Flora 263 (2020) 151535

Contents lists available at ScienceDirect

Flora
journal homepage: www.elsevier.com/locate/flora

Photochemical activity in early-developmental phases of Agave angustifolia T

subsp. tequilana under induced global warming: Implications to temperature
stress and tolerance
José Luis Aragón-Gastéluma,c,*, J. Efrain Ramírez-Beníteza, Enrique González-Durána,
Claudia González-Salvatierrab, Hugo M. Ramírez-Tobíasc, Joel Floresd,
Eduardo J. Gutiérrez-Alcántaraa, Evelyn Méndez-Guzmánc, Ramón Jarquín-Gálvezc
a
Facultad de Ciencias Químico-Biológicas, Universidad Autónoma de Campeche. Avenida Ing. Humberto Lanz Cárdenas y Fraccionamiento Ecológico Ambiental Siglo
XXIII. Colonia. Ex Hacienda Kalá, San Francisco de Campeche, Campeche, 24085, Mexico
b
Cátedras CONACYT-Tecnológico Nacional de México, Instituto Tecnológico de Chetumal, Avenida Insurgentes No. 330, Colonia David Gustavo Gutiérrez, Chetumal,
Quintana Roo, 77013, Mexico
c
Facultad de Agronomía y Veterinaria, Universidad Autónoma de San Luis Potosí, Km. 14.5 Carretera San Luis Potosí-Matehuala, Ejido Palma de la Cruz, Soledad de
Graciano Sánchez, S.L.P. 78321, Mexico
d
IPICYT-División de Ciencias Ambientales, Camino a la Presa San José No. 2055, Colonia Lomas 4a. Sección, San Luis Potosí, S.L.P. 78216, Mexico

A R T I C LE I N FO A B S T R A C T

Edited by: Hermann Heilmeier Agave species are a key component of the structure vegetation in the American drylands and have various
Keywords: anthropogenic uses. Agave angustifolia subsp. tequilana is an economically and culturally important Mexican
Anthocyanins species. Negative impacts on biodiversity are expected as a consequence of global warming; however, it has been
Asparagaceae documented that some Agave species are not affected by projected warming. Previous studies on some succulent
Chlorophyll fluorescence species under induced global warming have shown a certain level of tolerance to temperature stress. We eval-
Climate change uated the potential temperature stress tolerance of two early developmental phases of A. angustifolia subsp.
Leaf pigments tequilana, by means of the following photochemical parameters: maximum quantum efficiency of photosystem II
Open-top chambers
(Fv/Fm); effective quantum yield of PSII (ΦPSII); electron transport rate (ETR); non-photochemical quenching
(NPQ); and pigment content (total chlorophyll, carotenoids and anthocyanins) in two-year-old plants and five-
year-old plants under induced warming. Temperature and vapor pressure deficit (VPD) were higher in open-top
chamber (OTCs) induced warming treatment than control plots, reproducing current climatic conditions. Both
developmental phases showed high Fv/Fm values in both OTCs and control plots. Two-year-old plants displayed
higher NPQ values than five-year-old plants under warming. The ΦPSII and ETR values were low in both warming
treatments and developmental phases. Low anthocyanin content was found in both treatments and develop-
mental phases. Independently of warming, the two-year-old plants showed higher total chlorophyll and car-
otenoid content values than the five-year-old plants. Despite that induced warming affected the photochemical
activity of A. angustifolia subsp. tequilana, the high NPQ and pigment values indicate that this species has a high
tolerance to future global warming. Our results provide novel evidence of the high physiological tolerance that
could act in the early developmental phases of a key Agave species to cope the effects of global warming.

1. Introduction and semi-arid environments will be affected by both increased tem-

peratures and a decreased frequency and magnitude of rainfall events
Climate change is expected to pose a severe threat to biodiversity on (Stocker et al., 2014). The structure and functioning of semiarid eco-
our planet (Ureta et al., 2012). Future climate models predict that arid systems are strongly influenced by precipitation patterns, while

⁎
Corresponding author at: Facultad de Ciencias Químico-Biológicas, Universidad Autónoma de Campeche. Avenida Ing. Humberto Lanz Cárdenas y
Fraccionamiento Ecológico Ambiental Siglo XXIII. Colonia. Ex Hacienda Kalá, San Francisco de Campeche, Campeche, 24085, Mexico.
E-mail addresses: jlaragon@uacam.mx (J.L. Aragón-Gastélum), jeramire@uacam.mx (J.E. Ramírez-Benítez), eagonzal@uacam.mx (E. González-Durán),
cgonzalez@itchetumal.edu.mx (C. González-Salvatierra), hugo.ramirez@uaslp.mx (H.M. Ramírez-Tobías), joel@ipicyt.edu.mx (J. Flores),
ejgutier@uacam.mx (E.J. Gutiérrez-Alcántara), eeveelyn.daanieela@gmail.com (E. Méndez-Guzmán), ramon.jarquin@uaslp.mx (R. Jarquín-Gálvez).

