remote sensing
Article
The Efficiency of Foliar Kaolin Spray Assessed through
UAV-Based Thermal Infrared Imagery
Luís Pádua 1, * , Sara Bernardo 1 , Lia-Tânia Dinis 1 , Carlos Correia 1 , José Moutinho-Pereira 1
and Joaquim J. Sousa 2,3
Citation: Pádua, L.; Bernardo, S.;
Dinis, L.-T.; Correia, C.;
Moutinho-Pereira, J.; Sousa, J.J. The
Efficiency of Foliar Kaolin Spray
Assessed through UAV-Based
Thermal Infrared Imagery. Remote
Sens. 2022, 14, 4019. https://
doi.org/10.3390/rs14164019
Academic Editor: Konstantinos
X. Soulis
Received: 15 July 2022
Accepted: 16 August 2022
Published: 18 August 2022
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Remote Sens. 2022, 14, 4019. https://doi.org/10.3390/rs14164019
1 Centre for the Research and Technology of Agro-Environmental and Biological Sciences (CITAB),
University of Trás-os-Montes e Alto Douro, 5001-801 Vila Real, Portugal
2 School of Science and Technology, University of Trás-os-Montes e Alto Douro, 5000-801 Vila Real, Portugal
3 Centre for Robotics in Industry and Intelligent Systems (CRIIS), INESC Technology and Science (INESC-TEC),
4200-465 Porto, Portugal
* Correspondence: luispadua@utad.pt
Abstract: The water content in an agricultural crop is of crucial importance and can either be
estimated through proximal or remote sensing techniques, allowing better irrigation scheduling and
avoiding extreme water stress periods. However, the current climate change context is increasing the
use of eco-friendly practices to reconcile water management and thermal protection from sunburn.
These approaches aim to mitigate summer stress factors (high temperature, high radiation, and
water shortage) and improve the plants’ thermal efficiency. In this study, data from unmanned aerial
vehicles (UAVs) were used to monitor the efficiency of foliar kaolin application (5%) in a commercial
vineyard. Thermal infrared imagery (TIR) was used to compare the canopy temperature of grapevines
with and without kaolin and to compute crop water stress and stomatal conductance indices. The gas
exchange parameters of single leaves were also analysed to ascertain the physiological performance
of vines and validate the UAV-based TIR data. Generally, plants sprayed with kaolin presented a
lower temperature compared to untreated plants. Moreover, UAV-based data also showed a lower
water stress index and higher stomatal conductance, which relate to eco-physiological measurements
carried out in the field. Thus, the suitability of UAV-based TIR data proved to be a good approach to
monitor entire vineyards in regions affected by periods of heatwaves, as is the case of the analysed
study area.
Keywords: unmanned aerial vehicles; precision viticulture; remote sensing; geographical information
systems; summer stress; climate change
1. Introduction
Canopy and cluster sunburn is an unfortunate consequence of climate change in
European viticulture due to the rising incidence of extreme weather events (i.e., heatwaves)
throughout the growing season [1,2]. In Mediterranean wine-growing regions, which
were recently considered climate change hotspots, grapevine sunburn results from the
exposure to excessive photosynthetically active radiation (PAR), high temperature, and
ultraviolet (UV) radiation, hampering vines0 growth, yield, and grape quality potential [3,4].
The interaction of these environmental stress factors, together with canopy management
practices which modulate light interception, script the damage of sunburn in grapevines
throughout the summer season, characterised by leaf browning and shedding, decreased
source and sink capacity, and berry shrivelling [1,5].
Adapting Mediterranean viticulture to climatic challenges has driven the need for
the development and selection of eco-friendly strategies to bypass potential yield and
quality losses [6]. In the last few decades, the foliar application of white clay minerals,
such as kaolin (Al2 Si2 O5 (OH)4 ), has been considered an efficient measure to avoid
sunburn damage in several crops, improving the physiological performance of plants
https://www.mdpi.com/journal/remotesensing
Remote Sens. 2022, 14, 4019 2 of 18

and their ability to withstand adverse summer stress conditions [7–10]. A recent body
of literature has reported various consecutive effects driven by the reflective properties
of the kaolin-particle film in grapevines, ranging from a significant reduction in leaf
temperature, higher photosynthetic activity, and water use efficiency (WUE), to slight
changes in yield, berry size, and acidity traits [11–14]. For example, Dinis et al. [15] and
Bernardo et al. [16] demonstrated the leaf cooling effect promoted by the kaolin-particle
film in Touriga Nacional grapevines through single leaf temperature measurements in
the Douro Region (Portugal) in two consecutive growing seasons, along with improved
WUE and physiological performance. The benefits of this treatment were also observed
in several other crops, such as nuts [17,18], apple trees [19], coffee [20], wheat [21],
and papaya [22] plants. However, most studies regarding kaolin spraying effects in
vineyards showed increased efficiency of this protector under severe environmental
conditions [11,23]. In Italy, other authors have also found that kaolin particle-film can
increase the foliar reflection of PAR and UV radiation of Cabernet Sauvignon [24] and Pinot
Noir [13] under adverse stress conditions. More recently, the association between kaolin
treatment and the application of colourful shading nets has shown significant effects
in Sauvignon Blanc grapevines during warm summers in preserving vine water status,
delaying the technological maturity of grapes and preventing photo-inhibition [25].
Moreover, kaolin0 s effects in avoiding photo damage by optimizing chlorophyll and
xanthophyll cycles and improved heat dissipation mechanisms have also been observed
in Mediterranean-type climate regions [26,27].
Despite the effectiveness of kaolin coating to reduce heat load in red and white
grapevine varieties across several terroirs, the assessment of some parameters could be
optimised by emerging precision technologies applied in viticulture [28].
The employment of precision viticulture (PV) allows for monitoring of the spatial
vineyard variability [29]. This variability is reflected in the grapevine (Vitis vinifera
L.) biophysical parameters [30]. Thus, through the analysis of these parameters, it is
possible to identify the spatial variability and enable the implementation of the best
management for each part of the vineyard [31]. In this context, remote sensing platforms
are widely used in PV, providing data for decision support [32,33]. Among the different
remote sensing platforms, unnamed aerial vehicles (UAVs) showed a rapid proliferation,
especially in the last decade. UAVs stand out due to their great flexibility in the use
of several types of sensors and the high temporal and spatial resolution [34,35]. In
the context of viticulture, most applications and studies that use high resolution UAV
data use cost-effective RGB sensors to compute three-dimensional and orthorectified
information by means of photogrammetric processing. This approach allows for the
gathering of 2D orthorectified outcomes and 3D information, allowing the retrieval of
geometric-related grapevine features both at the plant and/or plot level (e.g., height,
width, length, area, and volume) [36–38]. Automatic or semi-automatic classification
of vineyards [39–41] through grapevine segmentation processes [37,42,43] are other
important results that this approach allows us to achieve. However, when the goal is
to estimate or determine plant-related biophysical parameters, the use of multispectral
or hyperspectral data may bring great advantages. It becomes possible to estimate
parameters such as leaf area index, vegetative vigour, chlorophyll concentration, and
yield [31,44–46]. Furthermore, the use of multi and hyperspectral data can also assist in
disease detection tasks [47,48]. All these advances provide knowledge at the plot level
that allows for management oriented to the optimization of resources while allowing for
the improvement of production and quality. By adding thermal data to this equation, a
different level will be reached, namely in terms of optimising the increasingly important
resource in the context of climate change: water. UAV-based thermal infrared imagery
(TIR), can be used in PV to obtain grapevine temperature and estimate water status
through the computation of specific indices [49–55].
 commercial vineyard from the Douro Demarcated Region was surveyed with the goal of
mapping the thermal variability within grapevines form vine rows with kaolin and
comparing them with adjacent rows without kaolin, serving as a control. The study
Remoteexplores an automatic approach to extracting grapevine TIR data, shows the3 ofleaf
Sens. 2022, 14, 4019 18

