1.9 ˘ 0.0b
168 11,128 ˘ 1245a 13.1 ˘ 1.7c 1431 ˘ 180d 2.0 ˘ 0.1c 2.1 ˘ 0.0a 1.8 ˘ 0.0b 2.0 ˘ 0.0c
0 14,085 ˘ 1853c 3.4 ˘ 0.5a 470 ˘ 65a 1.9 ˘ 0.0b 2.2 ˘ 0.1b 1.9 ˘ 0.0c 2.0 ˘ 0.0d
CaCl2
168 12,153 ˘ 1110b 2.5 ˘ 0.2a 303 ˘ 12a 1.7 ˘ 0.1a 2.0 ˘ 0.0a 1.8 ˘ 0.1a 1.9 ˘ 0.0a
0 12,277 ˘ 1332b 9.8 ˘ 1.5b 1140 ˘ 68c 1.9 ˘ 0.1b 2.1 ˘ 0.1a 1.8 ˘ 0.1b 2.0 ˘ 0.1c
Ca3 (PO3 )2
168 11,855 ˘ 1006b 10.6 ˘ 1.3b 1214 ˘ 71c 1.9 ˘ 0.0b 2.1 ˘ 0.1b 2.0 ˘ 0.1c 2.0 ˘ 0.1d
0 11,348 ˘ 1129a 9.3 ˘ 2.4b 974 ˘ 175b 2.1 ˘ 0.1c 2.2 ˘ 0.0b 1.8 ˘ 0.0b 2.1 ˘ 0.0d
CaO3 S2
168 11,941 ˘ 1557b 12.3 ˘ 1.6c 1408 ˘ 87d 2.0 ˘ 0.1c 2.1 ˘ 0.1b 1.9 ˘ 0.0b 2.0 ˘ 0.1d
1 DowFlake™ Xtra 83–87% calcium chloride. 2 TigerClaw Power-Cal 4% calcium phosphite. 3 CaTs® 6% calcium thiosulphate solution. 4 Treated with approximately 75 kg N¨ha´1 , but
this is not statistically comparable with other nitrogen treatments. 5 Treatment combinations not connected by the same letter are statistically different (p < 0.05) based on stepwise model
simplification of linear mixed model regression using Akaike Information Criterion to determine the minimally adequate model.
Agriculture 2019, 9, 63 7 of 23

Responses to foliar Ca varied by product. Specifically, by altering the amount of fruit retained
on the bush up until the time of harvest, foliar Ca treatments had different effects on the pre-drop
and post-drop yield estimates as well as average fruit weight at the time of harvest. This suggests
interactions of plant vigor and crop load in response to these foliar applications. The changes in crop
load that resulted from foliar Ca led to differential compensation in yield components. Plots with a
greater yield estimate prior to GFD were positively correlated with greater loss of fruit (r = 0.7152,
p < 0.001), but the yield estimate after GFD was not correlated (r = 0.2958, p = 0.2641) with the weight
of fruit lost during GFD.

3.2. Experiment 2
In the section of field without spring manure application, percent fruit drop and estimated loss of
yield were reduced to the greatest extent by CaCl2 treatment under high and low N rates and more
moderately reduced by Ca3 (PO3 )2 and CaO3 S2 (Table 2). Though no differences were noted between
control plots at different N rates, greater effects were seen at 0-N than at 168-N for estimated yield in
response to CaCl2 and for estimated yield, percent fruit drop and estimated yield loss in response to
CaO3 S2 . CaCl2 application resulted in statistically equivalent 74.0 and 80.9% reductions in percent fruit
drop, for 0-N and 168-N, respectively; and Ca3 (PO3 )2 application resulted in statistically equivalent
25.2 and 19.1% reductions at 0-N and 168-N, respectively. In contrast, the reduction in percent fruit
drop for CaO3 S2 was statistically greater at 0-N (29.0%) than at 168-N (6.1%).
Whereas the impacts on estimated yield loss substantially paralleled the effects on percent fruit
drop, the effects on estimated yield were more complex. In the absence of Ca applications, estimated
yield was significantly higher by 8.0%, 9.6%, and 8.5% for 0-N than 56-N, 112-N and 168-N, respectively,
these higher N rates having statistically equivalent yield estimate to one another. At 0-N, yield estimate
was increased by 15.8% by CaCl2 , was not altered by Ca3 (PO3 )2 and was decreased by 6.7% by CaO3 S2 .
At 168-N, yield estimate was increased by 9.2%, 6.5%, and 7.3% by CaCl2 , Ca3 (PO3 )2 , and CaO3 S2 ,
respectively. Further, CaCl2 -treated plots had 13.7% lower estimated yield at 168-N than 0-N, and
Ca3 (PO3 )2 plots showed no difference in estimated yield between 168-N and 0-N, but CaO3 S2 plots
had a 5.2% higher estimated yield at 168-N compared to 0-N. In summary, estimated yield was greater
for CaCl2 than the control at both N rates, and it was greater for Ca3 (PO3 )2 and CaO3 S2 than the
control at 168-N, but for CaO3 S2 it was lower than the control at 0-N. The statistical differences in fruit
weight found between Ca treatments and across N rates varied across sampling dates without a clearly
interpretable pattern.
Chemical nutrient analysis of fruit showed significantly greater Ca content for CaCl2 at 0-N
(0.06 ˘ 0.00 SE) and 168-N (0.07 ˘ 0.00 SE) than the control (both 0.04 ˘ 0.00 SE), with no significant
difference between N rates for either of the other two Ca treatments that were less effective in reducing
fruit drop. There was a strong negative association between green fruit Ca and percent fruit drop (R2 =
0.9878) for control and CaCl2 treatments under high and low N rates (Figure 1).
As for Experiment 1, complex responses to various combinations of Ca and N rates in Experiment
2 were seen across sampling periods, suggesting differential compensation of yield components to the
alterations in crop load that, in turn, resulted from reductions in fruit drop. Overall, there was lower
estimated yield at higher N rates without Ca treatment, the most effective Ca treatment increased
yield at both high and low N rates. Depending on the effectiveness of the Ca treatment, the resulting
increase in estimated yield was either equivalent across N rates, greater at the high N rate or greater at
the low N rate.
Across all treatments, estimated yield was negatively correlated with percent drop (r = ´0.5854,
p < 0.001) and positively correlated with average fruit weight (r = 0.4502, p = 0.0021). Therefore, over a
range of N rates, there was a typical, positive relationship between fruit size and yield, and plants that
dropped fewer fruit still effectively sized their fruit prior to harvest. This was observed as an increased
yield estimate rather than a reduced average fruit size. This was further demonstrated by a lack of
correlation between average fruit weight and percent drop (r = ´0.0399, p = 0.8081) and total yield loss
Agriculture 2019, 9, 63 8 of 23

estimate (r = ´0.2455, p = 0.1235). Higher yielding plants dropped a slightly lower percentage of fruit
but not a lower amount of fruit overall. There was no significant relationship between estimated yield
loss and the estimated yield before drop (r = ´0.0607, p = 0.7116), but there was a significant negative
correlation between percent drop and the pre-drop yield estimate (r = ´0.4280, p = 0.004). Therefore,
higher yielding plants appeared to lose a lower percentage of fruit while the overall pre-drop yield
estimate was not associated with the total amount of fruit that dropped. Though plants with a greater
Agriculture 2019, 9, x FOR PEER REVIEW 15 of 1
crop load before GFD tended to lose a greater percentage of their fruit, this did not result in a significant
relationship between estimated yield loss and the pre-drop yield estimate.