https://doi.org/10.1016/j.flora.2019.151535
Received 1 May 2019; Received in revised form 18 December 2019; Accepted 23 December 2019
Available online 31 December 2019
0367-2530/ © 2019 Elsevier GmbH. All rights reserved.
J.L. Aragón-Gastélum, et al.
vegetation dynamics in these areas are subject to severe drought stress
conditions as well as high temperatures and light intensity (Nobel,
2010), affecting both physiological and biochemical processes
(Hasanuzzaman et al., 2013). The Chihuahuan Desert is the largest hot
desert in North America (Archer and Predick, 2008), ranging from the
south-west of the United States to the Central Mexican Highlands.
Predictive models of global change indicate an increase of 1–3 °C in arid
and semi-arid landscapes worldwide by the end of the twenty-first
century (Stocker et al., 2014). This raises concerns, given that the
Chihuahuan Desert harbors a high richness of succulent plants
(Rzedowski, 1991), many of them protected by Mexican environmental
laws (Secretaría de Medio Ambiente y Recursos Naturales, 2010).
Agave species are native to the north and central sections of the
American continent, comprising more than 200 species, about 75% of
which are found in Mexico (García-Mendoza, 2007), and exerting a
determinant effect on the vegetation structure in diverse environments,
such as arid, semi-arid, temperate and tropical ecosystems (García-
Moya et al., 2011; Reyes-Agüero et al., 2000). Agave species are dis-
tributed over a wide elevational (0–3400 m.a.s.l.) range and are used as
a source of fiber, as well as for food, shelter, ornaments, and the pro-
duction of fermented beverages such as pulque, mezcal, bacanora and
tequila (Colunga García-Marín et al., 2007; García-Mendoza, 2007;
García-Moya et al., 2011). Additionally, some Agave species have been
proposed as a raw material for the production of second-generation
biofuels (Valenzuela, 2011) and the removal of heavy metals from soils
(Romero-González et al., 2007). An endemic Mexican Agave species
found in arid and semi-arid regions (Gentry, 1982), Agave angustifolia
subsp. tequilana is one of the most economically important Agave spe-
cies in the country because it is used as the basis for mezcal production
(Colunga García-Marín et al., 2007). Wild Agave angustifolia subsp. te-
quilana populations are being overexploited due to human population
pressure caused by increasing market demand for this beverage, re-
sulting in a decrease in the plant’s population size (Macías-Macías and
Valenzuela-Zapata, 2009).
Succulent plants have exceptional physiological (e.g. Crassulacean
Acid Metabolism or CAM) and ecological adaptations that allow them
to survive in unfavorable environmental conditions, such as high tem-
peratures, high levels of solar light, low air relative humidity (García-
Moya et al., 2011; Nobel, 2010). These strategies include anatomical
adaptations, efficient energy dissipation mechanisms and robust redox
protection systems (Nobel, 2010). However, these findings have been
reported for adult plants, neglecting the potential impact of these
stressful conditions in early developmental phases, during which suc-
culent species are often more sensitive to stress conditions than once
they have reached adult size (Flores et al., 2004).
Several methodologies have been proposed for evaluating the im-
pacts of climatic change on plants. Passive Open Top-Chambers (OTCs),
devices that do not require technological support, have been success-
fully used to assess plant responses to climate warming (a driver of
climate change) (Bokhorst et al., 2013; Hollister and Webber, 2000).
Various studies using OTCs have found differential responses in survival
rate (Aragón-Gastélum et al., 2017), germinability (Aragón-Gastélum
et al., 2018) and photosynthetic performance (Aragón-Gastélum et al.,
2014; Musil et al., 2009) to induced warming in various succulent
species. In CAM plants subjected to high-light stress, reductions in leaf
chlorophyll a and b content (Hazrati et al., 2016) and increases in
carotenoid and anthocyanin content (González-Salvatierra et al., 2010;
Jeon et al., 2006) have been found, along with higher photosynthetic
performance than non-stressed CAM plants.
In the case of Agave species, few studies have focused on assessing
variations in photoprotective pigments (see Graham and Nobel, 1996)
or photosynthetic performance. Moreover, studies assessing the impact
of global warming in the early developmental phases of Agave species
also remain scarce (see Pérez-Sánchez et al., 2015). Under induced
global warming treatments, Aragón-Gastélum et al. (2014) found de-
creased levels in the photosynthetic parameters of five-year-old
2
Flora 263 (2020) 151535
Echinocactus platyacanthus plants; in addition, Aragón-Gastélum et al.
(2017) documented a high mortality rate for two-year-old plants of
Ferocactus histrix, Stenocactus coptonogonus and E. platyacanthus. Both of
the above-mentioned studies suggest that future global warming could
severely threaten the persistence of wild populations of these cacti
species. It is possible that similar responses could be found in the early
developmental phases of Agave species.
The aim of the present research was to assess the effect of induced
global warming on photochemical activity and photoprotective pig-
ments in two-year-old and five-year-old A. angustifolia subsp. tequilana
plants. Our hypothesis is that induced global warming will negatively
impact on photochemical activity and pigment content in both of the
above-mentioned developmental phases of this species.
2. Materials and Methods
2.1. Plant material
Agave angustifolia subsp. tequilana (F.A.C. Weber) is a plant with
semelparous rosettes that commonly propagates asexually via bulbils
and rhizomatous offshoots (Gentry, 1982), with sexual propagation also
documented in this species (Ramírez-Tobías et al., 2012, 2014). While
A. angustifolia subsp. tequilana is primarily found in tropical deciduous
forest (García-Amaro, 2004) with a mean annual temperature > 18 °C
and an average annual precipitation > 900 mm, this species is also
found in arid and semi-arid environments (Ramírez-Tobías et al., 2012,
2014). The present study was conducted in the Altiplano Potosino, the
high plateau found in the state of San Luis Potosí, Mexico and the
southernmost section of the Chihuahuan Desert. The mean annual
temperature in this area is 17.8 °C, it can surpass 35 °C in summer
(June–September) and fall below −1 °C in winter (December–March).
The average annual precipitation is 341 mm, with the rainy season
occurring between June and October (Medina-García et al., 2005).
Seed capsules from A. angustifolia subsp. tequilana plants, grown at
the Institute for Research of Desert Areas at the Autonomous University
of San Luis Potosi, were collected during physiological maturity
(yellow-greenish capsules) in September 2011 and September 2014.
Vegetation is sclerophyllous scrub with species of Cactaceae such as E.
platyacanthus, F. histrix, Opuntia ficus-indica and Cylindropuntia spp. as
well as Fabaceae (Prosopis leavigata and Acacia farnesiana) with an an-
nual mean photosynthetic photon flux density (PPFD) of 1300 μmol
m−2s-1. The capsules were collected from at least 10 healthy vigorous
parent plants. Seeds were sown in plastic bags (one seed each) with an
11 l (22 cm diameter ×26 cm depth) capacity, in a mixture of landfield
soil (75%) and tezontle gravel (25%) of a sandy clay soil texture. The
plants were then grown in a glasshouse, under conditions comprising
temperatures of 25–30 °C, 40% relative humidity (RH) with similar
PPFD values as described above, for two (seedlings) and five (young
individuals) years. All bags were watered every week to field capacity
(100 ml per bag).
2.2. Experimental design
The experimental site was an abandoned agricultural field (22° 14´
11´´ N, 100° 51´ 46´´ W; 1844 m a.s.l.) in the southern section of the
Chihuahuan Desert, in the municipality of Soledad de Graciano Sanchez,
in the state of San Luis Potosí, Mexico. The vegetation is dominated by
sclerophyllous shrubs, with cacti and succulent monocots scattered
among these woody species. A 25 m × 25 m section was cleared and
then fenced with woven wire to a height of 2 m to prevent fauna from
accessing the experimental site. A 2.5 m-wide strip was established
from the edge of the landfield, with sixteen 5 m × 5 m plots (25 m2
each) drawn at the center. The OTC and control treatments were ap-
plied at the center of ten randomly selected plots, as described by
Aragón-Gastélum et al. (2014).
Devices enabling daytime passive heating by increasing air
J.L. Aragón-Gastélum, et al.
temperature, hexagonal OTCs are the most common and simplest tool
for assessing plant responses to climate warming in the field (Bokhorst
et al., 2013). The OTCs used in the present research were constructed
from six trapezoidal sheets (58 cm height × 1.22 m major side ×87 cm
minor side) and UV-resistant transparent acrylic (3 mm thick and wa-
velength transmission in the range 110 to < 280 nm) following the
design proposed by Marion (1996). On September 7th, 2016, six bags
containing three two-year-old plants and three five-year-old A. angu-
stifolia subsp. tequilana plants (one individual per bag) were placed at
random in the center of each all ten experimental plots, in a hexagonal
arrangement and with a spacing of 20 cm between them.
2.3. Measurement of environmental parameters
The variation of environmental factors, such as temperature, RH
and PPFD, during the experiment was determined. The temperature and
RH data for each experimental plot were registered using microclimatic
data-loggers (HOBO Pro v2, Onset Computer Corporation, Bourne,
Massachusetts, USA), every 30 min throughout the entire experiment
(from 1st October to 31st December 2016). A data-logger was installed
10 cm above the ground at the center of each experimental plot, giving
five randomly selected OTCs and five control plots. The daily mean,
minimum and maximum temperatures and daily mean relative hu-
midity of the air were calculated for both the OTC and control plots.
Based on determination of temperature and RH in both warming
treatments, we also calculated the mean and maximum daily vapour
pressure deficit (VPD). This variable was estimated in accordance with
Jones (2013) as VPD = es(T)-e, where es(T) is saturation vapour pressure,
which is a function of temperature following the Magnus equation de-
scribed by the following function:
es(T) = ƒ[a exp(bT/(c + T))]
Where es(T) is in Pascal (Pa), T is in °C and the empirical coefficients are:
a = 611.21, b = 18.678 – (T/234.5), c = 257.14. ƒ = 1.0007 + 3.46
× 10−8 P (atmospheric pressure in Pa). e is water vapour pressure and
was estimated as e = [es(T)] × RH. The VPD values were expressed as
Pascal (Pa). Daily rainfall (mm) and atmospheric pressure (P) data were
obtained from Soledad weather station (22° 11′ 00″ N, 100° 51′ 00″ W;
1882 m a.s.l.) at the National Institute of Forestry, Agriculture and
Livestock facilities in San Luis Potosí, Mexico.
2.4. Determination of the plants’ photochemical activity
Two rounds of chlorophyll fluorescence measurements were taken
from A. angustifolia subsp. tequilana leaves in both developmental
phases (namely from two-year-old and five-year-old plants) and in both
treatments (OTCs and control plots). The size of leaves in the two-year-
old and five-year-old plants was 10–12 cm and 25–30 cm respectively.
The measurements were carried out at both predawn (between 05:00
and 06:00 h – when the plants undergo both low daily temperature,
VPD and PPFD values) and noon (between 13:00 and 14:00 h – when
the plants undergo both high daily temperature, VPD and PPFD values),
using a portable photosynthesis fluorometer (LI-COR 6400-40 Leaf
Chamber, Lincoln, Nebraska, USA). We selected the biggest leaf in both
the two-year-old and five-year-old plants of A. angustifolia subsp. te-
quilana and then, we placed the central part inside of the photosynthesis
fluorometer leaf chamber to perform the chlorophyll fluorescence
measurements. All chlorophyll fluorescence measurements were done
in both the same leaf and place during the course of the experiment.
The measurements of photochemical activity in A. angustifolia subsp.
tequilana took place at time intervals of 18–22 days (October 8th,
October 31th, November 18th, December 7th and December 27 th).
We estimated the maximum quantum efficiency of PSII (Fv/Fm), the
effective quantum yield of PSII (ΦPSII), the electron transport rate (ETR)
and the non-photochemical quenching efficiency (NPQ). The first series
3
Flora 263 (2020) 151535
of chlorophyll fluorescence readings were taken on dark-adapted plants
at predawn at 0 μmol m−2s-1 of PPFD in order to assess Fv/Fm, which
was estimated as Fv/Fm = (Fm– F0)/Fm, where Fv = variable fluores-
cence, Fm = maximal fluorescence, and F0 = minimal fluorescence
(Maxwell and Johnson, 2000).
Values of ΦPSII were taken at noon and computed as ΦPSII = (Fm′ –
Ft Fm′ , where Fm′ is the maximum fluorescence when a saturating pulse
)/
(2000 μmol m−2 s-1, with 10% blue light) of actinic light is super-
imposed on the environmental light level and Ft is the fluorescence
emitted by leaves under steady-state illumination (Maxwell and
Johnson, 2000). The ambient PPFD values (recorded via a solar irra-
diation sensor in the fluorometer leaf clip) varied across of the ΦPSII
measures and were distributed as follows: October 8th =1219.3 μmol
m−2 s-1, October 31th =1309.7 μmol m−2 s-1, November 18th
=1483.5 μmol m−2 s-1, December 7th =1552.1 μmol m−2 s-1 and
December 27th =614.8 μmol m−2 s-1.
The ETR across the electron chain of chloroplasts was estimated as
ETR = ΦPSII × PPFD × 0.84 × 0.5. A reading of 0.84 is considered the
most common leaf absorbance coefficient for plants under a wide range
of environmental conditions, with 0.5 indicating that two photons are
used for exciting one electron, as an equal distribution of excitation was
assumed for both photosystems (Hernandez-Gonzalez and Briones-
Villareal, 2007).
Non-photochemical quenching efficiency (NPQ) was determined in
accordance with the equation NPQ = (Fmo – Fm′ )/ Fm′ , where Fmo is the
basal chlorophyll fluorescence emitted by plants in darkness at 0 μmol
m−2 s-1 and Fm′ is the maximum fluorescence emitted by chlorophyll
after a saturating pulse of actinic light is imposed at noon. These actinic
light values were according to the PPFD values in each treatment (OTC
and control plots) and ΦPSII measurements. Linearly related to heat
dissipation, NPQ lies on a scale of 0–infinity (Maxwell and Johnson,
2000) and is a parameter that has already been widely documented in
succulent species subject to different kinds of abiotic stress (Aragón-
Gastélum et al., 2014; Arroyo-Pérez et al., 2017; de la Rosa-Manzano
et al., 2016; González-Salvatierra et al., 2013; Ptushenko et al., 2013;
Samoilova et al., 2011).
2.5. Determination of photoprotective pigments
One feature of succulent species is their thick waxy cuticle, which
prevents excessive water loss and partially enables their high stress
tolerance (Nobel, 2010; Ogburn and Edwards, 2010). We assume that
there was no difference in photosynthetic pigments and anthocyanins in
A. angustifolia subsp. tequilana leaves at the beginning of the study
because the plants had been kept in optimal greenhouse conditions.
Thus, total chlorophylls, carotenoid and anthocyanin content were
determined at the mid-point of the photosynthetic experiment (after the
readings had been taken on November 18th) for the treated and control
groups for both developmental phases of our target species.
One leaf disk (0.05 g) was taken at noon from the mid-upper leaf
part considering both axial and abaxial face from of the same leaf used
in the chlorophyll fluorescence measurements (one leaf disk × each of
the three two-year-old and five-year-old individuals inside of the five
OTCs and control plots). The leaf samples were ground in liquid ni-
trogen, with crude extracts obtained with the addition of 1.5 ml of N,N-
dimethylformamide (DMF, Sigma-Aldrich, St. Louis, Missouri) and in-
cubated at room temperature (25 °C) for 24 h in darkness (Minocha
et al., 2009). Extracts were then centrifuged at 5000 × g for 5 min, with
the supernatant absorbance at 664, 647 and 480 nm measured using a
spectrophotometer (GENESYS 10uv, Thermo Electron Corporation).
Variations of total chlorophylls and carotenoids in both the OTCs and
control plots and both the two-year-old and five-year-old plants were
calculated according to Porra et al. (1989).
The extraction of anthocyanins was carried out in accordance with
Neff and Chory (1998). Samples of fresh leaves (0.05 g, similarly
sampling to the pigments described above) taken from both two-year-
J.L. Aragón-Gastélum, et al. Flora 263 (2020) 151535

