Enhancing cotton tolerance to cold and drought stress: A protective role of SikCOR413PM1 gene overexpression

doi:10.21203/rs.3.rs-2950312/v1

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Enhancing cotton tolerance to cold and drought stress: A protective role of SikCOR413PM1 gene overexpression

https://doi.org/10.21203/rs.3.rs-2950312/v1

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published 18 Nov, 2023

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This study explores the potential role of the SikCOR413PM1 gene, isolated from Saussurea involucrata (Kar Kir.), in enhancing cotton (Gossypium hirsutum) tolerance to cold and drought stress through transgenic methods. We found that overexpressing SikCOR413PM1 in cotton plants led to increased relative water content, reduced malondialdehyde content and relative conductivity, and elevated proline and soluble sugar levels as osmotic regulators. These findings suggest that SikCOR413PM1 can minimize cell membrane damage and boost plant stability under challenging conditions. Additionally, overexpression of this gene upregulated antioxidant enzyme-related genes in transgenic cotton plants, resulting in enhanced enzyme activity, lowered peroxide content, and reduced oxidative stress. SikCOR413PM1 overexpression also modulated the expression of stress-related genes (GhDREB1A, GhDREB1B, GhDREB1C, GhERF2, GhNAC3, and GhRD22). In field trials, transgenic cotton plants with SikCOR413PM1 overexpression displayed higher yields and increased environmental resistance. Our study concludes that SikCOR413PM1 can regulate stress-related genes, osmotic adjustment factors, and peroxide content while preserving cell membrane stability and improving cotton tolerance to low-temperature and drought conditions.

During their growth and development, plants are most commonly affected by abiotic stress factors such as cold, drought, and salt stress [1]. Previous studies have found that exposure of plants to a certain degree of abiotic stress in advance can improve their ability to better adapt to subsequent stress [2, 3].

To cope with environmental changes, plants have evolved various regulatory mechanisms under abiotic stress, including osmotic and oxidative stress, for self-protection. Stress can lead to increases in reactive oxygen species (ROS) in plants [4, 5, 6]. However, excessive ROS accumulation can cause oxidative damage to cells, inhibiting plant physiological processes [7, 8]. Under normal growth conditions, plants generally maintain a relatively low level of ROS, and ROS scavenging and production are usually balanced [9].

The main ROS components are O₂⁻ and H₂O₂, which are important signal transduction molecules mediating multiple stress tolerance [10]. H₂O₂ triggers the regulation of the plant antioxidant system through ABA and BRs to achieve cold stress regulation and improve cold tolerance [11, 12, 13, 14]. Numerous studies have shown that the antioxidant mechanism of inducing ROS scavenging is essential in resisting various stresses [15, 16, 17].

Plant responses to cold stress involve multiple levels of regulation, including metabolic changes such as the accumulation of sugars and amino acids and the C-receptor binding factor (CBF)/dehydration response element binding factor (DREB) pathway [18, 19]. CBF proteins can bind to C-receptor (CRT) cis-elements and activate the transcription of downstream cold-regulated (COR) genes to improve cold tolerance [12, 20, 21, 22].

The CBF transcription factor acts as a positive regulator of cold signals. The CBF-encoded domain binds to the conserved C-repeat/dehydration response element (CRT/DRE) motif on the COR gene promoter to activate the transcription and expression of the COR gene [12, 23, 24, 25]. Numerous studies have demonstrated that the expression of the COR gene is positively correlated with the cold resistance of plants [26, 27, 28, 29].

The COR413 family encodes plant-specific multi-transmembrane proteins [30]. Based on predicted intracellular localization, the COR413 protein family can be divided into two subclasses: chloroplast COR413-TM/IM (thylakoid membrane/intima) and plasma membrane COR413-PM [30]. Some studies have shown that COR413 family genes can improve plant stress resistance. For example, in Arabidopsis, AtCOR413IM can stabilize the chloroplast membrane under low-temperature stress [31, 32], while overexpression of COR413-TM1 in rice can enhance drought tolerance [33]. The gene encoding tomato chloroplast COR413 (SlCOR413IM1) also improves the drought tolerance of tobacco plants [34].

COR413-PM, located in the plasma membrane, can also improve plant stress resistance. Overexpression of SikCOR413PM1 isolated from Saussurea involucrata (Kar Kir.)enhanced the tolerance of transgenic tobacco to cold and drought stress [35]. Zhou et al. (2018) [36] found that overexpression of PsCOR413PM2 increased Arabidopsis tolerance to osmotic stress and improved its cold tolerance. Similarly, Zhang et al. (2021) [37] found that LeCOR413PM2 improved the cold tolerance of transgenic tomato [37]. Overexpression of the plasma membrane COR413PM gene isolated from drought-resistant plants in Arabidopsis thaliana can enhance tolerance to drought and low temperatures.

Cotton is one of the most important agricultural products in the world, and has been cultivated in over 80 countries according to statistics [38]. It is mainly grown in arid regions such as tropical and subtropical areas, but has poor tolerance to drought and cold stress [39, 40]. The cotton industry has been severely affected by drought, leading to a rapid decline in fiber production [41]. Furthermore, a combination of two or more abiotic stresses can cause much greater yield loss than one single stress [42]. Therefore, developing crops with tolerance to multiple stresses has significant practical significance for agricultural production. Many scientists have studied stress-related genes and used transgenic technology to improve plant stress resistance [43].

In our previous study, we have isolated the SikCOR413PM1 gene from Saussurea involucrata (Kar Kir.), whose overexpression was found to improve the stress resistance of transgenic tobacco [35]. This study further explored the role of SikCOR413PM1 gene overexpression in cotton. The results showed that SikCOR413PM1 cotton exhibited stronger stress tolerance compared to the wild-type in both greenhouse and field trials. The findings provide a theoretical and experimental basis for future research on this gene and the application of new cotton varieties.

Acquisition and identification of transgenic cotton plants with the SikCOR431PM1 gene

To investigate the influence of the SikCOR431PM1 gene on the cold and drought tolerance of cotton, we integrated the SikCOR431PM1 gene into cotton using the pollen tube channel method to obtain SikCOR431PM1 transgenic cotton. Ten transgenic cotton lines were screened and analyzed by qRT-PCR (Fig. 1A). The relative expression levels of OE-4, OE-5, and OE-7 were found to be higher and were further analyzed by RT-PCR. The results were consistent with those of qRT-PCR(Fig. 1B). Therefore, OE-4, OE-5, and OE-7 were selected to conduct further gene function research.

Growth of cotton seedlings overexpressing SikCOR413PM1 gene under low-temperature and drought stress conditions

Cotton seedlings were exposed to low temperature and 20% PEG6000 stress. Under normal conditions (25°C), the WT and transgenic lines grew well without treatment. However, after exposure to 4°C, the leaves of WT cotton seedlings showed severe wilting, and most of them became necrotic and damaged due to the low temperature. In contrast, the leaves of transgenic lines showed much less severe wilting and still had certain vitality (Fig. 2A). After treatment with 20% PEG6000, the leaves of some WT lines exhibited noticeable wilting and curling, while those of transgenic cotton showed only weak wilting. This pattern persisted for 15 days, during which time the WT lines exhibited more significant wilting and withering than the transgenic lines. After 20 days of treatment, some WT lines exhibited even more severe growth retardation and leaf wilting, whereas the transgenic lines showed less severe leaf wilting (Fig. 2B).

To further clarify whether overexpression of the SikCOR413PM1 gene can improve cell protection mediated by low-temperature and drought stress in cotton seedlings, we measured the survival rates of WT and transgenic lines (OE-4, OE-5, and OE-7) under low-temperature and 20% PEG6000 stress to evaluate their survival under these conditions. Under normal conditions, the survival rate of each cotton line was 100%. However, after exposure to 4°C, the survival rates of the WT and transgenic lines decreased to 23.3% (WT), 65.6% (OE-4), 73.3% (OE-5), and 80% (OE-7), respectively. After treatment with 20% PEG6000, the survival rates of the WT and transgenic lines were reduced to 33.3% (WT), 61.1% (OE-4), 53.3% (OE-5), and 68.9% (OE-7), respectively. Compared to the WT, the transgenic lines exhibited higher viability (Fig. 2C).