16

14

12 y = -393.3x + 29
R² = 0.9878
Fruit drop (%)

10

0
0.03 0.035 0.04 0.045 0.05 0.055 0.06 0.065 0.07
Ripe fruit calcium (% dry weight)

Figure
Figure 1.1.XYXY plots
plots withwith regression
regression equations
equations and coefficients
and coefficients of determination
of determination between
between percent fruit
percent
drop andfruit
ripe drop and ripe
fruit calcium forfruit calcium
‘Draper’ for ‘Draper’
highbush blueberryhighbush blueberry
treated (right) treated (right)
and non-treated and
(left) with
non-treated
foliar Ca under(left)
highwith foliar
and low Ca under
nitrogen highon
conditions and low nitrogen
a single farm site inconditions
Abbotsford,on a single
British farm
Columbia,
site in Abbotsford,
Canada in 2014. British Columbia, Canada in 2014.

No statistical comparison could be made between the plots used for Experiments 1 and 2 in 2014,
but there was a greater numerical percent fruit drop for the Ca control plots that received manure than
the Ca control plots at any rate of supplemental N in the absence of manure application. Moreover, the
effects of the Ca treatments were numerically smaller with manure than without manure, regardless the
N rate. Since some of the Ca treatments were less effective under higher N rates, plant fertility status,
and its effect on vigor, appeared to influence the ability to mitigate GFD with foliar Ca applications.

3.3. Experiment 3
As a result of an atypical season in 2015, with very early phenological development due to warm,
dry weather conditions, GFD was very minimal across the production region in Southwestern BC and
the neighboring areas of northwestern Washington. In contrast, though 2016 was also an abnormally
early season, climatic conditions were conducive to the most severe GFD experienced to-date. Due
to the absence of GFD in 2015, plant responses to foliar Ca applications did not show any statistical
differences (data not shown); in 2016, plant responses contrasted across replicated field sites, percent
fruit drop ranging from minimal at Site 3 to moderate at Site 1 to severe at Site 2 (Table 3).
Agriculture 2019, 9, 63 9 of 23

Table 3. Mean ˘ SE estimated fruit yield, pre-mature green fruit drop and fruit per cluster in ‘Draper’ highbush blueberry in response to foliar Ca application rates
and timings across three farm sites in the Fraser Valley, British Columbia, Canada in 2016.

Estimated Fruit Yield (kg¨¨ha´ 1 ) 7 Fruit Drop (%) Fruit Per Cluster
Product Timing Rate (ppm)
Site 1 Site 2 Site 3 Site 1 Site 2 Site 3 Site 1 Site 2 Site 3
SST 1 Mid 800 16,568 ˘ 1709ab 5252 ˘ 445b 7487 ˘ 2329ab 12.0 ˘ 0.8de 8.9 ˘ 3.4ab 2.8 ˘ 0.7b 5.8 ˘ 0.1ab 6.3 ˘ 0.3bc 5.9 ˘ 0.2a
1360 17,023 ˘ 2068a–c 7967 ˘ 576d 8809 ˘ 2959b–d 8.5 ˘ 3.3cd 5.6 ˘ 1.8a 0.8 ˘ 0.4a 6.3 ˘ 0.4bc 7.3 ˘ 0.3cd 7.0 ˘ 0.1b
Early 3
2720 19,784 ˘ 3229c–e 6655 ˘ 597c 8017 ˘ 2245a–c 3.5 ˘ 0.5ab 1.4 ˘ 0.3a 0.4 ˘ 0.1a 6.9 ˘ 0.1cd 8.0 ˘ 0.4d 7.0 ˘ 0.1b
1360 20,226 ˘ 2643de 8645 ˘ 894de 8149 ˘ 1745a–c 6.9 ˘ 0.7bc 4.7 ˘ 0.9a 0.9 ˘ 0.3a 7.0 ˘ 0.5cd 7.8 ˘ 0.7d 7.1 ˘ 0.6b
CaCl2 2 Mid 4
2720 17,956 ˘ 2354b-d 8340 ˘ 423de 8443 ˘ 2924a–d 6.2 ˘ 0.6a–c 1.7 ˘ 0.6a 0.7 ˘ 0.2a 6.7 ˘ 0.2cd 7.7 ˘ 0.3d 7.1 ˘ 0.3b
1360 16,043 ˘ 2153ab 6338 ˘ 337bc 7018 ˘ 2123a 14.1 ˘ 2.1e 25.2 ˘ 4.5cd 5.4 ˘ 0.9c 5.7 ˘ 0.2ab 4.8 ˘ 0.2a 5.9 ˘ 0.2a
Late 5
2720 14,977 ˘ 1931a 6626 ˘ 333c 8239 ˘ 2738a–c 16.4 ˘ 3.0ef 17.4 ˘ 3.9bc 3.0 ˘ 0.9b 5.5 ˘ 0.2a 5.5 ˘ 0.3ab 6.1 ˘ 0.2a
1360 20,429 ˘ 3212de 9945 ˘ 917f 9415 ˘ 2988cd 4.2 ˘ 0.6a–c 1.6 ˘ 0.5a 0.5 ˘ 0.1a 7.2 ˘ 0.3d 8.2 ˘ 0.6d 7.4 ˘ 0.2b
All 6
2720 21,245 ˘ 3593e 9312 ˘ 998ef 10,098 ˘ 3339d 2.1 ˘ 0.6a 0.4 ˘ 0.1a 0.2 ˘ 0.1a 7.0 ˘ 0.1cd 8.1 ˘ 0.7d 7.4 ˘ 0.1b
Control n/a 0 15,431 ˘ 3084ab 3950 ˘ 454a 8684 ˘ 3219a–d 18.8 ˘ 1.6f 32.5 ˘ 8.6d 5.3 ˘ 0.7c 5.4 ˘ 0.4a 4.4 ˘ 0.6a 6.2 ˘ 0.3a
1 Signature® 2 3 4
8% Calcium Silicate. TETRA Flake 77% calcium chloride. Three weekly early applications starting at mid-bloom stage. Three weekly middle applications starting at end
of petal fall stage. 5 Three weekly late applications starting during early green fruit stage. 6 Seven weekly applications spanning mid-bloom to green fruit stages. 7 Treatment combinations
not connected by the same letter are statistically different (p < 0.05) based on linear contrasts extracted from mixed regression models.
Agriculture 2019, 9, 63 10 of 23