protective effects of kaolin through time in the presence of environmental stress, and the
relationship of the With estimated UAV‐based parameters with eco‐physiological
this study, we demonstrate the added value of using UAV-based imagery,
measurements. namely TIR and RGB data, to monitor the effects of kaolin foliar applications. A com-
mercial vineyard from the Douro Demarcated Region was surveyed with the goal of
mapping the thermal variability within grapevines form vine rows with kaolin and com-
2. Materials and Methods
paring them with adjacent rows without kaolin, serving as a control. The study explores
2.1. Study Area and an
Experiment Characterization
automatic approach to extracting grapevine TIR data, shows the leaf protective effects
of kaolin through time in the presence of environmental stress, and the relationship of
A general overview of theUAV-based
the estimated vineyardparameters
analysedwithin eco-physiological
this study, located within the Douro
measurements.
Demarcated Region (São João da Pesqueira, Viseu, Portugal), is shown in Figure 1. It is a
2. Materials and Methods
commercial vineyard located in Quinta dos Aciprestes (41°12′28.9″N, 7°26′07.1″W) with
2.1. Study Area and Experiment Characterization
an area of approximately 5 ha,overview
A general and is ofcomposed
the vineyardof red grapevines
analysed (cv. Touriga
in this study, located within theNacional
Douro
grafted on 1103‐P rootstock). The (São
Demarcated Region plants
João are arranged
da Pesqueira, in Portugal),
Viseu, rows spaced is shown byin2.2 m,1.with
Figure It is a a
commercial vineyard located in Quinta the
dos Aciprestes ◦ 0 00 ◦
(41 12 28.9 N,it7 was 0
26 07.1 00 W) withat
distance between plants of one meter. Regarding kaolin application, applied
an area of approximately 5 ha, and is composed of red grapevines (cv. Touriga Nacional
a concentration of 5% over the whole canopy on 18 July 2018. Between the rows in which
grafted on 1103-P rootstock). The plants are arranged in rows spaced by 2.2 m, with a
kaolin was applied,distance
a set ofbetween
control rows
plants were
of one leftRegarding
meter. withoutthe any application.
kaolin application,From it was the 5 ha
applied
vineyard, a total of 3.9 ha were analysed (green and blue polygons in Figure 1), with in
at a concentration of 5% over the whole canopy on 18 July 2018. Between the rows 1.9
which kaolin was applied, a set of control rows were left without any application. From
ha containing kaolin‐treated grapevines and the remaining 2 ha, serving as a control,
the 5 ha vineyard, a total of 3.9 ha were analysed (green and blue polygons in Figure 1),
having untreated plants.
with 1.9 No other changes
ha containing in irrigation
kaolin-treated grapevinesorandtreatments
the remaining between
2 ha, servingtheastwoa
control, having untreated plants. No other changes in irrigation
trial setups were applied. Vine rows not marked in Figure 1 were not subjected to further or treatments between
the two trial setups were applied. Vine rows not marked in Figure 1 were not subjected
comparison. to further comparison.

Figure 1. Overview Figure

of the1. surveyed
Overview ofvineyard and
the surveyed the parts
vineyard analysed.
and the Background
parts analysed. Backgroundimage from
image from
Google Earth. Google Earth.

A weather contextualization is provided in Figure 2 (precipitation, evapotranspiration,

A weather and contextualization is provided
mean, minimum, and maximum in Figure
air temperatures) 2 (from
both monthly (precipitation,
November
evapotranspiration,2017
andto mean,
October minimum, and
2018) and daily maximum
(between airanalysed).
the period temperatures)
Data wereboth monthly
obtained from
the E-OBS gridded dataset v25.0e [56] by providing the central
(from November 2017 to October 2018) and daily (between the period analysed). Data latitude and longitude
coordinates of the vineyard.
were obtained from the E‐OBS gridded dataset v25.0e [56] by providing the central
latitude and longitude coordinates of the vineyard.
Remote Sens. 2022, 14, 4019 4 of 18
Remote Sens. 2022, 14, x FOR PEER REVIEW 4 of 18

Figure 2. Monthly mean weather for the study area between November 2017 and October 2018 (a)
Figure 2. Monthly
and daily mean
weather data forweather forofthe
the period thestudy area between
field study (b). MeanNovember 2017 and
(Tmean), minimum (TminOctober 2018
) and max‐
(a)imum
and daily
(Tmax)weather data for the
air temperatures, period of and
precipitation the field study (b). Mean
evapotranspiration (ET(T
0) mean ), minimum (Tmin ) and
values.
maximum (Tmax ) air temperatures, precipitation and evapotranspiration (ET0 ) values.
2.2. Remote Sensed Data
2.2. Remote Sensed Data
2.2.1. Data Acquisition
2.2.1. Data Acquisition
The vineyard was surveyed using two different unmanned aerial systems: the multi‐
The vineyard was surveyed using two different unmanned aerial systems: the multi-
rotor UAV Phantom 4 (DJI, Shenzhen, China), responsible for RGB data acquisition using
rotor UAV Phantom 4 (DJI, Shenzhen, China), responsible for RGB data acquisition using
its 12.4 MP sensor in the first flight campaign; and the fixed‐wing UAV eBee (senseFly SA,
its 12.4 MP sensor in the first flight campaign; and the fixed-wing UAV eBee (senseFly SA,
Lausanne, Switzerland) for TIR imagery acquisition using the thermoMAP sensor
Lausanne, Switzerland) for TIR imagery acquisition using the thermoMAP sensor (senseFly
(senseFly SA, Lausanne, Switzerland), allowing the data acquisition between 7500 nm and
SA, Lausanne, Switzerland), allowing the data acquisition between 7500 nm and 13,500 nm
13,500 nm with an image resolution of 640 × 512 and a temperature resolution of 0.1 °C.
with an image resolution of 640 × 512 and a temperature resolution of 0.1 ◦ C. The thermal
The thermal image‐based calibration is automatically performed in‐flight. Moreover, in
image-based calibration
the second flight is automatically
campaign, performed
the fixed‐wing UAV was in-flight. Moreover,
also used for RGBin theacquisition
data second flight
campaign, the fixed-wing UAV was
using a Canon IXUS 12 7 HS with 16.1 MP.also used for RGB data acquisition using a Canon IXUS
12 7 HS with 16.1 MP.
Flight campaigns were conducted using mission planning software for autonomous
Flight
flight: campaignson
DroneDeploy were conducted
an Android using mission
smartphone in theplanning software
case of the for autonomous
multi‐rotor UAV and
flight:
eMotion (senseFly SA, Lausanne, Switzerland), installed on a Windows PC, for theUAV
DroneDeploy on an Android smartphone in the case of the multi-rotor and
fixed‐
eMotion (senseFly SA, Lausanne, Switzerland), installed on a Windows
wing UAV. Two flight campaigns were conducted during the 2018 growing season, withPC, for the fixed-
wing UAV. Two
a temporal flight campaigns
separation of 19 days.were
Tableconducted
1 presentsduring the 2018 growing
the information related toseason, with
the flight
a parameters
temporal separation
and acquiredof 19 days. Table 1 presents the information related to the flight
imagery.
parameters and acquired imagery.
Table 1. Characteristics of the flight campaigns. Flight height is referred to the take‐off point for the
RGB1.sensor.
Table GSD: ground
Characteristics sample
of the flightdistance.
campaigns. Flight height is referred to the take-off point for the
RGB sensor. GSD: ground sample distance.
Survey Period 24 August 2018 12 September 2018
Sensor Period
Survey Type RGB 24 August 2018
TIR RGB12 SeptemberTIR
2018
Flight Height (m) 60 * 135 ** 135 ** 135 **
Sensor Type RGB TIR RGB TIR
No. ofHeight
Flight Images(m) 44560 * 2886
135 ** 67
135 ** 2464
135 **
Front/Side
No. of Images (%)
Overlap 80/70
445 90/70
2886 70/7067 90/70
2464
GSDOverlap
Front/Side (m) (%) 0.04
80/70 0.27
90/70 0.04
70/70 0.25
90/70
GSD (m)
* From the take‐off position; ** Following0.04 0.27
to the terrain elevation. 0.04 0.25
* From the take-off position; ** Following to the terrain elevation.
2.2.2. Data Processing
2.2.2. Data Processing
The photogrammetric processing of the UAV‐based imagery acquired from the two
The photogrammetric
sensors processing
in each flight campaign of theout
was carried UAV-based imagery acquired
using Pix4Dmapper Pro (Pix4Dfrom
SA,the two
Lau‐
sensors
sanne, in each flight campaign
Switzerland). was carried
It uses structure out using
from motion Pix4Dmapper
(SfM) Pro (Pix4D SA,
to generate high‐density Lau-
point
sanne,
cloudsSwitzerland).
that enable theIt computation
uses structure
of from motion
different (SfM) to generate
orthorectified outcomes.high-density point
The processing
applied
clouds to enable
that the datathe
starts by computing
computation a sparse orthorectified
of different point cloud, estimating
outcomes.3DThe
points from
processing
applied to the data starts by computing a sparse point cloud, estimating 3D points from
2D matching points in multiple images. Then, a geolocation correction is performed using
Remote Sens. 2022, 14, 4019 5 of 18

