Resistant Cotton Research Articles

GhODO1, an R2R3-type MYB transcription factor, positively regulates cotton resistance to Verticillium dahliae via the lignin biosynthesis and jasmonic acid signaling pathway

In plants, MYB transcription factors play diverse roles in growth, development, and response to abiotic and biotic stresses. However, the signaling processes of these transcription factors in defense against pathogen attacks remain largely unknown. This study isolated a novel R2R3-type MYB transcription factor GhODO1 from cotton (Gossypium hirsutum) and functionally characterized its positive role in tolerance to Verticillium dahliae. GhODO1 was induced by V. dahliae and jasmonic acid (JA) and transient expression of fused GhODO1-GFP in onion epidermal cells showed that GhODO1 protein was localized in the cell nucleus. Knockdown of GhODO1 significantly reduced the resistance of cotton to V. dahliae, whereas GhODO1 ectopic overexpression in Arabidopsis conferred enhanced resistance to V. dahliae. Lignin deposition was significantly decreased in GhODO1-silenced cotton plants after V. dahliae inoculation and mock treatment. The expression levels of genes and activities of enzymes involved in lignin biosynthesis were reduced in GhODO1-silenced cotton plants compared to the TRV:00. Yeast one-hybrid assays revealed that GhODO1 protein interacts with the promoters of lignin biosynthesis-related genes Gh4CL1 and GhCAD3, directly activates their expression, and enhances total lignin accumulation. Moreover, GhODO1 silencing compromised JA-mediated defense signaling and JA accumulation. These results show that GhODO1 is involved in cotton resistance to V. dahliae by involving the lignin biosynthesis and the JA signaling pathway.

Evolution and Stress Responses of CLO Genes and Potential Function of the GhCLO06 Gene in Salt Resistance of Cotton.

The caleosin (CLO) protein family displays calcium-binding properties and plays an important role in the abiotic stress response. Here, a total of 107 CLO genes were identified in 15 plant species, while no CLO genes were detected in two green algal species. Evolutionary analysis revealed that the CLO gene family may have evolved mainly in terrestrial plants and that biological functional differentiation between species and functional expansion within species have occurred. Of these, 56 CLO genes were identified in four cotton species. Collinearity analysis showed that CLO gene family expansion mainly occurred through segmental duplication and whole-genome duplication in cotton. Sequence alignment and phylogenetic analysis showed that the CLO proteins of the four cotton species were mainly divided into two types: H-caleosins (class I) and L-caleosins (class II). Cis-acting element analysis and quantitative RT–PCR (qRT–PCR) suggested that GhCLOs might be regulated by abscisic acid (ABA) and methyl jasmonate (MeJA). Moreover, transcriptome data and qRT–PCR results revealed that GhCLO genes responded to salt and drought stresses. Under salt stress, gene-silenced plants (TRV: GhCLO06) showed obvious yellowing and wilting, higher malondialdehyde (MDA) content accumulation, and significantly lower activities of superoxide dismutase (SOD) and peroxidase (POD), indicating that GhCLO06 plays a positive regulatory role in cotton salt tolerance. In gene-silenced plants (TRV: GhCLO06), ABA-related genes (GhABF2, GhABI5, and GhNAC4) were significantly upregulated after salt stress, suggesting that the regulation of salt tolerance may be related to the ABA signaling pathway. This research provides an important reference for further understanding and analyzing the molecular regulatory mechanism of CLOs for salt tolerance.

Cotton resistance to white mold (Sclerotinia sclerotiorum) evaluated by the oxalic acid method

The white mold (Sclerotinia sclerotiorum) is one of the most destructive cotton disease. Genetic resistance is one of the main strategies to control this disease. Thus, this study aimed to determine an appropriate period of exposition and concentration of oxalic acid to identify levels of physiological resistance to white mold in white fiber cotton genotypes. The study was implemented in two periods (August to October and November to January) in randomized complete block design, in a split-split-plot factorial in time, with five replications, where the plot factor was the concentration of oxalic acid (20; 40 mM) and the sub-plot factor was the 20 cotton genotypes, the sub-sub-plot factor was the time of exposure to oxalic acid (24h; 48h; 72h). The experimental units, or plots, were composed of test tubes with one cotton plant shoot partially immersed in oxalic acid solution. The biomass of the cotton shoots was evaluated at 24, 48, and 72 h. The loss of water caused by the exposure to oxalic acid indirectly indicates the level of cotton resistance to white mold. The results suggest that the most appropriate exposition time and concentration of oxalic acid for cotton evaluation of resistance to white mold were 20 mM at 48 hours, respectively. In these conditions, the cotton genotypes that presented the greatest average fresh mass were: UFU-14 A, UFU-14 B, UFU-14 F, UFU-14 H, and UFU-14 S, which could be used as sources of resistance to white mold in cotton breeding programs