old and five-year-old plants from inside and outside the OTCs were
ground in liquid nitrogen, resuspended in 1% HCl in methanol, and left
in darkness for 24 h at 4 °C. Solids were separated by centrifugation
(5000 × g for 5 min) and the absorbance of the supernatant then
measured at 530 nm using a spectrophotometer (Beckman Coulter
DU650). The total anthocyanin content was determined in accordance
with the equation Total Anthocyanins (TA) = A530 × dilution factor/
98.2, as proposed by Yang et al. (2007). All pigments described above
were expressed as μg mg−1 leaf dry weight (DW).

2.6. Statistical analysis

The environmental (mean, maximum and minimum daily air tem-

perature, RH, PPFD and VPD) variables corresponding to the experi-
mental plots were analyzed based on five replicates for each data logger
(five OTCs and five control plots) and compared via the application of
repeated measure ANOVA test. The differences in chlorophyll fluores-
cence (Fv/Fm, ΦPSII, ETR and NPQ) data throughout time were de-
termined by repeated measures ANOVA considering five replicates for
each OTC and control plots (n = 5). In addition, variations in the
fluorescence and pigments (total chlorophylls, carotenoids and antho-
cyanins) data between warming treatments (OTC and control plots) and
development phases (two-year-old and five-year-old plants) were
compared with a two-way hierarchical nested ANOVA (developmental
phases nested in treatments n = 5). Since normality and homo-
scedasticity were achieved only with pigment data, and fluorescence
data were not normal, comparisons were made with Tukey and
Newman-Keuls tests, respectively. All analyses described were con-
ducted and processed using STATISTICA version 7 software (StatSoft
Inc., Tulsa, OK, USA).

3. Results

3.1. Micro-environmental validation of open-top chambers

The maximum daily air temperature was significantly different

throughout the entire experiment (F (1,176) = 73. 353, P < 0.0001)
between OTCs (average =36.1 °C ± 0.7 SE) and controls (average
=28.1 °C ± 0.5 SE) (Fig. 1a). We found significant effects of both
treatments (F (1,176) = 73. 353, P < 0.0001) and time (F (2,352) =
120.178, P < 0.0001), while the interaction between both factors was
also significant (F (2,352) = 11.726, P < 0.0001). The maximum daily
air temperature values showed a decreasing pattern over time and they
were higher in the OTCs than in the control plots throughout the ex-
perimentation period. However, the difference between the two
warming treatments was different for various time periods; during some
days in November, the maximum daily air temperature values were
similar in both OTCs and control plots.
The minimum daily air temperature values did not differ between Fig. 1. Climatic variables (mean ± standard error, n = 5) measured in OTCs
the OTCs (average =7.0 °C ± 0.4 SE) and control plots (average =7.4 (solid symbols) and control plots (empty symbols). The upper panel shows the
°C ± 0.3 SE) (F (1,176) = 0.5274, P > 0.05). However, both time (F daily maximum air temperature values (triangles), daily mean air temperatures
(2,352) = 37. 2362, P < 0.0001) and the interaction between warming
(circles), and daily minimum air temperatures (squares). The central panel
and time (F (2,352) = 120. 1115, P < 0.0001) were significant shows the average relative humidity values. The lower panel shows the daily
(Fig. 1a). The minimum daily air temperature values decreased over maximum VPD values (triangles) and daily VPD (circles).
time, but they were slightly lower in control plots than OTCs at the end
of the experiment. The mean daily air temperature was higher in OTCs The total rainfall recorded during the experiment was 106.4 mm at
(17.9 °C ± 0.3) than in the control plots (15.9 °C ± 0.2; Fig. 1a), a the study site. Significant differences were observed in the RH values in
variable which significantly differed between treatments (F (1,176) = response to treatment (OTC vs. control plots; F (1,176) = 4.065, P =
20.632, P < 0.0001) and over time (F (2,352) = 59.401, P < 0.0001), 0.0453) and time (F (2,352) = 51.942, P < 0.0001), with the interac-
while the interaction between warming and time was also significant (F tion (F (2,352) = 24.685, P < 0.0001) between both factors also sig-
(2,352) = 161. 577, P < 0.0001). The mean daily air temperature va- nificant (Fig. 1b). Differences in RH between OTCs and control plots
lues decreased over time and they were higher in the OTCs than in the were generally smaller after beginning of November than before. The
control plots during the entire study period. However, the difference PPFD values were affected by time (F (4,232) = 21.422, P < 0.0001),
between OTCs and control plots also differed between various time but the warming (F (1,58) = 0.320, P = 0.5735) and the interaction
periods; during some days in November and December, the mean daily between both factors were not significant (F (4,232) = 1.004, P =
air temperature showed similar values in both warming treatments. 0.4059). The PPFD values decreased over time but this variable showed

