Investigation of salt tolerance in cotton germplasm by analyzing agro-physiological traits and ERF genes expression.

The development of genotypes that can tolerate high levels of salt is crucial for the efficient use of salt-affected land and for enhancing crop productivity worldwide. Therefore, incorporating salinity tolerance is a critical trait that crops must possess. Salt resistance is a complex character, controlled by multiple genes both physiologically and genetically. To examine the genetic foundation of salt tolerance, we assessed 16 F1 hybrids and their eight parental lines under normal and salt stress (15 dS/m) conditions. Under salt stress conditions significant reduction was observed for plant height (PH), bolls/plant (NBP), boll weight (BW), seed cotton yield (SCY), lint% (LP), fiber length (FL), fiber strength (FS), potassium to sodium ratio (K+/Na+), potassium contents (K+), total soluble proteins (TSP), carotenoids (Car) and chlorophyll contents. Furthermore, the mean values for hydrogen peroxide (H2O2), sodium contents (Na+), catalase (CAT), superoxide dismutase (SOD), peroxidase (POD), and fiber fineness (FF) were increased under salt stress. Moderate to high heritability and genetic advancement was observed for NBP, BW, LP, SCY, K+/Na+, SOD, CAT, POD, Car, TSP, FL, and FS. Mean performance and multivariate analysis of 24 cotton genotypes based on various agro-physiological and biochemical parameters suggested that the genotypes FBS-Falcon, Barani-333, JSQ-White Hold, Ghauri, along with crosses FBS-FALCON × JSQ-White Hold, FBG-222 × FBG-333, FBG-222 × Barani-222, and Barani-333 × FBG-333 achieved the maximum values for K+/Na+, K+, TSP, POD, Chlb, CAT, Car, LP, FS, FL, PH, NBP, BW, and SCY under salt stress and declared as salt resistant genotypes. The above-mentioned genotypes also showed relatively higher expression levels of Ghi-ERF-2D.6 and Ghi-ERF-7A.6 at 15 dS/m and proved the role of these ERF genes in salt tolerance in cotton. These findings suggest that these genotypes have the potential for the development of salt-tolerant cotton varieties with desirable fiber quality traits.

Drought and heat stress on cotton genotypes suggested agro-physiological and biochemical features for climate resilience.

This study aimed to investigate the impact of individual drought, heat, and combined drought and heat stress on twelve cotton genotypes, including eight tolerant and four susceptible genotypes. A field experiment was carried out by employing a randomized complete block split-plot design, with treatments (control, drought, heat, drought + heat), and cotton genotypes assigned to the main plots and sub-plots respectively. The results showed that the combined stress had a more severe impact on the yield and fiber quality of cotton genotypes compared to individual stresses. Among the studied genotypes, FB-Shaheen, FH-207, MNH-886, and White Gold exhibited superior performance in regard to agronomic and fiber quality characters under combined stress environments. Physiological parameters, including transpiration rate, stomatal conductance, relative water contents, and photosynthetic rate, were significantly reduced under combined stress. However, specific genotypes, MNH-886, FH-207, White Gold, and FB-Shaheen, demonstrated better maintenance of these parameters, indicating their enhanced tolerance to the combined stress. Furthermore, the accumulation of reactive oxygen species was more pronounced under combined stress compared to individual stressors. Tolerant genotypes showed lower levels of H2O2 and MDA accumulation, while susceptible genotypes exhibited higher levels of oxidative damage. Antioxidant enzyme activities, such as superoxide dismutase, peroxidase, and catalase, increased under combined stress, with tolerant genotypes displaying higher enzyme activities. Conversely, susceptible genotypes (AA-703, KZ 191, IR-6, and S-15) demonstrated lower increases in enzymatic activities under combined stress conditions. Biochemical traits, including proline, total phenolic content, flavonoids, and ascorbic acid, exhibited higher levels in resistant genotypes under combined stress, while sensitive genotypes displayed decreased levels of these traits. Additionally, chlorophyll a & b, and carotenoid levels were notably decreased under combined stress, with tolerant genotypes experiencing a lesser decrease compared to susceptible genotypes.

