Life 13 02067 v2
Life 13 02067 v2
Article
Assessing Drought Tolerance in a Large Number of Upland
Cotton Plants (Gossypium hirsutum L.) under Different
Irrigation Regimes at the Seedling Stage
Sadettin Çelik
Department of Forestry, Genç Vocational School, University of Bingol, Bingol 12500, Turkey;
[email protected]
Abstract: The cotton plant is important since it provides raw materials for various industry branches.
Even though cotton is generally drought-tolerant, it is affected negatively by long-term drought stress.
The trial was conducted according to the applied experimental design as a completely randomized
design (CRD) with three replications to determine a panel of 93 cotton genotypes’ genotypic responses
against drought under controlled conditions in 2022. All genotypes were watered with 80 mL−1 of
water (100% irrigation, field capacity) until three true leaves appeared, and then water stress was
applied at a limited irrigation of 75% (60 mL−1 ), 50% (40 mL−1 ), and 25% (20 mL−1 ) of the field
capacity. After the trial terminated at 52 days, the cv. G56, G44, G5, and G86 in RL; G1, G56, G44,
G86, G51, and G88 in RFW; advanced line G5, followed by the cv. G56, advanced line G44, G75,
and the cv. G90 in RDW; G44, followed by G86, the cv. G56, and elite lines G13 and G5 in NLRs
were observed as drought-tolerant genotypes, respectively, while G35, G15, G26, G67, and G56 in
SL; G15, G52, G60, G31, and G68 in SFW; G35, G52, G57, G41, and G60 in SDW show the highest
drought tolerance means, respectively. In conclusion, the commercial varieties with high means in
roots, namely G86, G56, G88, and G90, and the genotypes G67, G20, G60, and G57 showing tolerance
in shoots, are suggested to be potential parent plants for developing cotton varieties resistant to
drought. Using the cultivars found tolerant in the current study as parents in a drought-tolerant
variety development marker-assisted selection (MAS) plant breeding program will increase the
Citation: Çelik, S. Assessing Drought chance of success in reaching the target after genetic diversity analyses are performed. On the other
Tolerance in a Large Number of hand, it is highly recommended to continue the plant breeding program with the G44, G30, G19,
Upland Cotton Plants (Gossypium G1, G5, G75, G35, G15, G52, G29, and G76 genotypes, which show high tolerance in both root and
hirsutum L.) under Different shoot systems.
Irrigation Regimes at the Seedling
Stage. Life 2023, 13, 2067. https:// Keywords: drought; cotton; root; shoot; tolerance; water scarcity
doi.org/10.3390/life13102067
one of the most important abiotic stress factors that cause 73% of product loss in cotton [8].
In particular, in the USA, which is one of the leaders in world cotton production, the
production loss due to drought stress in the last 50 years is approximately 67% [9], while it
is 34% in Pakistan [10].
Although cotton has a higher tolerance to drought compared to other cultivated indus-
trial field crops, when it is exposed to long-term drought stress, undesirable effects such as
a decrease in yield, lower biomass, and stem weight, a slowdown in plant development,
a decrease in fiber quality, and small bolls may occur [11]. Drought adversely affects
Gossypium spp., along with other plants, by limiting plant height, leaf weight, the number
of nodes, transpiration rate, photosynthesis rate, and stomatal conductivity [11–15]. In
addition, when it is exposed to long-term drought stress, it closes its stomata, rolls its leaves,
tends to have osmotic regulation, and tries to reach water/moisture in the depths of the
soil [16], and the rate of photosynthesis is reduced [17].
A great deal of research is being conducted to understand the physiological, morpho-
logical, and metabolic reactions of plants against drought in roots [18,19]. Under drought
stress, the roots of the plants tend to go deeper and aim to reach underground water, and
these behaviors of the roots also play an important role in activating other mechanisms
related to drought [20]. Anjum et al. [21] indicated that a strong root system plays a very
critical role in physiological functions, carbohydrate storage, uptaking, and the absorption
of water and nutrients from the soil. In the first stage of drought, plants slow down the
elongation of the stem and develop the root system to reach deeper water [22].
The responses of the cotton plant to stress factors vary during different stages of
development. The seedling stage is a period of high sensitivity to environmental conditions
for many plant species, including cotton [23]. The seedling stage is not only adversely
affected by biotic stress factors but also negatively affected by abiotic stress factors, such
as drought [24] and salinity [25]. Brand et al. [26] discovered that low temperatures
at the seedling stage in cotton under chilling conditions are strongly associated with
low temperatures of the seedling stage diseases such as dumping off, which causes root
deterioration. Low temperatures at the seedling stage of cotton have a big effect on the
damage thresholds of pathogens such as Rhizoctonia and Pythium [27]. Furthermore,
seedling emergence rates and development under controlled conditions can determine
the sensitivity of varieties to cold weather [28]. When the cotton seeds emerged and were
exposed to sub-optimal temperatures in late spring, seedling growth was adversely affected,
and significant decreases were observed in yield [29]. When soil moisture tension generally
exceeds 30–50 centibars, drought stress occurs, depending on soil type [30].
To understand the root system responses under water scarcity conditions: a short-
term drought during the seedling period increased the cotton roots ability to reach deep
water but decreased the diameter of the roots [31]. It is revealed that the cotton roots were
affected by drought more than the root distribution [32]. Insufficient soil moisture reduces
the elongation of the roots [33,34], and a shortened root system is developed approximately
42–70 days after the germination of the cotton [32]. Root growth is a critical process to
be grown under sustainable temperatures [35,36]. Zahid et al. [37] stated that 22–30 ◦ C
is an optimal temperature for cotton root and shoot growth and an optimal root/shoot
(R/S) ratio, but a temperature between 32 and 40 ◦ C will restrict the distribution and
growth of the root/shoot system. A similar finding was declared by Koevoets et al. [38]
that the optimal temperature promotes an increase in the root/shoot ratio (R/S ratio);
nevertheless, a temperature above optimal temperature will decrease water absorption and
plant nutrients in the cotton plant root system, and as a results, the root system get weaker
against drought [39]. A decrease in soil water content by evaporation significantly reduces
root activities, but hydraulic lifting and transpiration by roots can increase groundwater
levels to levels close to the soil surface (to 0–40 cm soil depth) [40].
Various studies have been conducted to understand cotton plants’ genotypic responses
against drought. Studies such as developing cultivars that are tolerant to drought through
crossbreeding [41], hormonal regulation against drought and salinity stress [42], under-
Life 2023, 13, 2067 3 of 19
standing the drought tolerance mechanism of cotton and integrating it into variety de-
velopment breeding programs [43], drought stress generation and a determination of
tolerant genotypes in the cotton plant using Polyethylene Glycol (PEG) [44], determining
drought-stress-related traits in the seedling period [45], strategies for coping with drought
stress in cotton and their application to crop improvement programs [45], increasing the
drought tolerance of cotton genotypes by exposing them to salinity during the seedling
period [46], understanding the allelic genetic diversity of drought-resistant cotton geno-
types [47], and identifying cultivars carrying drought genes using Trait-assisted selection
(MAS) in cotton [48] have been carried out.
The research conducted shows that the seedling stage is the period during which
plant reactions can be observed the best [49]. This study aims to investigate the drought-
tolerant level of some advanced lines and some genetic stock cultivars of the cotton
(Gossypium hirsutum L.) species under artificial drought stress with limited irrigation meth-
ods during the seedling stage. In this study, unlike many studies, not only are cultivars
evaluated under drought stress conditions, but also advanced lines’ genetic potential
against drought is investigated.