4
J.L. Aragón-Gastélum, et al.
similar values between the OTCs and control plots.
Both mean and maximum daily VPD values were higher in OTCs
(1215.5 ± 62.5 and 4883.0 ± 241.0 Pa respectively) than in the con-
trol plots (798.9 ± 39.1 and 2643.4 ± 120.8 Pa respectively). Beyond
these differences between warming (F (2,352) = 31.8592, P < 0.0001,
and F (2,352) = 69.0025, P < 0.0001 respectively), these variables
changed time (F (2,352) = 53.0960, P < 0.0001 and F (2,352) =
77.5457, P < 0.0001 respectively) with a significant interaction be-
tween warming and time (F (2,352) = 11.9336, P < 0.0001 and F
(2,352) = 4.7846, P < 0.0001 respectively). The mean and maximum
VPD values decreased at the middle of the experiment and they were
higher in the OTCs than in the control plots; however, the difference
between the treatments was much less beginning of November com-
pared to other periods (Fig. 1c).
3.2. Photochemical activity under induced warming
The photochemical activity of A. angustifolia subsp. tequilana was
differentially affected by the warming treatments and the two devel-
opmental phases across time. The Fv/Fm values were higher than 0.7347
for almost all treatment combinations. Although the developmental
phase nested in the warming treatment was not significant (F(1,296) =
1.26, P = 0.2622), Fv/Fm did differ according to warming (F(1,56) =
12.55, P < 0.0001), time (F(4,224) = 19.76, P < 0.0001) and the in-
teraction between both factors (F(4,224) = 14.26, P < 0.0001)
(Fig. 2a). Thus, Fv/Fm was significantly higher in OTCs than control
plots on December 07, whereas differences at other dates were not
significant.
ΦPSII was affected by time (F(4,224) = 27.4315, P < 0.0001) and
the time × warming (F(4,224) = 5.1391, P < 0.0001) interaction;
however, we found no significant effects of warming (F(1,56) = 0.3567 P
= 0.5527) and developmental phase nested in the warming treatment
(F(1,296) = 1.1524, P = 0.3173). Whereas ΦPSII was higher for control
compared to OTC treatment at the start of the experiment, later on OTC
treatment showed higher ΦPSII values than control plots (Fig. 2b).
The ETR values were also affected by time (F(4,224) = 27.615,
P < 0.0001); however, warming (F(1,56) = 0.566, P = 0.4591), de-
velopmental phase nested in the warming treatment (F(1,296) =
0.2383, P = 0.7881), and the time × warming (F(4,224) = 2.215 P =
0.0685) interaction were all found not to be significant. This variable
showed higher values at the beginning of the experiment, and although
they decreased over time, the ETR values were similar in the two-year-
old plants and the five-year-old plants in both warming treatments
(Fig. 2c).
The NPQ values did differ for time (F(4,224) = 130.6258,
P < 0.0001) and the interaction between time and warming (F(4,224)
= 49.2444, P < 0.0001); however, warming (F(1,56) = 0.0003, P =
0.9858) and developmental phase nested in the warming treatment
(F(1,296) = 0.0851, P = 0.9184) were not significant. Thus, differ-
ential responses in the NPQ values in the experimental timeframe were
found. Although the NPQ values were similar at the beginning of the
experiment for both developmental phases inside of the OTCs and
control plots, the two-year-old plants displayed slightly higher NPQ
values than the five-year-old plants in the OTCs than the control plots at
the end of the experiment. Moreover, except for the first date, OTCs and
control plots responded differently to warming, although this response
was not consistent over the experimental period (Fig. 2d).
3.3. Pigment and antioxidant content
We found significant differences in the chlorophylls and carotenoids
in the A. angustifolia subsp. tequilana leaves (Table 1). Whereas the total
chlorophyll was affected by warming (F(1,56) = 4.0216, P = 0.0497), in
the case of the carotenoid content, the effect between OTC and control
plots was not significant (F(1,56) = 3.5885, P = 0.0633). Nevertheless,
both variables were affected by developmental phase nested in the
5
Flora 263 (2020) 151535
warming treatment (F (1,56) = 5.1648, P = 0.0085 and F (1,56) =
6.1568, P = 0.0038 respectively).
Independently of warming treatments (OTC vs control plots), the
two-year-old A. angustifolia subsp. tequilana plants showed higher total
chlorophyll and carotenoid content than the five-year-old plants.
Furthermore, the two-year-old A. angustifolia subsp. tequilana plants
inside OTCs also displayed higher values of these pigments than all
other treatment combinations (Table 1),
Finally, although the anthocyanin content did not differ between
warming (F(1,56) = 0.03047, P = 0.5831) and developmental phase
nested in the warming treatment (F (1,56) = 3.0606, P = 0.0547), in the
five-years-old A. angustifolia subsp. tequilana plants, the anthocyanin
values were lowest in control plots compared to all other treatment
combinations (Table 1).
4. Discussion
Scenarios of global warming for the mid to late twenty-first century
in arid and semi-arid ecosystems indicate a temperature increase of 1–3
°C (Stocker et al., 2014). We found that the mean daily air temperature
was 2.0 °C higher and RH was 3.3% lower in the OTCs than the control
plots. This temperature increase is within the above described scenarios
for arid and semi-arid ecosystems. In dryland landscapes, the mean
daily air temperature in OTCs have been found to be 1.9–5.0 °C higher
than the external environment (Aragón-Gastélum et al., 2014, 2017,
2018; Musil et al., 2005, 2009). At low RH, the atmospheric water
vapor is also low and consequently, water condensation in the soil is
reduced, negatively modifying water uptake via the plant’s superficial
roots (Matimati et al., 2013), which could be detrimental for the sur-
vival of succulent species (Aragón-Gastélum et al., 2014, 2017, 2018).
However, we found 100% survival in both the two-year-old and five-
year-old plants in the OTCs and control plots after three months, sug-
gesting that both developmental phases of A. angustifolia subsp. tequi-
lana have a high level of heat stress tolerance.
VPD is a combination of temperature and RH in a single value,
which indicates the difference between the saturation vapor pressure
and the actual vapor pressure of the air (Howell and Dusek, 1995). VPD
has an opposite relation to RH; when RH is high VPD is low; thus, it is
the relevant environmental parameter determining evapotranspiration.
In some well-watered crops, high VPD promotes a high evapo-
transpiration rate and consequently increases in the stomatal con-
ductance, performance and yield (Sinclair and Bennett, 1998); how-
ever, under unfavorable conditions such as drought, increases in VPD
limit the parameters above described (Bunce, 1981; Turner et al., 1984;
Comstock and Ehleringer, 1993). Under our global warming scenario,
we found lower RH values in the OTCs than the control plots, and
consequently higher VPD values in the OTCs than control plots were
registered; but this did not have a detrimental effect on survival in both
developmental phases of A. angustifolia subsp. tequilana. There are no
reports about VPD projections under global warming using OTCs;
therefore, our research is pioneer to include this parameter to forecast
population dynamics of an Agave species under future climatic condi-
tions.
In recent years, many studies have focused on assessing the impact
of expected climate change on arid and semi-arid environments.
Nevertheless, in succulent species, experimental physiological research
which takes photosynthetic parameters into account using OTCs to
forecast the impact of future global warming is still scare. Thus, we
hypothesized that induced global warming by OTCs would negatively
impact on photochemical activity and pigment content of two-year-old
and five-year-old A. angustifolia subsp. tequilana plants.
Using OTCs, decreases in the effective photosynthetic quantum
yields have been reported for lichen species from dry and humid sites
under temperature and precipitation restrictions in the arid South
African Succulent Karoo Biome (Maphangwa et al., 2012). Similarly,
Musil et al. (2009) reported a decline in both photochemical efficiency
J.L. Aragón-Gastélum, et al. Flora 263 (2020) 151535

Fig. 2. Variables of photochemical activity measured in the two developmental phases of Agave angustifolia subsp. tequilana under induced global warming. A)
Maximum quantum efficiency of PSII (Fv/Fm), B) Effective quantum yield of PSII (ΦPSII), C) Electron transport rate (ETR) and D) Non-photochemical quenching
(NPQ). Data shown are mean ± standard error, n = 5. Asterisks indicate significant differences between treatments and developmental phases in each time of
experimentation (P < 0.05).

