© The Authors, 2022, Published by the Universidad del Zulia*Corresponding author: mao.aguilar@correo.buap.mx
Keywords:
Carotenoids
Drought
Glycinebetaine
Osmolytes
Photosynthetic pigments
Interactive effect of moisture restriction and salicylic acid on biochemical responses in
Phaseolus coccineus
Efecto interactivo de la restricción de humedad y ácido salicílico sobre las respuestas bioquímicas
de Phaseolus coccineus
Efeito interativo da restrição de umidade e ácido salicílico nas respostas bioquímicas em Phaseolus
coccineus
Jesús Mao Aguilar-Luna
1*
Juan Manuel Loeza-Corte
2
Ernesto Díaz-López
3
Rev. Fac. Agron. (LUZ). 2022, 39(3): e223940
ISSN 2477-9407
DOI: https://doi.org/10.47280/RevFacAgron(LUZ).v39.n3.06
Crop Production
Associate editor: Dr. Jorge Vilchez-Perozo
University of Zulia, Faculty of Agronomy
Bolivarian Republic of Venezuela
1
Benemerita Autonomous
University of Puebla, Agroforest
Engineering. 73640. Mexico.
2
University of Cañada,
Agroindustrial Engineering. 68540.
Mexico.
3
Technological University of Tehuacan, Biofood Process
Engineering. 75859. Mexico.
Received: 30-06-2022
Accepted: 01-08-2022
Published:
22-08-2022
Abstract
The increase in water scarcity leads to consider the understanding of
staple crops under these conditions, coupled with this, the positive responses
of salicylic acid in different crops, may be an option in bringing to fruition
the cultivation of runner bean (Phaseolus coccineus). This study evaluated
the effect of salicylic acid (SA) on the biochemical responses in P. coccineus,
with humidity restriction in the periods from January to July 2019 and 2020,
at the Benemerita Autonomous University of Puebla, Mexico. The research
consisted of three levels of drought: 30, 60 and 100% soil moisture; ve
levels of SA: 0, 0.5, 1.0, 1.5 and 2.0 mM; and two levels of fertilization:
non-fertilizer and fertilizer [(00-60-30) at sowing + (30N) foliar nitrogen at
grain lling stage] for the two growing periods. The experimental design was
in factorial random blocks with ve replications. The results showed that the
foliar application with 1.5 mM of SA maintained the highest relative water
content in leaves (89.05%), as well as chlorophyll a, b and carotenoids (2.20,
1.11 and 0.90 µg.mL
-1
, respectively); of glycinebetaine (24.80 µmol.g
-1
DW)
and total soluble sugars (31.15 mg eq.glucose g
-1
DW), excluding proline.
The SA did not increase the protein fractions, even in plants with fertilizer;
but the positive effects of SA were greater in plants without hydric stress and
with fertilization.
This scientic publication in digital format is a continuation of the Printed Review: Legal Deposit pp 196802ZU42, ISSN 0378-7818.
Rev. Fac. Agron. (LUZ). 2022, 39(3): e223940. July - September. ISSN 2477-9407.
2-6 |
Resumen
El incremento en la escasez de agua lleva a considerar el
entendimiento de los cultivos básicos bajo estas condiciones, aunado
a esto, las respuestas positivas del ácido salicílico en diferentes
cultivos, puede ser una opción para llevar a buen término el cultivo
de frijol ayocote (Phaseolus coccineus). En este estudio se evaluó
el efecto del ácido salicílico (ÁS) en la respuesta bioquímica de P.
coccineus, con restricción de humedad en los periodos de enero a julio
de 2019 y 2020 en la Benemérita Universidad Autónoma de Puebla,
México. La investigación constó de tres niveles de sequía: 30, 60 y
100% de humedad del suelo; cinco niveles de ÁS: 0, 0.5, 1.0, 1.5 y 2.0
mM; y dos niveles de fertilización: sin fertilizante y con fertilizante
[(00-60-30) al momento de la siembra + (30N) nitrógeno foliar en
la etapa de llenado de grano] para los dos períodos de cultivo. El
diseño experimental fue factorial en bloques con cinco repeticiones.
Los resultados mostraron que la aplicación foliar con 1.5 mM de ÁS
mantuvo el mayor contenido relativo de agua en hojas (89.05%),
así como clorola a, b y carotenoides (2.20, 1.11 y 0.90 µg.mL
-1
,
respectivamente); de glicinabetaína (24.80 µmol.g
-1
en peso seco)
y azúcares solubles totales (31.15 mg eq.glucosa g
-1
en peso seco),
excluyendo prolina. El ÁS no incrementó las fracciones protéicas,
incluso en plantas con fertilización; pero los efectos positivos del ÁS
fueron mayores en plantas sin estrés hídrico y con fertilización.
