Effects of Temperature on the Growth and Anthocyanin Content of Echeveria agavoides and E. marcus

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Abstract Few studies have reported how temperature
influences growth and development of succulents, including
anthocyanin production, which could fetch better prices in
the market, and understanding the factors influencing such
pigments would benefit farmers. The present study investigated
the effect of temperature (10°C, 20°C, and 30°C) on the
growth, development, and anthocyanin concentrations in
Echeveria agavoides
and
E. marcus
. In
E.
agavoides
, similar
growth performance was observed at 10°C and 20°C based
on plant height and diameter. However, subjecting the
species to a high temperature of 30°C resulted in a decrease
in plant height. In
E. marcus
, optimal growth performance was
observed at 20°C. Different temperatures did not significantly
affect succulent quality and color hues. Only L* values were
significantly different among the Hunter’s Lab values. Similar
results were observed following anthocyanin and image
analyses, both of which were not significantly affected
by temperature. However, an intense red pigment was
observed at 20°C compared with the green pigment observed
at 10°C and 30°C based on the image analysis. The results
suggest that temperature influences growth, development,
and anthocyanin content of
Echeveria
succulents, and 20°C
could be the optimal temperature for the cultivation of the
species.
Additional key words: anthocyanin, CIELAB, image analysis,
segmentation, succulents, temperature
Introduction
Echeveria species are succulent species that belong to
the Crassulaceae family which has a worldwide distribution
with over 1,500 species with 33 genera and has been
characterized due to its rosette succulent leaves (Jimeno et
al. 2013). Because of its easy propagation and beauty of its
leaf rosettes and colorful flowers, this genus has been
growing in popularity among houseplant and botanical
collectors. This genus has over 130 species which have
diversity and has been found to be natives of Mexico to
Southern America and Northern Argentina (Eggli and Taylor
2002).
Although succulent crops have a high market demand
due to its water-sufficient trait (Sevilla et al. 2012), there are
many questions as to what appropriate environmental
conditions these ornamentals would thrive on and enhance
the plant quality (Rowley 1978). Succulents under this
genus are known for the development of gradient colors on
the margins of the lower or mature leaves of the plant
(Fischer and Schaufler 1981). The colors range from light
pink to red and even deep red hues that are already close
to brown or black. This change in color may be due to the
presence of the anthocyanin pigments (Welch et al. 2008).
One of the primary environmental factors that affect the
rate of plant development is temperature. It plays a
predominant role in the control and proper growth of
plants (Khodorva and Conti 2013). Different crop species
respond to temperature for their phenological growth such
their height, diameter, leaf structure, and color as well as
completion of their reproductive stages (Hatfield and
Prueger 2015).
*Corresponding author: Sang Yong Nam
Tel: +82-2-3399-1732
E-mail: namsy@syu.ac.kr
ORCID: htt
p
s://orchid.or
g
/0000-0003-0863-4721
Flower Res. J. (2019) 27(2) : 80-90
DOI htt
p
s://doi.or
g
/10.11623/fr
j
.2019.27.2.01
ISSN 1225-5009(Print)
ISSN 2287-772X
(
Online
)
O R IG IN AL AR TIC L E
Effects of Temperature on the Growth and Anthocyanin Content
of Echeveria agavoides and E. marcus
Raisa Aone M. Cabahug
1,2
, Young Jin Choi
1
, and Sang Yong Nam
1,2*
1Department of Environmental Horticulture, Sahmyook University, Seoul 01759, Korea
2Natural Science Research Institute, Sahmyook University, Seoul 01759, Korea
Received 2 November 2018; Revised 19 December 2018; Accepted 4 June 2019
Copyright © 2019 by The Korean Society for Floricultural Science

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Flower Res. J. (2019) 27(2) : 80-90 81
A defined range of maximum and minimum temperatures
for each species comes as a boundary for observable
growth. It is deemed that there must be a certain level in
which the plant will achieve its optimum growth (Hatfield
et al. 2011). Results of researches suggest that the level of
temperature is a key player in the assimilation of nutrients,
hormones and even pigments that would affect the plants’
growth rate (Adams et al. 2001). Aside from growth
parameters, studies of Rabino and Mancinelli (1986)
revealed that temperature affected the total amount of
anthocyanin in cabbage seedlings. Studies also showed that
temperature affects the biosynthesis of anthocyanin in
apples (Ubi et al. 2006), grapes (Yamane et al. 2006) and
also in roses (Lo Piero et al. 2005).