The low severity of drop in the control plots (5.3%) at Site 3 in Langley was observed for plants
with yield estimate of 8.7 t¨ha´1 and related to a high 6.2 fruit per cluster. Foliar Ca applications
reduced percent drop as low as 0.2%, Early, Mid, and All timings at both L-CaCl2 and H-CaCl2 having
statistically equivalent effects. The L-CaCl2 treatment at Late timing was equivalent to the control
while H-CaCl2 at Late timing and SST at Mid timing were statistically intermediate. With such a low
percent drop in general, no significant differences in estimated yield were detected between the control
and any of the Ca treatments. In contrast, there were significant increases in fruit per cluster, as high as
7.4, for Early, Mid, and All applications of L-CaCl2 and H-CaCl2 with no significant effects for either
rate at the Late timing or the SST treatment at Mid timing.
Under moderately severe drop of 18.8% in control plots at Site 1 in Abbotsford, almost all
treatments significantly reduced percent fruit drop. The lowest percent drop was 2.1% and was
observed for H-CaCl2 at All timing, which was equivalent to H-CaCl2 at Early and Mid timings as well
as L-CaCl2 at All timing. Early and Mid timings of L-CaCl2 were more modest in their effects. While
the impact of SST at Mid timing and L-CaCl2 at Late timing were even less pronounced, H-CaCl2 was
not significantly different from the control at Late timing. These differences in percent drop translated
to differences in estimated fruit yield with significant increases detected for H-CaCl2 at Early and All
timings and L-CaCl2 at Mid and All timings. Fruit per cluster was increased for all Early, Mid, and All
timings of CaCl2 application, but was not increased for SST or for the Late timing of CaCl2 application.
With a severe drop of 32.5% in control plots at Site 2, all treatments but Late L-CaCl2 significantly
decreased drop. The Early, Mid, and All applications of CaCl2 and Mid timing of SST were statistically
equivalent, but the decrease for Late timing of H-CaCl2 was more modest. To an even greater degree
than was seen under minimal and moderate drop conditions, there was a significant increase in
estimated yield for every foliar Ca treatment. L-CaCl2 and H-CaCl2 increased yield estimate to the
same extent at each timing. Only L-CaCl2 was more effective than either rate at Mid timing, which were
equivalent to L-CaCl2 and superior to H-CaCl2 at Early timing, the latter being equivalent to both rates
at Late timing. Only L-CaCl2 had as little an effect on estimated yield as SST at Mid timing, though this
treatment was still statistically different from the control. Fruit per cluster was significantly increased
by all treatments except for the Late timing applications; were equivalent for the two application rates
at Early, Mid, and All timings; and was statistically intermediate for SST at Mid timing, but this was
equivalent to L-CaCl2 at Early timing.
At the H-CaCl2 rate, some phytotoxicity was observed on leaves, especially following Early and
Mid applications, but there was no observable damage to either flowers or fruit. The lack of a negative
effect on fruit was verified in that the reduction in fruit drop was concurrent with increased yield
estimate and without negative effects on yield components such as fruit size. In fact, averaged across
three sites and two years, there were no significant differences in green or ripe fruit FW between the
control and any foliar Ca treatment. There were negative relationships between the percent fruit drop
and estimated yield across treatment combinations under moderate and high severity of GFD, and
there were positive relationships between fruit per cluster and estimated yield across all severities of
GFD (Figure 2).
All foliar treatments resulted in significant increases in percent Ca in green fruit samples on
both a FW and DW basis in 2015 (Table 4). Only the control was analyzed in 2016, making statistical
comparison impossible. Statistically greater increases in green fruit Ca were seen for H-CaCl2 compared
to L-CaCl2 at Mid, Late and All timings. H-CaCl2 at Early timing did not differ from L-CaCl2 at the
same timing or SST at Mid timing. Both L-CaCl2 and H-CaCl2 at All timing were higher than for
treatments of the same rates at any other timing, demonstrating generally greater increases in green
fruit Ca for seven applications compared with three.
Agriculture 2019, 9, 63 11 of 23
Agriculture 2019, 9, x FOR PEER REVIEW 15 of 24

35.0

30.0

25.0
y = -0.0046x + 43.947
Fruit drop (%) 20.0 R² = 0.5989
y = -0.0023x + 51.037
15.0 R² = 0.8525

10.0

5.0
y = -0.0012x + 12.406
0.0 R² = 0.2995
-2500 2500 7500 12500 17500 22500
-5.0
Estimated yield (kg . ha-1)
(a)

9.00
8.50
8.00
7.50 y = 0.0006x + 2.1512
Fruit per cluster

R² = 0.6788
7.00
y = 0.0005x + 2.4749
6.50 R² = 0.5501
y = 0.0003x + 1.2008
6.00 R² = 0.9264
5.50
5.00
4.50
4.00
0 5000 10000 15000 20000 25000
Estimated yield (kg . ha-1)
(b)
Figure 2. XY plots with regression equations and coefficients of determination between
Figure 2. XY plots with regression equations and coefficients of determination between plant responses
plant responses as affected by foliar Ca applications to ‘Draper’ highbush blueberry across
as affected bythree
foliar
farmCasites
applications
in the FrasertoValley,
‘Draper’ highbush
British Columbia,blueberry
Canada in across
2016 (a) three
Percentfarm
fruit sites
drop in the Fraser
Valley, British Columbia,
versus estimatedCanada
yield; (b)inFruit
2016per(a) Percent
cluster fruit
versus drop yield,
estimated versus● =estimated
Site 1 (dottedyield;
line), (b) Fruit per
cluster versus = Site 2 (dashed
◼ estimated yield,line),
‚= ◆ Site
= Site13(dotted
(solid line).
line),  = Site 2 (dashed line), ˛ = Site 3 (solid line).

