Engineered resistance and risk assessment associated with insecticidal and weeds resistant transgenic cotton using wister rat model.

Stacking multiple genes into cotton crop to cop up multiple biotic stresses such as insects and weeds is a promising tool to save crop from losses. Transgenic cotton variety, VH-289, with double Bt and cp4EPSPS genes under the control of 35S promoter was used for the expression analyses and biosafety studies. The transgenic cotton plants were screened through PCR amplification of fragments, 1.7 kb for Cry1Ac, 582 bp for Cry2A and 250 bp for cp4EPSPS; which confirmed the presence of all genes transformed in transgenic cotton. The Cry1Ac + Cry2A and cp4EPSPS proteins were quantified through ELISA in transgenic cotton plants. The Glyphosate assay performed by spraying 1900 mL per acre of glyphosate Roundup further confirmed complete survival of transgenic cotton plants as compared to the non-transgenic cotton plants and all weeds. Similarly, insect infestation data determined that almost 99% insect mortality was observed in controlled field grown transgenic cotton plants as compared to the non-transgenic control plants. Evaluation of effect of temperature and soil nutrients availability on transgene expression in cotton plants was done at two different cotton growing regions, Multan and Lahore, Pakistan and results suggested that despite of higher temperature in Multan field, an increased level of Cry and cp4EPSPS proteins was recorded due to higher soil organic matter availability compared to Lahore field. Before commercialization of any transgenic variety its biosafety study is mandatory so, a 90 days biosafety study of the transgenic cotton plants with 40% transgenic cottonseeds in standard diet showed no harmful effect on wister rat model when studied for liver function, renal function and serum electrolyte.

The application of ammonium sulphate and amino acid on cotton: effects on can improve growth, yield, quality and nitrogen absorption / Aplicação de sulfato de amônio e aminoácido no algodão: efeitos sobre o crescimento, o rendimento, a qualidade e a absorção de nitrogênio

A field study was carried out to determine the influence of foliage applied plant growth promoter and retardant in improving soil applied sulphur fertilizer use efficiency in cotton during two consecutive summers 2014 and 2015. Experimental trial comprised of three different sources of sulphur (ammonium sulphate, potassium sulphate and elemental sulphur) and foliar spray of plant growth promoter and growth retardant including tap water was taken as control. Among treatments soil applied ammonium sulphate with foliage applied amino acid produced maximum plant height, sympodial branches, pods per plant, seed cotton yield, fiber yield, biological yield, protein contents, oil contents and leaf nitrogen uptake as compared to the other treatments. Whereas, soil applied potassium sulphate with foliar spray of mepiquat chloride on cotton significantly improved the boll weight and leaf potassium uptake. We conclude that soil applied ammonium sulphate and foliage spray of amino acid was more effective in improving the productivity and quality attributes of cotton.

Foi realizado um estudo de campo para determinar a influência do promotor de crescimento das plantas e retardador da folhagem em algodão, para melhora da eficiência do uso de fertilizantes à base de enxofre aplicados no solo durante dois verões consecutivos (2014 e 2015). O ensaio experimental foi composto de três fontes diferentes de enxofre (sulfato de amônio, sulfato de potássio e enxofre elementar) e pulverização foliar do promotor de crescimento de plantas e retardador de crescimento, incluindo água da torneira que foi tomada como controle. Entre os tratamentos, o sulfato de amônio aplicado no solo com aminoácido aplicado na folhagem produziu o máximo na altura da planta, ramos simodiais, capulhos por planta, rendimento de algodão em caroço, rendimento de fibra, rendimento biológico, conteúdo de proteínas, conteúdo de óleo e absorção de nitrogênio nas folhas quando comparado a outros tratamentos. Enquanto o solo fertilizado com sulfato de potássio e aplicação foliar de cloreto de mepiquat no algodão melhorou, significativamente, o peso do capulho e a absorção de potássio nas folhas. Sulfato de amônio aplicado no solo e a aplicação foliar de aminoácidos foram mais eficazes na melhora dos atributos de produtividade e qualidade do algodão.