In plants, the presence of anthocyanin usually adds
beauty and colors to the ornamentals, however, in human
health, anthocyanin has an important role and considered
to be a source for dietary compounds and antioxidants
(Devi et al. 2012). Thus, the study aimed to determine the
effects of temperature levels on the growth, development,
and quality as well as the anthocyanin content of two
Echeveria species in controlled environments.
Materials and Methods
Planting materials
Two Echeveria species were chosen, namely E. agavoides
and E. marcus species, for the conduct of the study. These
succulent species were purchased from a succulent nursery
farm in Anseong Province in South Korea.
E. agavoides possesses mainly of a greener tone leaf
color while E. marcus has seen to be mainly on the blueish
green leaf color. Healthy and disease-free succulents
approximately sixty-days old grown from the said succulent
nursery and were grown at an average temperature of 18°C
(± 5°C). Experimental plants that were chosen were
standardized in both maturity and size. Succulents were
then transferred in the greenhouse of Sahmyook University,
Seoul, South Korea at the start of the experiment.
Experimental design, treatments and growth
conditions
The experiment was laid out in a completely randomized
design having three treatments with four replications (six
plants per replication), a total of seventy-two plants per
species. Three temperature levels were 10°C, 20°C, and 30°C
(± 1°C). All experimental plants were placed inside three
plant growth chambers (KGC-175 VH, Koenic Ltd., South
Korea). The relative humidity was set at 65%. There was a
14-hour light period and 10-hour dark period. The light
condition was set at 75 μmol･m
-2
s
-1
.
Data gathered
Plant height and diameter
The succulents’ height and diameter data were obtained
using a digital Vernier caliper (SR 44, Blue Bird Co.,
Japan). The plant height was taken from the base of the
soil to the highest part of the plant. The plant diameter was
taken from the largest possible measurement from each
opposing leaf tips. The data was taken at the termination of
the study which was about 6 weeks.
Visual quality rating
After the exposure of succulents to their respective
treatments, plants were subjected to the visual quality rating
(VQR). Twelve (12) respondents or judges were asked to
rate the representative plants per treatment and case study.
A modified visual score (Wang et al. 2005) was used with
the corresponding visual grading (1 - 5) where, 1: very
poor (leaves are chlorotic or irregular in color), 2: low
(leaves are light green and no red color), 3: good (leaves
are green with pinkish color), 4: very good (leaves are
green, with light red color), 5: excellent (leaves are rich
green with intense or distinguishable red color. Visual
quality rating (VQR) is a sensory analysis, which has been
widely used in horticultural crops such as fruits, vegetables,
and ornamental crops. The use of visual quality rating has
been adapted as a tool for measurement as a perception of
the human eye (Boumaza et al. 2009).

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82 Flower Res. J. (2019) 27(2) : 80-90
Hunter’s Lab
The color quality of leaves was determined using the
Hunter’s Lab. This was gathered with the use of a hand-held
spectrophotometer (Konica Minolta Spectrophotometer CM2600d,
Japan) which determines three color coordinates namely the
L* a* b* color space to indicate lightness hue and saturation
of colors. Lightness is indicated by the L* while the chromaticity
coordinates are represented by a* and b* values. The lightness
of the color is represented by the L* color value with a 0
to 100 value range. A higher positive value would indicate
a lighter color and a lower value indicates a darker color.
Positive a* values indicate the red direction while a negative
a* value indicates the green direction. On the other hand,
a positive b* value indicates a yellow direction while a negative
value indicates a blue direction.
One leaf of each plant was tagged to trace color
changes. The color value was measured by choosing the
area within the tagged leaf which were located at 1cm from
the margin of the top leaf surface (adaxial) and the
underside of the leaf (abaxial).