Palabras clave: carotenoides, sequía, glicinabetaína, osmolitos,
pigmentos fotosintéticos.
Resumo
O aumento da escassez de água leva-nos a considerar a
compreensão das culturas básicas sob estas condições, juntamente
com isto, as respostas positivas do ácido salicílico em diferentes
culturas, pode ser uma opção para levar à fruticação o cultivo de
feijão corredor (Phaseolus coccineus). Este estudo avaliou o efeito
do ácido salicílico (ÁS) sobre a resposta bioquímica do P. coccineus,
com restrição de umidade nos períodos de janeiro a julho de 2019 e
2020, na Universidade Autônoma Benemerita de Puebla, México. A
pesquisa consistiu em três níveis de seca: 30, 60 e 100% de umidade
do solo; cinco níveis de SA: 0, 0.5, 1.0, 1.5 e 2.0 mM; e dois níveis de
adubação: não fertilizante e fertilizante [(00-60-30) na semeadura +
(30N) nitrogênio foliar na fase de enchimento de grãos] para os dois
períodos de crescimento. O delineamento experimental foi em blocos
ao acaso fatorial com cinco repetições. Os resultados mostraram que
a aplicação foliar com 1.5 mM de ÁS manteve o maior teor relativo
de água nas folhas (89.05%), assim como clorola a, b e carotenóides
(2.20, 1.11 e 0.90 µg.mL
-1
, respectivamente); de glicinabetaína
(24.80 µmol.g
-1
de peso seco) e açúcares solúveis totais (31.15 mg
eq.glicose g
-1
de peso seco), excluindo prolina. O SA não aumentou as
frações protéicas, mesmo em plantas com fertilizante; mas os efeitos
positivos do SA foram maiores em plantas sem estresse hídrico e com
adubação.
Palavras-chave: carotenóides, seca, glicinabetaína, osmolitos,
pigmentos fotossintéticos.
Introduction
The runner bean [Phaseolus coccineus subsp. Polyanthus
(Grenm.) Maréchal, Mascherpa and Stainer] rank third in importance
within the genus Phaseolus, after P. vulgaris (L.) and P. lunatus (L.)
(Reyes-Matamoros et al., 2014; Morosan et al., 2017). P. coccineus
plants are sensitive to water deciency, which may be because it is a
semi-domesticated plant, and it presents the same sensitivity pattern
as P. vulgaris (Reyes-Matamoros et al., 2014).
When plants are exposed to humidity restriction, their cells protect
themselves by synthesizing and accumulating specic osmolytes
(betaine, soluble carbohydrates, glycine-betaine, proline, and
proteins such as osmotin) (Ozturk et al., 2020; Hossain et al., 2022);
to maintain cellular osmotic balance, which is disrupted under stress
conditions (Surabhi and Rout, 2020). Consequently, plants stressed
by humidity restriction exhibit a different biochemical response than
plants grown under optimal conditions.
To reduce hydric stress in plants, various growth regulators have
been used in the past by many researchers, e.g.: mepiquat chloride,
polyamines, brassinosteroids, jasmonate, 5-aminolevulinic acid and
salicylic acid (SA) (Kordi et al., 2013; Hossain et al., 2022). SA
is one of those that offers the best response, since it is a lipophilic
monohydroxybenzoic acid that regulates different physiological
processes in plants (Abdelaal et al., 2020); e.g.: positive effect on
both hypocotyl height and diameter, nutrient absorption, stomatal
closure, transpiration, chlorophyll, protein biosynthesis, proline
biosynthesis, antioxidative enzymes, plant defense and secondary
metabolism (Afshari et al., 2021; Gordillo-Curiel et al., 2021; Yan
et al., 2022). It improves the transpiratory rate, water use efciency,
stomatal conductance, internal CO
2
concentration, photosynthetic
assimilation rate and xation of enzymes such as RuBisCo (Galon et
al., 2022; Muhie, 2022).
The mode of action of SA depends on several factors, such as
the species, environmental conditions, concentration, and method of
application. Low SA concentrations have shown to exhibit benecial
effects in crops with hydric restriction, e.g.: a 10
-5
M SA solution in
Brassica juncea (L.), Zea mays (L.), Glycine max (L.) and other crops,
increased net photosynthesis (Muhie, 2022). A concentration of 10
-5
M SA in Cymbopogon exuosus (Nees ex Steud.) Watson, improved
proline content and production of essential oils (Idrees et al., 2010).