the ground control point (GCP) coordinates, acquired in the terrain using a global navi-
gation satellite system (GNSS) receiver. After this step, point cloud densification occurs,
by enlarging the number of 3D points. The point cloud density was set to high in all
projects. For the computation of the orthorectified outcomes, the dense point cloud was
interpolated based on the inverse distance weighting (IWD) method, and a noise filter and
surface smoothing were also applied in this step. However, depending on the source data,
different outcomes were computed. This way, when processing projects using RGB imagery,
orthophoto mosaics, digital surface models (DSMs), and digital terrain models (DTMs)
were generated. Moreover, crop surface models (CSM) were computed from the subtraction
of the DTM to the DSM as in (1). From the RGB orthophoto mosaic, the G% index [57] was
computed according to (2), being used for vegetation segmentation. The TIR data used in
this study can also be used to generate digital elevation models, but it heavily suffers from
smoothing effects due to lower resolution and a lack of textural information [58].

CSM = DSM − DTM (1)

Green
G% = (2)
Red + Green + Blue
From the acquired TIR imagery, the land surface temperature (LST) was computed.
The raw reflectance data was converted to degrees Celsius. However, temperature varies
along the day and epoch of the year and, for that reason, absolute values are useless for
multi-temporal data analysis [51]. Thus, to be able to compare results from different epochs,
it was necessary to normalise the data, which can be achieved by computing the crop water
stress index (CWSI) [59] and the stomatal conductance index (IG) [60]. While CWSI is related
to the detection of potential stress in agricultural crops, IG is directly related to stomatal
conductance [61]. The empirical estimation of these indices is presented in (3) and (4):

Tc − Twet
CWSI = (3)
Tdry − Twet

Tdry − Tc
IG = (4)
Tc − Twet
Both parameters rely on the use of upper and lower thermal limits (Twet and Tdry ),
which, respectively, correspond to the temperatures of a well-watered leaf and a non-
transpiring leaf, and which are used along with the actual grapevine canopy temperature
(Tc ). In this study, Twet and Tdry values were obtained as described in Matese and Di
Gennaro [62]. Twet values are obtained by watering grapevine leaves and immediately
measuring their temperature. Tdry is obtained by applying petroleum jelly to the leaves
directly exposed to the sun and its temperatures are measured after a few minutes. These
temperature references were acquired in the field using a handheld infrared thermometer
at the same time as the TIR flight was being conducted.
The vineyard segmentation techniques, published by Pádua et al. [42,63], were used to
isolate grapevine vegetation from soil and inter-row vegetation. As an output, the method
provides a mask of the pixels estimated as being grapevine. This mask was applied to the
LST, CWSI, and IG raster files. The open source geographical information system QGIS was
used for data processing of the photogrammetric outcomes and their subsequent analysis.

2.3. Eco-Physiological Measurements

2.3.1. Leaf Temperature
The average temperature of twenty sun-exposed and fully expanded leaves was mea-
sured at the midday period, using an infrared thermometer (Infratrace KM800S, Welwyn
Garden City, Hertfordshire, UK) with a 15◦ field view positioned approximately 1 m above
the foliar surface.
Remote Sens. 2022, 14, 4019 6 of 18

2.3.2. Leaf Gas Exchange

Leaf gas exchange measurements were performed using a portable infrared gas anal-
yser (LCpro+, ADC, Hoddesdon, UK), operated in the open mode, at the midday period
(14:00, GTM +1) in six sun-exposed and expanded leaves of the kaolin and untreated groups.
The net photosynthetic rate (A, µmol m−2 s−1 ), stomatal conductance (gs , mmol m−2 s−1 ),
transpiration rate (E, mmol m−2 s−1 ), and the ratio of intercellular to atmospheric CO2
concentration (Ci /Ca ) were estimated following the formulas proposed by von Caemmerer
and Farquhar [64]. The intrinsic water use efficiency was estimated as the ratio of A/gs to
exclude the potential effects of air humidity and temperature on transpiration [65].

2.3.3. Chlorophyll a Fluorescence

Chlorophyll a fluorescence emission was measured with a Pulse Amplitude Modula-
tion Fluorometer (mini-PAM, Photosynthesis Yield Analyzer; Walz, Effeltrich, Germany)
at midday (14:00 GTM +1) in six fully expanded leaves by two-step readings. The first
reading was recorded after a 35-s exposure to actinic light (1450 µmol m−2 s−1 ) in sunlight
exposed leaves to determine the light-adapted steady-state fluorescence yield (Fs ), followed
by exposure to a saturating light pulse (6000 µmol m−2 s−1 ) for 0.6 s to establish Fm ’. The
leaves were then shaded for 5 s with a far-red light source to determine F00 . Afterwards,
a leaf clip (DLC-8) was attached to the same leaf portion previously used to promote
dark acclimation. After 30 to 45 min, the maximum photochemical efficiency of PSII was
obtained by the relation Fv /Fm = (Fm − F0 )/Fm , where F0 corresponds to the minimum
fluorescence level excited by the very low intensity of the measuring light to keep PSII
reaction centres open, and Fm corresponds to the maximum fluorescence level elicited by a
pulse of saturating light (6000 µmol m−2 s−1 ), which closes all PSII reaction centres [66].
The non-photochemical quenching (NPQ) was then calculated according to Bilger and
Schreiber [66] and Genty et al. [67] as follows: NPQ = (Fm − Fm ’)/Fm ’.

2.3.4. Transient Chlorophyll a Fluorescence Analysis by JIP-Test Parameters

Briefly, six leaves of each sampling group were dark-adapted with clips for 30 min
before chlorophyll a fluorescence transient measurements at the midday period. Then, the
saturating pulse measurements were performed by 1 s illumination, providing a maximum
light intensity of 3000 µmol (photon) m−2 s−1 using a portable chlorophyll fluorimeter
OS-30p (Opti-Sciences Inc., Hudson, NH, USA). The fast fluorescence kinetics (F0 to Fm )
was recorded from 10 µs to 1 s. The fluorescence intensity at 50 µs were considered
as F0 . The biophysical parameters obtained were calculated according to the JIP test
equations [68,69], providing structural and functional information regarding photosystem
II (PSII): (i) specific energy fluxes per reaction centre (RC)–absorption (ABS/RC); electron
transport (ET0 /RC); trapping (TR0 /RC) and dissipation (DI0 /RC); (ii) phenomenological
energy fluxes per excited cross-section (CS)–absorption (ABS/CS); (iii) flux ratios or yields–
maximum quantum yield of primary photochemistry (ΦP0 ), electron transport probability
(Ψ0 ), and the quantum yield of electron transport (Ψ E0 ); (iv) performance index (PIABS ) on
an absorption basis, measuring the performance up to the photosystem I (PSI) end electron
acceptors.