G1 14 AD1 G. hirsutum L. Türkey G32 ARAS10 AD1 G. hirsutum L. Türkey G63 Fiona AD1 G. hirsutum L. Türkey
G2 15 AD1 G. hirsutum L. Türkey G33 ARAS11 AD1 G. hirsutum L. Türkey G64 Furkan AD1 G. hirsutum L. Türkey
G3 17 AD1 G. hirsutum L. Türkey G34 ARAS12 AD1 G. hirsutum L. Türkey G65 Gaia AD1 G. hirsutum L. Türkey
G4 18 AD1 G. hirsutum L. Türkey G35 ARAS13 AD1 G. hirsutum L. Türkey G66 Kartanesi AD1 G. hirsutum L. Türkey
G5 19 AD1 G. hirsutum L. Türkey G36 ARAS14 AD1 G. hirsutum L. Türkey G67 Lydia AD1 G. hirsutum L. Türkey
G6 21 AD1 G. hirsutum L. Türkey G37 ARAS15 AD1 G. hirsutum L. Türkey G68 Maraş 92 AD1 G. hirsutum L. Türkey
G7 22 AD1 G. hirsutum L. Türkey G38 ARAS16 AD1 G. hirsutum L. Türkey G69 May 344 AD1 G. hirsutum L. Türkey
G8 23 AD1 G. hirsutum L. Türkey G39 ARAS17 AD1 G. hirsutum L. Türkey G70 Maysos AD1 G. hirsutum L. India
G9 24 AD1 G. hirsutum L. Türkey G40 ARAS18 AD1 G. hirsutum L. Türkey G71 Nihal AD1 G. hirsutum L. Türkey
G10 26 AD1 G. hirsutum L. Türkey G41 ARAS19 AD1 G. hirsutum L. Türkey G72 SC-125 AD1 G. hirsutum L. Türkey
G11 27 AD1 G. hirsutum L. Türkey G42 ARAS20 AD1 G. hirsutum L. Türkey G73 SC-21 AD1 G. hirsutum L. Türkey
G12 28 AD1 G. hirsutum L. Türkey G43 ARAS23 AD1 G. hirsutum L. Türkey G74 SC-22 AD1 G. hirsutum L. Türkey
G13 30 AD1 G. hirsutum L. Türkey G44 ARAS24 AD1 G. hirsutum L. Türkey G75 SC-24 AD1 G. hirsutum L. Türkey
G14 31 AD1 G. hirsutum L. Türkey G45 ARAS26 AD1 G. hirsutum L. Türkey G76 SC-25 AD1 G. hirsutum L. Türkey
G15 37 AD1 G. hirsutum L. Türkey G46 ARAS27 AD1 G. hirsutum L. Türkey G77 SC-28 AD1 G. hirsutum L. Türkey
G16 69 AD1 G. hirsutum L. Türkey G47 ARAS31 AD1 G. hirsutum L. Türkey G78 SC-29 AD1 G. hirsutum L. Türkey
G17 ADN 513 AD1 G. hirsutum L. Türkey G48 ARAS32 AD1 G. hirsutum L. Türkey G79 SC-30 AD1 G. hirsutum L. Türkey
G18 ADN123 AD1 G. hirsutum L. Türkey G49 ARAS33 AD1 G. hirsutum L. Türkey G80 SC-36 AD1 G. hirsutum L. Türkey
G19 ADN413 AD1 G. hirsutum L. Türkey G50 ARAS34 AD1 G. hirsutum L. Türkey G81 SC-37 AD1 G. hirsutum L. Türkey
G20 ADN710 AD1 G. hirsutum L. Türkey G51 ARAS35 AD1 G. hirsutum L. Türkey G82 SC-38 AD1 G. hirsutum L. Türkey
G21 ADN741 AD1 G. hirsutum L. Türkey G52 ARAS43 AD1 G. hirsutum L. Türkey G83 SC-39 AD1 G. hirsutum L. Türkey
G22 ADN811 AD1 G. hirsutum L. Türkey G53 ARAS48 AD1 G. hirsutum L. Türkey G84 SC-41 AD1 G. hirsutum L. Türkey
G23 ARAS1 AD1 G. hirsutum L. Türkey G54 ASOS AD1 G. hirsutum L. Türkey G85 SC-42 AD1 G. hirsutum L. Türkey
Life 2023, 13, 2067 4 of 19
Table 1. Cont.
G24 ARAS2 AD1 G. hirsutum L. Türkey G55 Aşkabat 71 AD1 G. hirsutum L. Turkmenistan G86 STV373 AD1 G. hirsutum L. USA
G25 ARAS3 AD1 G. hirsutum L. Türkey G56 BA119 AD1 G. hirsutum L. Türkey G87 STV468 AD1 G. hirsutum L. USA
G26 ARAS4 AD1 G. hirsutum L. Türkey G57 BA151 AD1 G. hirsutum L. Türkey G88 STV498 AD1 G. hirsutum L. USA
G27 ARAS5 AD1 G. hirsutum L. Türkey G58 Beren AD1 G. hirsutum L. Türkey G89 SG 125 AD1 G. hirsutum L. USA
DeltaPine
G28 ARAS6 AD1 G. hirsutum L. Türkey G59 AD1 G. hirsutum L. USA G90 Text AD1 G. hirsutum L. USA
332
DeltaPine
G29 ARAS7 AD1 G. hirsutum L. Türkey G60 AD1 G. hirsutum L. USA G91 TMN 199 AD1 G. hirsutum L. Türkey
396
G30 ARAS8 AD1 G. hirsutum L. Türkey G61 Erşan92 AD1 G. hirsutum L. Türkiye G92 TMN108 AD1 G. hirsutum L. Türkey
G31 ARAS9 AD1 G. hirsutum L. Türkey G62 Famosa AD1 G. hirsutum L. USA G93 TMS 108 AD1 G. hirsutum L. Türkey
2.2. Method
The pattern of the trial is the applied experimental design as a completely randomized
design (CRD) with 3 replications. The experiment was conducted at Bingol University,
Genc Vocational School (Coordinates: 38 ◦ 442’58” N and 40 ◦ 320 11” E), climate chamber.
The soil used is prepared with peat, perlite, and soil in a ratio of 3:1:1. All three components
were sterilized with an autoclave at 121 ◦ C for 15 min and transferred to 200 mL plastic
pots. Triangular holes were made at the bottom of the plastic pots for the excess water
to be drained. four seeds were sowed in each pot at a depth of about 2 cm, and potting
soil was brought to the field capacity. For control purposes, STV373 (G86), BA119 (G56,
White gold), and TEX (G93) varieties adapted to the Southeastern Anatolia Region, where
the experiment was established, were used. In the experiment, three pots were used for
each genotype as the 3 replications. The soil temperature was raised to 15 ◦ C for seeds to
germinate, and since the experiment was established in winter, the cotton seeds, which are
likely to be dormant, were kept in a hot water bath device at 65 ◦ C for 15–30 min before
sowing to break the dormancy. Then, the ambient temperature was adjusted to be between
28 and 38 ◦ C, and the humidity was between 57 and 67% [50].
LED lights were adjusted to provide daylight at 2.500 lux for 14–15 h a day, and
each pot was given 0.2 g of pure nitrogen-containing urea fertilizer for effective growth
14 days after planting. From the starting point of the experiment, cotton genotypes were
irrigated with 100% irrigation every 10 days until true leaves bloomed. All genotypes
were fully watered with 80 mL-1 of water (100% irrigation, field capacity) until three true
leaves appeared. As soon as all the genotypes produced true leaves, the limited irrigation
was started. Then, water stress was applied at limited irrigations of 75% (60 mL−1 ), 50%
(40 mL−1 ), and 25% (20 mL−1 ) of the field capacity. The experiment was terminated on the
52nd day.
The cotton plants were taken out of their pots intact, the soil was washed using tap
water without damaging the roots, and parameters such as the Root Length (RL), Root
fresh weight (RFW), Root dry weight (RDW), Shoot length (SL), Shoot fresh weight (SFW),
Shoot dry weight (SDW) and the number of lateral roots (NLRs) of one plant from each
replication genotype were measured. RL was calculated by a direct measurement of fresh
taproots, RFW by direct weighting of taproots, RDW was measured after drying in the oven
for 24 hours at 60 ◦ C, NLRs by direct counting of roots before drying, SL by measuring with
a ruler, SFW by calculation with a scale with 0.5 mg precision, and SDW was measured
after drying in the oven for 24 hours at 60 ◦ C [50]
of the parameters was investigated (p < 0.05). The least significant difference (LSD) was
used to compare the means.
