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  <front>
    <journal-meta />
    <article-meta>
      <title-group>
        <article-title>Diurnal Variations in Leaf - Air Temperature and Vapor Pressure Deficit of Sunlit and Shaded Kenaf Leaves</article-title>
      </title-group>
      <contrib-group>
        <contrib contrib-type="author">
          <string-name>Ippolitos Gintsioudis</string-name>
          <xref ref-type="aff" rid="aff0">0</xref>
        </contrib>
        <contrib contrib-type="author">
          <string-name>Elpiniki Skoufogianni</string-name>
          <xref ref-type="aff" rid="aff0">0</xref>
        </contrib>
        <contrib contrib-type="author">
          <string-name>Dimitrios Bartzialis</string-name>
          <xref ref-type="aff" rid="aff0">0</xref>
        </contrib>
        <contrib contrib-type="author">
          <string-name>Kyriakos Giannoulis</string-name>
          <xref ref-type="aff" rid="aff0">0</xref>
        </contrib>
        <contrib contrib-type="author">
          <string-name>Nicholaos Danalatos</string-name>
          <xref ref-type="aff" rid="aff0">0</xref>
        </contrib>
        <aff id="aff0">
          <label>0</label>
          <institution>Department of Agriculture Crop Production and Rural Environment, University of Thessaly</institution>
          ,
          <country country="GR">Greece</country>
        </aff>
      </contrib-group>
      <fpage>574</fpage>
      <lpage>579</lpage>
      <abstract>
        <p>The microclimatic conditions within a crop canopy are differentiated from those outside resulting to the exposure of plant organs in different environmental conditions. In the present study, the diurnal variation of leaf temperature, air temperature and vapor pressure deficit (VPDis presented, of leaves inside (shaded) and outside (sunlit) a kenaf canopy, for a period of two days is presented. During the solar day, the VPD inside and outside the canopy showed no differences and maximized at 2.7 kPa. Inside the canopy air temperature showed statistically significant differences from the outside temperature the period from 6 am to 10 am and the leaf - air temperature comparison of sunlit leaves showed significant differences during 6 pm - 11 pm, while the leaf - air temperature comparison of shaded leaves showed significant differences, almost constantly, throughout the day.</p>
      </abstract>
      <kwd-group>
        <kwd>kenaf</kwd>
        <kwd>leaf temperature</kwd>
        <kwd>vapor pressure deficit (VPD)</kwd>
      </kwd-group>
    </article-meta>
  </front>
  <body>
    <sec id="sec-1">
      <title>1 Introduction</title>
      <p>
        Canopy temperature is primarily correlated with stomatal conductance, a
physiological process which affects photosynthesis rate, transpiration rate, plant
water use, leaf area index (LAI) and therefore crop yield (Lopes and Reynolds,
2010). Leaf temperature depends on air temperature, humidity, and wind speed
        <xref ref-type="bibr" rid="ref9">(Jones et al. 2009)</xref>
        factors which affect leaf transpiration. Thermal imaging provides
a good solution in screening leaf temperature, either in laboratory or field conditions,
and differences between leaf and air temperature can be used as a measure of plant
stress
        <xref ref-type="bibr" rid="ref8">(Jackson et al. 1977)</xref>
        . The canopy – air temperature differential has been used in
many studies to quantify crop water stress index (CWSI)
        <xref ref-type="bibr" rid="ref1 ref11 ref4 ref6">(Ahi et al. 2015; Erdem et
al. 2006; Carroll et al. 2017)</xref>
        . The infrared temperature measurements in most studies
are captured from the top of the canopy, considering the top canopy leaves as
representatives for the whole canopy. The microclimate inside and outside of a
canopy is different with the plant organs being exposed to different factors affecting
their physiological processes and growth. In crop modelling, data obtained from local
weather stations or national networks are used
        <xref ref-type="bibr" rid="ref10">(Jucker et al. 2018)</xref>
        and it is assumed
that every plant part in a field is exposed in the same conditions. This bias can have
implications when modelling plant responses to climate change. To reduce
inaccuracies, research of crop microclimate inside and outside of the canopy is
required.
      </p>
      <p>
        Kenaf (Hibiscus cannabinus L.) is a crop mainly cultivated for fiber and biomass
production. It is well adapted to Greek climatic conditions and it has a high potential
biomass productivity, about 22 t ha-1
        <xref ref-type="bibr" rid="ref2 ref5">(Danalatos &amp; Archontoulis 2010; Alexopoulou
et al. 2000)</xref>
        . Kenaf has the potential to be integrated as a new crop in Greece and
further research of plant – environment interactions are required.