2.4. Data Analysis

Statistical analyses of leaf temperature, leaf gas exchange, and chlorophyll a fluores-
cence were performed using the SPSS Statistics 20.0 software (IBM, New York, NY, USA).
After testing for ANOVA assumptions, statistical differences among kaolin-treated and
untreated leaves were evaluated by a two-way factorial ANOVA, followed by a post hoc
Tukey’s test and pairwise comparisons of the factors date (August and September) and
treatment (control and kaolin-treated). Different lower-case letters represent significant
differences (p < 0.05) between control and kaolin-treated leaves within each sampling date.
The asterisks represent significant differences (* p < 0.05) between sampling dates (August

Remote Sens. 2022, 14, 4019
cence were performed using the SPSS Statistics 20.0 software (IBM, New York, NY, USA).
After testing for ANOVA assumptions, statistical differences among kaolin‐treated and
untreated leaves were evaluated by a two‐way factorial ANOVA, followed by a post hoc
Tukey’s test and pairwise comparisons of the factors date (August and September) and
treatment (control and kaolin‐treated). Different lower‐case letters represent significant
differences (p < 0.05) between control and kaolin‐treated leaves within each sampling date. 7 of 18

The asterisks represent significant differences (* p < 0.05) between sampling dates (August
vs. September) within each treatment. The absence of letters and asterisks indicates no
significant
vs. September)difference.
within each treatment. The absence of letters and asterisks indicates no
significant
Regarding difference.
the UAV‐based data and given that two epochs are available, the temporal
Regarding
differences of thethevineyard’s
UAV-basedvegetative
data and given that two epochs
development are available,
were analysed. Thistheway,temporal
LST,
differences
CWSI, and IG of changes
the vineyard’s vegetative
were directly development
analysed werethe
to evaluate analysed.
temporal This way, LST,
effects CWSI,
in kaolin‐
and IGand
treated changes
in thewere directly
control analysed
plants. to evaluate
The estimated LSTthe temporal
was evaluated,effects in kaolin-treated
including all infor‐
mation and only including data from the estimated grapevine vegetation.information
and in the control plants. The estimated LST was evaluated, including all CWSI and and IG
onlyevaluated
were including at data
thefrom the estimated
grapevine grapevine
vegetation vegetation.
level. The grapevine CWSI and IG cover
vegetation were evaluated
for both
at the grapevine
treatments and itsvegetation
decline in level. The grapevine
area between the two vegetation
epochs werecover foranalysed.
also both treatments
The main and
its decline
reason in areathis
for doing between
analysistheistwo epochs
related were
to the also
fact analysed.
that The main
it is intended reason whether
to analyse for doing
this analysis isplants
kaolin‐treated related to the fact
present thatcanopy
a lower it is intended to analyse
projected whether
area decline kaolin-treated
than untreated plants.plants
present a lower canopy projected area decline than untreated plants.
The monitored vineyard was divided into eight polygons composed of kaolin‐treated
The monitored
vine rows and control vineyard was divided
rows (Table into eight
2 and Figure polygons
3). The composed
comparison plotsofwere
kaolin-treated
divided
vine rows and control rows (Table 2 and Figure 3). The comparison
according to the terrain slope and by the middle of the vine rows. Each polygon plots were divided
is labelled
according
with a lettertoaccording
the terraintoslope and
the set by the
of vine middle
rows and of theavine
with rows.according
number Each polygonto theislocation
labelled
with a letter according to the set of vine rows and with a number according
of the polygon in the vineyard: (1) for the upper part and (2) for the bottom part. There‐ to the location
of the
fore, polygonwith
polygons in the
thevineyard: (1) for
same letters arethe upper part
composed of and (2) forvine
the same the bottom part. 2).
rows (Table Therefore,
polygons with the same letters are composed of the same vine rows (Table 2).
Table 2. Area and number of rows used for comparing treatment plots.
Table 2. Area and number of rows used for comparing treatment plots.
No. of Rows
ID and Location Area (ha) No. of Rows Control
ID and Location Area (ha) Kaolin
A.1 0.35 Kaolin Control
11 14
A.2
A.1 0.31
0.35
11 14
A.2
B.1 0.31
0.57
B.1 0.57 13 15
B.2 0.59 13 15
B.2 0.59
C.1
C.1 0.53
0.53 1414 1010
C.2
C.2 0.46
0.46
D.1
D.1 0.62
0.62
D.2 0.52 2525 2121
D.2 0.52

Figure 3. Delimitation and identification of the compared treatment plots and the vineyard digital
terrain model. Orthophoto mosaic from the first flight campaign as background.

3. Results
This section presents the results provided by remote sensed data acquired using the
UAVs and by the eco-physiological parameters measured in the field. Kaolin treated plants
are compared with untreated plants on a general scale and with adjacent blocks (in the case
of UAV data). Values from both treatments are presented for each data acquisition period
(August and September) and changes among them are also compared.
 This section presents the results provided by remote sensed data acquired using the
UAVs and by the eco‐physiological parameters measured in the field. Kaolin treated
plants are compared with untreated plants on a general scale and with adjacent blocks (in
the case of UAV data). Values from both treatments are presented for each data acquisition
period (August and September) and changes among them are also compared.
Remote Sens. 2022, 14, 4019 8 of 18

3.1. UAV‐Based Results

3.1.Figure
UAV-Based4a presents
Results the LST computed for August and September flight campaigns. A
mean overall LST of 39.3the
Figure 4a presents °CLST
(SDcomputed
= 3.6 °C) for
wasAugust
observedand in August,flight
September whereas in September,
campaigns. A
this value
mean was 42.9
overall LST of °C39.3
(SD◦ C= (SD
3.1 °C).
= 3.6The
◦ C) maximum
was observed andin minimum LST temperature
August, whereas in September,were,
respectively,
this value was 51.2 °C◦and
42.9 C (SD27.6 °C◦in
= 3.1 C).August, and 55.3
The maximum and°Cminimum
and 35.4LST °C in September.
temperature A closer
were,
overview of an
respectively, 51.2 ◦
area treated
C and ◦
27.6 with kaolin and
C in August, and 55.3 ◦
another that
C and 35.4 ◦
was untreated
C in September. is A
presented
closer in
overview
Figure of an
4b. The area treated
vegetation with kaolin approach
segmentation and another that was
applied to untreated
the UAV‐based is presented in
data enabled
us Figure 4b. The
to estimate allvegetation segmentation
pixels belonging approachvegetation,
to grapevine applied to the UAV-based
and, therefore, data enabled veg‐
grapevine
us to estimate all pixels belonging to grapevine vegetation, and,
etation could be extracted. With regards to the mean temperature of the studied parts therefore, grapevine
vegetation could be extracted. With regards to the mean temperature of the studied
(polygons in Figure 1), when excluding non‐grapevine vegetation, it was 36.3 °C (SD = 2.0
parts (polygons in Figure 1), when excluding non-grapevine vegetation, it was 36.3 ◦ C
°C)(SD
in =August, and 45.0 °C (SD = 1.0 °C) in September. As for the mean vegetation tem‐
2.0 ◦ C) in August, and 45.0 ◦ C (SD = 1.0 ◦ C) in September. As for the mean vegetation
perature in kaolin
temperature in kaolin treated plants,
treated 36.236.2
plants, °C ◦(SD
C (SD= 1.8
= 1.8°C) was
◦ C) wasregistered
registered in Augustand
in August and 45.1
°C45.1
(SD C◦ = (SD
1.0 °C)
= 1.0inC)
◦ September.
in September. OnOn the
theother
other hand, controlplants
hand, control plants(no(no kaolin
kaolin application)
application)
showed
showed a atemperature
temperatureof of 36.7 ◦ C (SD = 1.8
36.7 °C 1.8 ◦°C) in August
C) in Augustand 45.1◦ C°C
and45.1 (SD(SD ◦ C)°C) in
= 1.0
= 1.0
in September.
September.