SA’s from 0.01 to 0.05 mM applications in Coffea arabica (L.)
increased its growth (Gordillo-Curiel et al., 2021). A concentration of
1.5 mM of SA in Ocimum basilicum (L.) favored the proline content
(Kordi et al., 2013). Applications of 0.05 M SA in Mentha arvensis
(L.) increased the content of chlorophylls a and b, and carotenoids
(Elhakem, 2019).
Worldwide, the works with SA to mitigate the humidity restriction
(hydric stress) in crops are very extensive; however, for P. coccineus
scarce information is available. Therefore, the objective of the present
study was to evaluate the effect of ve concentrations of SA in the
presence of two levels of fertilization with three levels of humidity
restriction on the biochemical responses in P. coccineus.
This scientic publication in digital format is a continuation of the Printed Review: Legal Deposit pp 196802ZU42, ISSN 0378-7818.
Aguilar-Luna et al. Rev. Fac. Agron. (LUZ). 2022, 39(3): e2239403-6 |
Materials and methods
Study location
The study was carried out on an agricultural plot with a tunnel-
type greenhouse at the Benemerita Autonomous University of Puebla
(latitude: 19°49’01’’N, longitude: 97°47’36’’W, altitude: 1,764
m); during January to July 2019 and 2020. The region’s climate is
temperate subhumid, in the two years, the climatic conditions were
similar, therefore, the monthly averages of the two years are shown
(gure 1). Experimental soil used had following characteristics, it
was Luvisol (FAO, 2015), with pH 6.83, 0.14 g.cm
-3
in bulk density,
0.50 Cmol(+) kg
-1
in cation exchange capacity, 0.34% organic matter,
0.19% total N, 0.80 mg.kg
-1
of P, 0.01 Cmol(+) kg
-1
of K, 0.03
Cmol(+) kg
-1
of Ca and 0.03 Cmol(+) kg
-1
of Mg.
Figure 1. Climatic conditions of the experimental site (monthly
averages, 2019 and 2020).
Biological material and treatments
The seeds were acquired from regional producers, washed with
running water, and disinfected before sowing with a fungicide
(N-Trichloromethylthio-4-cyclohexene-1,2-dicarboximide) at a dose
of 1.50 g.kg
-1
of seed. One seed per bag was sown, a black nursery
bag 30 cm high and 23 cm in diameter was used; the same ones that
were lled with reconstituted substrate, with soil from the region and
peat moss (3:1,
v
/
v
).
The experiments were developed in the dry season, with controlled
humidity conditions. The adaptation of the plants to drought stress
began 15 days after sowing and until the end of the experiment.
Following the method used by Reyes-Matamoros et al. (2014) three
humidity levels were established: without stress (100% humidity in
the soil), moderate stress (60% humidity in the soil) and severe stress
(30% humidity in the soil).
Five levels of SA (0, 0.5, 1.0, 1.5 and 2.0 mM) were evaluated,
which were applied to the foliage 15 days after sowing with a hand
sprinkler. In total, ve applications were made with intervals of 15
days. The SA concentrations were obtained from its molecular weight
(138.12 g.mol
-1
), each SA concentration was dissolved in 5% ethanol,
then it was lled with distilled water in a one-liter volumetric ask
and nally Tween 20 washing buffer was added, 0.5% as a surfactant.
Two levels of fertilization were also used: non fertilizer and
fertilizer [(00-60-30) edaphic fertilization at sowing + (30N) foliar
nitrogen applied in the grain lling stage]. Two pests appeared
during the development of the crop Liriomyza trifolii (Burgess) and
Bemicia tabaci (Gennadius), which were controlled with a systemic
insecticide: N-[1-(6-Chloro-pyridin-3-ylmethyl)-imidazolidin-2-
ylidene]nitramide. The experiment was presented in a factorial
randomized block design with ve replications. Each experimental
unit consisted of a bag with a plant.
Measurement of experimental variables
Relative water content (RWC). The second leaf of ve plants
selected at random by treatment was used; the fresh weight (FW)
was immediately recorded in them, then the leaves were soaked with
distilled water for 16 h in the dark to record the turgid weight (TW),
then they were dried in an oven at 70°C for 72 h to estimate the dry
weight (DW). The RWC was calculated using the following formula
(Chaimala et al., 2021):
RWC
(
%
)
=
FW DW
TW DW
(100) 1
Photosynthetic pigments. To determine the concentration
of pigments: chlorophyll a (CHLa), chlorophyll b (CHLb) and
carotenoids (CRT); the methodology of Lichtenthaler & Buschmann
(2001) was followed. The absorbance was read with a Perkin Elmer
Lambda 25 UV/Visible spectrophotometer, the optical density of the
solution was read at 662, 645 and 470 nm (CHLa, CHLb and CRT,
respectively). Finally, the concentration of pigments was calculated in
µg for each mL of extract
-1
, according to the equations:
Quantication of osmolytes. The samples for these analyze were
obtained during the grain lling stage (approximately 100 days
after sowing), ve samples were used per treatment, the data were
obtained, and the averages were calculated for:
Free Proline (PRO). It was determined according to the method of
Bates et al. (1973); the absorbance of the organic phase was read at
520 nm. The concentration of PRO was expressed as µmol.g
-1
of DW.