Sphingolipid Profile during Cotton Fiber Growth Revealed That a Phytoceramide Containing Hydroxylated and Saturated VLCFA Is Important for Fiber Cell Elongation.

Cotton fiber is a single-celled seed trichrome that arises from the epidermis of the ovule's outer integument. The fiber cell displays high polar expansion and thickens but not is disrupted by cell division. Therefore, it is an ideal model for studying the growth and development of plant cells. Sphingolipids are important components of membranes and are also active molecules in cells. However, the sphingolipid profile during fiber growth and the differences in sphingolipid metabolism at different developmental stages are still unclear. In this study, we detected that there were 6 classes and 95 molecular species of sphingolipids in cotton fibers by ultrahigh performance liquid chromatography-MS/MS (UHPLC-MS/MS). Among these, the phytoceramides (PhytoCer) contained the most molecular species, and the PhytoCer content was highest, while that of sphingosine-1-phosphate (S1P) was the lowest. The content of PhytoCer, phytoceramides with hydroxylated fatty acyls (PhytoCer-OHFA), phyto-glucosylceramides (Phyto-GluCer), and glycosyl-inositol-phospho-ceramides (GIPC) was higher than that of other classes in fiber cells. With the development of fiber cells, phytosphingosine-1-phosphate (t-S1P) and PhytoCer changed greatly. The sphingolipid molecular species Ceramide (Cer) d18:1/26:1, PhytoCer t18:1/26:0, PhytoCer t18:0/26:0, PhytoCer t18:1/h20:0, PhytoCer t18:1/h26:0, PhytoCer t18:0/h26:0, and GIPC t18:0/h16:0 were significantly enriched in 10-DPA fiber cells while Cer d18:1/20:0, Cer d18:1/22:0, and GIPC t18:0/h18:0 were significantly enriched in 20-DPA fiber cells, indicating that unsaturated PhytoCer containing hydroxylated and saturated very long chain fatty acids (VLCFA) play some role in fiber cell elongation. Consistent with the content analysis results, the related genes involved in long chain base (LCB) hydroxylation and unsaturation as well as VLCFA synthesis and hydroxylation were highly expressed in rapidly elongating fiber cells. Furthermore, the exogenous application of a potent inhibitor of serine palmitoyltransferase, myriocin, severely blocked fiber cell elongation, and the exogenous application of sphingosine antagonized the inhibition of myriocin for fiber elongation. Taking these points together, we concluded that sphingolipids play crucial roles in fiber cell elongation and SCW deposition. This provides a new perspective for further studies on the regulatory mechanism of the growth and development of cotton fiber cells.

Induction of polyploidy through colchicine in cotton (Gossypium herbaceum) and its conformity by cytology and flow cytometry analyses.

Cotton is one of the most important fibre crops in the world. An increase in ploidy level was observed in diploid cotton species namely Gossypium herbaceum in the experiment, through colchicine application. There were significant growth variations noticed during the induction of polyploidy in the cotton plants depending upon the concentration of colchicine, duration of the treatments and genotypes taken. An increase in the concentration of colchicine or the duration of the treatment had a retardation effect on seed viability in G. herbaceum. The hypocotyls of length between 4 and 8 mm were found to be most responsive to colchicine treatment. The root meristem of G. herbaceum, treated with 0.2 and 0.4% colchicine exhibited the most number of tetraploid cells during 16 h of treatment. The increase in the concentration of colchicine, along with an increase in duration of treatment led to the chromosomal abnormalities in the wild cotton species. Seed treatment for colchicine application, was the most efficient and reliable method when compared to Petri-plate application and cotton swabbing treatments for inducing polyploidy in diploid cotton.

Biochemical and Physiological Toxicity of Nanoparticles in Plant.