Under normal conditions, there was no significant difference in average fresh weight and dry weight between the WT and transgenic lines. However, after exposure to low temperatures, the wet weight of transgenic lines OE-4, OE-5, and OE-7 was 23.6%, 36.6%, and 41.3% higher than that of the WT, respectively (Fig. 2D), while the dry weight was 23.7%, 24.1%, and 28.1% higher (Fig. 2E). Under 20% PEG6000 stress, the fresh weight and dry weight were lower than in the untreated control group. Nevertheless, the OE-4, OE-5, and OE-7 lines exhibited 56.0%, 50.9%, and 72.5% higher fresh weight than the WT, and 56.5%, 44.7%, and 63.5% higher dry weight than the WT. These results demonstrate that overexpression of SikCOR413PM1 enhances the tolerance of transgenic cotton to low temperatures and drought at the seedling stage.

Overexpression of the SikCOR413PM1 gene enhanced the cold and drought tolerance of cotton seedlings.

The WT and transgenic cotton plants grown for 70 days exhibited robust growth at room temperature (25°C) without treatment. However, after 24 h of exposure to 4°C, the leaves at the top of the WT plants showed wilting, whereas there was no significant change in the transgenic plants. After 48 h of exposure to 4°C, the WT plants exhibited severe wilting at the top growth point, while the transgenic plants showed only slight leaf wilting and drooping (Fig. 3A). After 14 days of drought treatment, the WT plants showed obvious signs of wilting, and the leaves near the base became loose and yellow. In contrast, the leaves of transgenic cotton showed less server wilting. After 21 days of drought treatment, compared to the transgenic plants, the WT plants exhibited more severe growth retardation and leaf wilting and shedding, while the leaves of transgenic plants did not fall off, and the stems of all transgenic plants remained full throughout the treatment process (Fig. 3B).

Under normal conditions, there was no significant difference in average fresh weight and dry weight between the WT and transgenic plants. However, after exposure to low temperatures, the wet weights of transgenic plants OE-4, OE-5, and OE-7 were 19.4%, 24.2%, and 25.5% heavier than those of WT plants (Fig. 3C), and dry weights were 28.3%, 36.9%, and 41.1% heavier (Fig. 3D). Under drought stress, the fresh weight and dry weight were lower than in the untreated control group. Nevertheless, the fresh weight of transgenic plants OE-4, OE-5, and OE-7 was 30.1%, 37.2%, and 42.6% heavier than that of the WT, and the dry weight was 40.6%, 50.6%, and 60.4% heavier than that of the WT. These findings demonstrate that overexpression of the SikCOR413PM1 gene enhances the cold and drought resistance of transgenic cotton seedlings.

Figure 3 Phenotype analysis of WT and SikCOR413PM1 -expressing cotton plants under cold and drought stress.(A) Analysis of phenotype 70-d-old WT and transgenic plants OE-4, OE-5, and OE-7 under 4°C for 24 h and 48 h, respectively; (B) Analysis of phenotype 70-d-old WT and transgenic plants OE-4, OE-5, and OE-7 after drought treatment 14 and 21 days, respectively; (C) Fresh weight ; (D) Dry weight. The data represent the average of three independent biological replicates. Bars represent SDs. *P < 0.05 and **P < 0.01 indicate significant differences relative to WT cotton plants.

Overexpression of SikCOR413PM1 alleviated cell membrane damage and accumulated more osmoregulatory substances.

To further elucidate the possible physiological mechanism through which SikCOR413PM1 overexpression improves cell protection under low-temperature and drought stress in cotton, we assessed the malondialdehyde (MDA) content and relative conductivity of WT and transgenic plants (OE-4, OE-5, and OE-7) to evaluate their physiological response to low-temperature and drought stress conditions. Compared to that of the WT plants, the MDA content in the leaves of transgenic plants was significantly reduced (Fig. 4A). Under 4°C stress treatment, the MDA content of transgenic cotton was 31.7% (OE-4), 31.6% (OE-5), and 36.9% (OE-7) lower than that of WT cotton, respectively. Under drought treatment, the MDA content of transgenic cotton was 26.3% (OE-4), 27.5% (OE-5), and 35.8% (OE-7) lower than that of WT cotton, respectively (Fig. 4B). There was no significant difference in relative conductivity between transgenic lines and WT under normal conditions. However, after exposure to 4°C, the relative electrolyte leakage (REL) of the WT and transgenic plants increased by 49.9% (WT), 36.3% (OE-4), 33.9% (OE-5), and 39.4% (OE-7), respectively. After drought treatment, the REL of the WT and transgenic plants increased by 46.6% (WT), 33.2% (OE-4), 29.5% (OE-5), and 36.8% (OE-7), respectively.

Figures 4C and D(Fig. 4) show that before stress, the contents of proline and soluble sugar were low in both WT and SikCOR413PM1 transgenic plants, and there was no significant difference between them. However, the contents of these two osmolytes significantly increased after exposure to low temperature and drought stress. Under 4°C stress, the proline and soluble sugar contents of transgenic lines were 43.1% and 32.0% (OE-4), 40.8% and 38.6% (OE-5), and 55.1% and 42.9% (OE-7) higher than those of WT lines, respectively. After drought treatment, the proline and soluble sugar contents of transgenic lines were 41.8% and 36.9% (OE-4), 42.7% and 45.8% (OE-5), and 47.2% and 47.9% (OE-7) higher than those of the WT, respectively. In summary, overexpression of the SikCOR413PM1 gene in cotton can maintain the integrity of the cell membrane and promote the accumulation of osmotic protective agents under stress conditions.

Overexpression of the SikCOR413PM1 gene maintained high antioxidant enzyme activity and expression analysis of related genes.

In this study, we detected the contents of O²⁻ and H₂O₂, which can reflect the response of plants to stress in both WT and transgenic lines. The results showed that there was no significant difference among all lines under normal conditions. However, after exposure to low temperature and drought stress, there were significant differences between these lines. Compared to those in the WT lines, the contents of O²⁻ and H₂O₂ in transgenic lines were significantly reduced (Fig. 5A and B).

We also measured the activity levels of the antioxidant enzymes SOD, POD, CAT, and GST. Under normal conditions, there was no significant difference between the transgenic lines and the WT. However, under low-temperature and drought stress conditions, each stress treatment enhanced the antioxidant enzyme activity of all plants, and the transgenic lines had higher antioxidant enzyme activities than the WT lines. Consistently, the expression levels of the GhSOD, GhPOD, GhCAT, and GhGST coding genes in the transgenic lines were also significantly higher than those in the WT lines after stress treatment (Fig. 5C-J). In summary, overexpression of the SikCOR413PM1 gene increased the expression of antioxidant enzyme genes and enhanced the activity of corresponding antioxidant enzymes, thereby regulating the amount of ROS to minimize the effect of oxidative stress.

Figure 5 Analysis of ROS and antioxidant enzyme activity and expression of related genes in WT and transgenic SikCOR413PM1 plants under cold and drought stress.(A) O₂⁻ contents; (B) H₂O₂ contents; (C) SOD activity; (D) Expression level of GhSOD; (E) POD activity; (F) Expression level of GhPOD; (G) CAT activity; (H) Expression level of GhCAT; (I) GST activity; (J) Expression level of GhGST. Data for the WT are means of three replicates, whereas data for SikCOR413PM1 transgenic plants are means of three different lines. Bars represent SDs. *P < 0.05 and **P < 0.01 indicate significant differences relative to WT cotton plants.