Table 1 and ETR for the succulent Cephalophyllum spissum (Chenopodiaceae), a

Pigment concentration (mg g−1 DW) in the leaves taken from two develop- South African succulent species, following short (2 h) exposures to heat
mental phases of Agave angustifolia subsp. tequilana under induced global of increasing intensity in the temperature range 42–54 °C. Both studies
warming. Data shown are mean ± standard error, n = 5. conclude that under these induced global warming scenarios, the
Developmental phase Two-year-old plants Five-year-old plants thermal tolerance thresholds of the species were exceeded, and the
decreased photosynthetic activity could lead to metabolic impairment
Treatment/Variable OTC Control plots OTC Control plots and increased incidences of mortality.
Total chlorophyll 461.4 ± 445.6 ± 36.0 255.9 ± 396.7 ±
Commonly, Fv/Fm and ΦPSII are used as sensible indicators of a
30.7 16.1 37.1* plant’s photochemical activity (Maxwell and Johnson, 2000). A plant’s
Carotenoids 86.0 ± 5.7 79.9 ± 6.2 47.2 ± 3.0 75.0 ± 7.1* Fv/Fm and ΦPSII values oscillate between 0.80 and 0.83 if environmental
Anthocyanin 0.070 ± 0.097 ± 0.081 ± 0.064 ± stress is negligible, although these values fall as environmental stress
0.01 0.007 0.009 0.009
increases, indicating damage to the photosynthetic apparatus (Maxwell
* Asterisks indicate significant differences between developmental phase and Johnson, 2000). In terms of the photosynthetic parameters docu-
nested in each warming treatment (P < 0.05). mented in this research, our results reveal that both developmental
phases of A. angustifolia subsp. tequilana displayed optimal values in
potential photochemical efficiency (maximum quantum yield of PSII),