Leaf samples taken at the same time as green fruit sampling, rather than at the typical late July
sampling date recommended for tissue sampling [30], also showed significant differences in Ca on
both a FW and DW basis across treatments. Compared with the control in 2015, these increases were
significant for H-CaCl2 at the Late and All timings (on both a FW and DW basis) and for L-CaCl2 at
All timing (on only a DW basis). These findings likely relate to the proximity of application timings to
sampling dates as newly developed leaves unfold rapidly during this early part of the summer and
any potential increases in leaf Ca from earlier applications would have been on earlier sets of leaves
than those that were sampled as the first fully unfolded leaves on new shoots.
Averaged across 2015 and 2016, ripe fruit Ca was significantly higher for virtually all foliar Ca
treatments compared with the control on both a FW and DW basis (Table 5). The exceptions were
L-CaCl2 at Early timing on both a FW and DW basis and SST at Mid timing on only a DW basis. No
L-CaCl2 treatment resulted in higher Ca than SST when applied the same number of times. H-CaCl2
was higher than L-CaCl2 for Early, Mid, and All timings but not Late timing. H-CaCl2 and L-CaCl2
were equivalent to one another at Mid and Late timings. In contrast H-CaCl2 was higher than L-CaCl2
for All timing, and the additional applications for this treatment resulted in higher Ca than any of the
other timings for each of the rates.
Agriculture 2019, 9, 63 12 of 23

Late leaf samples, taken at the recommended sampling date for blueberry tissue analysis [30],
showed some significant differences in Ca content across treatments when 2015 and 2016 data were
averaged (Table 6). Only H-CaCl2 at Mid, Late, and All timings were significantly increased over
the control on both a FW and DW basis, but there were no significant differences between these
combinations of Ca rates and timings. Mid timing of SST and Early timing of both H-CaCl2 or L-CaCl2
had statistically equivalent leaf Ca content. Comparing the average treatment values for each of the
three locations (n = 10), there were negative relationships between percent fruit drop and ripe fruit Ca
on a FW basis that varied in strength from R2 = 0.2121, R2 = 0.3188, to R2 = 0.4336 for Sites 1, 2, and 3,
respectively, in 2016 (Figure 3). Also, there were positive associations between fruit per cluster and
ripe fruit Ca on a FW basis that varied in strength from R2 = 0.1266, R2 = 0.3985, to R2 = 0.3055 for Sites
1, 2, and 3, respectively.
Agriculture 2019, 9, x FOR PEER REVIEW 16 of 24

0.009

0.008
Ripe fruit calcium (% fresh weight)

y = -9E-05x + 0.0074
0.007 R² = 0.2121

0.006

y = -0.0003x + 0.007
0.005 y = -6E-05x + 0.0068 R² = 0.3188
R² = 0.4336
0.004

0.003
0.0 5.0 10.0 15.0 20.0 25.0 30.0 35.0
Fruit drop (%)
(a)
0.009

0.008
Ripe fruit calcium (% fresh weight)

y = 0.0006x + 0.0029
R² = 0.1266 y = 0.0008x + 0.001
0.007 R² = 0.3055

0.006

y = 0.0005x + 0.0028
0.005 R² = 0.3985

0.004

0.003
4.00 4.50 5.00 5.50 6.00 6.50 7.00 7.50 8.00 8.50
Fruit per cluster
(b)
Figure 3. XY plots with regression equations and coefficients of determination between
Figure 3. XYplant
plotsresponses
with regression equations and coefficients of determination between plant responses
and fruit Ca concentration as affected by foliar Ca applications to ‘Draper’
and fruit Ca highbush
concentration
blueberryas affected byfarm
across three foliar Cainapplications
sites to ‘Draper’
the Fraser Valley, highbush
British Columbia, blueberry across
Canada
in 2016
three farm sites (a) Percent
in the Fraser fruit dropBritish
Valley, versus Columbia,
ripe fruit calcium;
Canada(b) Fruit per cluster
in 2016 versusfruit
(a) Percent ripe fruit
drop versus ripe
fruit calcium;calcium, ● = per
(b) Fruit Site 1cluster
(dottedversus
line), ◼ ripe
= Sitefruit
2 (dashed line), ◆
calcium, ‚== Site
Site31(solid line).line),  = Site 2 (dashed
(dotted
line), ˛ =4.Site 3 (solid line).
Discussion

4.1. Correlation of Green Fruit Drop in ‘Draper’ Blueberry with Fruit Calcium and Plant Vigor
The correlation between reduced GFD and higher fruit Ca resulting from foliar applications in
the current study, relates to the physiology of plant Ca nutrition in numerous crop species. Since Ca
has little mobility in the phloem, its movement from the root system to the above-ground parts of the
Agriculture 2019, 9, 63 13 of 23

Table 4. Mean ˘ SE green fruit and early leaf calcium concentrations in ‘Draper’ highbush blueberry in response to foliar Ca products, application timings and rates
across three sites in the Fraser Valley, British Columbia, Canada in 2015 as well as for control plots in 2016.

Green Fruit Calcium (%) 7 Early Leaf Calcium (%)

Year Product Timing Rate (ppm) Dry Weight Fresh Weight Dry Weight Fresh Weight
SST 1 Mid 800 0.12 ˘ 0.01b 0.015 ˘ 0.001b 0.66 ˘ 0.05a–c 0.122 ˘ 0.009ab
1360 0.12 ˘ 0.01b 0.015 ˘ 0.001b 0.65 ˘ 0.05ab 0.120 ˘ 0.008a
Early 3
2720 0.12 ˘ 0.01b 0.015 ˘ 0.001b 0.63 ˘ 0.05a 0.116 ˘ 0.009a
2015 1360 0.12 ˘ 0.01b 0.015 ˘ 0.001b 0.64 ˘ 0.05ab 0.118 ˘ 0.009a
2 Mid 4 2720 0.14 ˘ 0.01c 0.017 ˘ 0.001c 0.67 ˘ 0.05a–c 0.123 ˘ 0.008a–c
CaCl2
1360 0.12 ˘ 0.01b 0.016 ˘ 0.001b 0.66 ˘ 0.04ab 0.120 ˘ 0.008a
Late 5 2720 0.14 ˘ 0.01c 0.019 ˘ 0.001c 0.72 ˘ 0.06cd 0.133 ˘ 0.011bc
1360 0.14 ˘ 0.00c 0.018 ˘ 0.001c 0.69 ˘ 0.06b–d 0.125 ˘ 0.010a–c
All 6 2720 0.19 ˘ 0.01d 0.023 ˘ 0.001d 0.75 ˘ 0.06d 0.134 ˘ 0.008c
0.11 ˘ 0.01a 0.014 ˘ 0.001a 0.63 ˘ 0.05a 0.116 ˘ 0.008a
2016 8 Control n/a 0 0.12 ˘ 0.00 0.017 ˘ 0.001 0.57 ˘ 0.03 0.214 ˘ 0.010
Average 8 0.11 ˘ 0.01 0.015 ˘ 0.001 0.60 ˘ 0.04 0.165 ˘ 0.009
1Signature® 8% Calcium Silicate. 2 TETRA Flake 77% calcium chloride. 3 Three weekly early applications starting at mid-bloom stage. 4 Three weekly middle applications starting at end
of petal fall stage. 5 Three weekly late applications starting during early green fruit stage. 6 Seven weekly applications spanning mid-bloom to green fruit stages. 7 Treatment combinations
not connected by the same letter are statistically different (p < 0.05) based on linear contrasts extracted from mixed regression models. 8 No statistical comparisons possible as analysis only
conducted for control in 2016.
Agriculture 2019, 9, 63 14 of 23

Table 5. Mean ˘ SE ripe fruit calcium concentration on a dry and fresh weight basis in ‘Draper’ highbush blueberry in response to foliar Ca products, application
timings and rates across three sites in the Fraser Valley, British Columbia, Canada in 2015 and 2016.