2
      </p>
    </sec>
    <sec id="sec-2">
      <title>Materials and Methods</title>
      <sec id="sec-2-1">
        <title>2.1 Experimental Area</title>
        <p>The experiment took place in a kenaf experimental field, located in Thessaly plain
of Greece (coordinates: 39°30'45" N, 22°28'03.1" E, altitude 170 m) during 14th (1st
day) and 15th (2nd day) of October 2013. The soil was characterized as clay. The
kenaf variety was Whitten and plant density 20 plants m-1. Emergence of kenaf dated
on 20/5/2013. Plot fertilization consisted on 200 kg N ha-1 (as ammonium nitrate) and
50 kg P ha-1 without any K fertilization applied, since soil analyses showed adequate
K concentration. Irrigation was applied weekly with the amount of applied water
being calculated as equal to maximum evapotranspiration.</p>
      </sec>
      <sec id="sec-2-2">
        <title>2.2 Temperature Measurement System</title>
        <p>A custom leaf temperature system was designed for automatic measurements and
data storage of leaf and air temperature, and air humidity. The main system
components were the microcontroller, the sensors, the power supply and a
datalogging system.</p>
        <p>The microcontroller was an ATmega2560 (Arduino Mega board) with 54 digital
input/output pins, 16 analog inputs, 256 KB of Flash Memory for storing code and
with 16MHz operating frequency. The microcontroller connects via a USB type port
to a computer for code upload and stored data download. Microcontroller’s code was
written in Arduino programming language with the Arduino IDE being used for the
communication between computer and microcontroller.</p>
        <p>
          For leaf temperature measurements, the infrared non-contact MLX90614 sensor
(Melexis, Concord, NH) was used. This type of sensors are appropriate for field use
and have been used in many plant leaf experiments
          <xref ref-type="bibr" rid="ref11 ref4 ref7">(Fisher &amp; Kebede 2010; Martinez
et al. 2017)</xref>
          . The sensor consists of an infrared-sensitive thermopile detector and a
signal conditioning chip integrated into a single unit. The on-board 17-bit
analog-todigital converter DAC transforms the analog temperature signal of the thermopile
detector into a digital signal, accessible to the microcontroller via the SMBus
communication protocol. The version of the sensor used had a10o Field of View
(FOV) and was factory calibrated for temperature measurements in the range of -40 –
85oC with 0.01oC resolution and ±0.5oC accuracy
          <xref ref-type="bibr" rid="ref12">(Melexis 2019)</xref>
          . A total of 7
infrared sensors were used.
        </p>
        <p>
          For air temperature and relevant humidity (%RH) measurements, the SHT15
sensors (SENSIRION AG) were used. The SHT15 contain a capacitive sensor
element for measuring relative humidity while temperature is measured by a
bandgap sensor. The integrated CMOSens® technology of the sensor further increase its
reliability and long-term stability. Both sensors are seamlessly coupled to a 14bit
analog to digital converter (ADC) and a serial interface circuit. The sensor
communicates with the microcontroller via the serial interface. The %RH accuracy of
the sensor is ±2% and the temperature accuracy ±0.3-0.4oC in the range of 5 - 40oC
          <xref ref-type="bibr" rid="ref13">(Sensirion 2015)</xref>
          . 6 SHT15 sensors were used.
        </p>
        <p>The power supply system was consisting of a 9-volt battery pack, capable to
operate the device for at least 48 hours, and for datalogging, the OpenLog module
(SparkFun Electronics) was used. The OpenLog module is an open source data
logger for microSD cards that works over serial connection.</p>
        <p>The leaf and air temperature sensors were coupled to form, a total of 6, sensor
heads. Each sensor head was attached to the end of a pole and positioned underneath
a healthy leaf, with the infrared sensor looking the underside of the leaf. The poles
were stabilized to the ground and attached to the plant stems to reduce changes in
plant leaf position due to wind streams. The fourth infrared sensor was placed
looking at the sky to be used as a correction factor in case an infrared sensor would
lose visual contact with the leaf. The system was often visually monitored to ensure
proper operation.</p>
      </sec>
      <sec id="sec-2-3">
        <title>2.3 Measurements</title>
        <p>Three sensor heads were placed underneath sunlit leaves on top of the kenaf
canopy (sunlit leaves) and another 3 underneath leaves inside the kenaf canopy
(shaded leaves), not visible by sun rays. In this stage of development, the canopy
Leaf Area Index (LAI) was ~ 3 ensuring good shading conditions inside canopy,
although proper attention was paid to the position selection.</p>
        <p>
          The measurements consisted of sunlit and shaded: a) leaf temperatures (Tleaf-sun,
Tleaf-shade), b) air temperatures (Tair-sun, Tair-shade) c) relevant humidity (RHsun, RHshade).