Analysis of stress-related gene expression in SikCOR413PM1 overexpressing plants

We analyzed the expression levels of several stress-related genes, including GhDREB1A, GhDREB1B, GhDREB1C, GhERF2, GhNAC3, and GhRD22. Under normal conditions, there was no significant difference in the expression levels of GhDREB1A, GhDREB1B, and GhDREB1C between the WT and transgenic lines. However, after low-temperature and drought stress treatment, the expression levels of these genes in transgenic lines were significantly higher than those in the WT lines. Under low-temperature stress, the expression of GhDREB1A, GhDREB1B, and GhDREB1C genes in transgenic lines was 16.7, 9.9, and 8.6 times (OE-4), 16.7, 9.5, and 9.1 times (OE-5), and 18.7, 13.1, and 10.2 times (OE-7) higher than those in the WT, respectively. Under drought stress, the expression of these genes in transgenic lines was 9.6, 17.7, and 5.5 times (OE-4), 9.1, 19.3, and 4.9 times (OE-5), and 11.0, 21.7, and 6.6 times (OE-7) higher than those in the WT, respectively (Fig. 6A-C). Under low-temperature stress, there was no significant difference in GhRD22 expression between transgenic and WT cotton. Under drought stress, the expression of GhRD22 was significantly upregulated and was 11.2-fold (OE-4), 13.1-fold (OE-5), and 15.1-fold (OE-7) higher than that of the WT, respectively (Fig. 6D). Additionally, the expression of the GhERF2 gene in WT and transgenic plants was significantly different under stress treatment. The expression of the GhERF2 gene in transgenic plants was 9.1 and 11.7 times (OE-4), 8.1 and 13.4 times (OE-5), and 13.8 and 15.9 times (OE-7) higher than that in the WT under low-temperature and drought stress, respectively (Fig. 6E). Under drought stress, there was no significant difference in the expression of the GhNAC3 gene between transgenic and WT cotton, but after cold stress treatment, the expression levels increased in each plant. The expression levels of transgenic lines OE-4, OE-5, and OE-7 were 10.7, 12.9, and 14.5 times higher than those of the WT (Fig. 6F). These results indicate that overexpression of SikCOR413PM1 helps increase the expression level of stress response genes and enhances the low-temperature and drought tolerance of cotton.

Agronomic traits of SikCOR413PM1 overexpressing cotton in the field

The plant height of transgenic cotton overexpressing SikCOR413PM1 increased slightly but not significantly, and the number of fruit branches per plant and bolls per plant increased compared with that of the WT (Table.S1). The SikCOR413PM1 transgenic lines had eight to nine fruit branches per plant, and the WT plants had only seven fruit branches. In addition, compared with WT plants, SikCOR413PM1 transgenic lines had an average of one or two bolls per plant. Although there was no significant difference in boll weight and lint weight between SikCOR413PM1 transgenic lines and WT plants, the fiber yield of SikCOR413PM1 transgenic cotton lines was moderately increased. The seed cotton yield and lint yield of SikCOR413PM1 cotton were significantly increased. This may result from multiple tolerance activities of SikCOR413PM1, which effectively reduce the yield loss caused by abiotic stresses such as low temperature and drought.

In addition to the greenhouse experiments for low-temperature and drought resistance, we also evaluated the drought resistance of cotton plants overexpressing SikCOR413PM1 in the field (Fig. 7). The single boll weight, single boll lint weight, and lint percentage did not significantly differ between OE-4 and OE-7 lines, but the plant height, fruit branch number per plant, boll number per plant, and seed cotton yield of transgenic OE-4 and OE-7 lines were significantly higher than those of the WT. Our results indicate that overexpression of SikCOR413PM1 can improve drought tolerance and yield of cotton under both normal and drought stress conditions in the field.

Plant materials and plant growth conditions

The cotton material (Gossypium hirsutum) used in this study was the seed of Xinjiang upland cotton Xinluzao 17, that was provided by Li Baocheng, the vice president of Xinjiang Academy of Agriculture and Reclamation Sciences. The tissue culture conditions for wild-type (WT) and transgenic cotton were a relative humidity of 60–70%, a photoperiod of 16h light and 8h dark cycle, and an average temperature of 25°C. The growth conditions for seedlings were in plastic pots containing a composite matrix of equal parts nutrient soil, vermiculite, and perlite (1:1:1), and a culture chamber with a light intensity of 500 µmol^− 2 s^− 1 at a temperature range of 22–28°C and relative humidity of 60–70%.

Real-time quantitative PCR for SikCOR413PM1 expression analysis

Total RNA was isolated using the RNAiso Plus kit (TaKaRa) and on-column DNase I treatment, following the product instructions. The first-strand cDNA was synthesized from the total RNA using oligo(dT) primers and PrimeScript® RTase (TaKaRa). The expression patterns of the SikCOR413PM1 gene and related stress response genes were determined by quantitative RT-PCR using the LightCycler 480 instrument (Roche Diagnostics, Switzerland) and SYBR® Premix Ex TaqTM (TaKaRa, China) following the supplier’s instructions and MIQE guidelines. Each reaction mixture (10 µL) contained 50 ng cDNA, 5 µL 2× SYBR® Green MasterMix reagent (Applied Biosystems), and 0.2 µM corresponding gene-specific primers (Table.S2). The qRT-PCR reaction program included pre-denaturation at 95°C for 60 s, denaturation at 95°C for 15 s, annealing at 61°C for 30 s, 40 cycles, extension at 72°C for 30 s, and pre-denaturation at 52°C for 40 s. The GhHis3 gene (AF024716) was used as the reference gene. Each analysis was repeated three times, and the relative gene expression was calculated using the ΔΔCt method [43].

Acquisition of transgenic cotton

In our previous studies, we have identified and sequenced the SikCOR413PM1 gene. Transgenic cotton plants containing the SikCOR413PM1 gene were obtained using the pollen tube pathway method, and the receptors without the exogenous gene were confirmed through molecular detection before the transformation. The receptor plants were transformed at the flowering stage until the cotton bolls opened. The seed cotton from the cotton bolls was harvested and labeled, and the seeds were separated into T0 seeds based on the treatment. The T0 seeds were planted in isolated and protected experimental fields during the second year. When the true leaves of the seedlings had expanded, kanamycin at a concentration of 2 g/L was used for screening. After the screening, the DNA of the plants to be tested was extracted for PCR detection, and the total RNA of the positive plants identified by PCR was extracted for qRT-PCR identification. The positive cotton plants screened and identified were routinely managed and grown to blossom and produce bolls. The transgenic cotton seeds were collected and sown on 1/2 MS medium containing kanamycin (80 µg/mL) for screening. The resistant seedlings were transferred to soil for growth, and the seeds were collected after maturation for continued screening up to the T4 generation as experimental materials.

Plant stress treatments

WT and SikCOR413PM1 overexpression transgenic cotton seedlings at 20 days old were subjected to 4°C treatment for 24, 48, and 72 h to observe the phenotype and were treated with 20% PEG6000 for 10, 15, and 20 days to observe the phenotype and to count the survival rate, wet weight, and dry weight. The 70-day-old WT and SikCOR413PM1 overexpression transgenic cotton seedlings were treated with low temperatures at 4°C for 24 and 48 h, respectively. Phenotypic changes were recorded, and cotton leaves in the same position were collected after 48 h of treatment. After 14 and 21 days of drought treatment, phenotypic observation was carried out, and samples were collected after 21 days to determine the fresh and dry weights. All collected samples were used for subsequent physiological index detection, enzyme activity detection, and related gene expression analysis.

Determination of physiological and biochemical indexes

The relative water content of leaves was measured using the weighing method [45]. The malondialdehyde content [46], relative conductivity [47], free proline, and soluble sugar [48] were determined using a spectrophotometer (UV-160A, Shimadzu Scientific Instrument, Japan).

Determination of antioxidant enzyme activity and active oxygen content

The activity of superoxide dismutase (SOD), peroxidase (POD), catalase (CAT), and glutathione s-transferase (GST) was determined after preparing the enzyme solution, following the instructions of the detection kit (Nanjing Jiancheng Bioengineering Institute). The quantitative determination of H₂O₂ and O₂⁻ was performed using the method of Benikhlef et al (2013)[49]. Each experimental plant was treated with at least three plants per treatment, and each index was repeated three times.