3. Results
3.1. Analysis of Variance (ANOVA)
In the experiment, the RL, RFW, RDW, NLRs, SL, SFW, and SDW of the cotton
germplasm panel were measured. As a result of the statistical analysis, it was observed that
there was a highly significant difference between the means for RL, RFW, RDW, NLRs, SL,
and SFW (p < 0.01). The statistical analysis conducted at a 95% confidence level (p < 0.05)
revealed that, according to the mean squares, there is a significant difference among the
genotypic means in terms of the shoot dry weight trait. Table 2 indicates the significance of
the population mean differences for each trait.
Figure 1
means above the overall mean and 43 genotypes with means below the overall mean. The
genotype with the highest frequency was 24.
A B
C D
E F
Figure 1. (A) Rooth length (RL), (B) root fresh weight (RFW), (C) root dry weight, (D) shoot length
(SL), (E) shoot fresh weight (SFW), (F) shoot dry weight (SDW), and (G) number of lateral roots
(NLRs) Frequency charts.
Life 2023, 13, 2067 7 of 19
Accordingly, the mean total shoot length (SL) of the genotypes was 12.47 cm. G35 (18.33 cm)
had the longest SL, and it was followed by the G15-G29 (16.67 cm) and G76 (16.33 cm)
breeding genotypes, respectively. The commercial cultivar G65 (6.33 cm) showed the
lowest SL means, followed by the elite lines G72-G32 (7.33 cm) and the local cotton variety
G22 (7.33) (Supplementary Table S1). Regarding the SL (Shoot length) trait, 31 genotypes
had means below the overall mean, and 62 genotypes had means above the overall mean
(Figure 1D).
The mean SFW of 93 cotton genotypes was 1.3280 grams. The highest SFW values were
recorded in the following genotypes: the advanced line G15 (1.853 g), G52 (1.815 g), and
the commercial variety G60 (1.805 g), respectively. These were followed by G31 (1.752 g),
G41 (1.736 g), G47 (1.724 g), G14 (1.700 g), and the commercial local variety G68 (1.691 g).
The lowest SFW value was measured in the advanced genotype G83 (0.686 g), followed
by G2 (0.716 g), G75 (0.739 g), and G91 (0.740 g), respectively (Supplementary Table S1).
Mostly of the cotton genotypes showed the expected responses under water scarcity. For
another drought-related trait, SFW (shoot fresh weight), 24 genotypes had the highest
means, while 68 genotypes had means above the population mean of 1.3280 g (Figure 1E).
The SDW mean of all the genotypes was recorded as 0.163 g. The highest SDW
value was recorded in G35 (0.349 g) and the lowest in the G65 (0.060 g) genotype. While
commercial cultivars such as G57 (0.337 g), G60 (0.294 g), and G66 (0.289 g) showed high
mean SDW values, the advanced lines G52 (0.345 g) and G41 (0.314) showed very high SDW
values. The second lowest SDW value was found for the commercial varieties G91 (0.078 g)
and G4 (0.078 g) and G20 (0.078 g), followed by G22 (0.081 g) and the cv. G90 (0.087 g)
(Supplementary Table S1). In the case of SDW (shoot dry weight), 34 genotypes had the
highest frequency, and the majority of the population had SDW values below the population
mean of 0.163 (Figure 1G).
In the current study, the G44 (38) RIL genotype had the greatest number of lateral roots
(NLRs), and it was followed by G86 (33), the commercial cv. G56 (32), and the advanced
lines G13 (30), G5 (29.33), and G49 (29). The mean of number of lateral roots of all the
genotypes was 19.16 (Supplementary Table S1). Regarding the NLRs (number of lateral
roots) characteristic, 21 genotypes had a mean of 15.556, 22 genotypes had a mean of 19.037,
and 26.778 was the mean for 18 genotypes. The population NLR mean was 19.159, with
28 genotypes having means below it and 68 genotypes having means above it (Figure 1G).
The scatter plot analysis was carried out to determine the relationship and relationship
type between every two variables of drought indicators. Yi [53] emphasized that the
closer they get to the linear line, the more the relationship between parameters increases.
According to the Figure 2 scatter plots and correlation diagram, while the strongest positive
correlation is seen between RL and RFW (0.823), followed by NLRs and RL (0.759), RFW and
NLRs (0.679), RFW–RDW (0.642), RL–RDW (0.568), and NLRs–RDW (0.494), the strongest
negative correlation took place between SL–RDW (−0.446), followed by RFW–SL (−0.327),
NLRs–SL (−0.318), NLRs–SFW (−0.219), and SDW–RFW (−0.143) (Table 3, Figure 2).
Similar correlations between the root–shoot traits of the cotton plant were identified by [54].
A principal component analysis (PCA) is a powerful multivariate statistical technique
widely used for identifying patterns of data, expressing the data, and highlighting their
similarities and dissimilarities [55]. A PCA simplifies complex datasets by transforming
them into a new coordinate system, where the data’s variance is maximized along the
principal components. This dimensionality reduction retains most of the relevant informa-
tion while eliminating redundant or noisy features, thus aiding in data compression and
visualization [56].
The graph (Figure 3) is the loading plot from the PCA analysis. The results of the
PCA are given in Figure 3. The PCA was conducted based on the morphological drought
parameters, and in the correlation circle, the first axes (F1) and the second axes (F2) represent
the discriminant function analysis (DFA), respectively, 47.6% and 21.4% of the variable.
Among the drought measurement parameters of the cotton plant’s aboveground portion,
the strongest positive correlation was found between shoot dry weight (SDW) and shoot
Life 2023, 13, 2067 8 of 19
fresh weight (SFW), while shoot length (SL) showed a positive correlation with both SFW
and SDW (Figure 3). Accordingly, as the SL increases, it appears that the SFW and SDW
values increase positively.
Figure 2. Scatter (XY) plot analysis of drought morphological traits using Minitab 19 [52] and
correlation diagram in JMP 17.00 [51]. version.
Figure 3. Correlation circle obtained by principal component analysis (PCA) of the variables factor
space of F1 and F2 using JMP 17.00 ver. [51] statistical analysis software.
Figure 4. Cluster analysis based on genotypes genotypic responses under drought stress using JMP
17.00 ver. [51] statistical analysis software.
4. Discussion
The responses of plant roots under drought stress have become the focus of researchers
in recent years [57,58]. The first plant organ that is exposed to water stress is the roots,
and by transmitting this stress to the rest of the plant, it goes through morphological,
Life 2023, 13, 2067 11 of 19
physiological, and metabolic changes, and the plant starts to activate its mechanisms to
cope with the water stress [18]. Root length (RL) growth has a great impact on the uptake
of nutrients and root system architecture [59,60]. It is a current trend in plant breeding
to select genotypes with the longest roots for drought tolerance [9,61]. It was seen in
this study that several advanced breeding lines had a high tolerance to drought in some
drought measurement parameters. The cv. G56 had the longest RL (27 cm), followed by
G44 (25.67 cm), G5 (24.33 cm), and G86 (23.67 cm) in this study (Supplementary Table S1).
Similar results were found by [50], which reported that taproots had a root length between
47.9 and 86.6 cm under full irrigation and between 48.3 and 82.5 cm under restricted
irrigation in their experiment. The higher root length means of [50] compared to our results
are believed to be dependent on the experimental conditions, trial materials, origins of the
plant varieties used, amount of water used, and duration of the experiment. In the study,
although the humidity and temperature in the greenhouse can increase excessively during
the day, the amount of soil mixture used was parallel to our study, and a longer taproot
system was observed.
Research has shown that immersing roots in depths (root length) where the moisture
content is high can compensate for the moisture losses that occur through evapotranspira-
tion [62]. In the current study, the G76 and G41 advanced lines produced a 5.33 cm RL as the
lowest RL, followed by G35, G14 (5.7), G79, G73, and G7 (7.3 cm) (Supplementary Table S1).
Our results are similar to [63], with the RL trait varying between 17.10 cm and 30.80 cm.