Novel insights into water-deficit-responsive mRNAs and lncRNAs during fiber development in Gossypium hirsutum

BackgroundThe fiber yield and quality of cotton are greatly and periodically affected by water deficit. However, the molecular mechanism of the water deficit response in cotton fiber cells has not been fully elucidated.ResultsIn this study, water deficit caused a significant reduction in fiber length, strength, and elongation rate but a dramatic increase in micronaire value. To explore genome-wide transcriptional changes, fibers from cotton plants subjected to water deficit (WD) and normal irrigation (NI) during fiber development were analyzed by transcriptome sequencing. Analysis showed that 3427 mRNAs and 1021 long noncoding RNAs (lncRNAs) from fibers were differentially expressed between WD and NI plants. The maximum number of differentially expressed genes (DEGs) and lncRNAs (DERs) was identified in fibers at the secondary cell wall biosynthesis stage, suggesting that this is a critical period in response to water deficit. Twelve genes in cotton fiber were differentially and persistently expressed at ≥ five time points, suggesting that these genes are involved in both fiber development and the water-deficit response and could potentially be used in breeding to improve cotton resistance to drought stress. A total of 540 DEGs were predicted to be potentially regulated by DERs by analysis of coexpression and genomic colocation, accounting for approximately 15.76% of all DEGs. Four DERs, potentially acting as target mimics for microRNAs (miRNAs), indirectly regulated their corresponding DEGs in response to water deficit.ConclusionsThis work provides a comprehensive transcriptome analysis of fiber cells and a set of protein-coding genes and lncRNAs implicated in the cotton response to water deficit, significantly affecting fiber quality during the fiber development stage.

Ghmyb102 Promotes Drought Resistance in Cotton (Gossypium Hirsutum L.) by Regulating Drought-Responsive Genes and Aba Biosynthesis

Ghmyb102 Promotes Drought Resistance in Cotton (Gossypium Hirsutum L.) by Regulating Drought-Responsive Genes and Aba Biosynthesis

“Zerox” increases relatively the resistance of cotton to root rot

“Zerox” increases relatively the resistance of cotton to root rot

Preparation and characterization of a self-dispersed and reactive TiO2/BiOBr photocatalyst for self-cleaning and ultraviolet resistant cotton fabrics

A novel and efficient approach is presented herein to develop self-cleaning and ultraviolet resistant cotton fabrics loaded with a self-dispersed and reactive TiO2/BiOBr photocatalyst.

Response of Xanthomonas citri pv. malvacearum Isolates to Cotton Differing in Susceptibility to the Bacterium and Their Predicted Type III Effectors

From 2015 to 2020, 342 isolates of Xanthomonas citri pv. malvacearum (Xcm) were obtained from cotton fields in Texas, including 64 isolates collected from symptomatic cultivars that were thought to be resistant to race 18 strains of Xcm (the predominant race in the United States). Symptoms on highly resistant cultivars prompted concern that a new race of Xcm was present. The 342 isolates were inoculated on a race 18 susceptible (DP 1747NR B2XF) and race 18 resistant (S295) cotton cultivar, and none of the isolates caused blight-type symptoms (water soaking and chlorosis) on S295, indicating that the B12 resistant gene was still beneficial for disease management. Four cultivars, varying in their field response to bacterial blight, were inoculated with each of 17 isolates of Xcm, and the incidence of plants exhibiting bacterial blight symptoms averaged 87% for DP 1747NR B2XF, 51% for partially susceptible NG 4936 B3XF, 16% for partially resistant DP 1646 B2XF, and 0% for S295. Xcm isolates from Texas (11), Georgia (1), and Oklahoma (1) were sequenced, and their type three effectors (T3Es) were predicted. All isolates (Georgia, Oklahoma, and Texas) had the same T3E proteins as previously identified Xcm race 18 isolates (tested for 25 genes), including XopJ. Race 1, 2, 3, and 12 of Xcm included in the comparisons were all missing the XopJ gene. Use of cultivars with the B12 gene is an effective strategy to manage bacterial blight of cotton.

