Agricultural Water Management 123 (2013) 93102

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Agricultural Water Management

journal homepage: www.elsevier.com/locate/agwat

Irrigation of an established vineyard with winery cleaning agent

solution (simulated winery wastewater): Vine growth, berry quality,
and soil chemistry
Kim P.M. Mosse a,b,c , Jungmin Lee d , Benjamin T. Leachman e , Sanjai J. Parikh f ,
Timothy R. Cavagnaro c , Antonio F. Patti a,b , Kerri L. Steenwerth g,
a

School of Applied Sciences and Engineering, Monash University, Churchill, Vic., 3842, Australia
Centre for Green Chemistry, Monash University, Vic., 3800, Australia
c
School of Biological Sciences and Australian Centre of Biodiversity, Monash University, Vic., 3800, Australia
d
United States Department of Agriculture, Agricultural Research Service, Horticulture Crops Research Unit worksite, 29603 U of I Ln, Parma, ID, 83660, USA
e
Department of Viticulture and Enology, University of California Davis, CA, 95616, USA
f
Department of Land, Air and Water Resources, University of California Davis, CA, 95616, USA
g
United States Department of Agriculture, Agricultural Research Service, Crops Pathology and Genetics Research Unit, 595 Hilgard Ln, Davis, CA, 95616, USA
b

a r t i c l e

i n f o

Article history:
Received 15 August 2012
Accepted 26 February 2013
Available online 16 April 2013
Keywords:
Shiraz
Vitis vinifera. L.
Saline efuent
Syrah
Wastewater reuse
Winery efuent

a b s t r a c t
The ability to use winery wastewater (WW) for irrigation purposes could be a benecial to the wine
industry. A major difculty in studying WW use is its inconsistent availability and composition. As such,
we applied four simulated WWs composed of salts from two main industrial cleaning agents, and a control to 12-year-old Syrah (clone 99, 3309C rootstock; Vitis riparia V. rupestris). Briey, the treatments
simulated wineries utilising a sodium (Na) based cleaner; a potassium (K) based cleaner; a K based cleaner
coupled with higher water use efciency, resulting in a higher K concentration; and a combination of elevated K levels with the presence of wine, to consider the potential synergistic/antagonistic effects of the
salts and organic matter. Soil salt concentrations increased consistently with the nature of the treatment
applied, with K and Na treatments causing increased soil K and Na, respectively. Analysis of salt accumulation at various depths indicated that Na was more mobile in soils; however, the addition of wine to
irrigation water enhanced K transport to the subsurface. Petiole concentrations of K and Na were approximately three-fold and nine-fold greater in the K and Na treatments than control. Attributes related to
berry and juice quality differed among treatments at both vraison (juice K and anthocyanin concentrations) and harvest (juice Na, juice K, total phenolics, berry weight, and harvest weights), although the
majority of these were slight, and therefore unlikely to have signicant impact on wine quality. Based on
the data collected, occasional irrigation with these simulated WWs appears to have had minimal impact
on established vines. Nonetheless, there is potential for greater impacts to occur over longer time periods
due to the perennial nature of grapevines and accumulation of salts in the soil.
Published by Elsevier B.V.

1. Introduction
Winery wastewater (WW) is produced in signicant volumes
around the world (Kumar and Kookana, 2006; Bustamante et al.,
2005), and may be reused as an irrigation water source. For
instance, it is estimated that 35 kL WW is generated per tonne
of grapes crushed in Australia (Kumar and Kookana, 2006). WW
contains relatively high levels of grape/wine residue, potassium (K)
and/or sodium (Na) salts that are introduced via winery cleaning

Corresponding author. Tel.: +1 5307527535; fax: +1 5307520382.

E-mail addresses: kerri.steenwerth@ars.usda.gov, ksteenwerth@ucdavis.edu
(K.L. Steenwerth).
0378-3774/$ see front matter. Published by Elsevier B.V.
http://dx.doi.org/10.1016/j.agwat.2013.02.008

operations (K concentrations up to 1000 mg L1 and a K:Na ratio of

3:1; Chapman, 2001; Bustamante et al., 2005; Quayle et al., 2010;
Sheridan et al., 2011). The potential for reuse of WW as a vineyard irrigation water source is uncertain and volume generated
is inconsistent, but determining the impacts of continued application of K, Na, and other WW components (Bustamante et al.,
2005) to grapevines merits further investigation (but see studies on other wastewater sources, e.g., Downton, 1977; McCarthy,
1981; McCarthy and Downton, 1981; Paranychianakis et al., 2004;
Hepaksoy et al., 2006; Tregeagle et al., 2006; Paranychianakis and
Angelakis, 2008). Projections of increased severity and incidence
of drought under global climate changes models (IPCC, 2001) will
likely increase incidence of WW reuse to meet water demands. Historically, the most commonly used cleaning agent has been sodium