Similar root length (RL) values were obtained by Pawar and Veena [64], with the highest
root length at 12.72, 12.17, and 12.88 cm and the lowest at 9.8, 9.85, and 10.5 cm with the
PEG 6000 drought application. At the same time, Pawar and Veena [64] revealed that the
root length increased when the PEG 6000 application was increased by 10% and decreased
after that level. This situation can be explained by the plant root tolerance mechanism of the
plant: the plant sends more photosynthesis products to the root in order to strengthen the
roots and uptake more water from the soil [65]. In a study, Zahid et al. [66] found a highly
significant correlation between genotypes and watering, and they found the root length
to be 5.161 cm. According to the current study, with mean root length of 27 cm, 5.161 cm
is highly low, and the irrigation period, the amount of water given in each irrigation, soil
mixture, infiltration and germination ability, and speed can be shown as the reasons why
it is found to be lower than our study. Wang et al. [67] reported that the distribution of
photosynthetic products in the roots of beans increased under drought stress, which did not
last long, but the photosynthetic products in the production organs decreased significantly.
It was observed that the plant tries to strengthen its root system under long-lasting drought
stress, thereby severely reducing its yield and economic added value as a result of giving
all its weight to the roots. Studies reported that a situation similar to the growth period of
wheat can be observed in cotton: moderate water stress causes photosynthetic products to
be transported to the roots by vascular bundles and has a similar positive effect on buds
and bolls [68]. It was reported that moderate water stress during the grain-filling period of
the wheat plant increases the transport of photosynthetic products to the roots, and this
provides a significant benefit to the wheat, as the wheat will have developed roots and thus
take more moisture and therefore more nutrients from the soil [69].
A higher root weight enables the uptake of more plant nutrients from the soil and
plays a key role in vegetative and generative development [70]. In the current study, the
RFW trait values changed between 0.123 g (G76) and 0.446 g (G1), with a population
mean of 0.260 g. The genotypes that were tolerant according to the root fresh weight
trait were the cv. G56 (0.427 g), G44 (0.424 g), the commercial variety G86 (0.411 g),
G51 (0.397 g), and G47 (0.148 g), respectively (Supplementary Table S1). Our findings are
similar to Iqbal et al. [71], Mvula et al. [63], and Jaafar et al. [72]. Similarly, Shah et al. [73]
obtained a root weight ranging from 0.049 to 0.155 g under water stress in a trial conducted
under normal and water-stressed conditions for 52 days. In a similar study, Akbar and
Hussain [74] obtained fresh root weights ranging from 0.222 g to 0.394 g under different
drought stresses. In Figure 1B, it is observed that 22 genotypes have a mean RFW of 0.267,
Life 2023, 13, 2067 12 of 19
which is slightly higher than the population mean. The fact that 57 genotypes produce a
mean higher than the population suggests genetic diversity within the population, with
some genotypes exhibiting superior root growth in terms of fresh weight. The genotypic
mean for the RDW is 14.27 (Figure 1B). The research showed that there is a positive
correlation between root weight and drought and that the genotypes with quantitatively
higher root weights had a higher tolerance to drought stress than the others. While the
specific context or objectives of the study are not provided, this trait likely relates to the
ability of genotypes to allocate resources to root growth.
Under long-term drought stress, the root dry weight increased and the leaf area
decreased in Chinese spinach [75]. In the current study of drought response measurement
root trait, root dry weight, the highest value was recorded in the advanced line G5 (0.127 g),
followed by the control variety G56 (0.112 g), G44 (0.099 g), G75 (0.092 g), and control variety
G90 (0.091 g), while lowest RDW values were recorded in the advanced line G76 (0.016 g),
followed by genotypes G35 (0.021 g), G20 (0.024 g), G61 (0.026 g), G70 (0.028 g), G59-G15
(0.029 g), and G67 (0.030 g) (Supplementary Table S1). The RDW varied from 0.79 g to
1.18 g [63]. Istiqomah et al. (2021) [70] found a very significant difference between the
applications in terms of dry root weight or total dry weight in corn plants. Similarly,
in their study investigating the responses of the germination stage root drought trait
to drought in cotton, Fathi-Sadabadi et al. [54] obtained dry root weights ranging from
0.00342 to 0.00685 g. In Figure 1C, the genotypes exhibit a wide range of RDW values, with
50 genotypes having means above the overall mean and 43 below. This variation could
have practical implications for breeding programs, as it suggests the presence of genotypes
with different root biomass production potentials. The variation in genotypic means for
RDW (Figure 1C) indicates that some genotypes are more efficient at producing dry root
biomass than others.
As the importance of NLRs traits observed in this current study indicates, the num-
ber and development of lateral roots are important indicators of drought tolerance. The
development of the root system, which allows plants to stand, hold on to the soil, give
yield, and grow and develop, plays an active role in the fight against agricultural drought.
Wang et al. [76] stated that many previous studies related to the crop–root system improve-
ment suggest that increasing the number of lateral roots (NLRs) could characterize plant
development and increase crop yield. Lateral root numbers and development are regulated
by many diverse hormones and their interactions [77,78]. Lateral root initiation (LRI) and
development is coordinated by many genes. In Arabidopsis thaliana, there are more than
seven genes associated with lateral roots [79,80]. The population mean of the number
of lateral roots (NLRs) changed from 5 to 38. The advanced line G44 (38) showed the
highest NLRs, and this value was followed by G86 (33), the cv. (control variety) G56 (32),
G13 (30), and G5 (29.33). The lowest NLR values ranged from 5 to 10. In the current
study, the lowest NLRs were recorded in G47 (5), followed by G76 (6.33), and G79 (7)
(Supplementary Table S1). Similarly, the NLR value showed variability between 29.67 and
47.10 [63]. McMichael et al. [81] obtained similar NLR values ranging from 4 to 50 at 30 ◦ C
within 7 days after germination. The reason for the higher LRN values may be associated
with not applying drought stress in the first 7 days and plenty of water and plant nutrients.
In a study with 120 exotic cotton genotypes provided, the NLRs mean changed from 3.5,
17.5, 25.97, and 49.75, respectively [81]. The genotypic means for NLRs vary significantly
(Figure 1G), with 28 genotypes below the population mean and 68 genotypes above it. This
trait might be related to drought tolerance or stress response, and the observed variation
suggests potential candidates for further research or breeding programs. Lateral roots
and numbers play a key role in plants reaching and absorbing water [82] and uptaking
plant nutrients such as phosphorus [83]. Hund et al. [84,85]. Gallardo et al. [86] reported
that enough water in soil accelerates root development, and Luo et al. [87] reported that
moderate water stress in the middle and upper layers of the soil strengthens the root
system in the early stage of cotton plant development and contributes significantly to roots
reaching deep waters. The number, development, and growth of lateral roots are critical for
Life 2023, 13, 2067 13 of 19
the development and yield of field crops [88,89]. In another study, Forde and Lorenzo [90]
stated that the lateral roots, which are important in plants with a taproot system, initially
act as taproots and take water and nutrients dissolved in water. To increase tolerance to
drought stress, it was seen that researchers should focus on the development of genotypes
of lateral root systems. It was seen that the drought-resistance RIL genotypes G44 (38),
G86 (33), and G56 (32) varieties, which have good adaptation in the Southeast Anatolia
region of Turkey, where cotton is mainly cultivated, develop better lateral root systems
under drought stress. Even though the current study breeding lines have developed good
taproot systems, they may not show high resistance against high temperatures compared
to commercial cultivars adapted in warm regions. It has been put forward that lateral root
numbers can have positive effects of vigorous growth under low temperatures in maize
plants. Considering the studies in the literature, it is seen that the number of lateral roots is
a drought trait that increases the tolerance of plants against drought stress.