6
J.L. Aragón-Gastélum, et al.
which is contrary to our hypothesis and to the studies in African suc-
culent species above described. Aragón-Gastélum et al. (2014) found
low Fv/Fm, ΦPSII and ETR values, as well as high NPQ values but 100%
survival in five-year-old E. platyacanthus (Cactaceae) plants using si-
milar OTCs during the dry season in the same study area. These authors
suggest that despite decreases in photosynthetic performance, E. pla-
tyacanthus shows a high tolerance under climatic variable conditions
such as global warming, which could be attributed to some factors such
as: both the high NPQ values and plasticity due to its widespread dis-
tribution in the Mexican arid and semi-arid environments. Although the
A. angustifolia subsp. tequilana developmental phases also showed si-
milar declines in the ΦPSII values at middle of the experimentation
under similar warming scenarios, increases in the Fv/Fm, ΦPSII, ETR, and
NPQ values in at least one developmental phase (two-year-old and five-
year-old plants) and/or warming treatment (OTCs and control plots)
could be indicators for high tolerance under global warming as sug-
gested by Aragón-Gastélum et al. (2014).
In our study, in both the OTCs and control plots and both devel-
opmental phases, the Fv/Fm values were higher than 0.74 in the A.
angustifolia subsp. tequilana plants, suggesting that this species has a
robust photosynthetic apparatus in order to deal with environmental
stress. It is important to highlight that the drastic decrease in the Fv/Fm
values found on December 7th for the two-year-old plants grown in the
control plots may be associated with the lower daily mean temperatures
(< 3 °C) registered in the surrounding environment during this day
than during remaining period of the study as well as the possible high
sensibility to low temperatures of this developmental phase, suggesting
that the micro-climate of the OTCs provides a buffer to low tempera-
tures. This micro-climate may have improved the photochemical re-
sponse of both the two-year-old and five-year-old plants in the OTCs
and could explain the significant Fv/Fm differences between warming
treatments for this variable documented in this research.
There is a positive relationship between ETR and plants’ ability to
assimilate CO2 (Kakani et al., 2008), because ETR (Baker and
Rosenqvist, 2004), and even ΦPSII (Maxwell and Johnson, 2000), are
involved in electron transfer via PSII. In this study, the ΦPSII values were
found to be low according to Maxwell and Johnson (2000) in both
developmental phases of A. angustifolia subsp. tequilana within the OTCs
and control plots for almost the entire experimentation period; never-
theless, these results did not have a negative impact on the survival in
the two-year-old and five-year-old A. angustifolia subsp. tequilana
plants.
The term NPQ refers to the mechanism used by plants to dissipate
excess heat energy when subject to high levels of environmental stress
and is, thus, a mechanism for avoiding photoinhibition (Maxwell and
Johnson, 2000). Regardless of the warming treatments assessed here,
the NPQ values in both developmental phases of A. angustifolia subsp.
tequilana were higher than in plants of Ariocarpus kotschoubeyanus
(Cactaceae) under nurse plants and in exposed area (Arroyo-Pérez et al.,
2017). These results support the premise about high tolerance in our
target species and could partially explain the high survival rate in the
two-year-old and five-year-old A. angustifolia subsp. tequilana plants in
the OTCs and control plots.
The NPQ energy dissipation mechanism has been mainly char-
acterized in ‘model’ Arabidopsis thaliana species and described in several
reviews (Demmig-Adams and Adams, 2006; Horton and Ruban, 2005;
Jahns and Holzwarth, 2012; Jahns et al., 2009; Murchie and Niyogi,
2011). This mechanism is controlled by specific xanthophyll cycle
pigments and the trans-thylakoid membrane pH gradient. The NPQ
mechanisms have been described in perennial plants such as winter
evergreens, forest understorey species and desiccation-tolerant plants
(García-Plazaola et al., 2012), comprising a sustained dissipation
strategy, a complementary lutein epoxide cycle and rapidly-induced
thermal energy dissipation, respectively. Our results in terms of dif-
ferential responses in the NPQ values in both two-year-old plants and
five-year-old plants of A. angustifolia subsp. tequilana suggest the
7
Flora 263 (2020) 151535
existence of a high level of diversity in the functioning of xanthophyll
cycle and NPQ mechanism between different developmental phases and
plant groups, moreover that succulents could present any of these
thermal energy dissipation mechanisms described above, although
further research is required.
We also found differences in both total chlorophyll and carotenoid
content between warming treatments. Photosynthetic pigments such as
chlorophylls, carotenoids and even anthocyanins are used as indicators
of plant responses to light stress. For instance, decreases of chlorophyll
in heat-stressed plants may be attributed to deficient chlorophyll
synthesis and / or its accelerated degradation (Li et al., 2010). In ad-
dition, carotenoids are essential for photoprotection and they have an
important role as a precursor in signaling during plant development
under both abiotic and biotic stress (Li et al., 2008). Therefore, re-
ductions in chlorophyll and increases in carotenoid concentrations in
plants subjected to high light intensities are expected (Ashraf and
Harris, 2013; de Carvalho Gonçalvez et al., 2001; Tan et al., 2000),
mainly due to the oversaturation of energy dissipation mechanisms and
an increase in oxidative stress-induced photooxidation.
In our induced global warming scenario (OTCs vs control plots), the
two-year-old and five-year-old A. angustifolia subsp. tequilana plants
suffered high light intensities because the PPFD values were high (ty-
pical condition in arid a semi-arid environment) in both the OTCs and
control plots throughout the entire experiment. Independently of
warming treatments, the two-year-old A. angustifolia subsp. tequilana
plants displayed higher total chlorophyll and carotenoid values than the
five-year-old plants; furthermore, the two-year-old A. angustifolia subsp.
tequilana plants inside OTCs also displayed higher values in these pig-
ments than in all other treatment combinations featured in this study.
These results are contrary with Musil et al. (2005), who found de-
creased values in chlorophyll a and b and carotenoid in both dwarf and
shrubby succulents inside OTCs than open spaces, suggesting that an-
thropogenic warming could significantly exceed the thermal thresholds
resulting in local extinctions of these species. In Agave species, studies
assessing variations in photoprotective pigments in response to climate
change remain scarce; nevertheless, Graham and Nobel (1996) did re-
port a decrease in the chlorophyll content per unit leaf area of Agave
deserti under high atmospheric CO2 concentrations. The declines in
chlorophylls and carotenoids described above are contrary to our re-
sults; thus, the higher content of these photosynthetic pigments in two-
year-old plants within OTCs than the five-year-old plants in both
warming treatments documented here could partially explain the high
tolerance in this early developmental phase of A. angustifolia subsp.
tequilana under future global warming conditions.
Several hypotheses have been postulated in explanation of the role
played by anthocyanins (Gould et al., 2000). Studies have reported that
anthocyanins provide a “sunscreen”, acting as “bulk” antioxidants and
protecting plant cells from high levels of light-associated oxidative
stress (Hormaetxe et al., 2005; Steyn et al., 2002). Increased antho-
cyanin concentrations occurring under unfavorable environmental
conditions have been documented in the literature (Albert et al., 2009;
González-Salvatierra et al., 2010; Hughes et al., 2010). Thus, increases
in anthocyanin concentrations in plants subjected to induced warming
are also expected. Nevertheless, we found low anthocyanin concentra-
tions in both developmental phases and warming treatments. It is
possible that a high anthocyanin concentration in the five-year-old and
two-year-old A. angustifolia subsp. tequilana plants is not required to
improve the tolerance in the early developmental phases of this species
and the high tolerance documented in this research could be also ex-
plained by a possible high acclimation under elevated temperature si-
milarly to other succulent species such as A. deserti (Asparagaceae) and
Ferocactus acanthodes (Cactaceae), which showed a high acclimation to
high temperatures and concomitantly, tolerance increased with in-
creasing age (Nobel, 1984). In future works, it is important to assess the
possible variations in anthocyanins content considering leaf anatomy
because in some succulents they can vary across the leaves and/or
J.L. Aragón-Gastélum, et al.
stems as dependent on environmental-stress intensity (González-
Salvatierra et al., 2010).
Global climate change scenarios consider both increases in atmo-
spheric CO2 level and temperature, as well as altered precipitation
patterns (Stocker et al., 2014). Studies suggest that Agave species may
cope well under climate change predicted (García-Moya et al., 2011;
Nobel, 2010). For instance, both young and adult plants of A. salmiana
and A. angustifolia subsp. tequilana are able to grow under low water
potential conditions (Peña-Valdivia and Sánchez-Urdaneta, 2009),
maintaining an active photosynthetic pathway (Pimienta-Barrios et al.,
2001), even after prolonged moisture restriction (Ramírez-Tobías et al.,
2014). Our findings support the assumption raised by García-Moya
et al. (2011) and Nobel (2010) because both developmental phases of
A. angustifolia subsp. tequilana showed an optimal potential photo-
chemical efficiency and relatively high photosynthetic pigment content