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K.P.M. Mosse et al. / Agricultural Water Management 123 (2013) 93102

hydroxide (NaOH) based. Due to concerns about high Na levels

entering environmental systems via the WW, many wineries are
shifting towards alternative K based cleaning technologies (Arienzo
et al., 2009).
Exposure of grapevines to high concentrations of irrigation
water Na can be toxic (1.85.6 dS m1 , depending upon rootstock;
Zhang et al., 2002), resulting in reduced transpiration and biomass
production (Shani and Ben-Gal, 2005). Most studies considering
salinity effects on grapevines have focused on NaCl, and as chloride (Cl) is also toxic to grapevines, separating effects of the two
ions is difcult (Tregeagle et al., 2006). Many plant species derive
salt tolerance from Na exclusion mechanisms (Munns, 1993), but
certain woody perennials (e.g., citrus) appear to employ Cl exclusion mechanisms (Storey and Walker, 1998). Various in vitro grape
cultivars can tolerate from 100 mM NaCl (cv. Cardinal) to 175 mM
NaCl (cv. Perlette) delivered via growth media, although survival
rates are much lower in comparison to growth in the absence of
NaCl (Singh et al., 2000). These Na concentrations are considerably
higher than what is typically found in WWs (up to 20 mM calculated; Mosse et al., 2011). Increased soil Na concentrations in WW
can also impact soil structure, causing clay dispersal, and thereby
negatively affect soil processes such as inltration and aggregation.
This alteration to soil structure, as well as the greater availability of
Na over a long time period may therefore impact vine growth and
berry quality, without necessarily inducing mortality.
Potassium is an essential nutrient for vine development, and is
involved in a variety of physiological processes, including enzyme
activation in photosynthesis and respiration (Bhandal and Malik,
1988) and maintenance of cellular osmotic potential in plants
(Salisbury and Ross, 1992). Potassium is mobile within the vine,
and mature leaves often reveal deciency symptoms (Mullins et al.,
1992). Due to the relatively high requirements in vineyards, K
is often applied as fertiliser, commonly as KCl, KNO3 , or K2 SO4
(Kasimatis and Christensen, 1976). As with many plant nutrients,
high K addition may negatively impact overall vine health and
wine quality (Jackson and Lombard, 1993; Mpelasoka et al., 2003;
Laurenson et al., 2011). Application of K can inuence the uptake
of other nutrients, with antagonistic uptake reported for calcium
(Ca), and magnesium (Mg) with K application (Morris and Cawthon,
1982; Wolf et al., 1983). In addition, K has similar dispersive effects
on clay particles as Na within soils, although the magnitude of this
dispersion is generally not as large (Arienzo et al., 2009).
The majority of vine K uptake is typically during the period
between bloom and vraison. After vraison, the uptake of K
decreases, and at the time of harvest, approximately 3070% of
the vines K is present in the berries, dependent upon the species
(Conradie, 1981; Williams and Biscay, 1991). Potassium as well
as Na uptake is further inuenced by both rootstock and scion
(Downton, 1977; Paranychianakis et al., 2004, 2006; Hepaksoy
et al., 2006; Paranychianakis and Angelakis, 2008), suggesting that
appropriate selection of varietals to receive WW may play a key role
in ensuring long term sustainability of the WW reuse. Increased
K application is associated with reduction in juice acidity (Morris
and Cawthon, 1982; Mpelasoka et al., 2003; Delgado et al., 2004),
which can result in decreased wine quality. Application of K to
grapevines has also been found to inuence anthocyanin levels and
color intensity (Morris et al., 1980; Delgado et al., 2004).
This is a study on the impacts of irrigating established Syrah
vines with simulated WWs containing main elemental components
of winery cleaning agents during a single growing season to identify
immediate effects on soil, grapevines, and grape quality. The WW
treatments focused primarily on the salts present in untreated WW
(Na and K) at industrially relevant concentrations (40 mM Na and
4080 mM K), as well as K in the presence of dilute wine to ascertain whether the additional organic matter present can ameliorate
potentially negative impacts of K application. The concern over

shifting from Na- to K-based cleaners has emerged as a high priority

issue for California wineries; therefore, presentation of this study
establishes necessary baseline information about WW application
in vineyards.
2. Materials and methods
2.1. Vineyard set-up
Studies were conducted at the University of California, Davis in
the Tyree vineyard, Yolo County, CA, USA (latitude 38.53 N, longitude 121.79 W). The region has a Mediterranean climate and
rainfall of approximately 450 mm annually, the majority of which
falls in the winter months. Soils were a Yolo silt loam (Fine-silty,
mixed, superactive, nonacid, thermic Mollic Xerouvent). Vines
were irrigated from June 21st to September 10th 2010. The study
site consisted of ve vine rows adjacent to each other, each comprised of 27 vines (1.8 m 2.4 m) of established (planted in 1998)
Syrah clone 99 on 3309C rootstock (Vitis riparia V. rupestris).
Prior to this trial, these vines had been used as a production block,
and were irrigated, pruned, and fertilized following CA commercial
practices. These experimental rows were divided into four plots,
each containing six vines (with buffer vines at the end of each row),
which were considered as eld replicates.
2.2. Irrigation treatments
Due to the inconsistent WW volume generated (a potential limitation for WW reuse) throughout the wine production process, a
simulated WW was composed of the main winery cleaning aid elements. The experimental plots were irrigated with the following
control and four simulated WW treatments:
a. Control (source water only): K concentrations were typically
12 mM, and Na concentrations 0.10.8 mM (tested externally at
A & L Western Agricultural Laboratories, Inc., Modesto, CA, USA).
Source water originated from Lake Berryessa, CA, USA (pH 8.3).
Other attributes of the source water included 0.06 mM nitrate,
1.1 104 phosphate, 0.3 mM sulfate, 0.023 mM boron, SAR 0.3,
EC 0.43 mmhos cm1 , 0.70 mM calcium, 1.98 mM magnesium,
0.14 mM carbonate, 4.2 mM bicarbonate, 0.25 mM chloride, and
393 ppm total dissolved solids.
b. High K: Elevated K in pre-vraison irrigation (80 mM K as K2 SO4 ;
The Kissner Group, Cambridge, Ontario, Canada), lowered to
40 mM K after vraison (pH 8.88.9). Levels of salt are often relatively higher in the lead up to harvest because wineries typically
clean and sanitize tanks, lines, and other equipment during this
period (Sheridan et al., 2011).
c. High K + W: Elevated K (80 mM K), then lowered to 40 mM K at
vraison in the presence of 2.5% red wine (Mission Bell Winery,
Madera, CA, USA). This treatment considered the effects of a combination of K2 SO4 and dilute wine (pH 3.81) to simulate organic
matter typically present in WW (Bustamante et al., 2005). Treatment pH was 6.56.6.
d. Low K: 40 mM K, as 20 mM K2 SO4 . WWs normally contain signicant amounts of Na and K with a K:Na ratio of 3:1 and K
concentrations of up to 1000 mg L1 (Chapman, 2001). Assuming a molar equivalent of KOH will be used when replacing
NaOH, this will result in 14.3 mM K in WW in addition to the
1000 mg L1 (25.6 mM) present from grape and wine origin.
Combined, this yields approximately 40 mM K. pH of the low K
treatment was 8.58.7.
e. Na: 40 mM Na, as 20 mM Na2 SO4 (The Kissner Group,
Cambridge). This treatment is a molar equivalent to the low K
treatment. pH of the Na treatment was 8.78.8.