Glycine betaine (GB). It was determined following the method of
Grieve & Grattan (1983); the absorbance of the solution was read at
365 nm. The concentration of GB was expressed as µmol.g
-1
of DW.
Total soluble sugars (TSS). They were quantied according to the
method of Dubois et al. (1956); the absorbance of the solution was
read at 490 nm. The TSS concentration was expressed as ˈmg glucose
equivalentˈ g
-1
of DW.
Protein fractions. The extraction of the fractions: albumin (ALB)
and globulins (GLB) was carried out using the method described
by Barba de la Rosa et al. (1992), using 50 g of dry seed our. For
the extraction of glutelins (GLT), the residue from the extraction of
prolamins (PRL) was dispersed in a 1M NaOH solution at a 1:15
solute/solvent ratio. The percentage of protein extracted in each
fraction was calculated, using the equation:
Protein
(
%
)
=
protein gram extracted in the fraction
protein gram in the flour
(100)
1
The protein content was calculated by the Kjeldahl method
according to the AOAC methodology (Latimer, 2012), using 6.25 as
the nitrogen to protein conversion factor. Protein quantication was
carried out by the method of Morr et al. (1985) with a kit (Sigma-
Aldrich-L1013). A standard curve was made with bovine serum
albumin (Sigma-Aldrich-A8531) at a concentration of 0.40 mg.mL
-1
;
the absorbance was measured at 600 nm.
This scientic publication in digital format is a continuation of the Printed Review: Legal Deposit pp 196802ZU42, ISSN 0378-7818.
Rev. Fac. Agron. (LUZ). 2022, 39(3): e223940. July - September. ISSN 2477-9407.
4-6 |
Statistical procedures
All the experiments were conducted using a factorial randomized
block design, with ve replications. The assignment of treatments
to the experimental units, as well as the data, were processed for a
regression analysis and multivariate statistical (correlations), using
Rstudio version 1.4.1717 (R Core Team, 2022).
Results and discussion
Relative water content
The application with 1.5 mM of SA, without fertilizer and
without stress (100% humidity in the soil), maintained the highest
RWC (89.05%) in the leaves; the lowest (25.36%) was in plants
with severe stress (30% humidity in the soil), with fertilizer and
with SA (0.5 mM) (gure 2). The RWC increased as the intensity
of drought decreased; in plants with fertilizer, with SA (1.5 mM)
and without stress, the RWC improved by 49.79%, with respect to
plants without fertilizer, with severe stress and without SA (0 mM).
Figure 2. Relative water content in P. coccineus under different
doses of salicylic acid and humidity restriction. A)
without stress, B) moderate stress, and C) severe stress.
The foliar application of SA can reduce the loss of water by
transpiration when there is hydric stress, since it regulates the
opening and closing of the stomata (Rao et al., 2012). In this
research, the foliar application of 1.5 mM of SA in P. coccineus
plants, improved the RWC. Similar results were obtained in Lippia
citriodora (L.) (Dianat et al., 2016), Z. mays (Farouk et al., 2018)
and in Calendula ofcinalis (L.) (Gholinezhad, 2020).
Photosynthetic pigments
The highest contents of CHLa, CHLb and CRT pigments (2.20,
1.11 and 0.90 µg.mL
-1
, respectively) were presented in plants
without stress (100% soil humidity), with fertilizer and with SA (1.5
mM); which contributed to the increase in chlorophyll contents by
47.27, 49.54 and 53.33%, with respect to plants without fertilizer,
with severe stress (30% of soil humidity) and without SA (0 mM)
(gure 3).
Photosynthetic efciency depends on pigments (CHLa, CHLb
and CRT), since they are fundamental in the photochemical stage
of photosynthesis (Muhie, 2022). The foliar application of 1.5 mM
of SA in P. coccineus, improved the chlorophyll contents. Similar
results were reported in M. arvensis (Elhakem, 2019), in Helianthus
annuus (L.) (Rehman et al., 2019) and in H. vulgare (Abdelaal et
al., 2020). This is because SA reacts as an antioxidant substance by
eliminating reactive oxygen species (Rehman et al., 2019).