Investigation of agronomic traits

The plant height, root length, and stem circumference of the selected WT and transgenic plants were measured using a tape and electronic vernier caliper. The plant height was measured from the root neck to the growth point, and the stem diameter was measured at 1 cm above the first leaf. The fresh weight and dry weight were measured using an electronic weighing scale.

Investigation on agronomic traits of cotton under field drought stress

Independent T0 transgenic plants were grown until maturity in the plot. Inbred line seeds (T1) were harvested and sown in the experimental field. Based on the results of PCR detection of the kanamycin resistance gene, T2 seeds were screened from positive T1 lines and transgenic homozygous plants. The homozygous transgenic lines were selected for selfing to obtain the T3 generation. This selection continued until the T4 progeny before the agronomic traits of the transgenic homozygous lines were stable. The performance of T4 homozygous transgenic and WT cotton lines was evaluated during the normal growing season of cotton in both normal and drought-prone areas.

The field traits under normal environmental conditions were studied in the experimental field of the Agricultural Biotechnology Key Laboratory of Shihezi University in Shihezi City, Xinjiang. One month after germination, when the plants had matured, irrigation was stopped for drought stress testing. In all field traits, plant growth followed a randomized complete block design with three replicates. Each transgenic cotton was planted in a test plot of about 10.25 m², with 240 plants per plot, and repeated three times. The plant spacing was 12.5 cm, and the row spacing was 30 cm. From sowing to harvest, all recommended plant protection measures were taken. Fifty plants were randomly selected from each replicate for important agronomic traits, including plant height, fruit branch number per plant, boll number per plant, boll weight, lint weight per boll, seed cotton yield per plant, lint yield per plant, lint percentage, seed cotton yield, and lint yield per plot. The plant height was determined as the height of the main stem at the boll opening stage. The vegetative branches and fruit branches of the main stem were artificially separated, and the number of fruit branches per plant was measured. Bolls per plant were collected and counted. All naturally opened bolls were collected from a plot and dried in an oven at 37°C. One hundred bolls were randomly selected for weight measurement. To determine the actual yield of cotton, all bolls before and after frost were collected in a small area and treated according to the above method.

Statistical analysis

GraphPad Prism 7.0 and SPSS software (version 11.0; SPSS, Chicago, IL, USA) were used for statistical analysis. The pictures were drawn using CorelDRAW X4 SP2 drawing software. The data were expressed as the mean ± standard deviation of three repeated experiments. The expression level at 0 h (control time point) was defined as 1.0. Dunnett’s multiple comparison test was used to determine the difference between WT and transgenic lines. All transgenic overexpression lines were statistically compared with WT lines, in which *P < 0.05 represents a significant difference, and **P < 0.01 represents an extremely significant difference.

Plants commonly exhibit physiological adaptation to water shortage, accumulation of various osmotic regulators and antioxidants, and production of ROS in response to drought and cold stress [22, 50]. Abiotic stresses are known to cause water deficits in plants [22, 51, 52]. Under cold and drought conditions, plants maintain cell water by accumulating osmotic solutes, regulating osmotic potential, and adjusting osmotic pressure [53, 54]. Plants often initiate defense mechanisms by increasing the concentration of sugars, proline, amino acids, and other substances in cells, reducing cell osmotic potential, and absorbing water from external sources [55].

In this study, the content of soluble sugar and proline increased under cold and drought stress, particularly in transgenic lines (Fig. 4C and Fig. 4D). Generally, the stronger the osmotic adjustment ability, the higher the tolerance and adaptability to osmotic stress. This finding suggests that overexpression of the SikCOR413PM1 gene enhances the environmental adaptability of cotton by improving the osmotic adjustment ability through accumulation of osmotic adjustment substances, thus improving cold and drought tolerance.

Under cold stress, plants stabilize cell membranes to prevent freezing-induced dehydration [56]. Environmental changes can affect cell membrane function, and the plasma membrane is the primary site of freezing injury [57, 58, 59]. Some studies have shown that certain soluble sugars and proline can stabilize proteins and cell structures, enhance cell membrane integrity, and improve frost resistance [60, 61, 62, 63]. In cotton, soluble sugars and proline protect cell membranes from oxidative damage caused by various abiotic stresses [64]. Proline, the most common osmotic solute found in many organisms, prevents water loss, reduces mechanical damage, and enhances stress tolerance [61, 65, 66]. The overexpression of the SikCOR413PM1 gene in plants resulted in a greater accumulation of soluble sugars and proline than in WT plants in cold and drought environments (Fig. 4C-D). We speculate that this accumulation is related to improvement of cell membrane stability.

The cell membrane is a primary target of adverse stress, and maintaining its integrity and stability under abiotic stress conditions is a critical for plant environmental stress tolerance [67]. Under low-temperature stress, the structure and function of plant cell membranes change, and membrane lipid peroxidation occurs, leading to increases in ROS, cell membrane permeability, and relative conductivity [68, 69]. The content of the membrane lipid peroxidation product, malondialdehyde (MDA), can reflect the degree of membrane lipid peroxidation and cell damage [68]. Relative conductivity indicates plasma membrane damage caused by freezing [69]. Both can be used to evaluate the cold tolerance of plants.

In comparison to that of WT plants, the MDA content in the leaves of transgenic plants was significantly lower (Fig. 4A), but there was no significant difference in relative conductivity between transgenic and WT plants. Both MDA content and relative conductivity had different degrees of improvement after post-treatment at 4°C and drought treatment under normal conditions (Fig. 4B). Therefore, this result further shows that lower MDA content and higher relative conductivity contribute to the stress tolerance of cotton by reducing the degree of plasma membrane damage. The overexpression of the SikCOR413PM1 gene in cotton can reduce the degree of plant plasma membrane damage. Plants can enhance freezing tolerance (FT) by improving antioxidant mechanisms [52, 70]. Thus, the decrease in MDA content in SikCOR413PM1 transgenic cotton also indicates a reduction in membrane lipid peroxidation and improved stress resistance of plants.

Several studies have investigated whether COR genes can be induced in response to water shortage and cold stress. Most Arabidopsis COR genes are induced in response to water shortage [71, 72]. Chen et al. (2011)[73] speculated that the COR25 gene is also induced by low temperature and drought to protect Brassica napus cells from dehydration, given that overexpression of the SikCOR413PM1 gene maintains cell membrane stability. Therefore, we speculate that the SikCOR413PM1 gene may also be induced by drought and cold stress. In summary, this study demonstrates that the transgenic SikCOR413PM1 gene protects cell membranes and reduces damage by affecting various osmotic regulators, MDA, and relative conductivity in cotton, thereby regulating plant tolerance to cold and drought stress.

Currently, research on COR413 family genes is limited, but studies have shown that overexpression of the LeCOR413PM2 gene located in the plasma membrane may enhance the tolerance of transgenic tomato to cold stress by protecting the membrane from cold damage [37]. Previous studies have also shown that the overexpression of the SikCOR413PM1 gene, which is located on the plasma membrane, improves drought and cold stress in tobacco and cotton by protecting cell membrane stability [35]. It has been predicted that COR413 family proteins located in the plasma membrane (COR413-PM) have a film-forming function [71]. Therefore, we speculate that COR413 family proteins located in the plasma membrane may play a role in maintaining cell membrane stability, which requires further investigation.

Under water shortage and extreme temperature stress, plants may experience cell dehydration, damage to photosynthesis, and oxidative damage to membranes and other cellular components [22]. Abiotic stresses can interfere with photosynthetic electron transport and produce ROS, which can cause serious damage to chloroplasts [74, 75, 76, 53]. The primary source of ROS production is organelles with high oxidative activity, such as mitochondria, chloroplasts, and peroxisomes, which generate ROS such as O₂⁻ and H₂O₂, leading to oxidative damage of proteins, DNA, and lipids [6, 8, 75]. Therefore, regulating ROS can protect cells from oxidative damage [78, 79].