K.P.M. Mosse et al. / Agricultural Water Management 123 (2013) 93102

Concentrated solutions corresponding to each treatment were

pre-mixed in 189 L (50 gallon) drums. The respective irrigation
treatments were diluted to specication using Dosatron injectors
(model D14MZ10, Dosatech, Deinze, Belgium). Dosatron dilution
ratios were adjusted, if necessary, after measuring the electrical conductivity (EC) of the respective irrigation treatments using
an Orion 4-Star Plus pH/Conductivity Portable Multiparameter
Meter (Thermo Scientic, USA). Sulphate served as a counterion (i.e., less growth restriction than Cl; Sols-Prez and Cabrera,
2007). Gypsum (two applications of 500 g per vine) was applied
as a solid beneath the drippers in all rows, as may occur in
an industrial application in California, to minimise soil dispersion due to high SAR/PAR waters (Rengasamy and Olsson, 1991;
Grifoen, 2001). This application will only occur during the initial
year of this study. Normal fertilization with K met the necessary
annual requirement. Weekly irrigation volumes delivered 70% of
the estimated potential evapotranspiration (ET) (Williams, 2001;
California Irrigation Management Information System, station #6
Davis, http://www.cimis.water.ca.gov/cimis/welcome.jsp) to each
vine, via 2 8 L h1 (2 gallons h1 ) drippers located on either side
of the vine trunk. A total of 603 L per vine was delivered prior to
vraison, and a total of 981 L per vine at the time of harvest.
2.3. Tissue sampling
Petiole and leaf samples were collected prior to irrigation commencement (June 21 st) and at 50% vraison (August 9th) and
harvest (September 10th). Fifty leaves and petioles were collected
from north and south sides of the vine rows in each block (i.e., 12
leaves per vine). The rst fully expanded leaf was collected; due
to hedging operations, this was not always possible, in which case
mature leaves were sampled. Leaves and petioles were dried at
50 C for 4872 h, and milled to 40 mesh prior to digestion.
Berry samples were collected at 50% vraison (vraison) and
harvest. Six clusters per vine were sampled, with eight individual berries randomly collected from each cluster (288 berries per
replicate). Fifty fresh berries were weighed and gently squeezed to
release juice. Juice was analysed for titratable acidity (TA), total
soluble solids (TSS), and pH (see below). The remaining berries
were frozen at 80 C prior to analysis of total phenolics, anthocyanins, tannins, and o-phthaldialdehyde (NOPA) (see below).
Harvest weights (kg, yield per plot) were determined by weighing all clusters on ve vines in each plot. Cane diameter (mm) and
length (mm) were measured between the 2nd and 3rd nodes using
a digital calliper.
2.4. Tissue digestion and nutrient analysis
Leaf and petiole tissue samples were digested using a wet ashing method (adapted from Campbell and Plank, 1998). Briey,
samples were weighed (0.5 0.001 g) into 50 mL glass tubes, and
pre-digested with 5 mL concentrated HNO3 at room temperature
(ca. 22 C) for 30 min. Samples were heated to 125 C for 90 min,
before the addition of 3 mL 30% H2 O2 , and further heating at 125 C
for 60 min, repeated once. The block temperature was increased
to 200 C, and the samples heated to dryness (ca. 60 min). After
cooling, 10 mL 10% HNO3 was added to the tubes, and samples
were diluted to 50 mL with DDI water (Barnstead Nanopure Ultrapure Water System, ThermoFisher Scientic Inc., Waltham, MA,
USA). These digested samples were analysed for Na, K, Ca, and Mg
(see below). For analysis of berry nutrient concentrations, frozen
(80 C) berries were ground, centrifuged to remove gross solids,
and then ltered through 0.45 m lter. The ltrate was analysed
for Na and K. All elemental analysis was performed using a Varian
SpectrAA (Varian, Palo Alto, CA, USA). Potassium and Na in digested
tissue samples was analysed using Atomic Emission Spectroscopy