Figure 4. Land surface temperature of the thermal infrared data acquired from an unmanned aerial
Figure 4. Land surface temperature of the thermal infrared data acquired from an unmanned
vehicle for the August and the September flight campaigns (a). Close‐up view of the points high‐
aerial vehicle for the August and the September flight campaigns (a). Close-up view of the points
lighted in (a) of the RGB orthophoto mosaic (b) and thermal infrared imagery filtered for grapevine
highlighted in (a) of the RGB orthophoto mosaic (b) and thermal infrared imagery filtered for
vegetation (c) in kaolin and control areas in August and September flight campaigns.
grapevine vegetation (c) in kaolin and control areas in August and September flight campaigns.

Regarding the UAV-based LST derived from TIR imagery (Figure 4a,c), an overall
increase in temperature of 3.6 ◦ C was verified between the two surveyed dates when
considering the whole vineyard. By considering only grapevine vegetation, this difference
increased to 7.7 ◦ C. When comparing kaolin-treated and untreated vines in August, a
difference of 0.66 ◦ C was reached, considering all pixels, and 0.42 ◦ C when observing only
grapevine vegetation. In September, these differences were inferior, being 0.15 ◦ C and
0.03 ◦ C, respectively, for the whole area and grapevine vegetation.
Remote Sens. 2022, 14, 4019 9 of 18

The temperature references to calculate CWSI and IG were 30 ◦ C and 40 ◦ C in August

and, 40 ◦ C and 47 ◦ C in the September flight for Twet and Tdry , respectively. The right side
of Figure 4c depicts the temperature using Twet and Tdry values to produce a false colour
representation in the filtered grapevine vegetation.
In August, the grapevine vegetation cover ratio was 34.4% (1.73 ha), compared to the
vineyard area. In September it represented 30.6% (1.54 ha), representing a decrease of 3.8%,
corresponding to 11% of vegetation area decline between the two analysed periods. In
what concerns the 3.9 ha of kaolin and control treatments (polygons in Figure 1), a higher
decrease (12.7%) was observed in plants without foliar kaolin application (from 0.69 ha in
the August survey to 0.60 ha in September), while the vegetative decline of kaolin-treated
plants was 11.6% (representing 0.65 ha in August and 0.58 ha in September).
As for CWSI (mean values for each period in Table 3), a mean overall increase of 0.08 is
verified from August to September, while IG presented a decrease of 0.11 when considering
grapevine vegetation. In comparing untreated and kaolin-treated plants, lower CWSI
values are verified in kaolin-treated plants in the two surveyed periods. An increase of 0.10
and 0.07 between periods is verified for treated and untreated kaolin plants, respectively.
As for IG, both treatments presented a decrease of 0.13 when analysing this index. The
mean values for each date and TIR-based metric are presented in Table 3.

Table 3. Mean temperature (◦ C), crop water stress index (CWSI), and IG values, and its standard
deviation (SD) in the two surveyed periods for the whole vineyard and for treated (kaolin) and
untreated (control) kaolin grapevines.

Period Area Analysed Temp. (SD) CWSI (SD) IG (SD)

Overall 36.3 (2.0) 0.46 (0.22) 0.55 (0.27)
Aug. Kaolin 36.2 (1.8) 0.45 (0.21) 0.56 (0.27)
Control 36.7 (1.8) 0.49 (0.20) 0.56 (0.27)
Overall 45.0 (1.0) 0.53 (0.25) 0.44 (0.28)
Sep. Kaolin 45.1 (1.0) 0.55 (0.25) 0.43 (0.28)
Control 45.1 (1.0) 0.56 (0.24) 0.43 (0.28)

3.2. Vineyard Spatial Variability

Section 3.1 presents a general overview of the UAV-based data among the flight
campaigns as it concerns the overall vineyard surface temperature, vegetation cover area,
and CWSI and IG values. Despite these spatio-temporal differences, when analysing the
overall perspective of these parameters, the spatial variability can be assessed in a finer
scale. The comparison of each parameter is presented in this section.
Results interpretation from the comparison of treatment plots is carried towards
considering high temperature and CWSI, along with lower IG, as being signs of higher
plant stress, while the inverse (lower plant temperature, CWSI, and higher IG) is considered
as fewer signs of plant stress. The differences among treatment plots regarding the kaolin-
treated plants and untreated plants can be derived from the different parameters estimated
from the UAV-based TIR imagery (Figure 5).
In August, a higher plot temperature (Figure 5a) was observed at A.1 (37.8 ◦ C for
kaolin and 37.6 ◦ C for the control). The highest CWSI (Figure 5b) was also verified in
this plot (0.60 and 0.62, respectively, for kaolin and control treatments). Lower values
of IG were observed at A.2. for kaolin treated plots and in A.1. for control plots, being
0.49 and 0.52, respectively. On the other hand, lower temperature and CWSI values were
verified at plot D.1, being 34.95 and 0.31 in the kaolin treated part, while the control part
presented 35.23 and a CWSI value of 0.32. The higher IG value is 0.59 in the plots B.2 and
C.2 for kaolin-treated plants and of 0.64 for in D.1 for control plants. As for September, the
maximum mean temperature was observed in plants in the plot B.2 (45.85 ◦ C for control
and 45.46 ◦ C for kaolin-treated plants). Higher mean CWSI values were registered in the
plot B.2 for kaolin-treated grapevines (0.63) and in the plot A.1 for control plants (0.73),
while IG minimum mean values were in the plot B.2 (0.32 for the control part and 0.34
 Remote Sens. 2022, 14, 4019 10 of 18

for the kaolin part of the plot). Lower temperature values are registered in plot A.2 for
kaolin-treated parts (43.19 ◦ C) and in D.1 for control plants (42.84 ◦ C). Regarding CWSI, the
control part of plot D.1 showed the lowest value (0.43), while kaolin-treated plants showed
Remote Sens. 2022, 14, x FOR PEER REVIEW 10 of 18
a value of 0.47 in plot D.2. These plots also showed higher IG values, being 0.58 for the
control treatment of D.1 and 0.50 for the kaolin-treated part of the plot.

Figure 5. Relative difference of the canopy temperature◦ (°C) (a) and of the estimated indices (b) from
Figure 5. Relative difference of the canopy temperature ( C) (a) and of the estimated indices (b) from
the grapevine vegetation in August and September in each plot. Data represents the average change
the grapevine vegetation in August and September in each plot. Data represents the average change
of kaolin‐treated plants relative to control vines of Touriga Nacional. CWSI: crop water stress index;
of kaolin-treated plants relative to control vines of Touriga Nacional. CWSI: crop water stress index;
IG: stomatal conductance index.
IG: stomatal conductance index.