The ROS signal is an adaptive regulation of plants to the stressful environment [53]. Plants have established an efficient ROS scavenging system that reduces ROS-induced damage through antioxidant enzymes and antioxidants [80]. Cell-protective enzymes mainly include superoxide dismutase (SOD), catalase (CAT), ascorbate peroxidase (APX), and glutathione reductase. Overexpression of these genes often significantly improves the ability of plants to scavenge ROS and thus improves their tolerance to abiotic stress [81, 82, 83, 84].

In this study, the content of O₂⁻ and H₂O₂, which can reflect the response of plants to stress, was detected in WT and transgenic lines. Compared with that in WT lines, the content of O₂⁻ and H₂O₂ in transgenic lines was significantly reduced (Fig. 5A and B), indicating that overexpression of the SikCOR413PM1 gene enhances plant ability to scavenge ROS. Moreover, the enzyme activity level of ROS scavenger antioxidant enzymes showed that all stress treatments enhanced the activity of antioxidant enzymes, particularly in the transgenic lines. The expression levels of GhSOD, GhPOD, GhCAT, and GhGST coding genes in transgenic lines were also significantly higher than those in WT lines after stress treatment (Fig. 5C–J). Therefore, the enhancement of ROS scavenging ability is closely related to ROS scavenger antioxidant enzymes. The overexpression of the SikCOR413PM1 gene increases the expression level of genes corresponding to antioxidant enzymes in transgenic cotton, thereby enhancing the activity of corresponding antioxidant enzymes that can regulate the amount of ROS to reduce oxidative stress. Therefore, the SikCOR413PM1 gene maintains high antioxidant enzyme activity and related gene expression, reducing oxidative stress in plants.

Previous studies have shown that cold-induced changes in antioxidant enzyme activity in the leaves of several cereal varieties are related to frost resistance [85, 86, 87]. Overexpression of SOD in tobacco and APX in tomato increased their tolerance to low temperature [88, 89]. The antioxidant enzyme defense system is closely related to plant frost resistance [75, 80, 90]. Cold acclimation induces the accumulation of ROS, activating the antioxidant enzyme defense system [91]. By increasing the activities of enzymes that scavenge ROS, such as guaiacol peroxidase, superoxide dismutase (SOD), and catalase (CAT), the contents of H₂O₂ and MDA were reduced before the drought, and the homeostasis of ROS was better maintained [92]. In the study of Chen et al (2015)[93], down-regulation of GbMYb5 resulted in a higher oxidative stress rate under drought conditions due to decreases in the activities of POD, SOD, and CAT enzymes. This further illustrates that the SikCOR413PM1 gene can reduce the oxidative stress of plants and improve their cold and drought resistance.

Proline protects cells from increased stress-related ROS levels [53]. In a previous study, transgenic wheat accumulated higher proline than WT plants and showed less membrane lipid peroxidation during drought, indicating that proline plays a role in reducing ROS damage during drought [94]. Cold acclimation induces rapid proline accumulation by changing the activity of related enzymes in the proline metabolic pathway, thereby improving frost resistance [95]. In this study, proline was used as an osmotic regulator to improve the cold and drought tolerance of cotton. The proline content of transgenic plants under drought and cold stress was higher than that of the WT plants. Based on research and analysis, we speculate that overexpression of the SikCOR413PM1 gene may increase proline accumulation, and proline can reduce the impact of ROS on cells for protection.

DREB family transcription factors have been shown to play an important role in cold adaptation-induced FT, and are transcription factors that induce the expression of cold and dehydration stress-regulated genes in plants [96, 97, 98]. The expression of DREB genes is enhanced under cold and drought conditions [99]. DREB1A, DREB1B, and DREB1C expression is strongly induced by cold stress [98]. The expression of the DREB1 gene can partly explain the resistance of plants to stress.

In this study, the expression levels of GhDREB1A, GhDREB1B, and GhDREB1C in transgenic lines were significantly higher than those in WT lines after low-temperature and drought stress treatment (Fig. 6A–C). The expression levels of GhRD22, GhERF2, and GhNAC3 also increased under the two stresses (Fig. 6D–F). These results indicate that GhDREB1A, GhDREB1B, and GhDREB1C are strongly induced by drought and cold, and the expression of the above five stress-related genes was increased in transgenic SikCOR413PM1 cotton plants compared with in WT. Therefore, overexpression of SikCOR413PM1 helps increase the expression level of stress response genes. The increase in the expression of the transgenic cotton DREB1 gene indicates that it has stronger tolerance than the WT under cold and drought conditions. In Arabidopsis, the DREB1 protein activates COR genes, such as COR15a [100], COR6.6 [101], and COR47 [102], and its protein plays a functional role in enhancing tolerance to cold and drought stress. Therefore, it can be speculated that stress increases the expression of the DREB1 gene, which may be related to the activation of the COR413 gene. In summary, the overexpression of SikCOR413PM1 promotes the expression level of stress genes in drought and cold environments, thereby improving plant stress resistance.

Overexpression of the SikCOR413PM1 gene in cotton was found to have positive effects on cotton growth under greenhouse conditions. To investigate its actual impact on cotton production, the effect of this gene on transgenic cotton was studied in the field. The results showed that transgenic cotton had significant increases in both seed cotton yield and lint yield, making it more resistant to yield loss under stress. Although the growth status of the two lines did not differ significantly under normal conditions, transgenic OE-4 and OE-7 lines had significantly better growth under drought conditions than the WT (Fig. 7). Moreover, the number of fruit branches per plant, number of bolls per plant, and seed cotton yield were also significantly higher in transgenic cotton (Table.S1). These results suggest that overexpression of the SikCOR413PM1 gene can improve the drought tolerance and yield of cotton. The findings provide a theoretical foundation for developing cotton varieties and demonstrate the potential for transgenic cotton to improve environmental adaptability and yield.

ABA: Abscisic Acid

BRs: Epibrassinolide

CAT: Catalase

CBF: C-receptor binding factor

COR: Cold-regulated

CRT: C-receptor

DREB: Dehydration response element binding factor

GST: Glutathione S-transferase

POD: Peroxidase

qRT-PCR: Quantitative Real-Time PCR

REL: Relative electrical conductivity

ROS: Reactive oxygen species

SOD: Superoxide Dismutase

MDA: Malondialdehyde

Availability of data and materials

We submitted the gene sequence at NCBI, and since the gene is not publicly available, only the GenBank accession number of the nucleotide sequence provided by NCBI is available: BankIt2706231

Acknowledgements

We are grateful to researcher Li Baocheng, Vice President of Xinjiang Academy of Agricultural Reclamation, for providing seeds of Xinjiang Land Cotton XinLuzao 17. In addition we would like to thank the reviewers and editors for their careful reading and helpful comments on this paper.

Funding

This work was supported by the Science and Technology Research Project in Key Fields of Shihezi University (KY2023GG03), the Independent Research project of Shihezi University (ZZZC202082B), and the High-level Talents Research Start-up Fund of Science and Technology Research and Development Program of Shihezi University (RCZK202045).

Authors information

Authors and Affiliations

College of Life Science , Shihezi University, Shihezi 832000,China

Mei Wang, Lepeng Wang, Xiangxue Yu, Jingyi Zhao, Zhijia Tian and Xinyong Guo

Medical College, Shihezi University, Shihezi 832000, China.

Li Zhang

Xinjiang Agricultural Development Group Crop Hospital Co. LTD, Tumusuk 844000, China.

Xiaohong Liu

Agricultural Science Institute of the seventh division of Xinjiang Corps, Kuitun 833200, China

Guoping Wang

Contributions

Mei Wang and Lepeng Wang were the experimental designers and study executors of the study, completed the data analysis, and wrote the first draft of the manuscript. Xiangxue Yu and Jingyi Zhao carried out the experimental results and data analysis. Zhijia Tian, Xiaohong Liu, and Guoping Wang participated in the experimental data inspection and data investigation. Xinyong Guo and Li Zhang were the project planners and principals, guiding the paper's writing and revision. All authors read and agreed to the final text.