Osmolyte accumulation
The highest content of PRO (31.77 µmol.g
-1
DW) was found in
plants without fertilizer, with severe stress and without SA (0 mM).
The lowest content of this (3.24 µmol.g
-1
DW) was in plants with
fertilizer, without stress (100% humidity in the soil) and 1.5 mM
of SA (best dose for RWC and photosynthetic pigments) (gure 4).
PRO increased with drought intensity by 8.8 times in plants without
fertilizer, without SA and severe stress; with respect to those plants
with fertilizer, with SA and without stress.
The accumulation of osmolytes (PRO, GB, TSS) in the
cytoplasm helps to maintain cellular osmotic balance in response to
abiotic stress (Ozturk et al., 2020). But with hydric stress, the plant
consumes many photosynthates to produce osmotic regulators and
reduce stress (Gholinezhad, 2020).
Figure 3. Concentration of chlorophyll a (CHLa, µg.mL
-1
),
chlorophyll b (CHLb, µg.mL
-1
) and carotenoids
(CRT, µg.mL
-1
); in P. coccineus under different doses
of salicylic acid and humidity restriction. Asterisks
indicate signicant differences in each level of water
stress, in plants with fertilizer and without fertilizer. *,
α<0.05; **, α<0.01; ***, α<0.001.
In this research, PRO increased with the application of SA (1.5
mM) in a condition of greater hydric stress, which was similar to
that obtained in C. exuosus (Idrees et al., 2010), in P. vulgaris
(Ghanbari et al., 2013) and in H. vulgare (Abdelaal et al., 2020).
The highest content of GB (24.80 µmol.g
-1
DW) was found
in plants with fertilizer, without stress and with SA (1.5 mM);
the lowest content of this (19.50 µmol.g
-1
DW) was obtained in
plants without fertilizer, severe stress and without SA. The GB
increased by decreasing the drought intensity by 1.2 times in the
plants with fertilizer, without stress and with SA; this with respect
to those plants without fertilizer, without SA and severe stress. In
this research, the GB was increased with the application of SA
(1.5 mM) in plants without water stress. Its presence under stress
conditions has been favorable in P. coccineus but not in P. vulgaris
(Morosan et al., 2017). Surabhi & Rout (2020) mentioned that high
concentrations of GB increase the permeability of the membrane,
the efciency of water use in conditions of drought stress and the
photosynthetic efciency.
This scientic publication in digital format is a continuation of the Printed Review: Legal Deposit pp 196802ZU42, ISSN 0378-7818.
Aguilar-Luna et al. Rev. Fac. Agron. (LUZ). 2022, 39(3): e2239405-6 |
Figure 4. Osmolyte levels: proline (PRO, µmol.g
-1
), glycine betaine
(GB, µmol.g
-1
) and total soluble sugars (TSS, mg.g
-1
);
in P. coccineus under different doses of salicylic acid
and humidity restriction. Asterisks indicate signicant
differences in each level of water stress, in plants with
fertilizer and without fertilizer. *, α<0.05; **, α<0.01;
***, α<0.001.
The signicant accumulation of PRO with a decrease in protein
fractions (ALB, GLB, PRL and GLT) in a severe stress condition,
suggests that the content of PRO may be related to the hydrolysis of
proteins (Goswami et al., 2020). In this research, the protein fractions
were affected by hydric stress because this type of stress reduces the
soluble carbohydrates of the leaves, amino acids, and total proteins
(Dianat et al., 2016). While the application of SA did not increase
the percentage of protein fractions. Regarding P. coccineus, Teniente-
Martínez et al. (2016) reported a protein content of 21.9% for the
species, while Jacinto-Hernández et al. (2019) indicated that with a
larger grain size there is also lower protein content, with an average
of 22.70%.
Conclusions
The humidity restriction in P. coccineus had negative effects on
the biochemistry of the crop. The foliar application of salicylic acid
improved the tolerance of the crop towards humidity restriction,
favoring physiological processes such as relative water content,
photosynthetic pigments, glycine betaine and total soluble sugars.
The positive effects of salicylic acid must be considered in crop
production, due to the erratic behavior of the climate worldwide.
Figure 5. Extracted protein (%) in each protein fraction: albumins
(ALB), globulins (GLB), prolamines (PRL) and glutelins
(GLT); in P. coccineus under different doses of salicylic
acid and humidity restriction. Asterisks indicate signicant
differences in each level of water stress, in plants with
fertilizer and without fertilizer. *, α<0.05; **, α<0.01; ***,
α<0.001.