Transgenerational impact of climatic changes on cotton production.

Changing climatic conditions are an increasing threat to cotton production worldwide. There is a need to develop multiple stress-tolerant cotton germplasms that can adapt to a wide range of environments. For this purpose, 30 cotton genotypes were evaluated for two years under drought (D), heat (H), and drought + heat stresses (DH) under field conditions. Results indicated that plant height, number of bolls, boll weight, seed cotton yield, fiber fineness, fiber strength, fiber length, K+, K+/Na+, relative water contents (RWC), chlorophyll a and b, carotenoids, and total soluble proteins got reduced under D and H and were lowest under DH, whereas superoxidase dismutase (SOD), H2O2, Na+, GOT%, total phenolic contents, ascorbate, and flavonoids got increased for consecutive years. Correlation studies indicated that there was a positive correlation between most of the traits, but a negative correlation with H2O2 and Na+ ions. PCA and clustering analysis indicated that MNH-786, KAHKSHAN, CEMB-33, MS-71, FH-142, NIAB-820, CRS-2007, and FH-312 consistently performed better than other genotypes for most traits under stress conditions. Identified genotypes can be utilized in the future cotton breeding program to develop high-yielding, climate change-resilient cotton.

Genetic basis and principal component analysis in cotton (Gossypium hirsutum L.) grown under water deficit condition.

Cotton is considered as the main crop in the agricultural sector of Pakistan. Water deficiency in this region in recent years has reduced the chances of high yields of cotton. Selection and creation of high-yielding varieties of cotton, even in water deficit conditions, is one of urgent tasks of today. For this purpose, 40 diverse genotypes of upland cotton were screened in normal and water deficit conditions in triplicate arrangement under split plot in a randomized complete block design. All the genotypes showed significant difference under both water regimes. Ten upland cotton accessions were screened out as water deficit tolerant (VH-144, IUB-212, MNH-886, VH-295, IR-3701, AA-802, NIAB-111, NS-121, FH-113, and FH-142) and five as water deficit sensitive (IR-3, CIM-443, FH-1000, MNH-147, and S-12) based on seed cotton yield and stress susceptibility index. These tolerant and sensitive genotypes were crossed in line × tester mating design. For further evaluation of genetic material, the seed of 50 F1 crosses and their 15 parents were field planted under normal and water deficit conditions during next cotton growing season. Traits related to yield under the study showed significant variations among the accessions and their half sibs. The results of the principal component analysis (PCA) exhibited that total variation exhibited by factors 1 and factor 2 were 55.55 and 41.95%, respectively. PCA transformed the variables into three factors, and only two factors (F1 and F2) had eigenvalue > 1. The degree of dominance revealed that all parameters were highly influenced by non-additive gene action under both water regimes. Furthermore, the line VH-295 and tester CIM-443 had better yield performance under water deficit stress. The cross-combinations, viz., VH-144 × S-12, NIAB-111 × IR-3, and VH-295 × MNH-147, were the best for yield contributing traits. These combinations may be helpful for germplasm enhancement on large scale under water scarcity. All the studied traits have non-additive types of gene action suggesting the usage of these genotypes in cotton hybrid development program against water deficit tolerance.

Heterologous expression of cry3Bb1 and cry3 genes for enhanced resistance against insect pests in cotton.