Corresponding author

Correspondence to Xinyong Guo

Ethics declarations

Ethics approval and consent to participate

The plant material Saussurea involucrata (Kar Kir.) used in this study was obtained from the Key Laboratory of Agricultural Biotechnology, College of Life Sciences, Shihezi University, Xinjiang, China. All the Saussurea involucrata (Kar Kir.) used in this experiment were laboratory histoculture seedlings and were not collected from endangered materials or species. All the procedures were followed in accordance with the relevant guidelines.

Consent for publication

Not applicable.

Competing interests

The authors declare that they have no competing interests.

Mboup M, Fischer I, Lainer H, Stephan W. Trans-species polymorphism and allele-specific expression in the CBF gene family of wild tomatoes. Mol Biol Evol. 2012;29(12):3641–52.
Walter J, Nagy L, Hein R, Rascher U, Beierkuhnlein C, Willner E, Jentsch A. Do plants remember drought? Hints towards a drought-memory in grasses. Environ Exp Bot. 2011;71:34–40.
Ramírez DA, Rolando JL, Yactayo W, Monneveux P, Mares V, Quiroz R. Improving potato drought tolerance through the induc-tion of long-term water stress memory. Plant Sci. 2015;238:26–32.
Shi Y, Ding Y, Yang S. Molecular regulation of CBF signaling in cold acclimation. Trends Plant Sci. 2018;23:623–37.
Yao D, Zhang X, Zhao X, Liu C, Wang C, Zhang Z, Zhang C, Wei Q, Wang Q, Yan H, Li F, Su Z. Transcriptome analysis reveals salt-stress-regulated biological processes and key pathways in roots of cotton (Gossypium hirsutum L). Genomics. 2011;98(1):47–55.
Sharma P, Jha AB, Dubey RS, Pessarakli M. Reactive oxygen species, oxidative damage, and antioxidative defense mechanism in plants under stressful conditions. J Bot.2012; 2012:217037.
Guler NS, Pehlivan N. Exogenous low-dose hydrogen peroxide enhances drought tolerance of soybean (Glycine max L.) through inducing antioxidant system. Acta Biol Hung. 2016;67(2):169–83.
Tripathy BC, Oelmüller R. Reactive oxygen species generation and signaling in plants. Plant Signal Behav. 2012;7(12):1621–33.
Hossain MA, Burritt DJ, Fujita M. Cross-stress tolerance in plants: molecular mechanisms and possible involvement of reactive oxygen species and methylglyoxal detoxification systems. In: Tuteja N, Gill SS, editors Abiotic stress response in plants. 2016;323–375.
Bhattacharjee S. Reactive oxygen species and oxidative burst: roles in stress, senescence and signal transduction in plants. Curr Sci. 2005;89:1113–21.
Baier M, Bittner A, Prescher A, Buer, Jvan. Preparing plants for improved cold tolerance by priming. Plant Cell Environ. 2019;42:782–800.
Chinnusamy V, Zhu J, Zhu JK. Cold stress regulation of gene expression in plants. Trends Plant Sci. 2007;12:444–51.
Lv B, Tian H, Zhang F, Liu J, Lu S, Bai M, Li C, Ding Z. Brassinosteroids regulate root growth by controlling reactive oxygen species homeostasis and dual effect on ethylene synthesis in Arabidopsis. PLoS Genet. 2018;14, e1007144.
Fang P, Yan M, Chi C, Wang M, Zhou Y, Zhou J, Shi K, Xia X, Foyer CH, Yu J. Brassinosteroids act as a positive regulator of photoprotection in response to chilling stress. Plant Physiol. 2019;180:2061–76.
Ning J, Li X, Hicks LM, Xiong L. A Raf-like MAPKKK gene DSM1 mediates drought resistance through reactive oxygen species scavenging in rice. Plant Physiol. 2010;152:876–90.
You J, Zong W, Hu H, Li X, Xiao J, Xiong L. A STRESS-RESPONSIVE NAC1-regulated protein phosphatase gene rice protein phosphatase18 modulates drought and oxidative stress tolerance through abscisic acid-independent reactive oxygen species scavenging in rice. Plant Physiol. 2014;166:2100–14.
Hazman M, Hause B, Eiche E, Nick P, Riemann M. Increased tolerance to salt stress in OPDA-deficient rice ALLENE OXIDE CYCLASE mutants is linked to an increased ROS-scavenging activity. J Exp Bot. 2015;66:3339–52.
Zhao C, Lang Z, Zhu J-K. Cold responsive gene transcription becomes more complex. Trends Plant Sci. 2015;20:466–8.
Ding Y, Shi Y, Yang S. Advances and challenges in uncovering cold tolerance regulatory mechanisms in plants. New Phytol. 2019;222:1690–704.
Winfield MO, Lu C, Wilson ID, Coghill JA, Edwards KJ. Plant responses to cold: Transcriptome analysis of wheat. Plant Biotechnol J. 2010;8(7):749–71.
Zhu J, Dong CH, Zhu JK. Interplay between cold-responsive gene regulation, metabolism and RNA processing during plant cold acclimation. Curr Opin Plant Biol. 2007;10(3):290–5.
Zhu JK. Abiotic stress signaling and responses in plants. Cell. 2016;167:313–24.
Liu J, Shi Y, Yang S. Insights into the regulation of C-repeat binding factors in plant cold signaling. J Integr Plant Biol. 2018;60(9):780–95.
Guo X, Liu D, Chong K. Cold signaling in plants: Insights into mechanisms and regulation. J Integr Plant Biol. 2018;60(9):745–56.
Lim CJ, Ali A, Park J, Shen M, Park KS, Baek D, Yun DJ. HOS15-PWR chromatin remodeling complex positively regulates cold stress in Arabidopsis. Plant Signal Behav. 2021;16(5):1893978.
Chen L, Zhong H, Ren F, Guo QQ, Hu XP, Li XB. A novel cold-regulated gene, COR25, of Brassica napus is involved in plant response and tolerance to cold stress. Plant Cell Rep. 2011;30(4):463–71.
Wathugala DL, Richards SA, Knight H, Knight MR. OsSFR6 is a functional rice orthologue of SENSITIVE TO FREEZING-6 and can act as a regulator of COR gene expression, osmotic stress and freezing tolerance in Arabidopsis. New Phytol. 2011;191(4):984–95.
Wan F, Pan Y, Li J, Chen X, Pan Y, Wang Y, Tian S, Zhang X. Heterologous expression of Arabidopsis C-repeat binding factor 3 (AtCBF3) and cold-regulated 15A (AtCOR15A) enhanced chilling tolerance in transgenic eggplant (Solanum melongena L). Plant Cell Rep. 2014;33(12):1951–61.
Yuan HM, Sheng Y, Chen WJ, Lu YQ, Tang X, Ou-Yang M, Huang X. Overexpression of Hevea brasiliensis HbICE1 Enhances Cold Tolerance in Arabidopsis. Front Plant Sci. 2017;8:1462.
Breton G, Danyluk J, Charron JB, Sarhan F. Expression profiling and bioinformatic analyses of a novel stress-regulated multispanning transmembrane protein family from cereals and Arabidopsis. Plant Physiol. 2003;132(1):64–74.
Okawa K, Nakayama K, Kakizaki T, Yamashita T, Inaba T. Identification and characterization of Cor413im proteins as novel components of the chloroplast inner envelope. Plant cell and environment. 2008;31(10):1470–83.
Okawa K, Inoue H, Adachi F, Nakayama K, Ito-Inaba Y, Schnell DJ, Uehara S, Inaba T. Targeting of a polytopic membrane protein to the inner envelope membrane of chloroplasts in vivo involves multiple transmembrane segments. J Exp Bot. 2014;65(18):5257–65.