95

(AES) at 766.5 nm and 589.0 nm, respectively. Caesium chloride

(CsCl) was added at a nal concentration of 0.1% to suppress ionisation. Calcium and Mg were analysed using atomic absorption
spectroscopy (AAS), at 422.7 nm and 285.2 nm, respectively. Strontium chloride (SrCl2 2H2 O) was added at a nal concentration of
0.1% to bind preferentially with phosphate if present. Samples were
diluted as necessary to ensure that readings were within the linear
response range.
2.5. Berry quality analysis
Titratable acidity (TA), pH, and total soluble solids (TSS) were
determined on the day of cluster/berry sampling, from freshly
pressed berries. TA (expressed as g tartaric acid L1 ) was determined by manual titration against 0.1 N NaOH, pH via Accumet
AP72 handheld probe (ThermoFisher Scientic Inc.) and TSS
( Brix) using a PAL-1 handheld refractometer (Atago, Japan). For
analysis of total phenolics, anthocyanins, tannins, and NOPA,
1015 frozen (80 C) berries were placed in a liquid nitrogen cooled mortar and pestle, and powdered. Samples were
ground to create a homogenate, which was used to make a 10%
(w/v) solution in 60% ethanol. This homogenate solution was
centrifuged, ltered through a 0.45 m lter, and the ltrate
analysed spectrophotometrically using the Spectronic-BioMate 3
UV-Visible Spectrometer (ThermoFisher Scientic Inc.). Total phenolics were determined using the Folin-Ciocalteu method (Slinkard
and Singleton, 1977), measuring absorbance at 765 nm. Total
anthocyanins were determined using the pH differential method
(Lee et al., 2005), measuring absorbance at 520 nm and 700 nm of
pH 1.0 and 4.5 solutions. Anthocyanins were expressed as malvidin3-glucoside, using extinction coefcient 28,000 L cm1 mol1 and
molecular weight 493.3 g mol1 . Tannin analysis was determined
using the methyl cellulose perceptible (MCP) assay (Sarneckis
et al., 2006), which measures the tannins precipitated by MCP at
280 nm. NOPA was analysed (Dukes and Butzke, 1998) by measuring absorbance at 335 nm following derivatisation of primary
amino groups with an o-phthaldialdehyde/N-acetyl-l-cysteine
(OPA/NAC) reagent.
2.6. Soil sampling and analysis
Soil samples were collected from 020 cm and 2040 cm depths
using a hand auger on August 11th (vraison) and September 17th
(harvest). Two soil cores (5 cm diam.) per depth were sampled from
the drip zone and directly underneath each emitter in two of the
four replicate blocks and combined (n = 2 per treatment). Due to
limited availability of sample vines, sampling extent was minimized to decrease frequency of soil disturbance in the drip zones.
Soils were air dried, and then extracted using the saturated paste
method (United States Salinity Laboratory, 1953). Briey, soil samples were sieved (2 mm), and then mixed with DDI H2 O until a
saturated paste was formed. Pastes were left for 12 h to equilibrate, and the saturated solution was then removed from the soil
under vacuum, and the pH and electrical conductivity (EC) measured (Thermo Orion 4 Star; Thermo Fisher Scientic Inc.). Samples
were acidied to pH 2.0 and stored at 4 C prior to ion chromatography (IC) analysis for K+ , Na+ , NH4 + , Ca2+ , and Mg2+ . Ion
chromatography analysis was performed using a Dionex ICS-2000
Ion Chromatograph tted with an IonPacCG17 column (Dionex,
Sunnyvale, CA, USA).
2.7. Statistical analysis
All statistical analysis was performed using SPSS 19.0 (Statistical Package for the Social Sciences, Chicago, IL, USA). Soil
cation concentrations were analysed using a three-way analysis of

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K.P.M. Mosse et al. / Agricultural Water Management 123 (2013) 93102

variance (ANOVA). Berry, leaf, and petiole parameters were all

analysed using an initial three-way mixed model, with time and
treatment as xed effects and block as a random effect; under
this model, block was not signicant under any conditions. A twoway mixed-model (as above without block) was subsequently
performed to consider time treatment interactions. To further
explore patterns in the data, a series of one-way ANOVAs were performed at each individual time-point, and, where the ANOVA was
signicant (P 0.05), Tukeys HSD (Honestly Signicant Difference)
test was used to determine between treatment differences.
3. Results and discussion
Application of Na- and K-rich simulated WWs as irrigation
waters over a single season affected nutrient concentrations of
leaves and petioles, as well as certain berry quality parameters.
The majority of these differences are slight, similar to Neilsen et al.
(1989) who examined municipal wastewater irrigation of young
Okanagan Riesling grapes.
3.1. Soil cations
Addition of Na and K treatments resulted in elevated salt concentrations in soil, consistent with the salt treatments applied
(Fig. 1, Table 1). Soil Na concentrations were approximately 24and 11-times higher (020 cm and 2040 cm, respectively) in the
Na treatment than the control at vraison, and 21- and 34-times
higher at harvest. Soil K concentrations varied among the three K
treatments (low K, high K, and high K + W), with vraison K treatments 32-, 31-, and 71-times higher in the 020 cm depth for low
K, high K and high K + W, respectively, and 2-, 4-, and 36-times
higher in the 2040 cm depth. Both K and Na levels showed a signicant time treatment depth interaction; soil Na levels were
comparatively higher in the 020 cm depth, at harvest, and in the
Na treatments. Soil K concentrations showed the same trends, being
comparatively higher in the 020 cm depth, at harvest, and in all
of the K treatments (Fig. 1). Quayle et al. (2010) also found that
K accumulation in a clayey soil (5060%) was restricted to the
upper soil depths after four years of irrigation with diluted and
undiluted WW from a working winery. K concentrations in the
WW were similar to the current study. The high K treatment had
lower soil K concentrations than the low K treatment. Potential
causes include K-xation in the interlayers of clay minerals (e.g.,
vermiculite), perhaps resulting from the high K treatment meeting threshold K values which result in layer collapse and xation,
and vine K uptakeSoil Ca and Mg concentrations were also affected
by irrigation treatments, although these effects were less distinct
(Fig. 1, Table 1). Soil Ca concentrations were 24-fold greater in the
K treatments than the control, at both vraison and harvest (Ca:
time treatment depth, P < 0.05). Soil Mg concentrations were 2to 5-fold greater in all treatments compared to the control, at both
time points. In the case of Mg, only the depth treatment and the
time treatment interaction terms were signicant.
Based primarily on cation valence and hydrated radius, the
predicted selectivity sequence, or lytropic series, for the cations
of interest to this study is as follows: Ca2+ > Mg2+ > K+ > Na+
(Helfferich, 1962). It is therefore largely expected that Mg and Ca
would not be readily displaced from soil cation exchange complexes by the monovalent Na and K ions; however, displacement
appears to have occurred. The above selectivity sequence does not
consider specic binding sites on mineral surfaces and therefore
some deviation from the predicted trends are not surprising. Cation
exchange on clay minerals that does not follow the above lytropic
series has been well documented (Jardine and Sparks, 1984; Endo
et al., 2002; Kopittke et al., 2006). The observed increase in Mg and