As increasing application of nanoparticles, nanoparticles have been becoming a new emerging environmental pollution that attracts a lot of attention from the scientific community and also regulatory agents. In the past decade, studying the toxicity and environmental impacts of nanoparticles is becoming a hot research field and more and more researches have been published using both plant and animal system. In this chapter, using oxidized metal nanoparticles as an example, we introduce a detailed protocol for performing research on biochemical and physiological toxicity of nanoparticles in plant. We employ a hydroponics system to study phytotoxicity of nanoparticles, which makes it easier to study the impact of nanoparticles. In this chapter, we majorly focus on plant respiration and photosynthesis, root vigor as well as oxidative stress. Oxidative stress is one major physiological response to different environmental pollution, in which we present a detailed method for detecting free radical oxygen species as well as the major molecules and enzymes associating with oxidative stress, including SOD and POD. Although we introduce the methods using cotton as an example, the protocols presented in this chapter can be used almost any plant species to test the biochemical and physiological toxicity of an environmental pollution.

Mapping-by-sequencing the locus of EMS-induced mutation responsible for tufted-fuzzless seed phenotype in cotton.

Cotton fiber mutants are valuable resources for studying functions of altered genes and their roles in fiber development. The n4t is a recessive tufted-fuzzless seed mutant created through chemical mutagenesis with ethyl methanesulfonate. Genetic analysis indicated that the tufted-fuzzless phenotype is controlled by a single recessive locus. In this study, we developed an F2 population of 602 progeny plants and sequenced the genomes of the parents and two DNA bulks from F2 progenies showing the mutant phenotype. We identified DNA sequence variants between the tufted-fuzzless mutant and wild type by aligning the sequence reads to the reference TM-1 genome and designed subgenome-specific SNP markers. We mapped the n4t locus on chromosome D04 within a genomic interval of about 411 kb. In this region, seven genes showed significant differential expression between the tufted-fuzzless mutant and wild type. Possible candidate genes are discussed in this study. The utilization of the n4t mutant along with other fiber mutants will facilitate our understanding of the molecular mechanisms of cotton fiber cell growth and development.

Cerium oxide nanoparticles improve cotton salt tolerance by enabling better ability to maintain cytosolic K+/Na+ ratio.

BACKGROUND: Salinity is a worldwide factor limiting the agricultural production. Cotton is an important cash crop; however, its yield and product quality are negatively affected by soil salinity. Use of nanomaterials such as cerium oxide nanoparticles (nanoceria) to improve plant tolerance to stress conditions, e.g. salinity, is an emerged approach in agricultural production. Nevertheless, to date, our knowledge about the role of nanoceria in cotton salt response and the behind mechanisms is still rare. RESULTS: We found that PNC (poly acrylic acid coated nanoceria) helped to improve cotton tolerance to salinity, showing better phenotypic performance, higher chlorophyll content (up to 68% increase) and biomass (up to 38% increase), and better photosynthetic performance such as carbon assimilation rate (up to 144% increase) in PNC treated cotton plants than the NNP (non-nanoparticle control) group. Under salinity stress, in consistent to the results of the enhanced activities of antioxidant enzymes, PNC treated cotton plants showed significant lower MDA (malondialdehyde, up to 44% decrease) content and reactive oxygen species (ROS) level such as hydrogen peroxide (H2O2, up to 79% decrease) than the NNP control group, both in the first and second true leaves. Further experiments showed that under salinity stress, PNC treated cotton plants had significant higher cytosolic K+ (up to 84% increase) and lower cytosolic Na+ (up to 77% decrease) fluorescent intensity in both the first and second true leaves than the NNP control group. This is further confirmed by the leaf ion content analysis, showed that PNC treated cotton plants maintained significant higher leaf K+ (up to 84% increase) and lower leaf Na+ content (up to 63% decrease), and thus the higher K+/Na+ ratio than the NNP control plants under salinity stress. Whereas no significant increase of mesophyll cell vacuolar Na+ intensity was observed in PNC treated plants than the NNP control under salinity stress, suggesting that the enhanced leaf K+ retention and leaf Na+ exclusion, but not leaf vacuolar Na+ sequestration are the main mechanisms behind PNC improved cotton salt tolerance. qPCR results showed that under salinity stress, the modulation of HKT1 but not SOS1 refers more to the PNC improved cotton leaf Na+ exclusion than the NNP control. CONCLUSIONS: PNC enhanced leaf K+ retention and Na+ exclusion, but not vacuolar Na+ sequestration to enable better maintained cytosolic K+/Na+ homeostasis and thus to improve cotton salt tolerance. Our results add more knowledge for better understanding the complexity of plant-nanoceria interaction in terms of nano-enabled plant stress tolerance.