Calcium-Dependent Protein Kinase GhCDPK28 Was Dentified and Involved in Verticillium Wilt Resistance in Cotton.

Verticillium dahliae is a soil-borne fungus that causes vascular wilt through the roots of plants. Verticillium wilt caused by V. dahliae is one of the main diseases in cotton producing areas of the world, resulting in huge economic losses. Breeding resistant varieties is the most economical and effective method to control Verticillium wilt. Calcium-dependent protein kinases (CDPKs) play a pivotal role in plant innate immunity, including regulation of oxidative burst, gene expression as well as hormone signal transduction. However, the function of cotton CDPKs in response to V. dahliae stress remains unexplored. In this study, 96, 44 and 57 CDPKs were identified from Gossypium hirsutum, Gossypium raimondii and Gossypium arboretum, respectively. Phylogenetic analysis showed that these CDPKs could be divided into four branches. All GhCDPKs of the same clade are generally similar in gene structure and conserved domain arrangement. Cis-acting elements related to hormones, stress response, cell cycle and development were predicted in the promoter region. The expression of GhCDPKs could be regulated by various stresses. Gh_D11G188500.1 and Gh_A11G186100.1 was up-regulated under Vd0738 and Vd991 stress. Further phosphoproteomics analysis showed that Gh_A11G186100.1 (named as GhCDPK28-6) was phosphorylated under the stress of V. dahliae. Knockdown of GhCDPK28-6 expression, the content of reactive oxygen species was increased, a series of defense responses were enhanced, and the sensitivity of cotton to V. dahliae was reduced. Moreover, overexpression of GhCDPK28-6 in Arabidopsis thaliana weakened the resistance of plants to this pathogen. Subcellular localization revealed that GhCDPK28-6 was localized in the cell membrane. We also found that GhPBL9 and GhRPL12C may interact with GhCDPK28-6. These results indicate that GhCDPK28-6 is a potential molecular target for improving resistance to Verticillium wilt in cotton. This lays a foundation for breeding disease-resistant varieties.

Genome-Wide Identification and Characterization of GhCOMT Gene Family during Fiber Development and Verticillium Wilt Resistance in Cotton

Caffeic acid O-methyltransferases (COMTs) play an essential role in lignin synthesis procession, especially in the plant’s phenylalanine metabolic pathway. The content of COMT genes in cotton and the relationship between their expression patterns have not been studied clearly in cotton. In this study, we have identified 190 COMT genes in cotton, which were classified into three groups (I, II and III), and mapped on the cotton chromosomes. In addition, we found that 135 of the 190 COMT genes result from dispersed duplication (DSD) and whole-genome duplication (WGD), indicating that DSD and WGD were the main forces driving COMT gene expansion. The Ka/Ks analysis showed that GhCOMT43 and GhCOMT41 evolved from GaCOMT27 and GrCOMT14 through positive selection. The results of qRT-PCR showed that GhCOMT13, GhCOMT28, GhCOMT39 and GhCOMT55 were related to lignin content during the cotton fiber development. GhCOMT28, GhCOMT39, GhCOMT55, GhCOMT56 and GhCOMT57 responded to Verticillium Wilt (VW) and maybe related to VW resistance through lignin synthesis. Conclusively, this study found that GhCOMTs were highly expressed in the secondary wall thickening stage and VW. These results provide a clue for studying the functions of GhCOMTs in the development of cotton fiber and VW resistance and could lay a foundation for breeding cotton cultivates with higher quantity and high resistance to VW.

The GhMYB36 transcription factor confers resistance to biotic and abiotic stress by enhancing PR1 gene expression in plants.

SummaryDrought and Verticillium wilt disease are two main factors that limit cotton production, which necessitates the identification of key molecular switch to simultaneously improve cotton resistance to Verticillium dahliae and tolerance to drought stress. R2R3‐type MYB proteins could play such a role because of their conserved functions in plant development, growth, and metabolism regulation, however, till date a MYB gene conferring the desired resistance to both biotic and abiotic stresses has not been found in cotton. Here, we describe the identification of GhMYB36, a gene encoding a R2R3‐type MYB protein in Gossypium hirsutum, which confers drought tolerance and Verticilium wilt resistance in both Arabidopsis and cotton. GhMYB36 was highly induced by PEG‐simulated drought stress in G. hirsutum. GhMYB36‐silenced cotton plants were more sensitive to both drought stress and Verticillium wilt. GhMYB36 overexpression in transgenic Arabidopsis and cotton plants gave rise to improved drought tolerance and Verticillium wilt resistance. Transient expression of fused GhMYB36‐GFP in tobacco cells was able to localize GhMYB36 in the cell nucleus. In addition, RNA‐seq analysis together with qRT‐PCR validation in transgenic Arabidopsis overexpressing GhMYB36 revealed significantly enhanced PR1 expression. Luciferase interaction assays indicated that GhMYB36 are probably bound to the promoter of PR1 to activate its expression and the interaction, which was further verified by Yeast one hybrid assay. Taken together, our results suggest that GhMYB36 functions as a transcription factor that is involved in drought tolerance and Verticillium wilt resistance in Arabidopsis and cotton by enhancing PR1 expression.