Ca concentrations in soil extracts following irrigation with K-rich

water can be attributed to soil mineralogy, the presence of external
(vs. internal) surface on clay minerals, surface potential (i.e., diffuse
double layer model), and ionic strength (Shainberg et al., 1980). In
addition to chemical exchange of monovalent cations for divalent
cations, the increase in Ca and/or Mg could also result from diffusion from soil micropores. The greater exchange ability (binding
afnity) of K, over Na observed in this study, is in agreement with
a recent study investigating the application of WWs with elevated
Na and K (Laurenson et al., 2011).
Examining the soil data by depth highlights a difference in the
mobility of K and Na (Fig. 1, Table 1). Elevated Na levels, from
Na treatment (vraison and harvest), are observed in both surface
(020 cm) and subsurface (2040 cm) samples. In high and low K
treatments, accumulation of K is observed only in surface samples, suggesting stronger interaction and binding of K in the soil.
When wine is added to the irrigation supply, there appears to be
facilitated transport of K, presumably through interaction with dissolved organic compounds (e.g., polyphenolics), as K accumulates
in both the surface and subsurface samples. This result suggests
that irrigation with organic-rich WW may reduce accumulation of
K in soils and could have important implications for the sustainability of such irrigation practices. The high K + W treatment also had
low juice K at harvest (Fig. 2).
3.2. Leaf and petiole nutrient analysis
The concentrations of Ca, K, and Mg in vine leaf and petiole tissue were comparable to previous reports (Tables 2 and 3; Downton,
1985; Stevens et al., 2011). Vine leaf and petiole Na concentrations
were considerably lower than what is often reported (Downton,
1985; Prior et al., 1992), but are comparable to some values
recorded in studies that show high variability in Na concentrations
(Walker et al., 2010; Bramley et al., 2011). Na in both leaf and petiole tissue and Ca in leaf tissue had a signicant time treatment
interaction (Tables 2 and 3). Petiole tissue showed differences in
concentrations of Ca at vraison and harvest, and in K and Na at
harvest only. Application of the high K treatment decreased petiole
Ca concentrations (ca. 2%) in comparison with the control, at both
vraison and harvest. Petiole K concentrations at harvest were 2to 3-fold higher in both low K and high K treatments than the control, although the low K and high K treatments did not differ from
each other. Petiole Na concentrations at harvest in the Na treatment
were approximately 4- to 7-fold higher than other treatments.
Although Na concentrations differed among treatments prior
to irrigation and at harvest, leaf Na concentration in the Na treatment was approximately 3-fold higher than the control at harvest
(Table 3). The Na treatment had the greatest impact on leaf and
petiole nutrient content (Tables 2 and 3), resulting in increased Na
concentrations in both petioles (0.092% DW in Na treatment, 0.013%
DW in control) and leaves (0.015% DW in Na treatment, 0.005%
DW in control) at harvest; grapevine Na uptake is typically proportional to the concentration of salt in the root zone (Stevens and
Walker, 2002). The magnitude of the increase was larger in petioles
than leaves, consistent with other studies suggesting that petiole
nutrient concentrations are a better indicator of vine nutrient status than leaf nutrient concentration (Downton, 1977; Hepner and
Bravdo, 1985). While Na accumulation patterns can vary dramatically among scions and rootstocks (e.g., 3309C is sensitive to salinity
where Ramsey is more tolerant; Zhang et al., 2002; Hepaksoy et al.,
2006; Tregeagle et al., 2006), the majority of Na uptake typically
occurs between vraison and harvest (Downton, 1977), as observed
in the current study. For comparison, Sultana grapevines irrigated
for six years with water ranging from 035 mM Na resulted in petiole Na concentrations of approximately 0.4% DW in the control to
1.62.8% DW in the 35 mM Na treatment (Prior et al., 1992). In

K.P.M. Mosse et al. / Agricultural Water Management 123 (2013) 93102

97

Table 1
Three-way ANOVA table for cation concentrations in soilwater extract from soils treated with simulated WWs and source water (control). One, two, and three stars indicate
P < 0.05, P < 0.01, and P < 0.001, respectively. n.s. = not signicant.
Ca2+ (cmol kg1 )
Time (ti)
Treatment (trt)
Depth (dpt)
ti trt
ti dpt
dpt trt
ti trt dpt

K+ (cmol kg1 )

Mg2+ (cmol kg1 )

***

n.s.