Sphingosine Promotes Embryo Biomass in Upland Cotton: A Biochemical and Transcriptomic Analysis.

Sphingolipids are essential membrane components and signal molecules, but their regulatory role in cotton embryo growth is largely unclear. In this study, we evaluated the effects of treatment with the sphingolipid synthesis inhibitor fumonisin B1 (FB1), the serine palmityl transferase (SPT) inhibitor myriocin, the SPT sphingolipid product DHS (d18:0 dihydrosphingosine), and the post-hydroxylation DHS product PHS (t18:0 phytosphingosine) on embryo growth in culture, and performed comparative transcriptomic analysis on control and PHS-treated samples. We found that FB1 could inhibit cotton embryo development. At the five-day ovule/embryo developmental stage, PHS was the most abundant sphingolipid. An SPT enzyme inhibitor reduced the fresh weight of embryos, while PHS had the opposite effect. The transcriptomic analysis identified 2769 differentially expressed genes (1983 upregulated and 786 downregulated) in the PHS samples. A large number of transcription factors were highly upregulated, such as zinc finger, MYB, NAC, bHLH, WRKY, MADS, and GRF in PHS-treated samples compared to controls. The lipid metabolism and plant hormone (auxin, brassinosteroid, and zeatin) related genes were also altered. Our findings provide target metabolites and genes for cotton seed improvement.

Soil applied potassium improves productivity and fiber quality of cotton cultivars grown on potassium deficient soils.

Cotton (Gossypium hirsutum L.) is considered as the most valuable cash crop of Pakistan. During last decade, its yield has been declined due to various biotic and abiotic factors. Among abiotic factors, improper use of fertilizers is considered very important specially regarding plant defense and yield. This study was conducted to evaluate the effect of different levels (0, 40, 80 and 120 kg ha-1) of K fertilizer (K2O) on different growth parameters of two commercial Bt cotton cultivars (CYTO-301 and IUB-2013) and one non-Bt cultivar (CYTO-142) during 2016 and 2017. Maximum plant height (124-134 cm), dry matter contents (915-1005%), fruiting point (441-462), bolls per plant (96-139), average boll weight (4.2-5.2 g) and seed cotton yield (2524-3175 kg ha-1) and minimum shedding (43-73%) were observed in plots receiving highest dose of K (120 kg ha-1). The CYTO-103 cultivar was found more responsive to K fertilizer as compared to rest of cultivars (CYTO-142 and IUB-2013). Concluding, ideal dose of fertilizer is very important (120 kg ha-1 in our case) for optimum growth and production of good quality fiber with enhanced seed cotton yield.

Genome-wide identification and expression analysis of the plant specific LIM genes in Gossypium arboreum under phytohormone, salt and pathogen stress.

Asiatic cotton (Gossypium arboreum) cultivated as 'desi cotton' in India, is renowned for its climate resilience and robustness against biotic and abiotic stresses. The genome of G. arboreum is therefore, considered as a valued reserve of information for discovering novel genes or gene functions for trait improvements in the present context of cotton cultivation world-wide. In the present study, we carried out genome-wide analysis of LIM gene family in desi cotton and identified twenty LIM domain proteins (GaLIMs) which include sixteen animals CRP-like GaLIMs and four plant specific GaLIMs with presence (GaDA1) or absence (GaDAR) of UIM (Ubiquitin Interacting Motifs). Among the sixteen CRP-like GaLIMs, eleven had two conventional LIM domains while, five had single LIM domain which was not reported in LIM gene family of the plant species studied, except in Brassica rapa. Phylogenetic analysis of these twenty GaLIM proteins in comparison with LIMs of Arabidopsis, chickpea and poplar categorized them into distinct αLIM1, ßLIM1, γLIM2, δLIM2 groups in CRP-like LIMs, and GaDA1 and GaDAR in plant specific LIMs group. Domain analysis had revealed consensus [(C-X2-C-X17-H-X2-C)-X2-(C-X2-C-X17-C-X2-H)] and [(C-X2-C-X17-H-X2-C)-X2-(C-X4-C-X15-C-X2-H)] being conserved as first and/or second LIM domains of animal CRP-like GaLIMs, respectively. Interestingly, single LIM domain containing GaLIM15 was found to contain unique consensus with longer inter-zinc-motif spacer but shorter second zinc finger motif. All twenty GaLIMs showed variable spatio-temporal expression patterns and accordingly further categorized into distinct groups of αLIM1, ßLIM1, γLIM2 δLIM2 and plant specific LIM (DA1/DAR). For the first time, response of GaDA1/DAR under the influence of biotic and abiotic stresses were studied in cotton, involving treatments with phytohormones (Jasmonic acid and Abscisic acid), salt (NaCl) and wilt causing pathogen (Fusarium oxysporum). Expressions patterns of GaDA1/DAR showed variable response and identified GaDA2 as a probable candidate gene for stress tolerance in G. arboreum.