The sky temperature was not used because no errors in infrared measurements were
detected. Measurements were logged every 5 mins and averaged in hourly means.
From air temperature and %RH of each SHT15 sensor, the VPD was calculated
based on the equations given in FAO 56
          <xref ref-type="bibr" rid="ref3">(Allen et al. 1998)</xref>
          .
3
        </p>
      </sec>
    </sec>
    <sec id="sec-3">
      <title>Results</title>
      <p>In Fig.1 the diurnal patterns of leaf and air temperatures, along with VPD, are
presented. During the 1st day, Tair-sun had a minimum value of 13.7oC, which
25
oC
reu20
tr
a
ep15
m
e
T10
5
0
35
30
4,0
3,5
3,0
2,5
1,5
1,0
0,5
aP 2,0
k</p>
      <sec id="sec-3-1">
        <title>Sunlit</title>
      </sec>
      <sec id="sec-3-2">
        <title>Shaded VPD</title>
        <p>Leaf
Air</p>
        <p>Leaf</p>
        <p>Air
Shaded
Sunlit
maximized at 27.9oC at 15th hour of the day, while the 2nd day the minimum and
maximum values were 16.1oC and 30.7oC. The respective values for Tair-shade were
15.1-26.2oC for the 1st day, and 17.3-29.0oC for the 2nd day. Maximum value of
Tairshade occurred one hour after Tair-sun.</p>
        <p>The mean hourly air and leaf temperature was compared for differences during the
2-day period. During the 1st day hourly Tair-shade to Tleaf-shade pair comparison showed
statistically significant differences (p ≤ 0.05) only during the 10th and between the
17th and the 22nd hour of the day. During the 2nd day the statistically significant
differences (p ≤ 0.05) showed at the 8th hour and during the 17th and the 22nd hour, as
on the 1st day. The pair comparison of hourly Tair-sun to Tleaf-sun showed constant
statistically significant differences (p ≤ 0.05) during both days, except hours 8, 9 and
19 to 23, of the 2nd day where no differences were found. During the experimental
period, the mean hourly air temperature was constantly higher than leaf temperature.
The range of Tair-shade - Tleaf-shade difference was 0.6-1.8oC during the 1st day and
0.73.2oC during the 2nd day. For Tair-sun and Tleaf-sun the range of their difference was
0.31.2oC and 0.8-2.1 for the 1st and 2nd day, respectively. Tleaf-shade range between
13.724.4oC and 14.8-27.1oC for the 1st and 2nd day, respectively, while Tleaf-sun range
between 12.5-26.6oC and 14.8-29.3oC.</p>
        <p>The hourly VPD means of shaded and sunlit leaves, throughout the day, were
compared for statistically significant differences. During solar day there were no
significant differences contrary to the period between 18th to 24th hour, of each of the
days. For both days, VPDsun was maximum at 15th hour of the day, while the
VPDshaded maximized one hour later. VPDsun maximum daily value was 1.6 and 2.7
kPa and VPDshaded maximum value was 1.4 and 2.4 kPa, for 1st and 2nd day,
respectively.</p>
      </sec>
    </sec>
    <sec id="sec-4">
      <title>4 Discussion</title>
      <p>Temperature and vapor pressure deficit (VPD) are important environmental
factors affecting crop growth and productivity by regulating leaf stomatal
conductance. In the present study VPD inside and on top of the canopy, showed no
significant differences throughout the solar day, apart from after 6 pm when the sun
is low on the horizon and the photosynthetic rate is low. The same pattern was
observed both in the inside and outside air temperature, and in both sunlit leaves
temperature and their air temperature. In both cases there was differentiation after 6
pm. On the contrary the shaded leaves had almost constantly statistically significant
different temperature from the temperature of their microclimate. These findings
suggest that leaf temperature varies within the day and depending on the canopy
height, and thus they should not be considered stable.</p>
      <p>Acknowledgments. This paper is part of PhD thesis of Ippolitos Gintsioudis and the
first author would like to thank the Laboratory of Agronomy and Applied Crop
Physiology of University of Thessaly for helping in the completion of this project.</p>
    </sec>
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