Ripe Fruit Ca (% dry Weight) 7 Ripe Fruit Ca (% Fresh Weight)

Product Timing Rate (ppm) 2015 2016 Average 2015 2016 Average
SST 1 Mid 800 0.04 ˘ 0.00ab 0.04 ˘ 0.00bc 0.04 ˘ 0.00ab 0.006 ˘ 0.000ab 0.007 ˘ 0.000c–e 0.007 ˘ 0.000b
1360 0.04 ˘ 0.00a 0.04 ˘ 0.00ab 0.04 ˘ 0.00a 0.006 ˘ 0.000a 0.005 ˘ 0.001a 0.006 ˘ 0.000a
Early 3
2720 0.04 ˘ 0.00a–c 0.05 ˘ 0.00cd 0.04 ˘ 0.00bc 0.007 ˘ 0.000a–c 0.007 ˘ 0.001c–f 0.007 ˘ 0.000bc
1360 0.04 ˘ 0.00bc 0.04 ˘ 0.00a–c 0.04 ˘ 0.00bc 0.007 ˘ 0.001a–c 0.006 ˘ 0.000a–c 0.007 ˘ 0.000b
2 Mid 4 2720 0.05 ˘ 0.00d 0.05 ˘ 0.00c–e 0.05 ˘ 0.00d 0.008 ˘ 0.000c 0.007 ˘ 0.000d–f 0.008 ˘ 0.000c
CaCl2
1360 0.04 ˘ 0.00b–d 0.04 ˘ 0.00bc 0.04 ˘ 0.00bc 0.007 ˘ 0.000bc 0.007 ˘ 0.000b–d 0.007 ˘ 0.000bc
Late 5 2720 0.05 ˘ 0.00cd 0.05 ˘ 0.00cd 0.05 ˘ 0.00cd 0.007 ˘ 0.000c 0.007 ˘ 0.000c–e 0.007 ˘ 0.000bc
1360 0.04 ˘ 0.00cd 0.05 ˘ 0.00de 0.05 ˘ 0.00d 0.007 ˘ 0.000bc 0.008 ˘ 0.000ef 0.007 ˘ 0.000c
All 6 2720 0.06 ˘ 0.00e 0.05 ˘ 0.00e 0.06 ˘ 0.00e 0.009 ˘ 0.000d 0.008 ˘ 0.000f 0.009 ˘ 0.000d
Control n/a 0 0.04 ˘ 0.00ab 0.04 ˘ 0.00a 0.04 ˘ 0.00a 0.006 ˘ 0.000a 0.006 ˘ 0.000ab 0.006 ˘ 0.000a
1 Signature® 2 3 4
8% Calcium Silicate. TETRA Flake 77% calcium chloride. Three weekly early applications starting at mid-bloom stage. Three weekly middle applications starting at end
of petal fall stage. 5 Three weekly late applications starting during early green fruit stage. 6 Seven weekly applications spanning mid-bloom to green fruit stages. 7 Treatment combinations
not connected by the same letter are statistically different (p < 0.05) based on linear contrasts extracted from mixed regression models. However, lack of numerical differences between
significantly different values at the appropriate number of significant figures may indicate a lack of biological relevance for these differences on a dry weight basis.
Agriculture 2019, 9, 63 15 of 23

Table 6. Mean ˘ SE late leaf sample concentration on a dry and fresh weight basis in ‘Draper’ highbush blueberry in response to foliar Ca products, application
timings and rates across three sites in the Fraser Valley, British Columbia, Canada in 2015 and 2016.

Late leaf Ca (% Dry Weight) 7 Late Leaf Ca (% Fresh Weight)

Product Timing Rate (ppm) 2015 2016 Average 2015 2016 Average
SST 1 Mid 800 0.81 ˘ 0.04ab 0.85 ˘ 0.03ab 0.83 ˘ 0.03ab 0.322 ˘ 0.015ab 0.365 ˘ 0.013ab 0.344 ˘ 0.011a–c
1360 0.81 ˘ 0.04ab 0.83 ˘ 0.04a 0.82 ˘ 0.03a 0.322 ˘ 0.015ab 0.353 ˘ 0.015a 0.338 ˘ 0.011ab
Early 3
2720 0.81 ˘ 0.04ab 0.88 ˘ 0.04a–c 0.84 ˘ 0.03a–c 0.323 ˘ 0.013ab 0.375 ˘ 0.015a–c 0.349 ˘ 0.011a–d
1360 0.80 ˘ 0.05ab 0.88 ˘ 0.03a–c 0.84 ˘ 0.03a–c 0.320 ˘ 0.019ab 0.375 ˘ 0.014a–c 0.348 ˘ 0.013a–d
2 Mid 4 2720 0.81 ˘ 0.04ab 0.92 ˘ 0.03c 0.87 ˘ 0.03bc 0.322 ˘ 0.015ab 0.393 ˘ 0.015c 0.357 ˘ 0.013b–d
CaCl2
1360 0.82 ˘ 0.04ab 0.89 ˘ 0.03bc 0.85 ˘ 0.03a–c 0.331 ˘ 0.016bc 0.380 ˘ 0.013bc 0.356 ˘ 0.011a–d
Late 5 2720 0.84 ˘ 0.03bc 0.90 ˘ 0.03bc 0.87 ˘ 0.02bc 0.338 ˘ 0.014bc 0.385 ˘ 0.013bc 0.362 ˘ 0.010cd
1360 0.83 ˘ 0.05ab 0.89 ˘ 0.03bc 0.86 ˘ 0.03a–c 0.331 ˘ 0.018bc 0.380 ˘ 0.012bc 0.355 ˘ 0.012a–d
All 6 2720 0.88 ˘ 0.04c 0.89 ˘ 0.03bc 0.88 ˘ 0.02c 0.351 ˘ 0.016c 0.379 ˘ 0.014a–c 0.365 ˘ 0.011d
Control n/a 0 0.78 ˘ 0.04a 0.85 ˘ 0.03ab 0.81 ˘ 0.02a 0.305 ˘ 0.017a 0.370 ˘ 0.012a–c 0.337 ˘ 0.012a
1 Signature® 2 3 4
8% Calcium Silicate. TETRA Flake 77% calcium chloride. Three weekly early applications starting at mid-bloom stage. Three weekly middle applications starting at end
of petal fall stage. 5 Three weekly late applications starting during early green fruit stage. 6 Seven weekly applications spanning mid-bloom to green fruit stages. 7 Treatment combinations
not connected by the same letter are statistically different (p < 0.05) based on linear contrasts extracted from mixed regression models.
Agriculture 2019, 9, 63 16 of 23