Compatibility of Bt biopesticides and adjuvants for Spodoptera frugiperda control.

Spodoptera frugiperda is a pest of economic importance for several crops with resistance reports to Bt crops and pesticides. Eco-friendly Bt biopesticides may be an alternative to chemical insecticides due to their selectivity and specificity. However, the efficacy of Bt biopesticides may be influenced by the association with other chemicals, such as adjuvants. This study evaluated the compatibility and toxicity of Bt biopesticides mixed with adjuvants for the control of S. frugiperda. The treatments included the association of Dipel SC and Dipel PM with adjuvants. Compatibility tests were used to evaluate the Bt mixture. Bt suspensions obtained from mixtures of Bt and adjuvants at 106 and 3 × 108 spores/mL-1 were used to evaluate S. frugiperda mortality and distilled water was used as the control. The addition of the adjuvant LI increased growth and sporulation, indicating compatibility with Bt biopesticides. The other adjuvants were toxic to reducing Bt growth and sporulation. Only the mixture of Bt with LI and Bt alone was effective to S. frugiperda. The addition of adjuvants to Bt biopesticide affect the Bt sporulation, growth and mortality.

Linking exogenous foliar application of glycine betaine and stomatal characteristics with salinity stress tolerance in cotton (Gossypium hirsutum L.) seedlings.

BACKGROUND: Glycine betaine (GB) plays a crucial role in plants responding to abiotic stresses. Studying the physiological response of cotton seedlings to exogenous GB under salt stress provides a reference for the application of GB to improve the resistance of cotton seedlings under salt stress. The purpose of this research is to examine the impacts of foliar-applied GB on leaf stomatal structure and characteristics, gas exchange and chlorophyll fluorescence characteristics and plant growth indicators of Gossypium hirsutum L. under NaCl stress conditions. RESULTS: Under the salinity of 150 mM, the four concentrations of GB are 0, 2.5, 5, and 7.5 mM, and the control (CK) was GB-untreated non-saline. Salt stress negatively affected leaf stomata as well as gas exchange and chlorophyll fluorescence and decreased plant growth parameters of cotton seedlings. The treatment with 5 mM GB significantly increased the evolution of photosynthetic rate (Pn), transpiration rate (Tr), intracellular CO2 concentration (Ci) and stomatal conductance (gs) compared to the GB-untreated saline treatment. The Exogenous foliar-applied GB has sustainably decreased the carboxylation efficiency (Pn/Ci) and water use efficiency (WUE). The concentration of 5 mM GB leads to a significant improvement of leaf stomatal characteristics. The leaf gas exchange attributes correlated positively with stomatal density (SD), stomatal length (SL) and stomatal with (SW). CONCLUSION: The overall results suggested that exogenous foliar supplementation with GB can effectively alleviate the damage of salt stress to cotton seedlings. The effect of applying 5 mM GB could be an optional choice for protecting cotton seedlings from NaCl stress through promoting the stomatal functions, photosynthetic activities and growth characteristics.

GhMYB4 downregulates lignin biosynthesis and enhances cotton resistance to Verticillium dahliae.