The Respiratory Burst Oxidase Homolog Protein D (GhRbohD) Positively Regulates the Cotton Resistance to Verticillium dahliae.

Verticillium wilt, mainly caused by a soil-inhabiting fungus Verticillium dahliae, can seriously reduce the yield and quality of cotton. The complex mechanism underlying cotton resistance to Verticillium wilt remains largely unknown. In plants, reactive oxygen species (ROS) mediated by Rbohs is one of the earliest responses of plants to biotic and abiotic stresses. In our previous study, we performed a time-course phospho-proteomic analysis of roots of resistant and susceptible cotton varieties in response to V. dahliae, and found early differentially expressed protein burst oxidase homolog protein D (GhRbohD). However, the role of GhRbohD-mediated ROS in cotton defense against V. dahliae needs further investigation. In this study, we analyzed the function of GhRbohD-mediated resistance of cotton against V. dahliae in vitro and in vivo. Bioinformatics analysis showed that GhRbohD possessed the conservative structural attributes of Rbohs family, 12 members of RbohD out of 57 Rbohs in cotton. The expression of GhRbohD was significantly upregulated after V. dahliae inoculation, peaking at 6 hpi, and the phosphorylation level was also increased. A VIGS test demonstrated that ROS production, NO, H2O2 and Ca2+ contents of GhRbohD-silenced cotton plants were significantly reduced, and lignin synthesis and callose accumulation were damaged, important reasons for the impairment of GhRbohD-silenced cotton’s defense against V. dahliae. The expression levels of resistance-related genes were downregulated in GhRbohD-silenced cotton by qRT-PCR, mainly involving the lignin metabolism pathway and the jasmonic acid signaling pathway. However, overexpression of GhRbohD enhanced resistance of transgenic Arabidopsis to V. dahliae challenge. Furthermore, Y2H assays were applied to find that GhPBL9 and GhRPL12C may interact with GhRbohD. These results strongly support that GhRbohD activates ROS production to positively regulate the resistance of plants against V. dahliae.

Influence of biopreparation ‘Zamin-M’ on cotton plants (gossypium hirsutum) under soil salinization in Uzbekistan

In this article, a microbial inoculant acting as a biofertilizer is considered as a biostimulant, consisting of three strains of local salt-tolerant strains of rhizobacteria Pseudomonas stutzeri SKB308, Bacillus subtilis SKB309, Bacillus megaterium SKB310. The authors have developed and introduced into agricultural practice an experimental sample of the biological product ‘Zamin-M’, which increases the resistance of cotton to stress conditions of soil salinity. It is proved that the biological product ‘Zamin-M’ increases soil fertility has a beneficial effect on the enzymatic activity of the soil and the development of a population of beneficial microorganisms. The bioproduct ‘Zamin-M’ is presented as a new stimulant in the practice of growing cotton in saline conditions. An agricultural technique for the use of a local biological product in agriculture has been developed. The ‘Zamin-M’ is included by the State Committee of the Republic of Uzbekistan in the list of agrochemicals and pesticides permitted for use in agriculture of the Republic of Uzbekistan (certificate No. 1A1005).

Enhancing the resilience of transgenic cotton for insect resistance.