In The
August, a higher
difference betweenplottreatments
temperature (Figurerevealed
in August 5a) wasthat observed
in six outat of
A.1the(37.8
eight°C for
kaolin
plots,and 37.6 temperature
a lower °C for the control). The highest
was observed CWSI (Figure
in kaolin-treated plants.5b) was
Only alsoA.1
plots verified
and A.2in this
plot (0.60 and
presented an 0.62,
inverserespectively,
behaviour. The for kaolin
same isand control
verified treatments).
for CWSI. As for IG,Lower values
control plots of IG
presented
were observed slightly higher
at A.2. values, treated
for kaolin with theplots
exception
and inbeing
A.1.plot
forC.2. As for
control September,
plots, the and
being 0.49
lower plant temperature is verified in six plots for kaolin-treated
0.52, respectively. On the other hand, lower temperature and CWSI values were verified plants, with D.1 and D.2
presenting a higher plant temperature for both treatments.
at plot D.1, being 34.95 and 0.31 in the kaolin treated part, while the control part presented
The mean vegetation decline within the treatments over the two periods was 12.2% for
35.23 and a CWSI value of 0.32. The higher IG value is 0.59 in the plots B.2 and C.2 for
kaolin-treated plots and 13.2% for control plots (Table 4). In an intra-plot comparison, both
kaolin‐treated
treatments present plantssimilar
and ofbehaviours
0.64 for ininD.1 forofcontrol
terms plants.
vegetative declineAs for
andSeptember,
growth. Only thein max‐
imum
plot mean
D.2 was temperature
a vegetativewas observed
increase in plants
observed. in thethe
Regarding plot B.2 (45.85
temporal °C forofcontrol
variation other and
45.46 °C for kaolin‐treated plants). Higher mean CWSI values
parameters, as seen in Section 3.1, an LST increase was observed between the two periods.were registered in the plot
B.2Infor kaolin‐treated
this case, the mean grapevines
grapevine (0.63) and inincrease
temperature the plotwasA.17.89 ◦
for control ◦
C (7.72 plants (0.73),
C for the plotwhile
IG parts
minimumtreatedmean with kaolin
valuesand were8.05in◦ C
theforplot
the B.2
control
(0.32part). Thecontrol
for the biggest part
differences
and 0.34 were for the
reached in plot B.2 for both treatments. CWSI presented a mean
kaolin part of the plot). Lower temperature values are registered in plot A.2 for kaolin‐ overall increase of 0.10,
beingparts
treated 0.09 in kaolin°C)
(43.19 treated
and in plots
D.1andfor0.11 in control
control plantsplots. All°C).
(42.84 the control
Regarding partsCWSI,
presentedthe con‐
an increase in CWSI, while the kaolin treated parts of plots A.1 and A.2 showed a decrease
trol part of plot D.1 showed the lowest value (0.43), while kaolin‐treated plants showed a
in this value. As for IG, which presented a mean overall decrease of 0.14, with control plots
value of 0.47ain
presenting plotofD.2.
mean These plots
0.16 decrease, and also showed
the kaolin plotshigher
showing IGavalues, being 0.58
mean decrease for the
of 0.11.
control
In thetreatment
case of thisof D.1 and
index, only0.50 for the kaolin‐treated
one kaolin-treated plot (A.2) part of the the
maintained plot.same mean plot
The difference between treatments in August
value as August, while all the other plots decreased its value. revealed that in six out of the eight
plots, a lower temperature was observed in kaolin‐treated plants. Only plots A.1 and A.2
3.3. Eco-Physiological
presented an inverse Parameters
behaviour. The same is verified for CWSI. As for IG, control plots
presented slightly higherparameters,
Leaf gas exchange values, with suchtheasexception
net photosynthetic
being plot rateC.2.
(A),Asstomatal conduc- the
for September,
tance
lower plant(g ), and transpiration rate (E) values, were mainly affected
s temperature is verified in six plots for kaolin‐treated plants, with D.1 and by kaolin treatment in D.2
August (Figure 6). At this stage, kaolin-treated vines showed simultaneously higher A
presenting a higher plant temperature for both treatments.
(10.10 µmol m−2 s−1 ), gs (119.85 mmol m−2 s−1 ), and E (3.39 mmol m−2 s−1 ) values than
The mean vegetation decline within the treatments over the two periods was 12.2%
the control ones. Conversely, the intrinsic water use efficiency (i WUE) and the ratio of
forintercellular
kaolin‐treated plots and CO
to atmospheric 13.2% for control plots (Table 4). In an intra‐plot comparison,
2 concentration (Ci /Ca ) parameters of kaolin-treated leaves
both treatments
increased aboutpresent
59.2% and similar behavioursininSeptember,
19%, respectively, terms of compared
vegetative to decline
untreated and growth.
vines.
Only in plot D.2 was a vegetative increase observed. Regarding the temporal variation of
other parameters, as seen in Section 3.1, an LST increase was observed between the two
periods. In this case, the mean grapevine temperature increase was 7.89 °C (7.72 °C for the
plot parts treated with kaolin and 8.05 °C for the control part). The biggest differences
were reached in plot B.2 for both treatments. CWSI presented a mean overall increase of
 between September to August mean plot values of kaolin‐treated plants with respect to the control
values. Temp: temperature; CWSI: crop water stress index; IG: stomatal conductance index.

Vegetative Decline (%) Temp. (°C) CWSI IG

Plot
Kaolin Control Abs. Diff. Kaolin Control Kaolin Control Kaolin Control
Remote A.1
Sens. 2022, 14,23.7
4019 29.2 5.4 6.00 8.12 −0.06 0.11 −0.03 11 of 18
−0.19
A.2 12.7 3.0 9.7 5.98 7.27 −0.05 0.04 0.00 −0.10
B.1 24.6 26.1 1.6 7.82 7.77 0.09 0.08 −0.13 −0.16
B.2 8.6 Table 4. Relative
10.6 2.0 difference8.79
between the8.92
two periods0.11
for the grapevine
0.15 vegetation area decline
−0.24 and
−0.28
C.1 15.9 estimated parameters
26.0 10.1 from8.07
the thermal infrared
8.44 imagery0.11in each plot.
0.14Data represents
−0.15 the difference
−0.19
between September to August mean plot values of kaolin-treated plants with respect to the control
C.2 9.7 8.7 1.0 8.69 8.74 0.16 0.15 −0.19 −0.22
values. Temp: temperature; CWSI: crop water stress index; IG: stomatal conductance index.
D.1 5.3 4.3 1.1 8.58 7.61 0.20 0.11 −0.09 −0.06
D.2 −2.9 Vegetative
−2.1Decline (%)0.8 7.86 Temp. (◦7.51
C) 0.12 CWSI 0.09 −0.08 IG −0.07
Plot
Kaolin Control Abs. Diff. Kaolin Control Kaolin Control Kaolin Control
3.3. Eco‐Physiological Parameters
A.1 23.7 29.2 5.4 6.00 8.12 −0.06 0.11 −0.03 −0.19
A.2 12.7 3.0
Leaf gas9.7
exchange parameters,
5.98
such as net−photosynthetic
7.27 0.05 0.04
rate (A), stomatal−conduct‐
0.00 0.10
B.1 24.6 26.1ance (gs), and1.6 transpiration
7.82 rate (E)7.77
values, were 0.09mainly 0.08
affected by−0.13
kaolin treatment
−0.16 in
B.2 8.6 10.6August (Figure2.0 6). At this8.79stage, kaolin‐treated
8.92 vines showed
0.11 0.15 simultaneously
−0.24 −higher
0.28 A
C.1 15.9 26.0(10.10 μmol m−2 s−1), gs (119.85
10.1 8.07 mmol 8.44m−2 s−1), 0.11
and E (3.390.14mmol m−2−0.15
s−1) values−0.19
than the
C.2 9.7 8.7 control ones.1.0Conversely, 8.69 8.74water use
the intrinsic 0.16efficiency
0.15 −0.19
(iWUE) and −0.22
the ratio of inter‐
D.1 5.3 4.3
cellular to atmospheric CO2 concentration (Ci/Ca) parameters of kaolin‐treated−leaves
1.1 8.58 7.61 0.20 0.11 − 0.09 0.06
in‐
D.2 −2.9 −2.1 0.8 7.86 7.51 0.12 0.09 −0.08 −0.07
creased about 59.2% and 19%, respectively, in September, compared to untreated vines.