KEY MESSAGE: GhMYB4 acts as a negative regulator in lignin biosynthesis, which results in alteration of cell wall integrity and activation of cotton defense response. Verticillium wilt of cotton (Gossypium hirsutum) caused by the soil-borne fungus Verticillium dahliae (V. dahliae) represents one of the most important constraints of cotton production worldwide. Mining of the genes involved in disease resistance and illuminating the molecular mechanisms that underlie this resistance is of great importance in cotton breeding programs. Defense-induced lignification in plants is necessary for innate immunity, and there are reports of a correlation between increased lignification and disease resistance. In this study, we present an example in cotton whereby plants with reduced lignin content also exhibit enhanced disease resistance. We identified a negative regulator of lignin synthesis, in cotton encoded in GhMYB4. Overexpression of GhMYB4 in cotton and Arabidopsis enhanced resistance to V. dahliae  with reduced lignin deposition. Moreover, GhMYB4 could bind the promoters of several genes involved in lignin synthesis, such as GhC4H-1, GhC4H-2, Gh4CL-4, and GhCAD-3, and impair their expression. The reduction of lignin content in GhMYB4-overexpressing cotton led to alterations of cell wall integrity (CWI) and released more oligogalacturonides (OGs) which may act as damage-associated molecular patterns (DAMPs) to stimulate plant defense responses. In support of this hypothesis, exogenous application with polygalacturonic acid (PGA) in cotton activated biosynthesis of jasmonic acid (JA) and JA-mediated defense against V. dahliae, similar to that described for cotton plants overexpressing GhMYB4. This study provides a new candidate gene for cotton disease-resistant breeding and an increased understanding of the relationship between lignin synthesis, OG release, and plant immunity.

Transcriptomic analyses show that 24-epibrassinolide (EBR) promotes cold tolerance in cotton seedlings.

In plants, brassinosteroids (BRs) are a class of steroidal hormones that are involved in numerous physiological responses. However, the function of BRs in cold tolerance in cotton has not been explored. In this study, cotton seedlings were treated with five concentrations (0, 0.05, 0.1, 0.2, 0.5 and 1.0 mg/L) of 24-Epibrassinolide (EBR) at 4°C. We measured the electrolyte leakage, malondialdehyde (MDA) content, proline content, and net photosynthesis rate (Pn) of the seedlings, which showed that EBR treatment increased cold tolerance in cotton in a dose-dependent manner, and that 0.2 mg/L is an optimum concentration for enhancing cold tolerance. The function of EBR in cotton cotyledons was investigated in the control 0 mg/L (Cold+water) and 0.2 mg/L (Cold+EBR) treatments using RNA-Seq. A total of 4,001 differentially expressed genes (DEGs), including 2,591 up-regulated genes and 1,409 down-regulated genes were identified. Gene Ontology (GO) and biochemical pathway enrichment analyses showed that EBR is involved in the genetic information process, secondary metabolism, and also inhibits abscisic acid (ABA) and ethylene (ETH) signal transduction. In this study, physiological experiments showed that EBR can increase cold tolerance in cotton seedlings, and the comprehensive RNA-seq data shed light on the mechanisms through which EBR increases cold tolerance in cotton seedlings.

Relationship between imidacloprid residues and control effect on cotton aphids in arid region.

In this case, the dissipation and residues of imidacloprid as well as its control efficacy against aphids (Aphis gossypii Glover) in cotton cropping system were reported. After the final spray at the rates of 10.5-42.5 g a.i. ha-1, the initial deposits were 0.59-2.25 mg kg-1 with half-lives of 2.12-2.84 days on leaves and 0.06-0.21 mg kg-1 with half-lives of 1.51-4.20 days in soil, respectively. The initial residues were significantly higher with longer persistence in the upper position of the leaf than in middle and lower positions. The different application dosages could induce a significant difference in the initial deposits, but not show consistent correlation with the dissipation rate. The repeated applications of imidacloprid could alter its residue levels and dissipation rates. The long-term residue concentrations of imidacloprid (60 days after the final application) reached to the nondetectable level in soil. Combined with the control efficacy results, it was considered that the recommended dose of imidacloprid on cotton could be used effectively and safe in this arid area from the view of crop protection and environmental contamination.