Transgenic technology played a crucial role in developing insect-resistant plants resulting in the reduced application of pesticides. This article reports the expression of two cry proteins (Cry3Bb1 and Cry3) in cotton for enhanced resistance against chewing insect pests. The aforementioned genes were synthetically developed and were cloned under appropriate regulatory sequences followed by transformation into Eagle-2 genotype (Gossypium hirsutum) of cotton through shoot apex-cut Agro-infiltration. The transgene integration was validated by polymerase chain reaction using primers flanking the aforementioned cry genes. Transgene expression was assessed by qRT-PCR using GADPH as a reference gene. The relative fold expression analyses revealed the highest expression of the transgene(s) in M1 plants, which is a 4.5-fold expression (Cry3 + Cry3Bb1) followed by M3 (fold expression, 3.0) (Cry3Bb1) and M2 (fold expression, 2.5) (Cry3) transformants of cotton. The confirmed transgenic plants were exposed to insect pests, pink bollworm (Pectinophora gossypiella), and army bollworm (Helicoverpa armigera). Bioassay results revealed that 60% mortality was observed against pink bollworm, and 75% mortality was observed against army bollworm in transgenic plants containing both Cry3Bb1 and Cry3 genes (M1 transgenic plants). In M2 transgenic plants containing only the Cry3Bb1 gene, the mortality was observed to be 40% in the pink bollworm population, whereas 45% mortality was observed in the army bollworm population. In the case of M3 transgenic plants containing single gene-Cry3, the mortality was 20% in the pink bollworm population, whereas 30% mortality was observed in the army bollworm population. Almost no mortality was observed in non-transgenic Eagle-2 control plants. Hence, the developed cotton transformants have improved resistance against chewing insect pests.

Genome-wide characterization and expression analysis of Erf gene family in cotton.

BACKGROUND: AP2/ERF transcription factors are important in a variety of biological activities, including plant growth, development, and responses to biotic and abiotic stressors. However, little study has been done on cotton's AP2/ERF genes, although cotton is an essential fibre crop. We were able to examine the tissue and expression patterns of AP2/ERF genes in cotton on a genome-wide basis because of the recently published whole genome sequence of cotton. Genome-wide analysis of ERF gene family within two diploid species (G. arboreum & G. raimondii) and two tetraploid species (G. barbadense, G. hirsutum) was performed. RESULTS: A total of 118, 120, 213, 220 genes containing the sequence of single AP2 domain were identified in G. arboreum, G. raimondii, G. barbadense and G. hirsutum respectively. The identified genes were unevenly distributed across 13/26 chromosomes of A and D genomes of cotton. Synteny and collinearity analysis revealed that segmental duplications may have played crucial roles in the expansion of the cotton ERF gene family, as well as tandem duplications played a minor role. Cis-acting elements of the promoter sites of Ghi-ERFs genes predict the involvement in multiple hormone responses and abiotic stresses. Transcriptome and qRT-PCR analysis revealed that Ghi-ERF-2D.6, Ghi-ERF-12D.13, Ghi-ERF-6D.1, Ghi-ERF-7A.6 and Ghi-ERF-11D.5 are candidate genes against salinity tolerance in upland cotton. CONCLUSION: Overwhelmingly, the present study paves the way to better understand the evolution of cotton ERF genes and lays a foundation for future investigation of ERF genes in improving salinity stress tolerance in cotton.

Characterization of SOD and GPX Gene Families in the Soybeans in Response to Drought and Salinity Stresses.

Plant stresses causing accumulation of reactive oxidative species (ROS) are scavenged by effective antioxidant defense systems. Therefore, the present study performed genome-wide identification of superoxide dismutase (SOD) and glutathione peroxidase (GPX) gene families in cultivated and wild soybeans, and 11 other legume species. We identified a total of 101 and 95 genes of SOD and GPX, respectively, across thirteen legume species. The highest numbers of SODs and GPXs were identified in cultivated (Glycine max) and wild (Glycine soja). A comparative phylogenetic study revealed highest homology among the SODs and GPXs of cultivated and wild soybeans relative to other legumes. The exon/intron structure, motif and synteny blocks were conserved in both soybean species. According to Ka/Ks, purifying the selection played the major evolutionary role in these gene families, and segmental duplication are major driving force for SODs and GPXs expansion. In addition, the qRT-PCR analysis of the G. max and G. soja SOD and GPX genes revealed significant differential expression of these genes in response to oxidative, drought and salinity stresses in root tissue. In conclusion, our study provides new insights for the evolution of SOD and GPX gene families in legumes, and provides resources for further functional characterization of these genes for multiple stresses.