Zhang C, Li C, Liu J, Lv Y, Yu C, Li H, Zhao T, Liu B. The OsABF1 transcription factor improves drought tolerance by activating the transcription of COR413-TM1 in rice. J Exp Bot. 2017;68(16):4695–707.
Ma X, Wang G, Zhao W, Yang M, Ma N, Kong F, et al. SlCOR413IM1, a novel cold regulation gene from tomato, enhances drought stress tolerance in tobacco. J Plant Physiol. 2017;216:88–99.
Guo X, Zhang L, Dong G, Xu Z, Li G, Liu N, Wang A, Zhu J. A novel cold-regulated protein isolated from Saussurea involucrata confers cold and drought tolerance in transgenic tobacco (Nicotiana tabacum). Plant science: an international journal of experimental plant biology. 2019;289:110246.
Zhou A, Liu E, Li H, Li Y, Feng S, Gong S, Wang J. PsCor413pm2, a Plasma Membrane-Localized, Cold-Regulated Protein from Phlox subulata, Confers Low Temperature Tolerance in Arabidopsis. Int J Mol Sci. 2018;19(9):2579.
Zhang L, Guo X, Zhang Z, Wang A, Zhu J. Cold-regulated gene LeCOR413PM2 confers cold stress tolerance in tomato plants. Gene. 2021;764:145097.
Lei W, Ben F. Analysis of the competitiveness of the cotton industry in Xinjiang based on the diamond model. UTMS J Econ. 2021;12:123–46.
Wendel JF, Grover CE. Taxonomy and evolution of the cotton genes, Gossypium. In: Fang DD, Percy RG, editors. Cotton. Volume 2. Madison: ASA, CSSA, and SSSA; 2015. pp. 25–44.
Penna JCV, Verhalen LM, Kirkham MB, McNew RW. Screening cotton genotypes for seedling drought tolerance. Genet Mol Biol. 1998;21:545–9.
Ullah A, Sun H, Yang X, Zhang X. Drought coping strategies in cotton: increased crop per drop. Plant Biotechnol J. 2017;15(3):271–84.
Mittler R. Abiotic stress, the field environment and stress combination. Trends Plant Sci. 2006;11(1):15–9.
Yamaguchi-Shinozaki K. Molecular mechanisms of abiotic stress responses: water and temperature stress conditions–overview. 2007; 52(6 Suppl):515–6.
Livak KJ, Schmittgen TD. Analysis of relative gene expression data using real-time quantitative PCR and the 2 (T) (-Delta Delta C) method. Methods. 2001;25:402–8.
Lara MV, Disante KB, Podesta FE, et al. Induction of a Crassulacean acid like metabolism in the C4 succulent plant, Portulaca oleracea L.: physiological ndmorphological changes are accompanied by specific modifications in phosphoenol pyruvate carboxylase. Photosyn Res. 2003;77:241–54.
Du ZY, Bramlage WJ. Modified thiobarbituric acid assay formeasuring lipid oxidation in sugar-rich plant tissue extracts. J Agric Food Chem. 1992;40:1566–70.
Lutts S, Kinet JM, Bouharmont J. NaCl-induced senescencein leaves of rice (Oryza sativa L.) cultivars differing in salinity resistance. Ann Bot. 1996;78:389–98.
Bates LS, Waldren RP, Teare ID. Rapid determination of free proline for water-stress studies. Plant Soil. 1973;1:205–7.
Benikhlef L, L’Haridon F, Aboumansour E, et al. Perception of soft mechanical stress in Arabidopsis leaves activates disease resistance. BMC Plant Biol. 2013;13:1–12.
Xiong L, Schumaker KS, Zhu JK. Cell signaling during cold, drought, and salt stress. Plant Cell. 2002;14(Suppl):165–83.
Steponkus PL. Role of the plasma membrane in freezing injury and cold acclimation. Annu Rev Plant Physiol. 1984;35:543–84.
Thomashow MF. Plant cold acclimation: freezing tolerance genes and regulatory mechanisms. Annu Rev Plant Physiol Plant Mol Biol. 1999;50:571–99.
Miller G, Suzuki N, Ciftci-Yilmaz S, Mittler R. Reactive oxygen species homeostasis and signaling during drought and salinity stresses. Plant Cell Environ. 2010;33:453–67.
Mitra J. Genetics and genetic improvement of drought resistance in crop plants.Curr. Sci (Bangalore). 2001;80:758–63.
Fang Y, Xiong L. General mechanisms of drought response and their application in drought resistance improvement in plants. Cell Mol Life Sci. 2015;72(4):673–89.
Steponkus PL, Webb MS. Freeze-induced dehydration and membrane destabilization in plants. In: Water and Life: Comparative Analysis of Water Relationships at the Organismic, Cellular and Molecular Level. 1992; 338–362.
Webb MS, Steponkus PL. Freeze-lnduced membrane ultrastructural alterations in rye (Sede cereale) leaves. Plant Physiol. 1993;101:955–63.
Uemura M, Joseph RA, Steponkus PL. Cold acclimation of Arabidopsis thaliana (effect on plasma membrane lipid composition and freeze-lnduced lesions). Plant Physiol. 1995;109:15–30.
Ouellet F. Cold acclimation and freezing tolerance in plants. Encycl Life Sci. 2007.
Guy CL. Cold acclimation and freezing stress tolerance: Role of protein metabolism. Annu Rev Plant Physiol Plant Mol Biol. 1990;41:187–223.
Ashraf M, Foolad MR. Roles of glycine betaine and proline in improving plant abiotic stress resistance. Environ Exp Bot. 2007;59:206–16.
Valliyodan B, Nguyen HT. Understanding regulatory networks and engineering for enhanced drought tolerance in plants. Curr Opin Plant Biol. 2006;9:189–95.
Janska´ A, Marsı´k P, Zelenkova´ S, Ovesna´ J. Cold stress and acclimation—what is important for metabolic adjustment? Plant Biol. 2010;12:395–405.
Khan MS, Ahmed D, Khan MA. Utilization of genes encoding osmoprotectants in transgenic plants for enhanced abiotic stress tolerance. Electron J Biotechnol. 2015;18:257–66.
Kovacs Z, Simon-Sarkadi L, Sovany C, Kirsch K, Galiba G, Kocsy G. Differential effects of cold acclimation and abscisic acid on free amino acid composition in wheat. Plant Sci. 2011;180:61–8.
Bartels D, Sunkar R. Drought and salt tolerance in plants.Critical Reviews in Plant Sciences. 2005; 24, 23–58.
Levitt J. Responses of Plants to Environmental Stresses: water, radiation, salt and other stresses. Volume II. New York: Academic Press; 1980. pp. 3–211.
Wang S, Liang D, Li C, Hao Y, Ma F, Shu H. Influence of drought stress on the cellular ultrastructure and antioxidant system in leaves of drought-tolerant and drought-sensitive apple rootstocks. Plant Physiol Biochem. 2012;51:81–9.
Jiang B, Shi Y, Zhang X, Xin X, Qi L, Guo H, Li J, Yang S. PIF3 is a negative regulator of the CBF pathway and freezing tolerance in Arabidopsis. Proc Natl Acad Sci USA. 2017;114(32):E6695–702.
Mahajan S, Tuteja N. Cold, salinity and drought stresses: an overview. Arch Biochem Biophys. 2005;444(2):139–58.
Ravindra KH. Molecular Cloning and Expression of cor (Cold-Regulated) Genes in Arabidopsis thaliana. Plant Physiol. 1990;93(3):1246–52.
Thomashow MF. Genes induced during cold acclimation in higher plants. Advances in low-temperature biology. 1993; 2, 183–210.
Chen Y, Jiang J, Chang Q, Gu C, Song A, Chen S, Dong B, Chen F. Cold acclimation induces freezing tolerance via antioxidative enzymes, proline metabolism and gene expression changes in two chrysanthemum species. Mol Biol Rep. 2014;41(2):815–22.
Niyogi KK. PHOTOPROTECTION REVISITED: Genetic and Molecular Approaches. Annual review of plant physiology and plant molecular biology. 1999; 50, 333–359.