Shoot length (SL) can be used as a criterion for the selection of drought-tolerant
plants [91,92]. As a result of this study, it has been observed that shoot length (SL) dur-
ing the seedling stage is one of the organs most affected by drought in cotton, and the
population mean obtained was 12.47, while the genotype means ranged from 18.33 cm
(G35) to 6.33 cm (G91) (Supplementary Table S1). The shoot length averages ranging from
35.20 cm to 43.37 cm, as reported by [63], resemble our findings. During the initial period
that drought stress is observed, it has been noted that the shoot length remains constant,
but in the advanced stages of drought, the shoot length starts to increase. This situation
can be explained by the fact that the plant absorbs more water from the soil, leading to in-
creased photosynthesis and the production of C6 H12 O6 (glucose). Similarly, Iqbal et al. [71]
reported that drought during the seedling stage negatively affects both the shoot and root
morphology of cotton, reducing the shoot length by 29%. A study indicated that drought
stress causes a significant reduction in shoot elongation, and the percentage of decreasing
SL ranged from 97.14 to 35.1 [66]. A reduction in the shoot or in the root length may be
caused by an imbalance of water [93]. Genotypes with a mean SL of 12.47 (Figure 1D) show
significant variability, with 31 genotypes below the population mean and 62 above it. This
trait may be important in the context of plant height and overall shoot development, which
can impact crop yield and plant architecture.
For shoot fresh weight (SFW) drought traits, an advanced line, G15, has the weight
with a 1.853 g value, followed by G52 (1.815 g) and the commercial variety G60 (1.805 g),
respectively. (Supplementary Table S1). Similar SFW trait results were obtained by Akbar
and Hussain [74], Lund and Elsayed [94], and Iftikhar et al. [95]. In Figure 1E, 24 genotypes
exhibit the highest means for SFW, with 68 genotypes exceeding the population mean of
13.280. This suggests that there are genotypes with a higher capacity for shoot growth and
biomass production, which could be advantageous in terms of yield potential.
In Figure 1G, the majority of genotypes have SDW values below the population mean.
This indicates that, in general, genotypes tend to have less dry shoot biomass compared to
the population mean. However, the presence of genotypes with higher SDW values could
be of interest for further study or breeding efforts. The best SDW value was recorded in
G35 (0.349 g) and the lowest in the G65 (0.060 g) genotype. While commercial cultivars
such as G57 (0.337 g), G60 (0.294 g) had lower SDW values. (Supplementary Table S1). In a
study, Ahmad et al. [96] obtained similar results while investigating the root–shoot traits
responses under water deficiency. The genotype G35, which shows the highest value in
shoot dry weight (SDW), also exhibits the highest value in the shoot length (SL) parameter
as a drought trait. Genotypes G22 and G91 have low means in the SL, SFW, and SDW
parameters, indicating their low tolerance to drought stress (Supplementary Table S1).
Basal et al. [50] highlighted traits such as SFW and SDW, due to their ease of measurement
and reliability, that could be used as selection criteria in measuring drought tolerance. The
genotype G35 has emerged as the most tolerant genotype in both traits, with a mean SL
of 18.33 cm and SDW of 0.349 g. Meanwhile, genotype G15 registers the second-highest
mean SL, and the same genotype gave the highest SFW mean (Supplementary Table S1).
Life 2023, 13, 2067 14 of 19
These values indicate that an increase in SL does not always correspond to the highest
SFW; however, there is a positive correlation between an increase in SL and SFW. A similar
situation is observed in the tolerant genotype G31 for the SL and SFW traits. On the contrary,
the genotype G91 has the lowest SL value. Generally, genotypes with higher SL means
have higher SFW means, and genotypes with lower SL means have lower SFW values
In the experiment, among plants that shade each other, those receiving less light
exhibit higher shoot length but lower shoot fresh and shoot dry weights compared to plants
receiving full light. This situation can be explained by the hypothesis that as the amount of
light increases, the plant enhances its photosynthetic rate and consequently increases its
biomass. Based on the conducted phenotypic observations, it has been observed that under
greenhouse conditions, cotton genotypes exhibit shorter shoot lengths compared to the
same duration of laboratory trials. However, it is evident that the experiment established in
greenhouse conditions has significantly longer root lengths and a higher lateral root count
compared to the laboratory drought trial. This suggests that when the plant experiences
actual water stress, it strengthens its roots and engages in coping mechanisms to deal with
water scarcity. This response aims to minimize the impact of drought and indicates the
activation of defense mechanisms to mitigate the effects of water stress.
The provided information presents valuable insights into the performance of vari-
ous genotypes concerning SFW and SDW traits, as well as their comparison to control
genotypes. Genotypes G60, G52, and G41 stand out for their high mean in both SFW and
SDW, indicating their potential as superior performers in terms of biomass production. A
noteworthy observation is made with genotype G52, which exhibits a positive correlation
between SFW and SDW. As the SFW increases, the SDW also increases proportionally. This
correlation suggests that as these plants put on more fresh weight, they also accumulate
more dry weight, indicating an efficient conversion of water and nutrients into biomass.
This is a desirable trait in agriculture as it signifies robust growth and resource utilization
efficiency (Supplementary Table S1). However, the genotype G60 presents an interesting
contrast. While it boasts the highest SFW value among the genotypes, it ranks sixth in
SDW. This discrepancy suggests that in the case of G60, an increase in fresh weight does
not necessarily lead to a corresponding increase in dry weight. This could be due to various
factors, such as differences in water content or the allocation of resources within the plant.
Further investigation is warranted to understand this specific genotype’s growth pattern
better. (Supplementary Table S1).
5. Conclusions
In this current study, the genotypes G44 and G56 exhibit common high values in all
four drought traits (NLRs, RL, RFW, and RDW). The genotypes G13 and G5 share high
values in NLRs and RL, while the genotype G5 coincides in terms of NLRs, RL, and RDW.
The genotypes G1, G56, G51, and G88 show the highest values in RFW, and the genotypes
G5, G56, G44, G75, and G90 consistently yield the highest means in the RDW traits. The
genotype G35 shows the highest mean values in both SL and SDW; the genotype G15
shares high values in both SL and SFW; and the genotypes G41, G52, and G60 have high
values in both the SFW and SDW parameters, demonstrating high drought tolerance. Using
the tolerant cultivars found in the current study as parents in drought-tolerant variety
development in the marker-assisted selection (MAS) plant breeding program will increase
the success of reaching the target after genetic diversity analyses are performed. It is also
possible for the genes responsible for this tolerance to transfer from the donor plants to the
recipient plants through hybridization.
References
1. Bardak, A.; Çelik, S.; Erdoğan, O.; Ekinci, R.; Dumlupinar, Z. Association Mapping of Verticillium Wilt Disease in a Worldwide
Collection of Cotton (Gossypium hirsutum L.). Plants 2021, 10, 306. [CrossRef]
2. Bolek, Y.; Tekerek, H.; Hayat, K.; Bardak, A. Screening of cotton genotypes for protein content, oil and fatty acid composition. J.
Agric. Sci. 2016, 8, 107–121. [CrossRef]
3. Çelik, S.; Bardak, A.; Erdoğan, O. Screening of upland cotton genotypes (Gossypium hirsutum L.) against cotton verticillium
(Verticillium dahliae Kleb.) Wilt. Bangladesh J. Bot. 2019, 48, 1185–1192. [CrossRef]
4. Bardak, A.; Fidan, M.S.; Dağgeçen, E.; Tekerek, H.; Çelik, S.; Parlak, D.; Hayat, K. Association Mapping for Gossypol Contents in
Cotton. J. Agric. Nat. 2017, 20, 236–240.
5. Riaz, M.; Farooq, J.; Sakhawat, G.; Mahmood, A.; Sadiq, M.A.; Yaseen, M. Genotypic variability for root/shoot parameters under
water stress in some advanced lines of cotton (Gossypium hirsutum L.). Genet. Mol. Res. 2013, 12, 552–561. [CrossRef]
6. Statista. 2022. Available online: http://www.statista.com/statistics/263055/cotton-production-worldwide-by-top-countries/
(accessed on 31 December 2022).
7. Mahmood, T.; Khalid, S.; Abdullah, M.; Ahmed, Z.; Shah, M.K.N.; Ghafoor, A.; Du, X. Insights into drought stress signaling in
plants and the molecular genetic basis of cotton drought tolerance. Cells 2020, 9, 105. [CrossRef] [PubMed]
8. Saranga, Y.; Paterson, A.H.; Levi, A. Bridging classical and molecular genetics of abiotic stress resistance in cotton. In Genetics and
Genomics of Cotton; Springer: New York, NY, USA, 2009; pp. 337–352. [CrossRef]
9. Comas, L.H.; Becker, S.R.; Cruz, V.M.V.; Byrne, P.F.; Dierig, D.A. Root traits contributing to plant productivity under drought.