***

***

***

***

***

***

**

***

n.s.

***

***

n.s.
n.s.

***

***

n.s.

***

***

**

***

n.s.

***

**

Colombard grafted to Ramsey rootstock, leaf Na concentrations

at harvest increased steadily over a six-year period, affected by the
timing of saline water application (35 mM Na). Leaf Na concentrations ranged between approximately 0.03% DW (control) and 0.08%
DW (saline irrigation during full bloom to vraison and vraison
to harvest, respectively) after the rst season, reaching concentrations of up to 0.6% DW (full bloom to vraison and vraison
to harvest irrigation periods) after six years of irrigation (Stevens
et al., 2011). Observed increases in Na concentrations in the grape
tissue are consistent with earlier reports, and differences in magnitude may be due to variation in rootstock/scion combinations,
which can affect Na accumulation (Paranychianakis and Angelakis,
2008).
Irrigation of vines with K-rich WWs had varied inuence on
vegetative tissue nutrient composition (Tables 2 and 3). Petiole K
concentrations were signicantly elevated in both high K and low
K treatments at harvest (0.4% in control, 1.1% in low K, 1.4% in high
K). This is consistent with previous work, in which application of

Na+ (cmol kg1 )

450 kg K ha1 to Concord (i.e. ve times lower than used here),

resulted in 3-fold elevated petiole K concentrations (from ca. 1.5%
in control to 5.0% in K treatment; Morris and Cawthon, 1982). Tissue
K accumulation with respect to application rate is greater than the
current study, and is likely due to differences in grape cultivar and
soil type. Interestingly, petiole K concentrations in the high K + W
treatment did not differ from the control. Given that the high K + W
treatment had the highest K concentrations of all treatments, and
resulted in higher concentrations of K in the soil, it suggests that
the organic matter present in the wine may have decreased the
amount of plant-available K (see Section 3.1) (Zhang et al., 2009).
Reduced petiole Ca concentration at vraison and harvest occurred
in the high K + W treatment (P < 0.05). Further, the K treatments
had elevated soil Ca concentrations at harvest (P < 0.05), suggesting
that selective Ca exclusion/K uptake mechanisms may have been
involved. Such antagonistic uptake between K and Ca has a been
reported previously with K application rates of 900 kg ha1 (Morris
et al., 1980).

Fig. 1. Cation concentrations in soilwater extract from soils treated with simulated WWs and source water (control). Data plotted are means and error bars represent the
standard error (n = 2).

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K.P.M. Mosse et al. / Agricultural Water Management 123 (2013) 93102

Fig. 2. Juice/berry quality parameters for Na, K, total phenolics, and anthocyanin at (a) vraison and (b) harvest following irrigation with WWs and source water (control).
Data plotted are means and error bars represent the standard error (n = 4).

3.3. Berry quality

At harvest, most Syrah berry qualities (Fig. 2), with the exception of Na, total phenolics and tannins, were comparable to
what has been previously reported (Butzke, 1998; Walker et al.,
1998; Mattivi et al., 2002; Ristic et al., 2007; Ginjom et al.,
2010), although it should be noted that scion/rootstock, vintage,

growing region, etc. can have signicant inuences on reported

values.
There were minor differences in juice inorganic composition
among the irrigation treatments at vraison and harvest (Fig. 2).
Overall, the treatments had greater impact on juice Na at harvest than at vraison. Juice Na concentrations at harvest were 1.7
fold greater in the Na than high and low K treatments, with no

K.P.M. Mosse et al. / Agricultural Water Management 123 (2013) 93102

99

Table 2
Leaf nutrient concentrations in vines irrigated with simulated WWs and source water (control). Values displayed are means, and values in parentheses are standard error
(n = 4). Means followed by the same letter are not signicantly different (P < 0.05) based on one-way ANOVA followed by Tukeys HSD test. One, two, and three stars indicate
P < 0.05, P < 0.01, and P < 0.001, respectively. n.s. = not signicant.
Time

Treatment

Ca2+ (% dw)

K+ (% dw)

Mg2+ (% dw)

Na+ (% dw)

Pre-Irrigation

Control
High K
High K + W
Low K
Na

0.98 (0.02)
1.02 (0.09)
1.22 (0.06)
1.20 (0.02)
1.18 (0.06)

n.s.

1.30 (0.44)
0.82 (0.01)
0.84 (0.05)
0.90 (0.02)
0.96 (0.02)

n.s.

0.47 (0.04)
0.51 (0.01)
0.49 (0.03)
0.48 (0.01)
0.47 (0.00)

n.s.

0.013 (0.001)
0.013 (0.001)
0.011 (0.000)
0.011 (0.000)
0.010 (0.000)

ab
b
ab
ab
a

Control
High K
High K + W
Low K
Na

1.71 (0.08)
1.69 (0.04)
1.59 (0.04)
1.75 (0.04)
1.59 (0.07)

n.s.

0.51 (0.04)
0.52 (0.03)
0.50 (0.03)
0.53 (0.02)
0.55 (0.03)

n.s.

0.80 (0.04)
0.84 (0.01)
0.68 (0.12)
0.83 (0.03)
0.74 (0.02)

n.s.

0.008 (0.001)
0.007 (0.000)
0.006 (0.000)
0.007 (0.000)
0.007 (0.001)

n.s.

Control
High K
High K + W
Low K
Na

2.38 (0.05)
2.24 (0.06)
2.32 (0.07)
2.15 (0.05)
2.11 (0.12)

n.s.