4. Discussion

4.1. Correlation of Green Fruit Drop in ‘Draper’ Blueberry with Fruit Calcium and Plant Vigor
The correlation between reduced GFD and higher fruit Ca resulting from foliar applications in
the current study, relates to the physiology of plant Ca nutrition in numerous crop species. Since Ca
has little mobility in the phloem, its movement from the root system to the above-ground parts of
the plant is primarily through the xylem, driven by transpiration [34]. Compared to other cationic
nutrients that can move in the phloem (e.g., K and Mg), Ca transport to fruit is often much lower than
to leaves, resulting in deficiencies in many crops e.g., [5–7]. Environmental conditions that result in
low transpiration, low uptake of Ca and high competition between floral and vegetative organs lead to
these disorders, especially in crops with separate floral and vegetative buds as in blueberry [20]. While
flowers and fruits are typically stronger sinks for phloem transported elements and carbohydrates
than shoot apices, the movement of xylem transported elements is dictated by transpirative demand.
A high transpiring organ (e.g., leaves with many active stomata) will draw more water from the roots
than a low transpiring organ (e.g., fruit with relatively few stomata that decrease in function as the fruit
develop) [34]. Competition for Ca as it moves by mass flow in the xylem is not a complete explanation
of the uptake and differential allocation of this mineral because water transport to the fruit does not
always correlate well with allocation of Ca, as seen in apple [13]. However, competition with other
ions such as K may be aggravated under low transpiration and high competition [35].
The association between severity of GFD and plant vigor evaluated in the current study through
comparison of Ca treatment under different rates of N management, is in accordance with findings
for Ca disorders in other crops. Specifically, several crops demonstrate an interaction between Ca
disorders and N management, with high levels of N resulting in increased vigor and often triggering
the Ca deficiency or exacerbating the related disorder [19]. In apple, for which these relations have
been most extensively studied, the competition between vigorous shoot growth and fruit Ca was
proposed more than sixty years ago [36]. This interaction is most likely due to competition between
growing vegetative shoot apices and developing fruit rather than inhibition of Ca uptake from the
roots by excessive N in the root zone [37]. Not just vigor of shoot growth but the timing of onset
for vigorous growth relative to fruit development is important in some Ca disorders in other crops.
Various rates and durations of Ca uptake to the fruit have been document in apple, ranging from
primarily in the first part of fruit growth to occurring with a decreasing linear trend that extends until
harvest [16,38]. In any case, Ca concentration in apple tends to reach its maximum soon after bloom
before dropping due to dilution as the fruit grow through cell enlargement [39]. Therefore, the key Ca
loading phase is early in development for many types of fruit and is highly influenced by variation in
transpiration [34], but the reasons for the decline in xylem transport of Ca into fruit after this phase are
still only partially understood, likely relating to a decline in xylem function [40].

4.2. Calcium Deficiency as a Genetic Pre-Disposition to Green Fruit Drop in ‘Draper’ Blueberry
The current finding that foliar Ca corrects GFD in ‘Draper’ builds on the current knowledge of Ca
nutrition in blueberry. In Oregon, a high leaf Ca and low fruit Ca were noted in a replicated survey
of tissue nutrient content across a range of cultivars [8], which validates that an interaction between
Ca and N is observed and may be due to competition between these plant organs for Ca in the xylem
sap. These authors explain the higher leaf Ca in ‘Draper’ as being related to the high vigor and many
growing shoot tips of ‘Draper’ in Oregon. The severity of GFD in Southwestern BC compared with
the near lack of GFD in Oregon indicates differential performance across environments. Specifically,
this belies marginal adaptation of ‘Draper’ to Southwestern BC in comparison to a highly adaptive
region like Oregon. Consequently, Ca uptake, adequacy of allocation to the fruit or both factors are in a
range of marginal sufficiency in this region, entering deficiency under certain field- and season-specific
conditions, subsequently resulting in GFD.
Agriculture 2019, 9, 63 17 of 23

In apple, the occurrence and severity of bitter pit is related, among many factors, to the specific
cultivar and where it is grown. Pre-disposition of ‘Draper’ to GFD is similar, a physiological condition
with an important genetic component, manifesting under certain environmental conditions. While
‘Draper’ is the first highbush blueberry commercial cultivar to manifest this condition, GFD has
also been observed in progenies of ‘Draper’ in breeding plots in the Willamette Valley of Oregon (C.
Finn, personal communication). As with apple bitter pit, GFD symptoms include decayed internal
tissues with no visible symptoms to the exterior. The internal browning is likely related to action of
degradation enzymes. Lack of damage to the exterior of the fruit is due to the ability of the soluble
fraction of fruit Ca to move within the fruit [41], leading to deficiencies in some parts and not in others.
Detailed comparison of skin, pulp and seed Ca levels have not been conducted to study a presumed
gradient in fruit Ca relating to GFD in ‘Draper’.
Therefore, with a genetic pre-disposition toward relatively greater allocation of Ca toward
vegetative structures at the expense of reproductive structures [8], ‘Draper’ responds to the
environmental conditions of Southwestern BC with a deficiency that can be characterized as an
environmental maladaptation. With Ca deficiency as a proximate cause of GFD, interacting with plant
vigor and presumably other factors, the ultimate cause of this maladaptation could be due to any
number of physiological mechanisms or disfunctions, likely relating to the high leaf/fruit Ca ratio
for the cultivar. For example, one could speculate that there is low expression of a cell membrane Ca
pump gene under environmental conditions conducive to GFD. As contributing factors, edaphic soil
properties and horticultural management practices may aggravate the effects of the climatic conditions
on plant phenology that reduce Ca uptake or result in greater competition for Ca allocation to the fruit.
Temperature, light intensity, precipitation, soil moisture, and relative humidity are all important
factors that affect transpiration. Climatic conditions in Southwestern BC are generally cooler, wetter
and more often overcast during spring phenological development of blueberry in comparison with
Oregon. In most years, Southwestern BC is likely to have higher relative humidity, wetter soils,
lower light levels, and cooler temperatures, indicating generally lower transpirative demand, in
comparison to Oregon with minimal GFD. High relative humidity decreases Ca uptake in either the
fruit or vegetative tissues of sweet pepper (Capsicum annuum L.), tomato, and apple [6,42,43]. Further,
well-watered plants have reduced transpiration, which could be made worse if leaf development is
relatively delayed very early after floral bud break, as observed in the field in the case of ‘Draper’.
Later in the season, when fruit development is in its first stage of cell division and the shoots are in a
rapid phase of elongation, excessive guttation on the leaves is sometimes observed in the field under
overcast conditions with high relative humidity, indicative of positive root pressure for xylem sap.
Tomato and apple demonstrate much reduced allocation of Ca to fruit under experimental conditions
of high relative humidity [44]. From these field observations, one or both potential environmental
factors could be involved in triggering GFD in ‘Draper’.
Parsing the effects of these putative environmental triggers would require detailed documentation
of fruit Ca loading over time and a comparative study of phenological and climatic factors across
regions with and without GFD. A companion survey of tissue and soil nutrient values across BC, WA,
and OR in 2015 and 2016 was conducted as a preliminary investigation of these fundamental issues
(unpublished data). Nevertheless, the current explanation of GFD is consistent with what is known of
Ca disorders in other fruit such as apple and pear, the severity of which tend to vary from year-to-year
and site-to-site by directly impacting plant parts, indirectly interfering with the interactions between
plant parts or otherwise responding to cultural management practices [20].