Anthocyanin analysis
A modified quantitative method for anthocyanin (Fuleki
and Francis 1968) was used in this study by gathering 1
inch from the tip of a tagged succulent leaf. One-gram
fresh-cut leaf samples were macerated using a mortar and
pestle. The macerated sample was added with 1 ml of 95
% ethanol and 1.5 N HCl (85:15) which served as the
extracting solvent. The mixed solution was transferred to a
separate container. Samples were then centrifuged at 13,000
rpm at 4°C using the Micro Refrigerated Centrifuge Smart
R17 (Hanil Science Co. Ltd., Seoul, South Korea). Samples
were then stored and refrigerated overnight at 4°C to
solidify the pulp residues at the bottom of the tubes after
the centrifuge process. This procedure was made in order
for the residues to remain at the base at the tube and easy
extraction of the liquid extract alone.
Samples were taken out of the refrigerator and were
fluids from the tubes without plant residues were placed in
a microplate that was then analyzed for a full-spectrum
UV/Vis absorbance at 535nm using the Fluostar Optima
Microplate Reader (BMG Labtech, Ortenberg, Germany). A
solution was placed at the primary or first plate entry.
Image analysis
Photos were taken using a digital single-lens reflex camera
(Canon 750D, Japan) with the same aperture, brightness, and
contrast at the same distance with a pixel size of 1080 p.
Individual images were cropped to show the succulents alone
without the pots and were processed using the Image-Pro
Premier ver. 9.3 (Media Cybernetics, Inc., USA). Smart
segmentation was applied to individual representative images
to determine the ratio of the colors green and red pigments.
Colored overlays of identified colors were presented as bases
for color identification.
Statistical analysis
Data gathering was done every two weeks for a month.
Aside from the Hunter’s Lab and anthocyanin content
analysis, growth and development parameters were also
collected. Statistical analyses were conducted using Statistical
Product and Service Solutions for Windows, version 16.0
(SPSS Inc., Japan). The data were analyzed using analysis of
variance (ANOVA), and the differences between the means
were tested using Duncan’s multiple range test (p < 0.05).
Results
Plant height and diameter
Based on the statistical analysis, plant height and
diameter was highly affected by temperature levels for both
species. Data for plant height is shown in Table 1 while
plant diameter is shown in Table 2.
Results revealed that succulent plant species of E.
agavoides that were subjected under high temperatures of
20°C and 30°C which had 42.36 mm and 42.33 mm. These
temperature levels did not significantly differ from each
other. Shortest plants were observed from the lowest
temperature level of 10°C which gave 41.72 mm. Different

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Flower Res. J. (2019) 27(2) : 80-90 83
results were observed with those of E. marcus wherein the
lowest and highest temperatures of 10°C and 30°C gave the
shortest plants with 40.30 mm and 41.81 mm. Tallest plants
were observed from those of 20°C having 42.54 mm which
significantly differed from other temperature levels.
In these results, it may be noted that despite belonging
to the same family, E. agavoides has a higher tolerance for
high temperatures. While E. marcus plant height may be
hindered by too low or too high temperatures.
E. agavoides species plant diameter was significantly
affected by different temperature levels. Results showed that
plants that were exposed to 20°C and 30°C were not
significantly different from each other having a plant
diameter of 94.46 mm and 93.40 mm, respectively. Shortest
plant diameter was measured from those plants subjected to
10°C with 79.42 mm. For E. marcus, it was observed that
those grown under 20°C highly gave the largest plants with
79.94 mm. Results also suggested that a low temperature of
10°C and a high temperature of 30°C were not significantly
different from each other which had 73.72 mm and 72.53
mm, respectively.
A similar trend can be observed for both parameters of
each succulent species. Although with different significant
levels, it was also noted that 20°C was consistent with the
results for growth parameters. It may be said that E.
agavoides is sensitive to colder temperature while E.
marcus may is sensitive to both lower temperature and too
high temperatures.
Visual quality rating
The results for the visual quality rating of Echeveria
species in response to temperature levels are shown in
Table 3. Results revealed that temperature levels highly
affected the visual quality rating of Echeveria species.