Apel K, Hirt H. Reactive oxygen species: metabolism, oxidative stress, and signal transduction. Annu Rev Plant Biol. 2004;55:373–99.
Asada K. Production and scavenging of reactive oxygen species in chloroplasts and their functions. Plant Physiol. 2006;141:391–6.
Tripathy BC, Oelmüller R. Reactive oxygen species generation and signaling in plants. Plant Signal Behav. 2012;7:1621–33.
Fang Y, Liao K, Du H, Xu Y, Song H, Li X, Xiong L. A stress-responsive NAC transcription factor SNAC3 confers heat and drought tolerance through modulation of reactive oxygen species in rice. J Exp Bot. 2015;66(21):6803–17.
Nuruzzaman M, Sharoni AM, Kikuchi S. Roles of NAC transcription factors in the regulation of biotic and abiotic stress responses in plants. Front Microbiol. 2013;4:248.
Ahmad P, Jaleel CA, Salem MA, Nabi G, Sharma S. Roles of enzymatic and nonenzymatic antioxidants in plants during abiotic stress. Crit Rev Biotechnol. 2010;30(3):161–75.
Matsumura T, Tabayashi N, Kamagata Y, Souma C, Saruyama H. Wheat catalase expressed in transgenic rice can improve tolerance against low temperature stress. Physiol Plant. 2010;116(3):317–27.
Badawi GH, Kawano N, Yamauchi Y, Shimada E, Sasaki R, Kubo A, Tanaka K. Over-expression of ascorbate peroxidase in tobacco chloroplasts enhances the tolerance to salt stress and water deficit. Physiol Plant. 2004;121(2):231–8.
Eltayeb AE, Kawano N, Badawi GH, Kaminaka H, Sanekata T, Shibahara T, Inanaga S, Tanaka K. Overexpression of monodehydroascorbate reductase in transgenic tobacco confers enhanced tolerance to ozone, salt and polyethylene glycol stresses. Planta. 2007;225(5):1255–64.
Kim YH, Kim CY, Song WK, Park DS, Kwon SY, Lee HS, Bang JW, Kwak SS. Overexpression of sweetpotato swpa4 peroxidase results in increased hydrogen peroxide production and enhances stress tolerance in tobacco. Planta. 2008;227(4):867–81.
Dai F, Huang Y, Zhou M, Zhang G. The influence of cold acclimation on antioxidative enzymes and antioxidants in sensitive and tolerant barley cultivars. Biol Plant. 2009;3:257–62.
Janda T, Szalai G, Rios-Gonzalez K, Veisz O, Pa´ldi E. Comparative study of frost tolerance and antioxidant activity in cereals. Plant Sci. 2003;64:301–6.
Solte´sz A, Tı´ma´r I, Vashegyi I, To´th B, Kell}os T, Szalai G, Va´gu´jfalvi A, Kocsy G, Galiba G. Redox changes during cold acclimation affect freezing tolerance but not the vegetative/ reproductive transition of the shoot apex in wheat. Plant Biol. 2011;13:757–66.
Gupta AS, Heinen JL, Holaday AS, Burke JJ, Allen RD. Increased resistance to oxidative stress in transgenic plants that overexpress chloroplastic Cu/Zn superoxide dismutase. Proc Natl Acad Sci USA. 1993;90(4):1629–33.
Wang YJ, Wisniewski M, Meilan R, Cui MG, Webb R, Fuchigami L. Overexpression of cytosolic ascorbate peroxidase in tomato confers tolerance to chilling and salt stress. J Am SocHortic Sci. 2005;130:167–73.
Turan O, Ekmekci Y. Activities of photosystem II and antioxidant enzymes in chickpea (Cicer arietinum L.) cultivars exposed to chilling temperatures. Acta Physiol Plant. 2011;33:67–78.
Zhang Y, Luo Y, Hou YX, Jiang H, Chen Q, Tang HR. Chilling acclimation induced changes in the distribution of H2O2 and antioxidant system of strawberry leaves. Agric J. 2008;3:286–91.
Ben Abdallah M, Methenni K, Nouairi I, Zarrouk M, Youssef NB. Drought priming improves subsequent more severe drought in a drought-sensitive cultivar of olive cv. Chétoui Scientia horticulturae. 2017;221:43–52.
Chen T, Li W, Hu X, Guo J, Liu A, Zhang BA, Cotton MYB. Transcription Factor, GbMYB5, is Positively Involved in Plant Adaptive Response to Drought Stress. Plant Cell Physiol. 2015;56(5):917–29.
Vendruscolo EC, Schuster I, Pileggi M, Scapim CA, Molinari HB, Marur CJ, Vieira LG. Stress-induced synthesis of proline confers tolerance to water deficit in transgenic wheat. J Plant Physiol. 2007;164:1367–76.
Ruiz JM, Sanchez E, Garcia PC, Lopez-Lefebre LR, Rivero RM, Romero L. Proline metabolism and NAD kinase activity in greenbean plants subjected to cold-shock. Phytochemistry. 2002;59:473–8.
Gilmour SJ, Zarka DG, Stockinger EJ, Salazar MP, Houghton JM, Thomashow MF. Low temperature regulation of the Arabidopsis CBF family of AP2 transcriptional activators as an early step in cold-induced COR gene expression. The Plant journal: for cell and molecular biology. 1998;16(4):433–42.
Liu Q, Kasuga M, Sakuma Y, Abe H, Miura S, Yamaguchi-Shinozaki K, Shinozaki K. Two transcription factors, DREB1 and DREB2, with an EREBP/AP2 DNA binding domain separate two cellular signal transduction pathways in drought- and low-temperature-responsive gene expression, respectively, in Arabidopsis. Plant Cell. 1998;10(8):1391–406.
Shinwari ZK, Nakashima K, Miura S, Kasuga M, Seki M, Yamaguchi-Shinozaki K, Shinozaki K. An Arabidopsis gene family encoding DRE/CRT binding proteins involved in low-temperature-responsive gene expression. Biochem Biophys Res Commun. 1998;250(1):161–70.
Kidokoro S, Watanabe K, Ohori T, Moriwaki T, Maruyama K, Mizoi J, Myint Phyu Sin Htwe N, Fujita Y, Sekita S, Shinozaki K, Yamaguchi-Shinozaki K. Soybean DREB1/CBF-type transcription factors function in heat and drought as well as cold stress-responsive gene expression. The Plant journal: for cell and molecular biology. 2015;81(3):505–18.
Steponkus PL, Uemura M, Joseph RA, Gilmour SJ, Thomashow MF. Mode of action of the COR15a gene on the freezing tolerance of Arabidopsis thaliana. Proc Natl Acad Sci USA. 1998;95(24):14570–5.
Wang H, Datla R, Georges F, Loewen M, Cutler AJ. Promoters from kin1 and cor6.6, two homologous Arabidopsis thaliana genes: transcriptional regulation and gene expression induced by low temperature, ABA, osmoticum and dehydration. Plant Mol Biol. 1995;28(4):605–17.
Maruyama K, Sakuma Y, Kasuga M, Ito Y, Seki M, Goda H, Shimada Y, Yoshida S, Shinozaki K, Yamaguchi-Shinozaki K. Identification of cold-inducible downstream genes of the Arabidopsis DREB1A/CBF3 transcriptional factor using two microarray systems. The Plant journal: for cell and molecular biology. 2004;38(6):982–93.

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Enhancing cotton tolerance to cold and drought stress: A protective role of SikCOR413PM1 gene overexpression

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Plant materials and plant growth conditions

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Investigation of agronomic traits

Investigation on agronomic traits of cotton under field drought stress

Statistical analysis

Discussion

Abbreviations

Declarations

References

Additional Declarations

Supplementary Files

Status:

Journal Publication

Version 1