Figure
Figure 6. 6.Leaf
Leafgas
gasexchange
exchangeparameters
parametersofofTouriga
Touriga Nacional
Nacional control
control and
and kaolin‐treated
kaolin-treated grapevines
grapevines atat
midday (14:00 GTM+1). Stomatal conductance (gs, mmol−m −2 s−1), net CO2 assimilation rate (A, μmol
2 s−1 ), net CO assimilation rate (A, µmol
midday (14:00 GTM+1). Stomatal conductance (gs , mmol m 2
m
−
−2 s−1), intrinsic water use efficiency (A/gs, μmol
2 − 1 mol
−
−1), intercellular CO
1 2 concentration (Ci), ratio
m s ), intrinsic water use efficiency (A/gs , µmol mol ), intercellular CO2 concentration (Ci ), ratio
of intercellular to atmospheric CO2 concentration (Ci/Ca), and transpiration rate (E, mmol m−2 s−1).
ofData
intercellular to ±atmospheric CO2 concentration −2 s−1 ).
are mean SD of six replicates. Different(C i /Ca ), andletters
lower‐case transpiration ratesignificant
represent (E, mmol mdifferences
Data are mean
between ± SD of(control
treatments six replicates. Different lower-case
vs. kaolin‐treated) within each letters
month.represent significant
* p < 0.05 differences
represent significant
between treatments (control vs. kaolin-treated) within each month. * p
differences between sampling months (August vs. September) within each treatment. significant
< 0.05 represent
differences between sampling months (August vs. September) within each treatment.
Figure 7 shows no significant changes in PSII photochemistry between kaolin‐treated
andFigure 7 shows
untreated noinsignificant
vines changes
both sampling in PSII
dates. photochemistry
However, between
most of the kaolin-treated
parameters analysed
and
from August to September were greatly reduced in control leaves compared to analysed
untreated vines in both sampling dates. However, most of the parameters the treated
from August
ones. to September
Specifically, were greatly
Fv/Fm decreased reduced
50.3% in control
in control leaves
vines and compared
37.4% in to the treated
treated plants,
ones. Specifically, Fv /Fm decreased 50.3% in control vines and 37.4% in treated plants,
while NPQ increased around 120.8% and 164.1% in untreated and kaolin-treated vines,
respectively. Moreover, chlorophyll transient analysis (Table 5) shows a significantly higher
relative change of the kaolin-treated to control leaves in some specific energy fluxes per
reaction centre, particularly DI0 /RC and ET0 /RC in September. Likewise, the flux ratio of
the electron transport probability (Ψ0 ) increased in kaolin-treated vines around 315.71%
in September.

Remote Sens. 2022, 14, 4019
while NPQ increased around 120.8% and 164.1% in untreated and kaolin‐treated vines,
respectively. Moreover, chlorophyll transient analysis (Table 5) shows a significantly
higher relative change of the kaolin‐treated to control leaves in some specific energy fluxes
per reaction centre, particularly DI0/RC and ET0/RC in September. Likewise, the flux ratio
12 of 18
of the electron transport probability (Ψ0) increased in kaolin‐treated vines around 315.71%
in September.

Figure 7. Midday chlorophyll fluorescence variables of Touriga‐Nacional grapevines in control and

Figure 7. Midday chlorophyll fluorescence variables of Touriga-Nacional grapevines in control and
kaolin‐treated leaves throughout the experiment. Parameters measured: maximum (Fv/Fm) quantum
kaolin-treated leaves
efficiency of PSII throughout
and the experiment.
non‐photochemical Parameters
quenching (NPQ). measured:
Data aremaximum
mean ± SD (Fvof
/Fsix
m ) quantum
replicates.
efficiency of PSII and non-photochemical quenching (NPQ). Data are mean ± SD
Different lower‐case letters represent significant differences (p < 0.05) between treatments of six replicates.
within
Different lower-case
each month. letters
* denote representdifferences
significant significant differences (p < 0.05) months
(p < 0.05) between between(August
treatmentsvs.within each
September)
month.
within *the
denote significant differences (p < 0.05) between months (August vs. September) within
treatment.
the treatment.
Table 5. Relative difference of the JIP parameters deduced from chlorophyll a fluorescence OJIP
transients
Table in August
5. Relative and September
difference of the JIPinparameters
the leaves deduced
of Touriga‐Nacional treated with
from chlorophyll 5% kaolin.OJIP
a fluorescence Data
are mean ± SD of six replicates. * Indicates a significant (p < 0.05) relative change in the
transients in August and September in the leaves of Touriga-Nacional treated with 5% kaolin. Data arekaolin‐treated
group respecting to control.
mean ± SD of six replicates. * Indicates a significant (p < 0.05) relative change in the kaolin-treated
group respecting to control. Period
Parameter
August September
Period
F0
Parameter 13.32 1.16
August September
ABS/RC 20.62 4.15
TRF00/RC 13.32
0.48 1.16
−9.64
ABS/RC 20.62 4.15
DI0/RC
TR0 /RC
31.86
0.48 −51.27
9.64
*
ET
DI 0/RC
0 /RC
2.57
31.86 80.73* *
51.27
P0
ET0φ/RC −4.22
2.57 1.81*
80.73
ϕP Ψ00 −4.22
2.37 1.81
315.71 *
Ψ0 2.37 315.71 *
Ψ E0 2.52 27.5
Ψ E0 2.52 27.5
φ
ϕ D0
D0 15.87
15.87 27.5
27.5
PIPI ABS
ABS 30.05
30.05 67.72
67.72

4.4.Discussion
Discussion
Theacquired
The acquiredthermal
thermaldata
dataenabled
enabledusustotoobserve
observethatthatananoverall
overallincrease
increase inin temper‐
tempera-
ature occurred in between the two surveyed periods. Maximum
ture occurred in between the two surveyed periods. Maximum temperatures are observed temperatures are ob‐
inserved in non‐vegetated
non-vegetated areasexposed
areas directly directly toexposed to sunlight.
sunlight. In contrast, In minimum
contrast, minimum temper‐
temperatures are
atures are
reached reached inareas
in shadowed shadowed areas
close to close toplants,
grapevine grapevine
where plants,
there where there were
were possible leakspossible
in the
leaks in the
irrigation irrigation
system. Thesesystem. These can
malfunctions malfunctions
be seen in can be seen
inter-row in inter‐row
parts parts of
of the vineyard landthe
vineyard
surface land surface
temperature temperature
in Figure 4a, withinmore
Figure 4a, withinmore
emphasis emphasis
September, this in September,
being one advan-this
being
tage of one
usingadvantage
UAV-based of using UAV‐based
TIR data. TIR data.
Furthermore, Furthermore,
the data processingthe data processing
approach followedap‐ a
proach followed
traditional a traditional
photogrammetric photogrammetric
alignment using GCPsalignment
to align RGBusingandGCPs to align
TIR data, RGB and
allowing us
toTIR data, estimate
directly allowingtheus grapevine
to directlycanopy
estimate thethe
from grapevine canopy
RGB imagery. fromthe
Thus, theuse
RGB imagery.
of TIR data
Thus,
only the use ofwhich
is avoided, TIR data only
could leadis to
avoided, which could lead
the misinterpretation to the misinterpretation
of grapevine values or couldof
require more laborious and computer-intensive data coregistration approaches [70,71].
To assess if foliar kaolin applications caused a higher vegetation decrease over time,
the vegetation cover ratio of the projected area was evaluated in both periods. The typi-
cal grapevine temporal and phenological dynamics demonstrated a natural overall area
decrease between both August and September. The obtained grapevine cover area is
within the values reported in other studies [42] and the vegetative decline was expected
to occur as plants are close to harvest. If prior periods (with a higher vegetative develop-
Remote Sens. 2022, 14, 4019 13 of 18