under extreme conditions such as induced global warming. The present
research suggests that both the two-year-old plants and the five-year-
old plants of A. angustifolia subsp. tequilana have a high tolerance, de-
spite that the early developmental phases in succulent species are
particularly sensitive to abiotic (Flores et al., 2004) and anthro-
pogenically-induced (Aragón-Gastélum et al., 2017; Godínez-Álvarez
et al., 2003) stress conditions.
5. Conclusions
Our results indicate that both developmental phases of A. angusti-
folia subsp. tequilana showed high tolerance to thermal stress under
induced global warming, displaying an optimal potential photo-
chemical efficiency as well as high NPQ values and pigment content.
The present study provides evidence that high physiological tolerance
could help to cope with the potential effects of global warming in early
developmental phases of A. angustifolia subsp. tequilana, one of the most
economically important Agave species.
CRediT authorship contribution statement
José Luis Aragón-Gastélum: Conceptualization, Formal analysis,
Investigation, Methodology, Project administration, Validation,
Visualization, Writing - original draft, Writing - review & editing. J.
Efrain Ramírez-Benítez: Formal analysis, Visualization, Writing -
original draft, Writing - review & editing. Enrique González-Durán:
Visualization, Writing - review & editing. Claudia González-
Salvatierra: Formal analysis, Methodology, Writing - original draft.
Hugo M. Ramírez-Tobías: Funding acquisition, Conceptualization,
Project administration, Resources, Supervision, Validation. Joel Flores:
Conceptualization, Supervision, Validation. Eduardo J. Gutiérrez-
Alcántara: Visualization, Writing - review & editing. Evelyn Méndez-
Guzmán: Methodology. Ramón Jarquín-Gálvez: Funding acquisition.
Declaration of Competing Interest
The authors declare that we have no conflict of interest with the
results of this manuscript.
Acknowledgments
We thank Enrique Jurado and an anonymous reviewer for their
suggestions to improve the manuscript. This research was supported by
the Secretaría de Educación Pública (Ministry of Public Education)
Programa de Mejoramiento del Profesorado (Program for the
Improvement of Higher Education Teachers) (No. 511-6/17-1968 and
511-6/18-8491).
Appendix A. Supplementary data
Supplementary material related to this article can be found, in the
8
Flora 263 (2020) 151535
online version, at doi:https://doi.org/10.1016/j.flora.2019.151535.
References
Albert, N.W., Lewis, D.H., Zhang, H., Irving, L.J., Jameson, P.E., Davies, K.M., 2009.
Light-induced vegetative anthocyanin pigmentation in Petunia. J. Exp. Bot. 60,
2191–2202. https://doi.org/10.1093/jxb/erp097.
Aragón-Gastélum, J.L., Badano, E., Yáñez-Espinosa, L., Ramírez-Tobías, H.M., Rodas-
Ortiz, J.P., González-Salvatierra, C., Flores, J., 2017. Seedling survival of three en-
demic and threatened Mexican cacti under induced climate change. Plant Species
Biol. 32, 92–99. https://doi.org/10.1111/1442-1984.12120.
Aragón-Gastélum, J.L., Flores, J., Jurado, E., Ramírez-Tobías, H.M., Robles-Díaz, E.,
Rodas-Ortiz, J.P., Yáñez-Espinosa, L., 2018. Potential impact of global warming on
seed bank, dormancy and germination of three succulent species from the
Chihuahuan Desert. Seed Sci. Res. 1–7. https://doi.org/10.1017/
s0960258518000302.
Aragón-Gastélum, J.L., Flores, J., Yáñez-Espinosa, L., Badano, E., Ramírez-Tobías, H.M.,
Rodas-Ortíz, J.P., González-Salvatierra, C., 2014. Induced climate change impairs
photosynthetic performance in Echinocactus platyacanthus, an especially protected
Mexican cactus species. Flora 209, 499–503. https://doi.org/10.1016/j.flora.2014.
06.002.
Archer, S.R., Predick, K.I., 2008. Climate change and ecosystems of the Southwestern
United States. Rangelands 30, 23–28. https://doi.org/10.2111/1551-501x(2008)
30[23:ccaeot]2.0.co;2.
Arroyo-Pérez, E., Flores, J., González-Salvatierra, C., Matías-Palafox, M.L., Jiménez-
Sierra, C., 2017. High tolerance to high-light conditions for the protected species
Ariocarpus kotschoubeyanus (Cactaceae). Conserv. Physiol. 5https://doi.org/10.1093/
conphys/cox042. cox042.
Ashraf, M., Harris, P.J.C., 2013. Photosynthesis under stressful environments: an over-
view. Photosynthetica 51, 163–190. https://doi.org/10.1007/s11099-013-0021-6.
Baker, N.R., Rosenqvist, E., 2004. Applications of chlorophyll fluorescence can improve
crop production strategies: an examination of future possibilities. J. Exp. Bot. 55,
1607–1621. https://doi.org/10.1093/jxb/erh196.
Bokhorst, S., Huiskes, A., Aerts, R., Convey, P., Cooper, E.J., Dalen, L., Erschbamer, B.,
Gudmundsson, J., Hofgaard, A., Hollister, R.D., Johnstone, J., Jónsdóttir, I.S.,
Lebouvier, M., Van de Vijver, B., Wahren, C.-H., Dorrepaal, E., 2013. Variable tem-
perature effects of Open Top Chambers at polar and alpine sites explained by irra-
diance and snow depth. Glob. Chang. Biol. 19, 64–74. https://doi.org/10.1111/gcb.
12028.
Bunce, J.A., 1981. Comparative responses of leaf conductance to humidity in single at-
tached leaves. J. Exp. Bot. 32, 629–634. https://doi.org/10.1093/jxb/32.3.629.
Colunga García-Marín, P., Zizumbo-Villarreal, D., Martínez-Torres, J., 2007. Tradiciones
en el aprovechamiento de los agaves mexicanos: una aportación a la protección legal
y conservación de su diversidad biológica y cultural. In: Colunga García-Marín, P.,
Larqué-Saavedra, A., Eguiarte, L.E., Zizumbo-Villarreal, D. (Eds.), En lo ancestral hay
futuro: del tequila, los mezcales y otros agaves. Centro de Investigación Científica de
Yucatán, A.C., Mérida, MEX, pp. 229–248.
Comstock, J., Ehleringer, J., 1993. Stomatal response to humidity in common bean
(Phaseolus vulgaris): implications for maximum transpiration rate, water-use effi-
ciency and productivity. Funct. Plant Biol. 20, 669–691.
de Carvalho Gonçalvez, J.F., Marenco, R.A., Vieira, G., 2001. Concentration of photo-
synthetic pigments and chlorophyll fluorescence of mahogany and tonka bean under
two light environments. Braz. J. Plant Physiol. 13, 149–157. https://doi.org/10.
1590/S0103-31312001000200004.
de la Rosa-Manzano, E., Flores, J., Delgado-Sánchez, P., 2016. Effects of spine-shading on
aspects of photosynthesis for three cactus species. Bot. Sci. 94, 301–310. https://doi.
org/10.17129/botsci.458.
Demmig-Adams, B., Adams, W.W., 2006. Photoprotection in an ecological context: the
remarkable complexity of thermal energy dissipation. New Phytol. 172, 11–21.
https://doi.org/10.1111/j.1469-8137.2006.01835.x.
Flores, J., Briones, O., Flores, A., Sánchez-Colón, S., 2004. Effect of predation and solar
exposure on the emergence and survival of desert seedlings of contrasting life-forms.
J. Arid Environ. 58, 1–18. https://doi.org/10.1016/S0140-1963(03)00127-7.
García-Amaro, E., 2004. Modificaciones Al Sistema De Clasificación Climática De Köppen,
5th ed. Universidad Nacional Autónoma de México, Instituto de Geografía, MEX.
García-Mendoza, A.J., 2007. Los agaves en México. Ciencias 87, 14–23.
García-Moya, E., Romero-Manzanares, A., Nobel, P.S., 2011. Highlights for Agave pro-
ductivity. GCB Bioenergy 3, 4–14. https://doi.org/10.1111/j.1757-1707.2010.
01078.x.
García-Plazaola, J.I., Esteban, R., Fernández-Marín, B., Kranner, I., Porcar-Castell, A.,
2012. Thermal energy dissipation and xanthophyll cycles beyond the Arabidopsis
model. Photosyn. Res. 113, 89–103. https://doi.org/10.1007/s11120-012-9760-7.
Gentry, H.S., 1982. Agaves of Continental North America. University of Arizona Press,
Arizona, USA.
Godínez-Álvarez, H., Valverde, T., Ortega-Baes, P., 2003. Demographic trends in the
Cactaceae. Bot. Rev. 69, 173–201. https://doi.org/10.1663/0006-8101(2003)
069[0173:DTITC]2.0.CO;2.
González-Salvatierra, C., Andrade, J.L., Escalante-Erosa, F., García-Sosa, K., Peña-
Rodríguez, L.M., 2010. Antioxidant content in two CAM bromeliad species as a re-
sponse to seasonal light changes in a tropical dry deciduous forest. J. Plant Physiol.
167, 792–799. https://doi.org/10.1016/j.jplph.2010.01.001.
González-Salvatierra, C., Andrade, J.L., Orellana, R., Peña-Rodríguez, L.M., Reyes-García,
C., 2013. Microambiente lumínico y morfología y fisiología foliar de Bromelia karatas
(Bromeliaceae) en una selva baja caducifolia de Yucatán, México. Bot. Sci. 91, 75–84.
Gould, K.S., Markham, K.R., Smith, R.H., Goris, J.J., 2000. Functional role of
J.L. Aragón-Gastélum, et al.
anthocyanins in the leaves of Quintinia serrata A. Cunn. J. Exp. Bot. 51, 1107–1115.
https://doi.org/10.1093/jexbot/51.347.1107.
Graham, E.A., Nobel, P.S., 1996. Long-term effects of a doubled atmospheric CO2 con-
centration on the CAM species Agave deserti. J. Exp. Bot. 47, 61–69. https://doi.org/
10.1093/jxb/47.1.61.
Hasanuzzaman, M., Nahar, K., Fujita, M., 2013. Extreme temperature responses, oxidative
stress and antioxidant defense in plants. In: Vahdati, K., Leslie, C. (Eds.), Abiotic
Stress-Plant Responses and Applications in Agriculture. InTech, London, UK, pp.
169–205.
Hazrati, S., Tahmasebi-Sarvestani, Z., Modarres-Sanavy, S.A.M., Mokhtassi-Bidgoli, A.,
Nicola, S., 2016. Effects of water stress and light intensity on chlorophyll fluorescence
parameters and pigments of Aloe vera L. Plant Physiol. Biochem. 106, 141–148.
https://doi.org/10.1016/j.plaphy.2016.04.046.
Hernandez-Gonzalez, O., Briones-Villareal, O., 2007. Crassulacean acid metabolism
photosynthesis in columnar cactus seedlings during ontogeny: the effect of light on
nocturnal acidity accumulation and chlorophyll fluorescence. Am. J. Bot. 94,
1344–1351. https://doi.org/10.3732/ajb.94.8.1344.
Hollister, R.D., Webber, P.J., 2000. Biotic validation of small open-top chambers in a
tundra ecosystem. Glob. Chang. Biol. 6, 835–842. https://doi.org/10.1046/j.1365-
2486.2000.00363.x.
Hormaetxe, K., Becerril, J.M., Fleck, I., Pintó, M., García-Plazaola, J.I., 2005. Functional
role of red (retro)-carotenoids as passive light filters in the leaves of Buxus semper-
virens L.: increased protection of photosynthetic tissues? J. Exp. Bot. 56, 2629–2636.
https://doi.org/10.1093/jxb/eri255.