0.35 (0.01)
0.33 (0.01)
0.32 (0.01)
0.34 (0.01)
0.26 (0.05)

n.s.

1.04 (0.03)
1.04 (0.03)
1.05 (0.05)
1.07 (0.03)
1.03 (0.05)

n.s.

0.005 (0.000)
0.007 (0.001)
0.006 (0.000)
0.005 (0.000)
0.015 (0.002)

a
a
a
a
b

Time (ti)
Treatment (trt)
ti trt

***
n.s.
*

Vraison

Harvest

Two-way ANOVA

***
n.s.
n.s.

***
n.s.
n.s.

***
***
***

Table 3
Petiole nutrient concentrations in vines irrigated with simulated WWs and source water (control). Values displayed are means, and values in parentheses are standard
error (n = 4). Means followed by the same letter are not signicantly different (P < 0.05) based on one-way ANOVA followed by Tukeys HSD test. One, two, and three stars
indicate P < 0.05, P < 0.01, and P < 0.001, respectively. n.s. = not signicant.
Time

Treatment

Ca2+ (% dw)

K+ (% dw)

Mg2+ (% dw)

Na+ (% dw)

Pre-Irrigation

Control
High K
High K + W
Low K
Na

0.93 (0.06)
1.02 (0.08)
0.99 (0.02)
0.94 (0.03)
0.96 (0.09)

n.s.

2.22 (0.13)
2.26 (0.46)
2.53 (0.10)
1.85 (0.09)
2.60 (0.10)

n.s.

1.10 (0.20)
1.01 (0.17)
1.19 (0.09)
0.87 (0.33)
1.27 (0.08)

n.s.

0.013 (0.001)
0.015 (0.001)
0.013 (0.001)
0.012 (0.000)
0.012 (0.001)

n.s.

Control
High K
High K + W
Low K
Na

1.23 (0.02)
1.09 (0.06)
1.20 (0.02)
1.19 (0.04)
1.16 (0.02)

b
a
ab
ab
ab

0.85 (0.24)
1.24 (0.13)
0.77 (0.03)
1.29 (0.31)
1.08 (0.13)

n.s.

1.89 (0.08)
1.98 (0.09)
1.96 (0.05)
1.93 (0.04)
1.83 (0.04)

n.s.

0.012 (0.002)
0.009 (0.001)
0.012 (0.001)
0.012 (0.001)
0.016 (0.002)

n.s.

Control
High K
High K + W
Low K
Na

1.55 (0.04)
1.20 (0.10)
1.40 (0.06)
1.38 (0.03)
1.37 (0.05)

b
a
ab
ab
ab

0.40 (0.08)
1.40 (0.25)
0.93 (0.19)
1.10 (0.06)
0.63 (0.02)

a
c
abc
bc
ab

2.52 (.010)
2.24 (0.05)
2.49 (0.08)
2.30 (0.08)
2.36 (0.05)

n.s.

0.013 (0.001)
0.022 (0.002)
0.019 (0.001)
0.020 (0.002)
0.092 (0.009)

a
a
a
a
b

Time (ti)
Treatment (trt)
ti trt

***
**
n.s.

Vraison

Harvest

Two-way ANOVA

difference among remaining treatments. As with leaf and petiole

Na status, juice Na concentrations were at the very low end of
the documented spectrum (Walker et al., 2010), but the relative
differences among treatments remain of interest. Juice K concentrations varied among treatments, with the relative concentrations
at harvest being high K + W < control < low K < high K < Na (in the
order of low to high; 1.3-fold difference). Application of K leads to
greater availability of soil K, and therefore higher levels of plant
K uptake can occur. The elevated juice concentrations of K in the
Na treatment suggests that a different mechanism is involved, in
which K is selectively taken up in greater amounts to counteract
the effects of greater Na uptake. There are a range of transport
proteins involved in uptake of Na and/or K, and it has been suggested that plants may be able to regulate specic K/Na uptake in
order to maintain Na:K homeostasis (Schachtman and Liu, 1999).
At vraison, juice K concentrations were higher in the low K treatment compared with the control. Whilst elevated juice K following
K application is to be expected (Morris and Cawthon, 1982; Ruhl
et al., 1992), that the effect was only seen in the lower K treatment
is surprising. However, these differences did not persist at the time

***
*
n.s.

***
n.s.
n.s.

***
***
***

of harvest. In general, berry K levels were comparable to previous

reports (Walker et al., 1998).
Total phenolic concentrations in this study (42256062 mg kg1
at vraison and 48937997 mg kg1 at harvest as epicatechin
equivalents) were typically higher than what has been reported in
other studies (Fig. 2). Other studies reported total phenolic concentrations of 15001800 mg kg1 catechin equivalents (Italy; Mattivi
et al., 2002) and 12502130 mg kg1 as tannic acid equivalents
in Syrah grapes (South Africa; Ozden et al., 2010). Syrah wine
(n = 16) studies in France and Australia reported total phenolic concentrations of 16502590 mg L1 as gallic acid (Landrault et al.,
2001; Ginjom et al., 2010). The phenolic concentrations reported
here and elsewhere may reect differences in extraction techniques, sample form, and standard used to express these values. The
relative differences among treatments remain valid nonetheless, as
the same extraction conditions were utilised across all treatments
tested in this study. Total phenolics were signicantly elevated in
the Na treatment at harvest, although the specic phenolic groups
(tannins and anthocyanins) were not affected (Fig. 2). In contrast,
no relationship between juice Na and phenolic concentration was