For E. agavoides, 10°C and 20°C were not significantly
different from each other with the highest visual quality
rating of 3.67 and 3.75, respectively, and these ratings are
described as very good quality. The results for 10°C and
20°C, however, did not significantly differ from each other.
These were followed by those treated with 30°C with a
visual quality rating of 2.08 described as a low quality
which was significantly lower compared to the two
previously mentioned temperature levels.
For E. marcus, succulents that were exposed to 20°C had
a rating of 3.75 followed by those treated under 10°C which
had a visual quality rating of 3.67. The lowest visual quality
rating was observed from in those grown under 30°C with
2.42 described as low quality.
T
able 1. Plant height (mm) of Echeveria species in response to different temperature levels at 6 weeks after treatment.
Temperature levels E. agavoides E. marcus
10°C 41.72 b
z
40.30 b
20°C 42.36 a 42.54 a
30°C 42.33 a 41.81 b
F-test
y
** **
z
Mean separation within columns by Duncan’s multiple range test at p = 0.05.
y
NS, *, **Non-significant, significant, highly significant at p = 0.05 and p = 0.01, respectively.
T
able 2. Plant diameter (mm) of Echeveria species in response to different temperature levels at 6 weeks after treatment.
Temperature levels E. agavoides E. marcus
10°C 79.42 b
z
73.72 b
20°C 94.46 a 79.94 a
30°C 93.40 a 72.53 b
F-test
y
** **
z
Mean separation within columns by Duncan’s multiple range test at p = 0.05.
y
NS, *, **Non-significant, significant, highly significant at p = 0.05 and
p
= 0.01, respectively.

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84 Flower Res. J. (2019) 27(2) : 80-90
Hunter’s Lab
Statistical results of the analysis showed that only L*
value was significantly affected by the different temperature
levels. Results of the Hunter’s Lab of Echeveria species are
shown on two separate tables below.
Succulent plants that were grown under 20°C gave the
highest value for the brightness of color or hunter L* value
with 47.36. These were then followed by those plants
exposed with 10°C with 40.31 for hunter L* value and 30°C
with an L* hunter value of 37.95. However, these two
treatments did not significantly differ from each other for E.
agavoides species in an adaxial portion of the leaves.
Hunter a* was significantly affected by the temperature
which had a similar trend to those of L* value. However,
b* values were not significantly different from each other
(Table 4).
On the abaxial portion of the tagged leaves, the highest
hunter L* value was taken from those of 10°C and 20°C
which did not significantly differed from each other. These
were then followed by those succulents grown under 30°C
with a Hunter L* value of 35.72 which means that it had a
darker color value compared to other temperature levels.
For E. marcus, similar results were also observed
whereby only hunter L* values were significantly affected
by the temperature levels (Table 5). Results revealed that
low and high temperatures had a higher lightness value
with 42.12 for 10°C and 40.51 for 30°C which did not
significantly differ with each other for the top portions of
the leaves. For the bottom portions, 10°C and 20°C did not
significantly differ from each other with 34.34 and 34.25
hunter *L values. Hunter a* was significantly affected by the
temperature which had a similar trend to those of L* value.
However, b* values were not significantly different from
each other.
Anthocyanin analysis
Results of the anthocyanin analysis of Echeveria species
in response to different temperature levels are shown in
Table 6. The average anthocyanin content was significantly
affected by temperature levels for both E. agavoides and E.
marcus species.
Results showed that exposure of E. agavoides plants to
low temperatures of 10°C had the highest anthocyanin
content with an amount of 0.92 µg/g FW. This result,
however, was significantly the same with plants that were
subjected to 20°C with 0.89 µg/g FW. The high temperature
of 30°C gave the lowest anthocyanin content among
temperature levels with an amount of 0.54 µg/g FW which
significantly differed from the other temperature levels.
Succulent plants of E. marcus were also significantly
affected with the use of different temperature levels on its
anthocyanin content. E. marcus species which were grown
under 20°C had the highest anthocyanin content amounting
to 0.63 µg/g FW which was significantly higher compared
with those of 10°C and 30°C with more or less the same
T
able 3. Visual quality rating of Echeveria species in response to different temperature levels at 6 weeks after treatment.