ment) were analysed, this decline would be higher [63], as the survey period in August
already corresponded to an advanced ripening stage. This study shows that kaolin does
not negatively affect the plants when analysing grapevine vegetation area. In fact, it was
quite the opposite, a lower overall vegetative decline was observed (12.7% in control and
11.6% in kaolin areas) and, in the direct comparison of most treatment plots (Table 4), five
kaolin-treated plots presented a lower vegetative decline than control plots. One treatment
plot (D.2) presented a growth in both treatments, being higher in the kaolin part. In the
present study, kaolin treatment showed no significant leaf cooling effect in August and
September. Yet, most of the literature reports significant decreases in leaf temperature
with kaolin application in grapevines, which could be associated with the grain size of the
mineral, efficiency of foliar coverage, concentration, number of applications, and weather
conditions [72,73]. Nonetheless, once applied as a suspension over the whole canopy,
kaolin functions as a whitish barrier with reflective properties, screening radiation and
heat loading, thus avoiding sunburn damage in Mediterranean crops and improving the
physiological performance of vines [15,18].
Regarding the UAV-based parameters estimated from the grapevine vegetation (Figure 5),
the differences among treatments in August revealed that in six out of the eight plots, a
lower temperature was observed in kaolin-treated plants, only plots A.1 and A.2 presented
an inverse behaviour. This result is in agreement with several data collected by hand
infrared thermometers in different varieties sprayed with a kaolin suspension on vineyards
located in Australia [74], Portugal [9], Italy [13], and the United States [72]. A similar
response was verified for CWSI. As for IG, control plots presented slightly higher values,
with the exception being plot C.2. In September, a lower plant temperature was verified in
six plots for kaolin-treated plants, with D.1 and D.2 presenting a higher plant temperature
for both treatments.
The two indices estimated from the TIR data showed a better performance in kaolin-
treated areas, presenting lower signs of water stress (CWSI) and higher stomatal conduc-
tance (IG). Generally, both indices decrease in between flight campaigns. When observing
the eco-physiological measurements on single leaves (Figure 6) through an IRGA, the
stomatal conductance (gs ), net photosynthesis (A), and transpiration rate (E) presented
similar trends. This positive effect was mainly observed in August, as reported in Touriga
Nacional and Touriga Franca leaves grown in Mediterranean wine-growing regions [11].
Though no relevant changes were found in the gs values in September, the iWUE was
higher in kaolin-treated leaves at this stage, suggesting a prolonged effect of kaolin coating
on improving plant performance under adverse environmental conditions. Nonetheless,
the most physiological effects of kaolin treatment were mainly observed in August, pos-
sibly due to the occurrence of some precipitation after kaolin application that dissolves
easily in water. By comparing the temporal differences (Table 4), higher temperatures
are registered in September. Moreover, between the two survey periods, the effectiveness
of kaolin protective film decreased. This can be seen in the sections of the orthophoto
mosaics related to September data (presented in Figure 4b) in which the kaolin treated
plants showed a higher colour resemblance to the control plants. This could also be an
effect of strong winds and precipitation (Figure 2b) that occurred in between the two survey
periods. Moreover, in the same region and grapevine variety, Tosin et al. [28] revealed
that kaolin’s protective effect lasted for 20 days after its application and stopped about
60 days after. Regarding chlorophyll a fluorescence transient analysis (Table 5), the specific
energy flux data combined with the quantum yield analysis showed higher values in
kaolin-treated leaves in both studied dates, excepting the yield or primary photochemistry
(φP0 ) in August and the trapping energy flux per reaction centre (TR0 /RC) in September.
This response was recently demonstrated by Bernardo et al. [16], indicating an apparent
antenna size reduction and lower inactivation of reaction centres at the beginning of the
experiment. This photoprotective response might explain the higher performance index
(PIABS) found in treated plants, as previously observed in Dinis et al. [8].
Remote Sens. 2022, 14, 4019 14 of 18

Regarding in-field data acquisition and the UAV-based approaches, both have their
strengths and weaknesses. Manual leaf temperature retrieval enables the precise gathering
of temperature in a specific spot in the vegetative wall but has the drawback of being
laborious and time-consuming for medium to large-scale assessments. On the other hand,
UAV-based thermal infrared imagery can cover more area in a faster manner, retrieving
temperature from the whole vineyard, but plant temperature can be influenced by soil
temperature. In fact, when comparing both of these two approaches, several studies
have shown that there is a high correlation (above 0.9 R2 ) when comparing temperature
driven from UAV-based TIR imagery with in-ground handheld infrared thermometers or
cameras [75–79]. Therefore, the temperature can be deemed correct, which, allied to the
automatic self-calibration applied by the sensor before each flight line, ensures that the
radiometric conditions of the imagery are comparable through space and time.
In future work, leaf density differences between control and kaolin-treated plants should
also be assessed by using vegetation indices computed with multispectral or hyperspectral
data. However, it should be noted that due to kaolin foliar application, grapevine spectral
properties can change, which could prevent the use of typical vegetation indices. Thus, kaolin
could employ a noisy effect in the spectral reflectance since it is applied throughout the leaves.
This could also be an advantage to evaluate if a new kaolin foliar application is required. The
combined effect of kaolin application with other irrigation strategies should also be considered
to improve water management in Mediterranean vineyards.

5. Conclusions
This study shows the suitability of using UAV-based thermal infrared data in precision
viticulture research. By performing UAV flight campaigns, it was not only possible to obtain
thermal information about the whole vineyard but also to filter it to exclude non-grapevine
information and to refine the analysis towards the plants. Thus, by comparing this with
field observations, it is demonstrated that by using such a type of remote sensed data,
more area can be assessed in the same period of time, obtaining more information and
providing more complete decision support. Several temperature acquisitions on the same
day could also help in understanding thermal gaps between treated-kaolin plants and
untreated plants. Moreover, the CWSI and IG indices calculated from the data provided
using UAV are in accordance with gas exchange parameters measured in Touriga Nacional
leaves (i WUE, gs , and A), indicating a general improved physiological performance of
kaolin-treated leaves under the current conditions, whose effects were mainly noticed at
the beginning of the experiment.
In a water stress scenario, it is crucial to deliver data quickly, as a delay of some days
can have negative consequences for grapevine development and berry composition, as
well as economic impacts for winegrowers. Thus, quicker ways for UAV-based water
stress estimation are needed, which can be based on cloud processing pipelines or even
in-the-field processing through a mobile office with results being provided the same day,
enabling winegrowers to quickly act upon stressed areas.

Author Contributions: Conceptualization, L.P., J.M.-P. and J.J.S.; methodology, L.P. and S.B.; software,
L.P.; validation, L.P., L.-T.D., C.C., J.M.-P. and J.J.S.; formal analysis, L.P. and S.B.; investigation, L.P.
and S.B.; resources, L.P., S.B., L.-T.D., C.C., J.M.-P. and J.J.S.; data curation, L.P.; writing—original draft
preparation, L.P. and S.B.; writing—review and editing, L.-T.D., C.C., J.M.-P. and J.J.S.; visualization,
L.P. and S.B.; supervision, L.-T.D., J.M.-P. and J.J.S.; project administration, L.-T.D., C.C., J.M.-P.
and J.J.S.; funding acquisition, J.J.S. All authors have read and agreed to the published version of
the manuscript.
Funding: This research activity was supported by National Funds by FCT—Portuguese Foundation
for Science and Technology, under the project UIDB/04033/2020 and by the project “DATI—Digital
Agriculture Technologies for Irrigation efficiency”. PRIMA—Partnership for Research and Innovation
in the Mediterranean Area, (Research and Innovation activities), financed by the states participating
in the PRIMA partnership and by the European Union, through Horizon 2020.
Remote Sens. 2022, 14, 4019 15 of 18

Data Availability Statement: The data presented in this study are available on request from the
corresponding author.
Acknowledgments: We acknowledge the E-OBS dataset from the EU-FP6 project UERRA (http://www.
uerra.eu) and the Copernicus Climate Change Service, and the data providers in the ECA&D project
(https://www.ecad.eu). We would also like to thank to “Real Companhia Velha” for the collaboration
and efforts in making the vineyard0 s facilities available for the research and particularly to Rui Soares
and Sérgio Soares for their valuable collaboration. We also thank ADVID for the encouragement given
in carrying out this study.
Conflicts of Interest: The authors declare no conflict of interest. The funders had no role in the design
of the study; in the collection, analyses, or interpretation of data; in the writing of the manuscript, or
in the decision to publish the results.

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