100

K.P.M. Mosse et al. / Agricultural Water Management 123 (2013) 93102

detected in wine produced from Syrah exposed to differing salinity conditions (Australia; Walker et al., 2000). Increased phenolic
concentration is typically associated with increased wine quality,
and therefore the application of Na salts over one growth season appears to not be problematic from this perspective. In the
same vein, the ratio of TSS to TA increased in Cabernet Sauvignon on Ruggeri rootstock that was irrigated with saline water
for one growth season, suggesting an increase in berry quality; phenolic concentrations were not measured (Hepaksoy et al.,
2006).
Anthocyanin was elevated in high K + W treatment at vraison
compared to high K and low K; at harvest, high K + W anthocyanin levels were not signicantly different from other irrigation
regimes (Fig. 2). This distinction in anthocyanin values among treatments at vraison but not at harvest suggests a slight delay in
color development due to these two treatments (high K and low
K), denoting that either the 2.5% wine provided constituents that
encouraged coloration, or that the differences were exacerbated
by the imprecise nature of the vraison time-point and was not
the result of berry size difference (not signicantly different at
vraison). Tannin concentrations did not differ among at vraison 352429 mg kg1 ) and harvest (405619 mg kg1 , indicating
that one season of irrigation with simulated WWs did not affect
this phenolic group. These tannin concentrations at harvest were
within the lower levels of a previously reported tannin range for
Syrah wines (4004900 mg epicatechin L1 , n = 1107; Smith et al.,
2010).
All fruit maturity indexes (TA, pH, and TSS) were not altered
due to these WW treatments, at either sampling time-points
(Fig. 2). In comparison, juice pH and TSS (slight increase compared to control) were the only quality factors altered in young
Okanagan Riesling grapes irrigated for three years with municipal wastewater (Neilsen et al., 1989). Grape NOPA concentrations
were >140 mg kg1 across all treatments at harvest and were adequate for a healthy fermentation (Agenbach, 1977; Butzke, 1998),
and the WW treatments did not negatively inuence grape NOPA.
At harvest, berry weight was higher in the Na treatment than in
the low K treatment (1.1-fold), and harvest weights were 1.2-fold
lower in the low K treatment in comparison with K + W, control
and Na treatments. The magnitudes of these differences are very
slight, and it is difcult to reach any rm conclusions at this stage.
There was no difference in cane diameter and length, measured at
harvest.
The ndings in the current preliminary study indicate that little to no impact on grapes occurred after one season of irrigation
with these simulated WW designed to address changes in industrial cleaning regimes. Clearly, other WW components such as
inorganic nitrogen, organic carbon species (e.g., alcohols, organic
acids), chemical oxygen demand (COD), macronutrients (e.g., calcium, magnesium) and micronutrients (e.g., zinc, boron) (Chapman,
2001; Bustamante et al., 2005; Quayle et al., 2010; Sheridan et al.,
2011), and the timing of such inputs (e.g., pulse of K at crush;
Sheridan et al., 2011) potentially could inuence vine growth.
Nor did it address the effects of shifting the irrigation levels,
though grapevine sensitivity to salts increases with increasing
water decit and decreasing irrigation level (i.e., 100% ET, 75%
ET, 50% ET) (Paranychianakis et al., 2004; Paranychianakis and
Angelakis, 2008). In Soultanina grafted to various rootstocks, leaf
injuries were greater in vines irrigated at 50% ET than 75% and 100%
ET, suggesting that despite less Na applied overall in the 50% ET
treatment, this water decit and resultant water stress exacerbated
effects of the salt stress. These ndings indicate that the current
studys irrigation level (i.e., 70% ET) may not have been sufciently
severe enough to incur physical symptoms of vine stress due to
high salt application (see McCarthy, 1981, McCarthy and Downton,
1981).

4. Conclusions
In an established twelve-year old Syrah vineyard, most of the
simulated WWs did not inuence the majority of vine growth
and juice/berry quality measurements, relative to the control, after
a single growth season except for WW containing 40 mM Na.
This suggests that application of these simulated WW treatments
focusing on shifts in salt composition and concentration due to
changes in industrial cleaning regimes does not have large immediate impacts on vine health or wine quality over a single growing
season. As grapevines are a long-lived perennial species with signicant root nutrient reserves (Schreiner and Scagel, 2006; Kodur
et al., 2010a,b), this study will continue over an extended time
period to ascertain whether winter rainfall is adequate to leach
increased salts from the soil prole, and whether new growth is
affected in subsequent growing seasons. Finally, generation of WW
often occurs during the grapevines dormant period, and thus its
application to other cropping systems is a necessity to respond to
future projections of limited water resources for irrigation.

Acknowledgements
This project was nancially supported by Australian grape
growers and winemakers investment body, the Grape and Wine
Research and Development Corporation, Australian Government,
the Cooperative Research Centre for Irrigation Futures, United
States Fulbright Postgraduate Scholarship, Monash Research Accelerator Scheme, the Kearney Foundation of Soil Science (number
2009.011), and USDA-Agricultural Research Service (ARS) CRIS #
530621220-004-00D, and # 535821000-041-00D. The authors
wish to thank Dr. Stuart Pettygrove (UC Davis) and Ms. Maya
Buelow (UC Davis) for valuable discussions and eld assistance in
this study, and Constellation Wines U.S. for their generous donation of the wine used in this study. Mention of trade names or
commercial products in this publication is solely for the purpose
of providing specic information and does not imply recommendation or endorsement by the U.S. Department of Agriculture.

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