Temperature levels E. agavoides E. marcus
10°C 3.67 a
z
3.67 b
20°C 3.75 a 3.75 a
30°C 2.08 b 2.42 c
F-test
y
** **
z
Mean separation within columns by Duncan’s multiple range test at p = 0.05.
y
NS, *, **Non-significant, significant, highly significant at p = 0.05 and p = 0.01, respectively.
V
isual quality score (Wang et al. 2005):
1 : very poor (leaves are chlorotic or irregular in color).
2 : low (leaves are light green and no red color).
3 : good (leaves are green with pinkish color).
4 : very good (leaves are green, with light red color).
5 : excellent (leaves are rich green with intense or distinguishable red color).

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Flower Res. J. (2019) 27(2) : 80-90 85
average amount of 0.49 µg/g FW. Statistical analysis also
revealed that the use of 10°C was significantly different
from those results of 30°C. Thus, the use of too low
temperatures or too high temperatures is found to reduce
the anthocyanin content of the species.
It may be evident that higher anthocyanin content is
taken from those with E. agavoides compared to those of
E. marcus by a few percentages. This may be due to the
fact that E. agavoides may be more sensitive to temperature
levels, thus more production of the anthocyanin has been
taken. On the other hand, E. marcus has been known as
a succulent that is more tolerant of cooler or hotter
climates.
Image analysis
The image analysis was done to determine the ratio of red
and green pixels and to quantify these data from a raw image
taken at the termination of the study. The image was subjected
to the smart segmentation and histogram of the image.
Smart segmentation of images coupled with the original
T
able 4. Average Hunter’s Lab values (L*, a*, b*) using a spectrophotometer of E. agavoides leaves at adaxial and abaxial leaf orientatio
n
in response to different temperature levels at 6 weeks after treatment.
Temperature levels Adaxial leaf Abaxial leaf
L*
x
a* b* L* a* b*
10°C 40.31 b
z
1.81 15.23 47.41 a 1.23 11.88
20°C 47.36 a 0.51 12.34 47.41 a 0.57 12.31
30°C 37.95 b 0.80 12.06 35.72 b 3.73 10.26
F-test
y
** NS NS ** NS NS
z
Mean separation within columns by Duncan’s multiple range test at p = 0.05.
y
NS, *, **Non-significant, significant, highly significant at p = 0.05 and p = 0.01, respectively.
x
L*, a*, b* represent lightness from 100 (white) to 0 (black), redness (negative values indicate green, positive values indicate red),
yellowness (negative values indicate blue, positive values indicate yellow), respectively.
T
able 5. Average Hunter’s Lab values (L*, a*, b*) using a spectrophotometer of E. marcus leaves at adaxial and abaxial leaf orientation
in response to different temperature levels at 6 weeks after treatment.
Temperature levels Adaxial leaf Abaxial leaf
L*
x
a* b* L* a* b*
10°C 42.12 a
z
-0.59 10.95 34.34 b 1.16 10.32
20°C 34.18 b -1.77 11.23 34.25 b -3.20 10.74
30°C 40.51 a -0.36 12.05 42.68 a -1.19 13.00
F-test
y
** NS NS * NS NS
z
Mean separation within columns by Duncan’s multiple range test at p = 0.05.
y
NS, *, **Non-significant, significant, highly significant at p = 0.05 and p = 0.01, respectively.
x
L*, a*, b* represent lightness from 100 (white) to 0 (black), redness (negative values indicate green, positive values indicate red),
yellowness (negative values indicate blue, positive values indicate yellow), respectively.
T
able 6. Anthocyanin content (µg/g FW) of Echeveria species in response to temperature levels at 6 weeks after treatment.
Temperature levels E. agavoides E. marcus
10°C 0.92 a
z
0.49 b
20°C 0.89 a 0.63 a
30°C 0.54 b 0.49 b
F-test
y
** **
z
Mean separation within columns by Duncan’s multiple range test at p = 0.05.
y
NS, *, **Non-significant, significant, highly significant at p = 0.05 and
p
= 0.01, respectively.