Unraveling Heat Tolerance in Upland Cotton (Gossypium hirsutum L.) Using Univariate and Multivariate Analysis.

The ever-changing global environment currently includes an increasing ambient temperature that can be a devastating stress for organisms. Plants, being sessile, are adversely affected by heat stress in their physiology, development, growth, and ultimately yield. Since little is known about the response of biochemical traits to high-temperature ambiance, we evaluated eight parental lines (five lines and three testers) and their 15 F1 hybrids under normal and high-temperature stress to assess the impact of these conditions over 2 consecutive years. The research was performed under a triplicate randomized complete block design including a split-plot arrangement. Data were recorded for agronomic, biochemical, and fiber quality traits. Mean values of agronomic traits were significantly reduced under heat stress conditions, while hydrogen peroxide, peroxidase, total soluble protein, superoxide dismutase, catalase (CAT), carotenoids, and fiber strength displayed higher mean values under heat stress conditions. Under both conditions, high genetic advance and high heritability were observed for seed cotton yield (SCY), CAT, micronaire value, plant height, and chlorophyll-a and b content, indicating that an additive type of gene action controls these traits under both the conditions. For more insights into variation, Pearson correlation analysis and principal component analysis (PCA) were performed. Significant positive associations were observed among agronomic, biochemical, and fiber quality-related traits. The multivariate analyses involving hierarchical clustering and PCA classified the 23 experimental genotypes into four groups under normal and high-temperature stress conditions. Under both conditions, the F1 hybrid genotype FB-SHAHEEN × JSQ WHITE GOLD followed by Ghuari-1, CCRI-24, Eagle-2 × FB-Falcon, Ghuari-1 × JSQ White Gold, and Eagle-2 exhibited better performance in response to high-temperature stress regarding the agronomic and fiber quality-related traits. The mentioned genotypes could be utilized in future cotton breeding programs to enhance heat tolerance and improve cotton yield and productivity through resistance to environmental stressors.

Genome-wide identification and evolution of Dof transcription factor family in cultivated and ancestral cotton species.

The DNA-binding with one finger (Dof) proteins are transcription factors involved in many biological processes in plants. To predict the evolutionary pattern, a genome-wide in-silico analysis of Dof TFs family in diploid (Gossypium arboreum and Gossypium raimondii) and allotetraploid (Gossypium hirsutum and Gossypium barbadense) cotton species were carried out. In G. arboreum, we have identified 58 non-redundant genes encoding Dof proteins renamed as GaDof (G. arboreum Dof), 55 Dof genes were identified in G. raimondii (GrDof), 89 were predicted ffrom G. hirsutum (GhDof) and the highest, 110 Dof genes were identified in G. barbadense (GbDof). The phylogenetic analysis, physical location, gene structure, conserved domain analyses were also investigated for G. arboreum, G. raimondii, and G. hirsutum. The gene expression pattern in G. hirsutum, at different growth stages, revealing the probable involvement of some GhDof genes in growth and development. These genes may improve seed germination and growth in cotton.

Salt stress induces physiochemical alterations in rice grain composition and quality.

Salinity has drastic effects on plant growth and productivity and is one of the major factors responsible for crop yield losses throughout the agricultural soils of the world. The mechanisms of salinity tolerance in plants are regulated by a set of inherent multigenes and prevalent environmental factors, which bring about a myriad of metabolic changes in each plant part. The stress-induced metabolic changes in the rice plant have been intensively studied, but extensively in plant parts such as stem, leaf, and root. However, little information exists in the literature about such stress-induced architectural and physiological changes in rice grain, a premier staple food of a large proportion of human population. Thus, the current review comprehensively describes the effects of salinity stress on rice grain composition including changes in carbohydrate, protein, fat, and mineral contents. Elucidation of salinity induced changes in rice grain composition would help to understand whether or not a nutritious and healthy staple food is available to human population from rice grown under saline environments.