The highest content of TSS (31.15 mg eq.glucose g
-1
DW) was
obtained in plants with fertilizer, without stress and with SA (1.5
mM); the lowest content of this (25.11 mg eq.glucose g
-1
DW) was in
plants without fertilizer, severe stress and without SA. TSS increased
by decreasing stress by 1.2 times in plants with fertilizer, without
stress and with SA; the above with respect to plants without fertilizer,
without SA and severe stress.
TSS increased when there was less hydric stress, which differs
from that obtained in Ziziphus mauritiania (Lam) (Gadi & Laxmi,
2012) and Saposhnikovia divaricata (Turcz.) Schischk. (Men et al.,
2018). In P. coccineus, Morosan et al. (2017) found no differences
in the TSS of plants subjected to different stress; but a sugar content
of 9.45 to 11.60% was associated with a sweeter taste (Jacinto-
Hernández et al., 2019).
Protein fraction
In plants without stress, with fertilizer and without SA (0 mM),
higher values of ALB, GLB, PRL and GLT were obtained (23.54,
11.28, 0.54 and 0.61%, respectively). The SA did not increase the
percentage of protein fractions, even in plants with fertilizer (gure
5). On the contrary, 2.0 mM of SA and severe stress (30% of soil
humidity), caused a decrease of 19.71, 35.06, 25.64 and 20.83% in
the protein fractions of plants without fertilizer; and 17.71, 25.88,
20.37 and 18.03% in plants with fertilizer; all this with respect to
plants without SA and without stress.
This scientic publication in digital format is a continuation of the Printed Review: Legal Deposit pp 196802ZU42, ISSN 0378-7818.
Rev. Fac. Agron. (LUZ). 2022, 39(3): e223940. July - September. ISSN 2477-9407.
6-6 |
Literature cited
Abdelaal, K.A.A., Attia, K.A., Alamery, S.F., El-Afry, M.M., Ghazy, A.I.,
Tantawy, D.S., & Hafez, Y.M. (2020). Exogenous application of proline
and salicylic acid can mitigate the injurious impacts of drought stress on
barley plants associated with physiological and histological characters.
Sustainability, 12:1-15. http://dx.doi.org/10.3390/su12051736
Afshari, F., Nakhaei, F., Mosavi, S., & Seghatoleslami, M. (2022). Physiological
and biochemical responses of Stevia rebaudiana Bertoni to nutri-priming
and foliar nutrition under water supply restrictions. Industrial Crops and
Products, 176: 114399. https://doi.org/10.1016/j.indcrop.2021.114399
Barba de la Rosa, A.P., Gueguen, J., Paredes-López, O., & Viroben, G. (1992).
Fractionation procedures, electrophoretic characterization, and amino
acid composition of amaranth seed proteins. Journal of Agricultural and
Food Chemistry, 40:931-936. https://doi.org/10.1021/jf00018a002
Bates, L.S., Waldren, R.P., Teare, I.D., Bates, L.S., Waldern, R.P., & Teare, I.
(1973). Rapid determination of free proline for water-stress studies. Plant
Soil, 39:205-207. https://doi.org/10.1007/BF00018060
Chaimala, A., Jogloy, S., Vorasoot, N., Holbrook, C.C., Kvien, C.K., &
Laohasiriwong, S. (2021). The variation of relative water content, SPAD
chlorophyll meter reading, stomatal conductance, leaf area, and specic
leaf area of Jerusalem artichoke genotypes under different durations
of terminal drought in tropical region. Journal of Agronomy and Crop
Science, 00:1-15. https://doi.org/10.1111/jac.12561
Dianat, M., Saharkhiz, M.J., & Tavassolian, I. (2016). Salicylic acid mitigates
drought stress in Lippia citriodora L.: Effects on biochemical traits and
essential oil yield. Biocatalysis and Agricultural Biotechnology, 8:286-
293. https://doi.org/10.1016/j.bcab.2016.10.010
Dubois, M., Gilles, K.A., Hamilton, J.K., Rebers, P.A., & Smith, F. (1956).
Colorimetric method for determination of sugars and related substances.