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86 Flower Res. J. (2019) 27(2) : 80-90
images for three temperature levels and their corresponding
histogram results is presented on Fig. 1 for E. agavoides
and Fig. 2 for E. marcus.
The results for smart segmentation for the red and green
pixels including the ratio of pixels in a raw image is shown
in Table 7. Results suggest that the smart segmentation
results of green and red pixel ratio were significantly affected
by the different temperature levels for both Echeveria
species.
E. agavoides succulents subjected to 20°C gave the
highest red pixel count of 936.6 which accounts to 33.45%
of the total pixel in an image. However, these results did
not significantly differ from those of 10°C with a 532.9-pixel
count which accounts for 30.31% of the total pixel. The
lowest recorded number of a pixel was found in the plant
that was exposed to 30°C which has 37.3-pixel count
accounting for only 0.04% of the total image. This means
that there are more prominent green pixels compared to
the red ones. An increase of green pixels was noticed
when there was a lower temperature for succulents.
Temperature level Segmented image Histogram
10°C
20°C
30°C
Fig. 1. Smart segmentation analysis showing original image, processed segmentation image and histogram graph determining area rati
o
of green and red pigments using number of pixels of E. agavoides in response to different temperature levels.

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Flower Res. J. (2019) 27(2) : 80-90 87
Temperature level Segmented image Histogram
10°C
20°C
30°C
Fig. 2. Smart segmentation analysis showing original image, processed segmentation image and histogram graph determining area rati
o
of green and red pigments using number of pixels of E. marcus in response to different temperature levels.
T
able 7. Red and green pigment ratio and color percentages of using smart segmentation as an image analysis tool in response to
different temperature levels of Echeveria species at 6 weeks after treatment.
Temperature levels Green pixels Red pixels Sum of pixels Green pixel (%) Red pixel (%)
E. agavoides
10°C 1225.3 ± 4.09
y
532.9 ± 6.23 1758.2 69.69 b
z
30.31 a
20°C 1863.3 ± 5.07 936.6 ± 5.69 2799.9 66.55 b 33.45 a
30°C 87008.9 ± 4.08 37.31 ± 7.32 87046.2 99.96 a 0.04 b
E. marcus
10°C 930.3 ± 2.82 388.0 ± 4.16 1318.3 70.57 b 29.43 a
20°C 970.4 ± 2.35 632.5 ± 8.30 1602.8 60.54 b 39.46 a
30°C 5562.6 ± 4.49 52.9 ± 7.80 5615.6 99.06 a 0.94 b
y
Mean ± SE (Standard error).
z
Mean separation within columns by Duncan’s multiple range test at
p
= 0.05.

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88 Flower Res. J. (2019) 27(2) : 80-90
Succulent species of E. marcus pixel count and percentage
of red and green pixel ratio was significantly affected by
temperature levels. Among temperature levels that were
used to grow E. marcus plants, the highest pixel count was
taken from those that were subjected to 20°C with a pixel
count of 632.5 which accounts for 39.46%. This result was
not significantly different from those of 10°C with a red
pixel count of 388.0 which accounts for 29.43%. The
lowest amount of pixel count of 52.9 only was taken from
those succulents that were grown under 30°C which
accounts for 0.94% of the total segmented image.
It was noted that the green pixel increases when the
temperature is too low or when too high for E. marcus.
However, higher temperatures would significantly increase
green pixel compared to lower temperatures.
Discussion
Many environmental factors act singly or interact to affect
plant productivity (Haferkamp 1988) thus, basic knowledge
on the changes or manipulation of this environmental factor
must be taken to advantage to provide an optimum and
conducive growing conditions for the crop. Temperature is
considered one of the vital component as an environmental
factor for plant growth and development.