Effects of biochar and biofertilizer on cadmium-contaminated cotton growth and the antioxidative defense system.

Consistent use of large amounts of fertilizers, pesticides, and mulch can cause the accumulation of harmful substances in cotton plants. Among these harmful substances, cadmium (Cd), an undegradable element, stands out as being particularly highly toxic to plants. The objective of this study was to evaluate the ability of biochar (3%) and biofertilizer (1.5%) to decrease Cd uptake, increase cotton dry weight, and modulate the activities of photosynthetic and peroxidase (POD), superoxide dismutase (SOD), catalase enzyme (CAT) in cotton (Gossypium hirsutum L.) grown in Cd-contaminated soil (0, 1, 2, or 4 mg Cd kg-1 soil) in pots. These studies showed that, as expected, exogenous Cd adversely affects cotton chlorophyll and photosynthesis. However, biochar and biofertilizer increased cotton dry weight by an average of 16.82% and 32.62%, respectively. Meanwhile, biochar and biofertilizer decreased the accumulation of Cd in cotton organs, and there was a significant reduction in the amount of Cd in bolls (P < 0.05). Biochar and biofertilizer have a positive impact on cotton chlorophyll content, net photosynthesis, stomatal conductance, transpiration rate, and intercellular CO2 concentration. Thus, the addition of biochar and biofertilizer promote cotton growth. However, biochar and biofertilizer increased the SOD activity of leaves (47.70% and 77.21%), CAT activity of leaves (35.40% and 72.82%), SOD activity of roots (33.62% and 39.37%), and CAT activity of roots (36.91% and 60.29%), respectively, and the addition of biochar and biofertilizer decreased the content of MDA and electrolyte leakage rate. Redundancy analyses showed that biochar and biofertilizer also improved SOD and POD activities by reducing the heavy metal-induced oxidative stress in cotton and reducing Cd uptake in cotton organs. Therefore, biochar and biofertilizer have a positive effect on the growth of cotton.

Effects of controlled-release urea combined with fulvic acid on soil inorganic nitrogen, leaf senescence and yield of cotton.

A split-plot field experiment was conducted in 2018-2019 to study the effects of nitrogen fertilizer types and fulvic acid (FA) rates on soil nitrogen and cotton growth. The nitrogen fertilizers included controlled-release urea (CRU) and urea, which were applied combined with three FA rates (90, 180 and 270 kg ha-1). The main plot was the nitrogen fertilizer type, and the subplot was the FA rate. The results showed that the lint yield of the FA180 treatment was 5.2-8.6% higher than the FA90 and FA270 treatments. Moreover, moderate FA application markedly improved the cotton leaf SPAD value (chlorophyll relative value), photosynthesis and chlorophyll fluorescence parameters compared with low and high FA rates. Replacing urea with CRU significantly increased the soil inorganic nitrogen and nitrogen use efficiency and also improved cotton fiber quality parameters. Meanwhile, the boll weight and seed yield of the CRU treatments were 1.5-8.4% and 3.3-19.1% higher, respectively, than the urea treatments. The interaction between nitrogen type and FA rate had a positive effect on cotton growth. Thus, the application of CRU combined with 180 kg ha-1 FA on cotton can not only improve the fiber quality and delay leaf senescence but also increase the yield and economic benefit.

Histone Deacetylase Inhibitor SAHA Improves High Salinity Tolerance Associated with Hyperacetylation-Enhancing Expression of Ion Homeostasis-Related Genes in Cotton.

Histone acetylation plays an important role in regulation of chromatin structure and gene expression in terms of responding to abiotic stresses. Histone acetylation is modulated by histone deacetylases (HDACs) and histone acetyltransferases. Recently, the effectiveness of HDAC inhibitors (HDACis) for conferring plant salt tolerance has been reported. However, the role of HDACis in cotton has not been elucidated. In the present study, we assessed the effects of the HDACi suberoylanilide hydroxamic acid (SAHA) during high salinity stress in cotton. We demonstrated that 10 µM SAHA pretreatment could rescue of cotton from 250 mM NaCl stress, accompanied with reduced Na+ accumulation and a strong expression of the ion homeostasis-related genes. Western blotting and immunostaining results revealed that SAHA pretreatment could induce global hyperacetylation of histone H3 at lysine 9 (H3K9) and histone H4 at lysine 5 (H4K5) under 250 mM NaCl stress, indicating that SAHA could act as the HDACi in cotton. Chromatin immunoprecipitation and chromatin accessibility coupled with real time quantitative PCR analyses showed that the upregulation of the ion homeostasis-related genes was associated with the elevated acetylation levels of H3K9 and H4K5 and increased chromatin accessibility on the promoter regions of these genes. Our results could provide a theoretical basis for analyzing the mechanism of HDACi application on salt tolerance in plants.