Analytical Chemistry, 28:350-356. https://doi.org/10.1021/ac60111a017
Elhakem, A.H. (2019). Impact of salicylic acid application on growth,
photosynthetic pigments and organic osmolytes response in Mentha
arvensis under drought stress. Journal of Biological Sciences, 19:372-
380. https://doi.org/10.3923/jbs.2019.372.380
FAO. (2015). Base referencial mundial del recurso suelo. Organización de la
Naciones Unidas para la Alimentación y la Agricultura (FAO). Rome,
Italy. https://www.fao.org/soils-portal/soil-survey/clasicacion-de-suelos/
base-referencial-mundial/es/
Farouk, S., Arafa, S.A., & Nassar, R.M.A. (2018). Improving drought tolerance in
corn (Zea mays L.) by foliar application with salicylic acid. International
Journal of Environment, 7:104-123. http://www.curresweb.com/ije/
ije/2018/104-123.pdf
Gadi, B.R., and Laxmi, V. (2012). Effect of salicylic acid and moisture
stress on sugar content and sucrose synthase activity in Ziziphus
seedlings. Biochemical and Cellular Archives, 12:21-23. http://
www.connectjournals.com/achivestoc2.php?fulltext=1378201H_2.
pdf&&bookmark=CJ-033216&&issue_id=01&&yaer=2012
Galon, L., Pawelkiewicz, R., Müller, C., da Silva, M.D., Orestes, S.C., Barretta,
F.M., de Oliveira R.E.R., Agazzi, L.R., Aspiazú, I., & Forte, C.T. (2022).
Morphophysiological changes in cleareld oilseed rape as a result of the
application of ALS-herbicides and weed competition. Journal of Plant
Diseases and Protection, 129:993-1003. https://doi.org/10.1007/s41348-
022-00607-6
Ghanbari, A.A., Mousavi, S.H., Gorji, A.M., & Rao, I. (2013). Effects of water
stress on leaves and seeds of bean (Phaseolus vulgaris L.). Turkish
Journal of Field Crops, 18:73-77. http://www.eld-crops.org/assets/pdf/
product5211c60894006.pdf
Gholinezhad, E. (2020). Impact of drought stress and stress modiers on water use
efciency, membrane lipidation indices, and water relationship indices
of pot marigold (Calendula ofcinalis L.). Brazilian Journal of Botany,
43:747-759. https://doi.org/10.1007/s40415-020-00651-2
Gordillo-Curiel, A., Rodríguez-Larramendi, L.A., Salas-Marina, M.Á., & Rosales-
Esquinca, M.Á. (2021). Effect of salicylic acid on the germination and
initial growth of coffee (Coffea arabica L. var. Costa Rica 95). Revista
de la Facultad de Agronomía de la Universidad del Zulia, 38(1):43-59.
https://doi.org/10.47280/RevFacAgron(LUZ).v38.n1.03
Goswami, B., Rankawat, R., & Gadi, B.R. (2020). Physiological and antioxidative
responses associated with drought tolerance of Lasiurus sindicus Henr.
endemic to Thar desert, India. Brazilian Journal of Botany, 43:761-773.
https://doi.org/10.1007/s40415-020-00666-9
Grieve, C.M., and Grattan, S.R. (1983). Rapid assay for the determination of water
soluble quaternary ammonium compounds. Plant and Soil, 70:303-307.
https://doi.org/10.1007/BF02374789
Hossain, A., Pamanick, B., Venugopalan, V.K., Ibrahimova, U., Rahman, M.A.,
Siyal, A.L., Maitra, S., Chatterjee, S., & Aftab, T. (2022). Emerging roles
of plant growth regulators for plants adaptation to abiotic stress-induced
oxidative stress. In: Aftab, T., Naeem, M. (Eds). Emerging Plant Growth
Regulators in Agriculture. Academic Press, UK. pp: 1-72. https://doi.
org/10.1016/B978-0-323-91005-7.00010-2
Idrees, M., Khan, M.M.A., Aftab, T., Naeem, M., & Hashmi, N. (2010). Salicylic
acid-induced physiological and biochemical changes in lemongrass
varieties under water stress. Journal of Plant Interactions, 5:293-303.
https://doi.org/10.1080/17429145.2010.508566
Jacinto-Hernández, C., Coria-Peña, M., Contreras-Santos, G., Martínez-López,
L., Zapata-Martelo, E., y Ayala-Carrillo, M.R. (2019). Azúcares totales
y proteína en frijol nativo de la región Triqui Alta, Oaxaca. Revista
Mexicana de Ciencias Agrícolas, 10:1667-1674. https://doi.org/10.29312/
remexca.v10i7.2114
Kordi, S., Saidi, M., & Ghanbari, F. (2013). Induction of drought tolerance in
sweet basil (Ocimum basilicum L) by salicylic acid. International Journal
of Agricultural and Food Research, 2:18-26. https://doi.org/10.24102/
ijafr.v2i2.149
Latimer, G.W. (2012). Ofcial methods of analysis of AOAC international. 19th
ed. Gaithersburg, Maryland, USA.