4.3. Mitigation of Green Fruit Drop in ‘Draper’ Blueberry Using Corrective Foliar Calcium Applications
In BC, GFD is observed in fields with a broad range of soil Ca (data not shown), with some
severely affected fields having very high levels of soil Ca based on established nutrient management
recommendations for the Pacific Northwest [30]. For this reason, and since blueberry is a calcifuge
that is highly efficient at taking up Ca under low pH conditions [11], soil applications are unlikely to
Agriculture 2019, 9, 63 18 of 23

impact the incidence of GFD. Whereas application of lime or gypsum to the soil may benefit overall
crop health in the long-term management of soil pH of blueberry plantings where pH is dropping
below the recommended range, soil applications of these fertilizers demonstrated little effect on fruit
Ca and no effect on fruit yield, size or firmness in studies in Michigan [45].
As seen in other crops, loading of Ca into blueberry fruit occurs primarily when shoot growth is
limited, and low transpiring fruit accumulate less Ca than high transpiring leaves overall [18]. This
results in a high DW leaf/fruit Ca ratio, which increases several-fold as the season progresses [26].
While Ca is more likely than more mobile elements to be affected by foliar application due to its
immobility within the plant, affecting a substantial change in fruit Ca via exogenous treatment is
challenging for this same reason. That is, without translocation of Ca from leaves to fruit by the
phloem, effective correction of acute fruit deficiencies requires movement of exogenous Ca into fruit
tissues, primarily through stomata, as described in apple [46]. This is problematic because fruit
stomata are only functional for a short period of time during early fruit development, the waxy cuticle
decreasing their functionality as it develops [47]. In apple, uptake of exogenous Ca occurs in very
small in quantities (<1 ug¨cm´2 ) during foliar management of bitter pit, necessitating as many as ten or
eleven applications to be even partially successful in some varieties [38,48]. Notwithstanding, uptake
of exogenous Ca is known to occur by first penetrating the cuticle and then moving apoplastically
through the flesh [49]. Evidence for this ability is based on Ca concentration changes within different
parts of the apple fruit during maturation, and even post-harvest [50]. With the availability of the
mobile, physiologically-active forms of water-soluble Ca increasing as the fruit develop [41], the small
quantity of Ca that effectively penetrates the fruit surface can redistribute to portions of the fruit that
are deficient.
In blueberry, fruit Ca levels impact fruit quality [51], but attempts to use foliar Ca to increase
fruit Ca concentration, usually for the purpose of improving fruit quality, have been met with varying
degrees of success. In one study in Michigan, a 0.08% Ca solution was applied to ‘Bluecrop’, ‘Blueray’,
and ‘Ivanhoe’, but fruit Ca and quality parameters such as firmness were not affected [28]. In
comparison, Experiment 3 used 0.136% and 0.272% Ca, which were 1.7ˆ and 3.4ˆ higher, respectively,
and were effective at increasing ripe fruit Ca significantly for all timing treatments except for L-CaCl2
at Early timing (Table 5). In Chile, increases in Ca levels in fruit skin and seeds were seen at a low rate
of 0.0475%, but increases in fruit pulp Ca required 0.09% or 0.18% to be detected [52]. Experiment 3,
which did not compare Ca concentrations in different parts of the fruit, used rates of CaCl2 that were
1.5ˆ and 3.0ˆ higher, respectively, than the lowest amount needed by these researchers to affect an
increase in Ca content in the pulp in addition to the skin and seeds. In another Chilean study, increases
in fruit Ca were achieved following two applications at 0.06% Ca but not 0.036% Ca or 0.006% Ca [26].
The rates in Experiment 3 were 2.3ˆ 4.5ˆ higher.
In contrast, work in Oregon used a range of Ca products (chloride, silicate, acetate, and chelate)
with 3–5 applications from late bloom to early green fruit development (1–4 weeks before harvest) on
‘Spartan’, ‘Draper’, ‘Liberty’, and ‘Legacy’ [53]. The highest rate of CaCl2 in this study was applied at
0.09% Ca in 748 L water¨Ha´1 , and the researchers saw no significant effect on fruit Ca or fruit quality.
While the concentrations used for Experiment 3 were maintained as constants, the amount of spray
volume applied to experimental plots varied from 371 to 914 L water¨Ha´1 , depending on the field,
year, and stage of plant development. Comparing the total amount of Ca applied would require a
nuanced comparison of the plant sizes used in these studies, but with applications being made in the
current study to the point of runoff, it is the actual concentration of the spray solution that contacts
the fruit surfaces that is most relevant to uptake. Therefore, the concentrations applied in the current
study were 1.5ˆ and 3.0ˆ higher than the highest rate applied in Oregon, which largely explains the
difference in results. Further, while the timings of application in Oregon started at approximately the
same time as the effective Mid timing used in BC, the Oregon study did not use a surfactant, based on
supplier recommendations (B. Strik, personal communication), which may also have resulted in lower
uptake of Ca across fruit tissues.
Agriculture 2019, 9, 63 19 of 23

Foliar Ca treatment has long been used as a standard production practice by apple growers
to decrease bitter pit [54]. Foliar applications are effective in reducing bitter pit, and this is often
associated with a measurable increase in fruit Ca but not in all studies [55,56]. Determining significant
differences in fruit Ca is often problematic in experimental bitter pit research due to a high degree of
between-plant and within-plant variation in mean Ca [57,58]. Fruit Ca was determined in the current
study on both a FW and DW basis to remove the impact of differences in fruit size across samples
based on the same approach taken in apple research [59]. However, the results on a FW and DW basis
showed very similar effects across treatments in the current study.
As seen in bitter pit control [59], CaCl2 applications demonstrated risk of phytotoxicity (i.e., leaf
marginal necrosis) in blueberry when applied at the higher rate in the current study. More phytotoxicity
is seen in apple in response to early applications [60], which was also observed in the current study. In
Michigan, a very low rate (0.08% Ca) resulted in similar phytotoxicity [28], but no such damage was
observed at more than twice this rate (0.18% Ca) in Chile [52], a rate that was intermediate to the low
and high rates used in Experiment 3.
In summary, foliar application at sufficiently early stages of fruit development is important but so
is use of the correct material. Likewise, early applications are most effective for bitter pit control in
apple [59] and CaCl2 is generally the most effective material [60]. Moreover, the amount of exogenous
Ca uptake by fruit is not just related to the permeability of the fruit tissues at the time of application
but to the quantity of spray volume that contacts the fruit tissues and its concentration at the time of
contact [61]. For these reasons, the success of the current study in increasing fruit Ca and mitigating
GFD in ‘Draper’ is attributed to five factors: (1) use of a Ca material that has been demonstrated to
affect high uptake in other crops; (2) use of high concentrations of Ca, afforded by the use of a basic
fertilizer product (CaCl2 ) rather than a formulated product with a restricted label rate; (3) application
during early stages of fruit development when stomata are most functional and the cuticle least
developed; (4) repeated application, three being substantially as effective as seven; and (5) use of a
surfactant in tank mixes to decrease the surface tension of water droplets as they contacted fruit tissue,
extending the period of time over which Ca was absorbed. Further qualification of this final point is
that applications were made to the point of run-off under slow drying conditions that were conducive
to optimal foliar uptake.