Front. Plant Sci. 2013, 4, 442. [CrossRef] [PubMed]
10. Dawn News. Cotton Production Plummets 34pc. Available online: http://www.dawn.com/news/1240448 (accessed on
19 February 2016).
11. Parida, A.K.; Dagaonkar, V.S.; Phalak, M.S.; Umalkar, G.V.; Aurangabadkar, L.P. Alterations in photosynthetic pigments, protein
and osmotic components in cotton genotypes subjected to short-term drought stress followed by recovery. Plant Biotechnol. Rep.
2007, 1, 37–48. [CrossRef]
12. Asati, R.; Tripathi, M.K.; Tiwari, S.; Yadav, R.K.; Tripathi, N. Molecular Breeding and Drought Tolerance in Chickpea. Life 2022,
12, 1846. [CrossRef]
13. Zhang, X.; Yao, D.; Wang, Q.; Xu, W.; Wei, Q.; Wang, C.; Li, F. mRNA-seq analysis of the Gossypium arboreum transcriptome
reveals tissue selective signaling in response to water stress during the seedling stage. PLoS ONE 2013, 8, e54762. [CrossRef]
14. Ranjan, A.; Nigam, D.; Asif, M.H.; Singh, R.; Ranjan, S.; Mantri, S.; Pandey, N.; Trivedi, I.; Rai, K.M.; Jena, S.N.; et al. Genome-wide
expression profiling of two accessions of G. herbaceum L. in response to drought. BMC Genom. 2012, 13, 94. [CrossRef] [PubMed]
15. Bowman, M.J.; Park, W.; Bauer, P.J.; Udall, J.A.; Page, J.T.; Raney, J.; Scheffler, B.E.; Jones, D.C.; Campbell, B.T. RNA-Seq
transcriptome profiling of upland cotton (Gossypium hirsutum L.) root tissue under water-deficit stress. PLoS ONE 2013, 8, e82634.
[CrossRef] [PubMed]
16. Hejnák, V.; Tatar Atasoy, G.D.; Martinková, J.; Çelen, A.E.; Hnilička, F.; Skalický, M. Growth, and photosynthesis of upland and
Pima cotton: Response to drought and heat stress. Plant Soil Environ. 2015, 62, 507–514. [CrossRef]
17. Chastain, D.R.; Snider, J.L.; Collins, G.D.; Perry, C.D.; Whitaker, J.; Byrd, S.A. Water deficit in field-grown Gossypium hirsutum
primarily limits net photosynthesis by decreasing stomatal conductance, increasing photorespiration, and increasing the ratio of
dark respiration to gross photosynthesis. Plant Physiol. 2014, 171, 1576–1585. [CrossRef] [PubMed]
18. Kang, J.; Peng, Y.; Xu, W. Crop Root Responses to Drought Stress: Molecular Mechanisms, Nutrient Regulations, and Interactions
with Microorganisms in the Rhizosphere. Int. J. Mol. Sci. 2022, 23, 9310. [CrossRef]
19. Ahmad, I.; Mian, A.; Maathuis, F.J. Overexpression of the rice AKT1 potassium channel affects potassium nutrition and rice
drought tolerance. J. Exp. Bot. 2016, 67, 2689–2698. [CrossRef]
20. Španić, V.; Ižaković, M.; Marček, T. Wheat germination and seedlings under PEG-induced conditions. Agron. Glas. Agron. J. 2017,
79, 99–109. [CrossRef]
21. Anjum, S.A.; Xie, X.; Wang, L.; Saleem, M.F.; Man, C.; Lei, W. Morphological, physiological and biochemical responses of plants
to drought stress. Afr. J. Agric. Res. 2011, 6, 2026–2032. [CrossRef]
22. Kul, R.; Ekinci, M.; Turan, M.; Ors, S.; Yildirim, E. How abiotic stress conditions effects plant roots. Plant Root. 2020, 6–10.
[CrossRef]
Life 2023, 13, 2067 16 of 19
23. Gutterman, Y. Survival Strategies of Annual Desert Plants; Springer: Berlin/Heidelberg, Germany, 2002; Available online: https:
//link.springer.com/book/10.1007/978-3-642-55974-7 (accessed on 3 January 2023).
24. Tarnawa, Á.; Kende, Z.; Sghaier, A.H.; Kovács, G.P.; Gyuricza, C.; Khaeim, H. Effect of Abiotic Stresses from Drought, Temperature,
and Density on Germination and Seedling Growth of Barley (Hordeum vulgare L.). Plants 2023, 12, 1792. [CrossRef]
25. Aydinşakir, K.; Büyüktaş, D.; Nazmi, D.; Karaca, C. Impact of salinity stress on growing, seedling development and water
consumption of peanut (Arachis hypogaea cv. NC-7). Akdeniz Univ. J. Fac. Agric. 2015, 28, 77–84.
26. Brand, D.; Wijewardana, C.; Gao, W.; Reddy, K.R. Interactive effects of carbon dioxide, low temperature, and ultraviolet-B
radiation on cotton seedling root and shoot morphology and growth. Front. Earth Sci. 2016, 10, 607–620. [CrossRef]
27. Brown, E.A.; McCarter, S.M. Effect of a seedling disease caused by Rhizoctonia solani on subsequent growth and yield of cotton.
Phytopathology 1976, 66, 111–115. [CrossRef]
28. Singh, B.; Norvell, E.; Wijewardana, C.; Wallace, T.; Chastain, D.; Reddy, K.R. Assessing morphological characteristics of advanced
cotton lines from different breeding programs for low temperature and drought tolerance. J. Agron. Crop. Sci. 2018, 4, 467–476.
[CrossRef]
29. Bradow, J.M.; Bauer, P.J. Germination and seedling development. In Physiology of Cotton; Stewart, J.M., Oosterhuis, D.M.,
Heitholt, J.J., Mauney, J.R., Eds.; Springer: Dordrecht, The Netherlands, 2010; pp. 48–56.
30. Perry, C.; Barnes, E.; Munk, D.; Fisher, K.; Bauer, P. Cotton İrrigation Management for Humid Regions; Cotton Incorporated: Cary,
NC, USA, 2012.
31. Pace, P.F.; Cralle, H.T.; El-Halawany, S.H.; Cothren, J.T.; Senseman, S.A. Drought-induced changes in shoot and root growth of
young cotton plants. J. Cotton Sci. 1999, 3, 183–187.
32. Malik, R.S.; Dhankar, J.S.; Turner, N.C. Influence of soil water deficits on root growth of cotton seedlings. Plant Soil 1979,
53, 109–115. [CrossRef]
33. Ball, R.A.; Oosterhuis, D.M.; Mauromoustakos, A. Growth dynamics of the cotton plant during water-deficit stress. J. Agron. 1994,
86, 788–795. [CrossRef]
34. Prior, S.A.; Rogers, H.H.; Runion, G.B.; Kimball, B.A.; Mauney, J.R.; Lewin, K.F.; Nagy, J.; Hendrey, G.R. Free-air carbon dioxide
enrichment of cotton: Root morphological characteristics. J. Environ. Qual. 1995, 24, 678–683. [CrossRef]
35. Mai, W.; Xue, X.; Feng, G.; Yang, R.; Tian, C. Can optimization of phosphorus input lead to high productivity and high phosphorus
use efficiency of cotton through maximization of root/mycorrhizal efficiency in phosphorus acquisition? Field Crops Res. 2018,
216, 100–108. [CrossRef]
36. Gavelienė, V.; Jurkonienė, S.; Jankovska-Bortkevič, E.; Švegždienė, D. Effects of Elevated Temperature on Root System Develop-
ment of Two Lupine Species. Plants 2022, 11, 192. [CrossRef]
37. Zahid, K.R.; Ali, F.; Shah, F.; Younas, M.; Shah, T.; Shahwar, D.; Wu, W. Response and tolerance mechanism of cotton Gossypium
spp.to elevated temperature stress: A review. Front. Plant Sci. 2016, 7, 937. [CrossRef] [PubMed]
38. Koevoets, I.T.; Venema, J.H.; Elzenga, J.T.M.; Testerink, C. Roots withstanding their environment: Exploiting root system
architecture responses to abiotic stress to improve crop tolerance. Front. Plant Sci. 2016, 7, 1335. [CrossRef] [PubMed]
39. Luo, H.; Xu, H.; Chu, C.; He, F.; Fang, S. High temperature can change root system architecture and intensify root interactions of
plant seedlings. Front. Plant Sci. 2020, 11, 160. [CrossRef] [PubMed]
40. Chiatante, D.; Di Iorio, A.; Maiuro, L.; Scippa, S.G. Effect of water stress on root meristems in woody and herbaceous plants
during the first stage of development. Plant Soil 1999, 217, 159–172. [CrossRef]
41. Kıvılcım, N.; Şahin, A.; Ekşi, İ.; Özbek, N.; Yolcu, S.; Naza, İ.; Bilgen, F.; İmamoğlu, A.; Sezener, V.; Gençyılmaz, E.; et al.