Lichtenthaler, H.K., and Buschmann, C. (2001). Chlorophylls and carotenoids:
measurement and characterization by UV-Vis spectroscopy. In: Wrolstad,
R.E., Acree, T.E., An, H., Decker, E.A., Penner, M.H., Reid, D.S.,
Schwartz, S.J., Shoemaker, C.F., Sporns, P. (eds). Current protocols in
food analytical chemistry. John Wiley and Sons, New York, USA. Unit
F4.3. https://doi.org/10.1002/0471142913.faf0403s01
Men, Y., Wang, D., Li, B., Su, Y., & Chen, G. (2018). Effects of drought stress
on the antioxidant system, osmolytes and secondary metabolites of
Saposhnikovia divaricata seedlings. Acta Physiologiae Plantarum,
40:191. https://doi.org/10.1007/s11738-018-2762-0
Morosan, M., Al Hassan, M., Naranjo, M.A., López-Gresa, M.P., Boscaiu, M.,
& Vicente, O. (2017). Comparative analysis of drought responses in
Phaseolus vulgaris (common bean) and P. coccineus (runner bean)
cultivars. The EuroBiotech Journal, 1:247-252. https://doi.org/10.24190/
ISSN2564-615X/2017/03.09
Morr, C., German, B., Kinsella, J., Regenstein, J., Buren, J.V., Kilara, A.,
Lewis, B., & Mangino, M. (1985). A collaborative study to develop a
standardized food protein solubility procedure. Journal of Food Science,
50:1715-1718. https://doi.org/10.1111/j.1365-2621.1985.tb10572.x
Muhie, S.H. (2022). Optimization of photosynthesis for sustainable crop
production. CABI Agriculture and Bioscience, 3, 50. https://doi.
org/10.1186/s43170-022-00117-3
Ozturk, M., Unal, B.T., García-Caparrós, P., Khursheed, A., Gul, A., &
Hasanuzzaman, M. (2020). Osmoregulation and its actions during
the drought stress in plants. Physiologia Plantarum, 1-15. https://doi.
org/10.1111/ppl.13297
R Core Team (2022). R: A language and environment for statistical computing.
R Foundation for Statistical Computing, Vienna, Austria. https://www.R-
project.org/
Rao, S.R., Qayyum, A., Razzaq, A., Ahmad, M., Mahmood, I., & Sher, A. (2012).
Role of foliar application of salicylic acid and L-tryptophan in drought
tolerance of maize. The Journal of Animal & Plant Sciences, 22:768-772.
http://thejaps.org.pk/docs/v-22-3/42.pdf
Rehman, A., Safeer, M., Qamar, R., Mohsin-Altaf, M., Sarwar, N., Farooq, O.,
Mazher-Iqbal, M., and Ahmad, S. (2019). Exogenous application of
salicylic acid ameliorates growth and yield of sunower (Helianthus
annuus L.) in saline soil. Agrociencia, 53:207-217. http://www.colpos.
mx/agrocien/Bimestral/2019/feb-mar/art-6.pdf
Reyes-Matamoros, J., Martínez-Moreno, D., Rueda-Luna, R., & Rodríguez-
Ramírez, T. (2014). Efecto del estrés hídrico en plantas de frijol (Phaseolus
vulgaris L.) en condiciones de invernadero. Revista Iberoamericana
de Ciencias, 1:191-203. http://www.reibci.org/publicados/2014/
julio/2200132.pdf
Surabhi, G.K., and Rout, A. (2020). Glycine betaine and crop abiotic stress
tolerance: an update. In: Roychoudhury, A., Tripathi, D.K. (Eds).
Protective chemical agents in the amelioration of plant abiotic stress:
biochemical and molecular perspectives. John Wiley & Sons, NJ, USA.
pp: 24-52. https://doi.org/10.1002/9781119552154.ch2
Teniente-Martínez, G., González-Cruz, L., Cariño-Cortes, R., & Bernardino-
Nicanor, A. (2016). Caracterización de las proteínas del frijol ayocote
(Phaseolus coccineus L.). Investigación y Desarrollo en Ciencia y
Tecnología de Alimentos, 1:1-6. http://www.fcb.uanl.mx/IDCyTA/les/
volume1/1/1/1.pdf
Yan, X., Liu, J., Wu, K.X., Yang, N., Pan, L.B., Ying, C., Liu, Y. & Zhong-Hua,
T. (2022). Comparative analysis of endogenous hormones and metabolite
proles in early-spring owering plants and unowered plants revealing
the strategy of blossom. Journal of Plant Growth Regulation, 41:2421-
2434. https://doi.org/10.1007/s00344-021-10452-w