Horton, P., Ruban, A., 2005. Molecular design of the photosystem II light-harvesting
antenna: photosynthesis and photoprotection. J. Exp. Bot. 56, 365–373. https://doi.
org/10.1093/jxb/eri023.
Howell, T.A., Dusek, D.A., 1995. Comparison of vapor-pressure-deficit calculation
methods—southern high plains. J. Irrig. Drain. Eng. 121, 191–198. https://doi.org/
10.1061/(ASCE)0733-9437(1995)121:2(191).
Hughes, N.M., Reinhardt, K., Feild, T.S., Gerardi, A.R., Smith, W.K., 2010. Association
between winter anthocyanin production and drought stress in angiosperm evergreen
species. J. Exp. Bot. 61, 1699–1709. https://doi.org/10.1093/jxb/erq042.
Jahns, P., Holzwarth, A.R., 2012. The role of the xanthophyll cycle and of lutein in
photoprotection of photosystem II. Biochimica et Biophysica Acta (BBA) -
Bioenergetics 1817, 182–193. https://doi.org/10.1016/j.bbabio.2011.04.012.
Jahns, P., Latowski, D., Strzalka, K., 2009. Mechanism and regulation of the violaxanthin
cycle: the role of antenna proteins and membrane lipids. Biochim. Biophys. Acta.
Bioenergetic 1787, 3–14. https://doi.org/10.1016/j.bbabio.2008.09.013.
Jeon, M.-W., Ali, M.B., Hahn, E.-J., Paek, K.-Y., 2006. Photosynthetic pigments, mor-
phology and leaf gas exchange during ex vitro acclimatization of micropropagated
CAM Doritaenopsis plantlets under relative humidity and air temperature. Environ.
Exp. Bot. 55, 183–194. https://doi.org/10.1016/j.envexpbot.2004.10.014.
Jones, H.G., 2013. Plants and Microclimate: A Quantitative Approach to Environmental
Plant Physiology, 3rd ed. Cambridge University Press.
Kakani, V.G., Surabhi, G.K., Reddy, K.R., 2008. Photosynthesis and fluorescence re-
sponses of C4 plant Andropogon gerardii acclimated to temperature and carbon di-
oxide. Photosynthetica 46, 420–430. https://doi.org/10.1007/s11099-008-0074-0.
Li, T.Y., Zhang, Y., Liu, H., Wu, Y., Li, W., Zhang, H., 2010. Stable expression of
Arabidopsis vacuolar Na+/H+ antiporter gene AtNHX1, and salt tolerance in trans-
genic soybean for over six generations. Chinese Sci. Bull. 55, 1127–1134. https://doi.
org/10.1007/s11434-010-0092-8.
Li, F., Vallabhaneni, R., Yu, J., Rocheford, T., Wurtzel, E.T., 2008. The maize phytoene
synthase gene family: overlapping roles for carotenogenesis in endosperm, photo-
morphogenesis, and thermal stress tolerance. Plant Physiol. 147, 1334–1346. https://
doi.org/10.1104/pp.108.122119.
Macías-Macías, A., Valenzuela-Zapata, A.G., 2009. El tequila en tiempos de la mundia-
lización. Comercio exterior 59, 459–472.
Maphangwa, K.W., Musil, C.F., Raitt, L., Zedda, L., 2012. Experimental climate warming
decreases photosynthetic efficiency of lichens in an arid South African ecosystem.
Oecologia 169, 257–268. https://doi.org/10.1007/s00442-011-2184-9.
Marion, G.M., 1996. Temperature enhancement experiments. In: Molau, U., Mølgaard, P.
(Eds.), International Tundra Experiment (ITEX) Manual, 2nd ed. Danish Polar Center,
Copenhagen, Denmark, pp. 17–22.
Matimati, I., Musil, C.F., Raitt, L., February, E., 2013. Non rainfall moisture interception
by dwarf succulents and their relative abundance in an inland arid South African
ecosystem. Ecohydrology 6, 818–825. https://doi.org/10.1002/eco.1304.
Maxwell, K., Johnson, G.N., 2000. Chlorophyll fluorescence—a practical guide. J. Exp.
Bot. 51, 659–668. https://doi.org/10.1093/jexbot/51.345.659.
Medina-García, G., Díaz-Padilla, G., Loredo-Osti, C., Serrano-Altamirano, V., Cano-García,
M., 2005. . Estadísticas Climatológicas Básicas Del Estado De San Luís Potosí (Periodo
1961-2001). Centro de Investigación Regional Noreste, San Luis Potosí, MEX.
Minocha, R., Martinez, G., Lyons, B., Long, S., 2009. Development of a standardized
methodology for quantifying total chlorophyll and carotenoids from foliage of
hardwood and conifer tree species. Can. J. For. Res. 39, 849–861. https://doi.org/10.
1139/x09-015.
Murchie, E.H., Niyogi, K.K., 2011. Manipulation of photoprotection to improve plant
photosynthesis. Plant Physiol. 155, 86–92. https://doi.org/10.1104/pp.110.168831.
Musil, C.F., Schmiedel, U., Midgley, G.F., 2005. Lethal effects of experimental warming
approximating a future climate scenario on southern African quartz-field succulents:
a pilot study. New Phytol. 165, 539–547. https://doi.org/10.1111/j.1469-8137.
Flora 263 (2020) 151535
2004.01243.x.
Musil, C.F., Van Heerden, P.D.R., Cilliers, C.D., Schmiedel, U., 2009. Mild experimental
climate warming induces metabolic impairment and massive mortalities in southern
African quartz field succulents. Environ. Exp. Bot. 66, 79–87. https://doi.org/10.
1016/j.envexpbot.2008.11.008.
Neff, M.M., Chory, J., 1998. Genetic interactions between phytochrome A, phytochrome
B, and cryptochrome 1 during Arabidopsis development. Plant Physiol. 118, 27–35.
https://doi.org/10.1104/pp.118.1.27.
Nobel, P.S., 1984. Extreme temperatures and thermal tolerances for seedlings of desert
succulents. Oecologia 62, 310–317.
Nobel, P.S., 2010. Desert wisdom/agaves and Cacti: CO2, Water, Climate Change.
iUniverse, New York, USA.
Ogburn, R.M., Edwards, E.J., 2010. The ecological water-use strategies of succulent
plants. In: Kader, J.-C., Delseny, M. (Eds.), Advances in Botanical Research Vol. 55.
Academic Press, London, UK, pp. 179–225.
Peña-Valdivia, C.B., Sánchez-Urdaneta, A.B., 2009. Effects of substrate water potential in
root growth of Agave salmiana Otto ex Salm-Dyck seedlings. Biol. Res. 42, 239–248.
https://doi.org/10.4067/S0716-97602009000200013.
Pérez-Sánchez, R.M., Flores, J., Jurado, E., González-Salvatierra, C., 2015. Growth and
ecophysiology of succulent seedlings under the protection of nurse plants in the
Southern Chihuahuan Desert. Ecosphere 6, 36. https://doi.org/10.1890/ES14-
00408.1.
Pimienta-Barrios, E., Robles-Murguia, C., Nobel, P.S., 2001. Net CO2 uptake for Agave
tequilana in a warm and a temperate environment. Biotropica 33, 312–318. https://
doi.org/10.1111/j.1744-7429.2001.tb00181.x.
Porra, R.J., Thompson, W.A., Kriedemann, P.E., 1989. Determination of accurate ex-
tinction coefficients and simultaneous equations for assaying chlorophylls a and b
extracted with four different solvents: verification of the concentration of chlorophyll
standards by atomic absorption spectroscopy. Biochimica et Biophysica Acta (BBA) -
Bioenergetics 975, 384–394. https://doi.org/10.1016/S0005-2728(89)80347-0.
Ptushenko, V.V., Ptushenko, E.A., Samoilova, O.P., Tikhonov, A.N., 2013. Chlorophyll
fluorescence in the leaves of Tradescantia species of different ecological groups: in-
duction events at different intensities of actinic light. Biosystems 114, 85–97. https://
doi.org/10.1016/j.biosystems.2013.08.001.
Ramírez-Tobías, H.M., Peña-Valdivia, C.B., Aguirre-Rivera, J.R., 2014. Respuestas
bioquímico-fisiológicas de especies de Agave a la restricción de humedad. Bot. Sci.
92, 131–139.
Ramírez-Tobías, H.M., Peña-Valdivia, C.B., Aguirre-Rivera, J.R., Reyes-Agüero, J.A.,
Sánchez-Urdaneta, A.B., Valle-G, S., 2012. Seed germination temperatures of eight
Mexican Agave species with economic importance. Plant Species Biol. 27, 124–137.
https://doi.org/10.1111/j.1442-1984.2011.00341.x.
Reyes-Agüero, J.A., Aguirre-Rivera, J.R., Peña-Valdivia, C.B., 2000. Biología y apro-
vechamiento de Agave lechuguilla Torrey. Bol. Soc. Bot. Mex. 67, 75–88.
Romero-González, J., Parra-Vargas, F., Cano-Rodríguez, I., Rodríguez, E., Ríos-Arana, J.,
Fuentes-Hernández, R., Ramírez-Flores, J., 2007. Biosorption of Pb (II) by Agave te-
quilana Weber (agave azul) biomass. Rev. Mex. Ing. Quim. 6, 295–300.
Rzedowski, J., 1991. Diversidad y orígenes de la flora fanerogámica de México. Acta Bot.
Mex. 3–21.
Samoilova, O.P., Ptushenko, V.V., Kuvykin, I.V., Kiselev, S.A., Ptushenko, O.S., Tikhonov,
A.N., 2011. Effects of light environment on the induction of chlorophyll fluorescence
in leaves: a comparative study of Tradescantia species of different ecotypes.
Biosystems 105, 41–48. https://doi.org/10.1016/j.biosystems.2011.03.003.
Secretaría de Medio Ambiente y Recursos Naturales, 2010. Norma oficial mexicana NOM-
059-ECOL-2010: protección ambiental-especies nativas de México de flora y fauna
silvestres. In: Ambiente, P.F. (Ed.), Categorías de riesgo y especificaciones para su
inclusión, exclusión o cambio. Lista de especies en riesgo. Diario Oficial de la
Federación, Ciudad de México, MEX, pp. 1–78.
Sinclair, T.R., Bennett, J.M., 1998. Water. In: Sinclair, T.R., Gardner, F.P. (Eds.),
Principles of Ecology in Plant Production. CAB International, pp. 103–120.
Steyn, W.J., Wand, S.J.E., Holcroft, D.M., Jacobs, G., 2002. Anthocyanins in vegetative
tissues: a proposed unified function in photoprotection. New Phytol. 155, 349–361.
https://doi.org/10.1046/j.1469-8137.2002.00482.x.
Stocker, T.F., Qin, D., Plattner, G.K., Tignor, M., Allen, S.K., Boschung, J., Nauels, A., Xia,
Y., Bex, V., Midgley, P.M., 2014. Climate Change 2013: The Physical Science Basis.
Cambridge University Press, New York, USA.
Tan, Y.-X., Jiang, J.-H., Wu, H.-L., Cui, H., Yu, R.-Q., 2000. Resolution of kinetic system of
simultaneous degradations of chlorophyll a and b by PARAFAC. Anal. Chim. Acta
412, 195–202. https://doi.org/10.1016/S0003-2670(99)00813-2.
Turner, N.C., Schulze, E.D., Gollan, T., 1984. The responses of stomata and leaf gas ex-
change to vapour pressure deficits and soil water content. Oecologia 63, 338–342.
https://doi.org/10.1007/BF00390662.
Ureta, C., Martínez-Meyer, E., Perales, H.R., Álvarez-Buylla, E.R., 2012. Projecting the
effects of climate change on the distribution of maize races and their wild relatives in
Mexico. Glob. Change Biol. 18, 1073–1082. https://doi.org/10.1111/j.1365-2486.
2011.02607.x.
Valenzuela, A., 2011. A new agenda for blue agave landraces: food, energy and tequila.
Glob. Change Biol. Bioenergy 3, 15–24. https://doi.org/10.1111/j.1757-1707.2010.
01082.x.
Yang, Z., Fan, G., Gu, Z., Han, Y., Chen, Z., 2007. Optimization extraction of anthocyanins
from purple corn (Zea mays L.) cob using tristimulus colorimetry. Eur. Food Res.
Technol. 227, 409. https://doi.org/10.1007/s00217-007-0735-4.