Development of Drought-Tolerant Cotton Varieties by Crossbreeding; Project Report. TAGEM Project no: 97/03/01/004; Directorate of
Agricultural Research, Nazilli Cotton Research Institute: Aydın, Turkey, 2005.
42. Riemann, M.; Dhakarey, R.; Hazman, M.; Miro, B.; Kohli, A.; Nick, P. Exploring jasmonates in the hormonal network of drought
and salinity responses. Front. Plant Sci. 2015, 6, 1077. [CrossRef] [PubMed]
43. Fang, Y.; Xiong, L. General mechanisms of drought response and their application in drought resistance improvement in plants.
Cell. Mol. Life Sci. 2015, 72, 673–689. [CrossRef]
44. Zhang, X.Y.; Liu, C.L.; Wang, J.J.; Li, F.G.; Ye, W.E. Evaluation to the drought tolerance of cotton by PEG water-stress. J. Cotton Sci.
2007, 19, 205–209. [CrossRef]
45. Li, H.M.; Liu, S.D.; Ge, C.W.; Zhang, X.M.; Zhang, S.P.; Chen, J.; Shen, Q.; Ju, F.Y.; Yang, Y.F.; Li, Y.; et al. Association Analysis of
Drought Tolerance and Associated Traits in Upland Cotton at the Seedling. Int. J. Mol. Sci. 2019, 20, 3888. [CrossRef]
46. Ullah, A.; Sun, H.; Yang, X.; Zhang, X. Drought coping strategies in cotton: Increased crop per drop. Plant Biotechnol. J. 2017,
15, 271–284. [CrossRef]
47. Javaid, A.; Awan, F.S.; Azhar, F.M.; Khan, I.A. Assessment of allelic diversity among drought-resistant cotton genotypes using
microsatellite traits. Genet. Mol Res. 2017, 16, 28549206. [CrossRef]
48. Ulloa, M.; De Santiago, L.M.; Hulse-Kemp, A.M.; Stelly, D.M.; Burke, J.J. Enhancing Upland cotton for drought resilience,
productivity, and fiber quality: Comparative evaluation and genetic dissection. Mol. Genet. Genom. 2020, 295, 155–176. [CrossRef]
[PubMed]
49. Uniyal, R.C.; Nautiyal, A.R. Seed germination and seedling extension growth in Ougeinia dalbergioides Benth. underwater and
salinity stress. New For. 1998, 16, 265–272. [CrossRef]
Life 2023, 13, 2067 17 of 19
50. Basal, H.; Smith, C.W.; Thaxton, P.S.; Hemphill, J.K. Seedling drought tolerance in upland cotton. Crop Sci. 2005, 45, 766–771.
[CrossRef]
51. JMP®. Version <17>. SAS Institute Inc.: Cary, NC, USA 1989–2021. Available online: https://www.jmp.com/en_us/home.html
(accessed on 1 March 2022).
52. Minitab, LLC. Minitab. 2021. Available online: https://www.minitab.com (accessed on 5 September 2022).
53. Yi, M. A Complete Guide to Scatter Plots. 2022. Available online: https://chartio.com/learn/charts/what-is-a-scatter-plot/
(accessed on 11 July 2022).
54. Fathi-Sadabadi, M.; Ranjbar, G.; Zangi, M.R.; Kazemitabar, S.K.; Najafi-Zarrini, H. Assessment of Salinity Tolerance in the Selected
Genotypes of Cotton. Agrotech. Ind. Crops 2020, 2, 65–73. [CrossRef]
55. Mishra, N.; Sun, L.; Zhu, X.; Smith, J.; Prakash Srivastava, A.; Yang, X.; Pehlivan, N.; Esmaeili, N.; Luo, H.; Shen, G.; et al.
Overexpression of the Rice SUMO E3 Ligase Gene OsSIZ1 in Cotton Enhances Drought and Heat Tolerance, and Substantially
Improves Fiber Yields in the Field under Reduced Irrigation and Rainfed Conditions. Plant Cell Physiol. 2017, 58, 735–746.
[CrossRef] [PubMed]
56. Jolliffe, I.T. Principal Component Analysis; Wiley Online Library: Barcelona, Spain, 2002.
57. Gupta, A.; Rico-Medina, A.; Caño-Delgado, A.I. The physiology of plant responses to drought. Science 2020, 368, 266–269.
[CrossRef]
58. Yamaguchi, M.; Sharp, R.E. Complexity and coordination of root growth at low water potentials: Recent advances from
transcriptomic and proteomic analyses. Plant Cell Environ. 2010, 33, 590–603. [CrossRef] [PubMed]
59. Tian, H.; De Smet, I.; Ding, Z. Shaping a root system: Regulating lateral versus primary root growth. Trends Plant Sci. 2014,
19, 426–431. [CrossRef]
60. Sun, C.H.; Yu, J.Q.; Hu, D.G. Nitrate: A crucial signal during lateral root development. Front. Plant Sci. 2017, 8, 485. [CrossRef]
61. Paez-Garcia, A.; Motes, C.M.; Scheible, W.R.; Chen, R.; Blancaflor, E.B.; Monteros, M.J. Root traits and phenotyping strategies for
plant improvement. Plants 2015, 4, 334–355. [CrossRef]
62. Nguyen, T.T.T.; Klueva, N.; Chamareck, V.; Aarti, A.; Magpantay, G.; Millena, A.C.M.; Pathan, M.S. Saturation mapping of
QTL regions and identification of putative candidate genes for drought tolerance in rice. Mol. Genet. Genom. 2004, 272, 35–46.
[CrossRef] [PubMed]
63. Mvula, J.; Bokosi, J.M.; Kabambe, V.; Banda, M.H.P. Screening cotton (Gossypium hirsutum L.) genotypes for drought tolerance
under screen house conditions in Malawi. J. Plant Breed. Crop. Sci. 2018, 10, 48–57.
64. Pawar, K.N.; Veena, V.B. Evaluation of Cotton Genotypes for Drought Tolerance using PEG6000 Water Stress by Slanting Glass
Plate Technique. Int. J. Curr. Microbiol. App. Sci. 2020, 9, 32033212. [CrossRef]
65. Babu, A.G.; Patil, B.C.; Pawar, K.N. Evaluation of cotton genotypes for drought tolerance using PEG-6000 water stress by slanting
glass plate technique. Bioscan 2014, 9, 1419–1424.
66. Zahid, Z.; Khan, M.K.R.; Hameed, A.; Akhtar, M.; Ditta, A.; Hassan, H.M.; Farid, G. Dissection of drought tolerance in upland
cotton through morpho-physiological and biochemical traits at seedling stage. Front. Plant Sci. 2021, 12, 627107. [CrossRef]
67. Wang, L.; Wang, P.C.; Zhang, T.; Zhang, H.Y.; Ding, S.Y. Effect of short-term drought and rewatering during the pod-setting stage
on leaf photosynthesis and yield of the soybean. Acta Ecol. Sin. 2009, 29, 3328–3334.