Fumonisin B1-Induced Changes in Cotton Fiber Elongation Revealed by Sphingolipidomics and Proteomics.

Sphingolipids are essential biomolecules and membrane components, but their regulatory role in cotton fiber development is poorly understood. Here, we found that fumonisin B1 (FB1)-a sphingolipid synthesis inhibitor-could block fiber elongation severely. Using liquid chromatography tandem mass spectrometry (LC-MS/MS), we detected 95 sphingolipids that were altered by FB1 treatment; of these, 29 (mainly simple sphingolipids) were significantly increased, while 33 (mostly complex sphingolipids) were significantly decreased. A quantitative analysis of the global proteome, using an integrated quantitative approach with tandem mass tag (TMT) labeling and LC-MS/MS, indicated the upregulation of 633 and the downregulation of 672 proteins after FB1 treatment. Most differentially expressed proteins (DEPs) were involved in processes related to phenylpropanoid and flavonoid biosynthesis. In addition, up to 20 peroxidases (POD) were found to be upregulated, and POD activity was also increased by the inhibitor. To our knowledge, this is the first report on the effects of FB1 treatment on cotton fiber and ovule sphingolipidomics and proteomics. Our findings provide target metabolites and biological pathways for cotton fiber improvement.

Responses of leaf gas exchange attributes, photosynthetic pigments and antioxidant enzymes in NaCl-stressed cotton (Gossypium hirsutum L.) seedlings to exogenous glycine betaine and salicylic acid.

BACKGROUND: Application of exogenous glycine betaine (GB) and exogenous salicylic acid (SA) mitigates the adverse effects of salinity. Foliar spraying with exogenous GB or SA alleviates salt stress in plants by increasing leaf gas exchange and stimulating antioxidant enzyme activity. The effects of foliar application of exogenous GB and SA on the physiology and biochemistry of cotton seedlings subjected to salt stress remain unclear. RESULTS: Results showed that salt stress of 150 mM NaCl significantly reduced leaf gas exchange and chlorophyll fluorescence and decreased photosynthetic pigment quantities and leaf relative water content. Foliar spray concentrations of 5.0 mM exogenous GB and 1.0 mM exogenous SA promoted gas exchange and fluorescence in cotton seedlings, increased quantities of chlorophyll pigments, and stimulated the antioxidant enzyme activity. The foliar spray also increased leaf relative water content and endogenous GB and SA content in comparison with the salt-stressed only control. Despite the salt-induced increase in antioxidant enzyme content, exogenous GB and SA in experimental concentrations significantly increased the activity of glutathione reductase, ascorbate peroxidase, superoxide dismutase, catalase and peroxidase, and decreased malondialdehyde content under salt stress. Across all experimental foliar spray GB and SA concentrations, the photochemical efficiency of photosystem II (FV/FM) reached a peak at a concentration of 5.0 mM GB. The net photosynthetic rate (Pn) and FV/FM were positively correlated with chlorophyll a and chlorophyll b content in response to foliar spraying of exogenous GB and SA under salt stress. CONCLUSIONS: We concluded, from our results, that concentrations of 5.0 mM GB or 1.0 mM SA are optimal choices for mitigating NaCl-induced damage in cotton seedlings because they promote leaf photosynthesis, increase quantities of photosynthetic pigments, and stimulate antioxidant enzyme activity. Among, 5.0 mM GB and 1.0 mM SA, the best performance in enhancing endogenous GB and SA concentrations was obtained with the foliar application of 1.0 mM SA under salt stress.