Based on the study, results revealed that for E. agavoides,
succulents grown in 10°C had the shortest and smallest in
diameter compared to those of 20°C and 30°C. A review done
by Hatfield and Prueger (2015) stated that the rate of
phenological development in warm temperatures, especially
in controlled environment studies, has found to increase
height, weight, diameter, and other growth parameters. This
is because most plant species require a higher optimum
temperature for vegetative development such as corn
(Warrington and Kanemasu 1982), sweet orange (Ribeiro et
al. 2012) and pineapple (Friend and Lydon 1979) among
others. Studies of Gent and Enoch (1983), using a mathematical
model, revealed that increased temperature or warm
environments coupled with nonstructural carbohydrate has a
corresponding increased dry matter. They added that with
reaching the high optimum temperature levels will also
increase rates of photosynthesis, growth and maintains
respiration. This was the same case in Pineapple wherein the
use of 30/20°C presented higher carbon metabolism,
photosynthetic rates, higher shoot growth and increased root
growth rates compared to lower temperature levels. Went
(1953) further discussed the effects of temperatures on plant
growth of which he stated that very low and very high
temperatures affect the physio-chemical process involved in
its development and at times may cause injury effects.
According to Pennisi et al. (2016), there are significant
negative effects that may harm plants in low temperatures
including damaged foliage, wilted stature, produced misshapen
new growth, discolored foliage and had a portion or the
whole plant dies. They also added that the effects of low
temperature may also be unseen by the naked eye but may
manifest in the later part of growth and development
through delayed flowering or stunted growth. This may be
due the interruption and damage along pathways of water,
nutrients and other important caused by freezing of plant
cells. On the other hand, high temperatures or heat stress,
plants are said to go through three mechanisms including
excessive membrane fluidity, disruption of protein function
and turnover, and metabolic imbalances (Farrell 2015). With
these mechanisms occurring in the plant, the net photosynthesis
is first to be inhibited (Allakhverdiev et al. 2008). In the
case of E. agavoides, low temperature at 10°C prompted
delayed or stunted growth in succulents.
Results showed that growing E. agavoides in temperatures
lower than 20°C had the highest anthocyanin content. It is
an established theory that temperature affects the gene
expression of enzymes involved in producing anthocyanin
(Christie et al. 1994; Leyva et al. 1995; Shvarts et al. 1997).
Rehman et al. (2017) reported that exposure to high
temperatures of Malus profusion induced anthocyanin
inhibition and activated its degradation. Proponents of the
study explain that exposure to high temperatures of 33 -
25°C increased the expression of anthocyanin repressors
MYBs and MpMYB15 and reduced MpVHA-B1 and

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Flower Res. J. (2019) 27(2) : 80-90 89
MpVHA-B2 transcriptors which are involved in the vascular
transportation of anthocyanin. These were evident in the
studies of Mori et al. (2007) which suggested that the
exposure of grapes in high temperature of a maximum of
35°C reduced the total anthocyanin content to less than half
of the control (20 - 25°C).
In the studies of Lo Piero et al. (2005) with red oranges,
it was concluded that low temperatures could also reduce
the expression of anthocyanin, especially during long exposures.
On the other hand, Solecka et al. (1999) reported that
anthocyanins have increased when subjecting oilseed rape
leaves to low temperatures which were explained to be
involved in the process of protecting mesophyll cells against
cold environments. This may be the reason for which there
was a high anthocyanin content for plants treated at 10°C
that was comparable to those of 20°C in E. agavoides.
The anthocyanin content in plants are generally the
reason for the change in color in plants and have been
specifically studied in ornamental plants as changes of color
in the foliage or inflorescence can increase visual quality.
The use of Hunter’s Lab is able to differentiate the lightness
of the color and the difference between two opposing
colors such as that or red/green and yellow/blue (Konica
Minolta 2018). Results showed that there was no significant
difference between a* and b* values both adaxial and
abaxial leaf. This may be because the difference was
exiguous between the hues, however, this was not true to
the intensity (lightness or deepness) of the colors which
was found to be significantly different between temperature
levels. Studies of Shisa and Takano (1964) suggested that
the effects of temperature on the epidermal cells have
affected the lightness or deepness of red coloration of rose
flower petals.
Acknowledgment
This work is supported by Succulents Export Innovation
Model Development towards Chinese Market (514006-03-1-
HD040)’, Ministry of Agriculture, Food and Rural Affair and
Sahmyook University Research Fund.
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