68. Shan, L.; Zhang, S.Q. Is possible to save large irrigation water? The situation and prospect of water-saving agriculture in China.
Chin. J. Nat. 2006, 28, 71–74. [CrossRef]
69. Proffitt, A.P.B.; Berliner, P.R.; Oosterhuis, D.M. A Comparative Study of Root Distribution and Water Extraction Efficiency by
Wheat Grown Under High-and Low-Frequency Irrigation 1. J. Agron. 1985, 77, 655–662. [CrossRef]
70. Istiqomah, N.; Fidiyawati, E.; Prayitno, A. Total root dry weight per soil volume in maize by application of organic fertilizer
reducing the use of N, P and K fertilizer on terraced land. In IOP Conference Series: Earth and Environmental Science; IOP Publishing:
Bogor, Indonesia, 2021; Volume 648, p. 012189.
71. Iqbal, A.; Dong, Q.; Wang, X.; Gui, H.; Zhang, H.; Zhang, X.; Song, M. High Nitrogen Enhance Drought Tolerance in Cotton
through Antioxidant Enzymatic Activities, Nitrogen Metabolism and Osmotic Adjustment. Plants 2020, 9, 178. [CrossRef]
72. Jaafar, K.S.; Mohammed, M.A.; Mohammed, S.M. Screening for drought tolerance in cotton (Gossypium hirsutum L.) using in vitro
technique. J. Dryland Agric. 2021, 7, 52–59.
73. Shah, A.R.; Khan, T.M.; Sadaqat, H.A.; Chatha, A.A. Alterations in Leaf Pigments in Cotton (Gossypium hirsutum) Genotypes
Subjected to Drought Stress Conditions. Int. J. Agric. Biol. 2011, 13, 902–908.
74. Akbar, M.; Hussain, S.B.H. Assessment of drought tolerant cotton genotypes based on seedling & physiological attributes at
different moisture levels. Pure Appl. Biol. 2019, 8, 93–107.
75. Liu, F.; Stützel, H. Biomass partitioning, specific leaf area, and water use efficiency of vegetable amaranth (Amaranthus spp.) in
response to drought stress. Sci. Hortic. 2004, 102, 15–27. [CrossRef]
76. Wang, Y.; Meng, Z.; Liang, C.; Meng, Z.; Wang, Y.; Sun, G.; Zhu, T.; Cai, Y.; Guo, S.; Zhang, R.; et al. Increased lateral root
formation by CRISPR/Cas9-mediated editing of arginase genes in cotton. Sci. China Life Sci. 2017, 60, 524–527. [CrossRef]
[PubMed]
77. Shkolnik-Inbar, D.; Bar-Zvi, D. ABI4 mediates abscisic acid and cytokinin inhibition of lateral root formation by reducing polar
auxin transport in Arabidopsis. Plant Cell 2010, 22, 3560–3573. [CrossRef] [PubMed]
Life 2023, 13, 2067 18 of 19
78. Nishiyama, R.; Watanabe, Y.; Fujita, Y.; Le, D.T.; Kojima, M.; Werner, T.; Vankova, R.; Yamaguchi-Shinozaki, K.; Shinozaki, K.;
Kakimoto, T.; et al. Analysis of cytokinin mutants and regulation of cytokinin metabolic genes reveals important regulatory roles
of cytokinins in drought, salt and abscisic acid responses, and abscisic acid biosynthesis. Plant Cell 2011, 23, 2169–2183. [CrossRef]
[PubMed]
79. Benková, E. Local, efflux-dependent auxin gradients as a common module for plant organ formation. Cell 2003, 115, 591–602.
[CrossRef] [PubMed]
80. De Smet, I.; Vanneste, S.; Inzé, D.; Beeckman, T. LRI or the birth of a new meristem. Plant Mol. Biol. 2006, 60, 871–887. [CrossRef]
81. McMichael, B.L.; Quisenberry, J.E.; Upchruch, D.R.; Upchruch, D.R. Lateral root development in exotic cottons. Environ. Exp. Bot.
1987, 27, 499–502. [CrossRef]
82. Varney, G.T.; Canny, M.J. Rates of water-uptake into the mature root-system of maize plants. New Phytol. 1993, 123, 775–786.
[CrossRef]
83. Lambers, H.; Shane, M.W.; Cramer, M.D.; Pearse, S.J.; Veneklaas, E.J. Root structure and functioning for efficient acquisition of
phosphorus: Matching morphological and physiological traits. Ann. Bot. 2006, 98, 693–713. [CrossRef] [PubMed]
84. Hund, A.; Richner, W.; Soldati, A.; Fracheboud, Y.; Stamp, P. Root morphology and photosynthetic performance of maize inbred
lines at low temperature. Eur. J. Agron. 2007, 27, 52–61. [CrossRef]
85. Hund, A.; Fracheboud, Y.; Soldati, A.; Stamp, P. Cold tolerance of maize seedlings as determined by root morphology and
photosynthetic traits. Eur. J. Agron. 2008, 28, 178–185. [CrossRef]
86. Gallardo, M.; Turner, N.C.; Ludwig, C. Water relations, gas exchange and abscisic acid content of Lupinus cosentinii leaves in
response to drying different proportions of the root system. J. Exp. Bot. 1994, 45, 909–918. [CrossRef]
87. Luo, H.; Zhang, H.; Han, H.; Hu, Y.; Zhang, Y.; Zhang, W. Effects of water storage in deeper soil layers on growth, yield, and
water productivity of cotton (Gossypium hirsutum L.) in arid areas of northwestern. Irrig. Drain. 2014, 63, 59–70. [CrossRef]
88. Barber, S.A.; Silberbush, M. Plant root morphology and nutrient uptake. Roots Nutr. Water Influx Plant Growth 1984, 49, 65–87.
89. Russel, R.S. Plant Root Systems, Their Fimction, and İnteraction with the Soil; McGraw-Hill Books: London, UK, 1977.
90. Forde, B.; Lorenzo, H. The nutritional control of root development. Plant Soil 2001, 232, 51–68. [CrossRef]
91. Iqbal, K.; Azhar, F.M.; Khan, I.A.; Ullah, E. Assessment of cotton (Gossypium hirsutum) germplasm under water stress condition.
Int. J. Agric. Biol. 2010, 12, 251–255.
92. Chaturvedi, G.S.; Anuradha, S.; Bahadur, R. Screening techniques for evaluating crop germplasm for drought tolerance. Plant
Arch. 2012, 12, 11–18.
93. Simonneau, T.; Habib, R.; Goutouly, J.P.; Huguet, J.G. Diurnal changes in stem diameter depend upon variations in water content:
Direct evidence in peach trees. J. Exp. Bot. 1993, 44, 615–621. [CrossRef]
94. Lund, M.; Elsayed, H. Rapid assessment of water severe stress tolerant cotton (Gossypium hirsutum L.) genotypes based on
seedling and morph-physiological traits at two moisture levels. Ann. Plant. Sci. 2019, 8, 3651–3659.
95. Iftikhar, M.S.; Talha, G.M.; Shahzad, R.; Jameel, S.; Aleem, M.; Iqbal, M.Z. Early response of cotton (Gossypium hirsutum L.)
genotype against drought stress. Int. J. Biosci. 2019, 14, 537–544.
96. Ahmad, A.; Aslam, Z.; Bellitürk, K.; Ibrahim, M.U.; Nadeem, M.; Yilmaz, K.; Ali, U.; Akbar, G. Morphological study of cotton
(Gossypium hirsutum L.) varieties against drought tolerance under greenhouse conditions. Pure Appl. Biol. 2020, 9, 696–703.
Disclaimer/Publisher’s Note: The statements, opinions and data contained in all publications are solely those of the individual
author(s) and contributor(s) and not of MDPI and/or the editor(s). MDPI and/or the editor(s) disclaim responsibility for any injury to
people or property resulting from any ideas, methods, instructions or products referred to in the content.