4.4. Future Research Directions for Green Fruit Drop in Blueberry

For decades, the research in apple and tomato have indicated that low Ca predisposes these
crops to bitter pit and blossom-end rot, respectively, but that there are other factors which require
consideration [62,63]. While mitigation of GFD with foliar Ca applications is highly effective and
straight-forward, investigation into additional factors contributing to GFD in ‘Draper’ highbush
blueberry should be part of an integrated approach to combatting this economically devastating
disorder for commercial growers. Specifically, the antagonistic relationships of Ca with K and Mg seen
in apple bitter pit and strawberry tip burn, respectively, are due to the ability of these other cations to
move freely via the phloem and compete with Ca, influencing allocation to different organs [14,64].
Moreover, in apple, the role of K, Mg, Zn, and Mn, and ratios between these cations and Ca, have been
known for more than two decades though their role is still not fully understood [65].
Climatic maladaption resulting in poor carbohydrate partitioning is also a potential area of future
research relating to the underlying physiological cause of GFD in ‘Draper’. Unbalance partitioning
plays a role in poor Ca allocation and contributes to bitter pit in apple [66]. In ‘Draper’, shade
studies have not, however, demonstrated a significant effect on severity of GFD [67]. Alternatively,
the use of plant growth regulators (PGRs) to modify the uptake of Ca, or enhance its allocation
to fruit, is established as strategy for decreasing bitter pit in apple. For example, PGRs such as
Prohexadion-Calcium (a gibberellic acid inhibitor), 1-methylcyclopropene (1-MCP), or diphenylamine
(DPA) have been used in apple [68,69]. The potential negative effect of the types of PGRs on fruit set
make this avenue of investigation problematic.
Agriculture 2019, 9, 63 20 of 23

Seeds are important for the accumulation of fruit Ca [70], and ‘Draper’ does demonstrate
problematic pollination under some conditions, perhaps related to a squat floral morphology or
some other cause of low attractiveness to honey bees (Apis mellifera L.) [67]. However, dissection
of fruit during extensive field observations across the region has demonstrated that dropped fruit
consistently contain numerous developing seeds, and the pattern of GFD severity across fields and
years does not present as being related to poor pollination or low fruit set. Consequently, the role of
enhanced pollination as an additional GFD mitigation strategy is unlikely to be worthwhile.
In apple, Ca is remobilized in the xylem sap from storage in tissues such as bark prior to significant
root growth [35]. The same is seen in blueberry, and it is possible that Ca levels in these storage
structures is low for ‘Draper’ in BC. Fall foliar Ca applications to increase the amount that is stored by
the plant during the dormant season, and that will be available for remobilization in the spring, is a
potential strategy for reducing GFD pre-disposition. Thus far, our initial attempts to reduce GFD by
increasing plant Ca through fall foliar applications in the previous year have not been effective (data
not shown), and a subsequent study by researchers in northwestern WA has shown no effect of fall Ca
on GFD in the following year [71].
Finally, the genetics of predisposition to GFD is the most important avenue of future research
because the disorder is not just a feature of ‘Draper’. It appears to be a heritable condition in highbush
blueberry. ‘Draper’ is being used in many northern highbush breeding programs around the world
due to its ability to transmit high fruit quality to its progeny. A few cultivars with ‘Draper’ as a parent
(e.g., ‘Calypso’) have already been commercialized and are being widely planted in regions where
GFD is observed. Therefore, evaluating the genetic tendency for GFD is an important objective for
breeding new cultivars of blueberry.

5. Conclusions
The principal findings of this study are that GFD is a genetic disorder seen in ‘Draper’ highbush
blueberry that is: (1) correlated with low fruit Ca and high plant vigor; and (2) corrected via foliar
applications of Ca, which: (a) require specific rates and timings to be effective; (b) result in increased
yield components; and (c) correlate with higher fruit Ca. Successful mitigation of GFD requires a Ca
product that is readily absorbed by plant tissues, and applications should be made at relatively high
concentrations compared to rates used in other studies on blueberry but ranging from approximately
75 to 150% of the rates typically used in treatment of bitter pit in apple. Leaf phytotoxicity was
observed at the higher rate used in the current study, but the lower rate was as effective in reducing
GFD in most cases. Three applications starting at mid-bloom were effective, as were three applications
starting at the end of bloom, but commercial growers may choose to make applications across the
range of these effective dates or apply more frequently than every seven days to ensure maximal Ca
uptake. Application during slow evaporative conditions increases Ca uptake, and caution with Ca
materials and rates must be taken to avoid phytotoxicity.

Author Contributions: For this study, all three authors were involved in designing the experiments, interpreting
their results and producing this manuscript. Further, M.S. assisted E.M.G. in application for funding and collection
of field data while C.M. assisted E.M.G. in coordination and implementation of field trials. E.M.G. performed the
statistical analysis and drafted this manuscript, and C.M. and M.S. provided insight into discussion of results
during manuscript preparation and revision.
Funding: This research was funded by the British Columbia Blueberry Council, Sidhu Growers Ltd., the
Lower Mainland Horticultural Improvement Association, the Investment Agriculture Foundation of British
Columbia grant number INN208, and Agriculture and Agri-Food Canada’s Growing Forward II initiative grant
number AIP-P002.
Acknowledgments: The authors thank Rudy Janzen, Rajinder and Harinder Cheema and Ajit and Paul Sanga
for hosting trial sites as well as Michael Dossett, Lisa DeVetter, Bernadine Strik, Tom Baumann, and Karina
Sakalauskas for providing advice during the course of this study. The authors also thank Brookside Laboratories
Inc. for performing all chemical nutrient analyses for this study.
Conflicts of Interest: The authors declare no conflict of interest.
Agriculture 2019, 9, 63 21 of 23

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