The efficacy of Bt crystal proteins has been compromised due to their extensive utilization in the field. The second-generation Bt vegetative insecticidal proteins could be the best-suited alternative to combat resistance build-up due to their broad range affinity with midgut receptors of insects. The codon-optimized synthetic vegetative insecticidal proteins (Vip3Aa) gene under the control of CaMV35S promoter was transformed into a locally developed transgenic cotton variety (CKC-01) expressing cry1Ac and cry2A genes. Transformation efficiency of 1.63% was recorded. The highest Vip3Aa expression (51.98-fold) was found in MS3 transgenic cotton plant. Maximum Vip3Aa protein concentration (4.23µg/mL) was calculated in transgenic cotton plant MS3 through ELISA. The transgenic cotton plant (MS3) showed one copy number on both chromatids in the homozygous form at chromosome 8 at the telophase stage. Almost 99% mortality of H. armigera was recorded in transgenic cotton plants expressing double crystal proteins pyramided with Vip3Aa gene as contrasted to transgenic cotton plant expressing only double crystal protein with 70% mortality. The results obtained during this study suggest that the combination of Bt cry1Ac, cry2A, and Vip3Aa toxins is the best possible alternative approach to combat chewing insects.

GhPLP2 Positively Regulates Cotton Resistance to Verticillium Wilt by Modulating Fatty Acid Accumulation and Jasmonic Acid Signaling Pathway.

Patatin-like proteins (PLPs) have non-specific lipid acyl hydrolysis (LAH) activity, which can hydrolyze membrane lipids into fatty acids and lysophospholipids. The vital role of PLPs in plant growth and abiotic stress has been well documented. However, the function of PLPs in plant defense responses against pathogens is still poorly understood. Here, we isolated and identified a novel cotton (Gossypium hirsutum) PLP gene GhPLP2. The expression of GhPLP2 was induced upon treatment with Verticillium dahliae, the signaling molecules jasmonic acid (JA) and ethylene (ETH) in cotton plants. Subcellular localization revealed that GhPLP2 was localized to the plasma membrane. GhPLP2-silenced cotton plants were more susceptible to infection by V. dahliae, while the overexpression of GhPLP2 in Arabidopsis enhanced its resistance to V. dahliae, which was apparent as mild symptoms, and a decrease in the disease index and fungal biomass. The hypersensitive response, deposition of callose, and H2O2 accumulation triggered by V. dahliae elicitor were reduced in GhPLP2-silenced cotton plants. The overexpression of GhPLP2 in Arabidopsis resulted in the accumulation of linoleic acid (LA, 18:2) and α-linolenic acid (ALA, 18:3) and facilitated the biosynthesis of JA and JA-mediated defensive responses. GhPLP2 silencing in cotton plants consistently reduced the accumulation of linoleic acid (LA, 18:2) and α-linolenic acid (ALA, 18:3) and suppressed the biosynthesis of JA and the defensive responses mediated by JA. These results indicate that GhPLP2 is involved in the resistance of cotton to V. dahliae by maintaining fatty acid metabolism pools for JA biosynthesis and activating the JA signaling pathway.

Improvement in the Design and Methods of Calculation of Parameters of Vibration Multifaceted Gridirons of Natural Fibre Cleaners

The article outlines the operating concept of the recommended multifaceted gridirons on an elastic bearing support, analyses the excursion of the gridiron, justifies the characteristics and gives the results of comparative production tests. As well as the effective scheme and principle of work of cotton cleaner from large impurities are presented based on the research of fluctuations in the parameters of a grid-iron on an elastic support system. The obtained results of the recommended cotton cleaner were provided. Experiment results showed that the cleaning purificatory effect increases on average by up to 8.11% in comparison with the existing option of a fire grate; mechanical damage to seeds decreases by up to 1.09%, and free fibre in the seed-cotton decreases two times, to 0.113%. With an increase in the resistance of cotton from 19.7 Н up to 60 Н (average value), the movement of the gridiron increases from 0.65·10-3 m up to 3.2·10-3 m. This is due to the fact that in the interaction of raw cotton with vibroisolating conical grates, cotton-yarn is additionally shaken, increasing their direction of movement due to the taper rate of the gridirons, which leads to an increase in the cleaning effect.

Transcriptome Analysis of a Cotton Cultivar Provides Insights into the Differentially Expressed Genes Underlying Heightened Resistance to the Devastating Verticillium Wilt.

Cotton is an important economic crop worldwide. Verticillium wilt (VW) caused by Verticillium dahliae (V. dahliae) is a serious disease in cotton, resulting in massive yield losses and decline of fiber quality. Breeding resistant cotton cultivars is an efficient but elaborate method to improve the resistance of cotton against V. dahliae infection. However, the functional mechanism of several excellent VW resistant cotton cultivars is poorly understood at present. In our current study, we carried out RNA-seq to discover the differentially expressed genes (DEGs) in the roots of susceptible cotton Gossypium hirsutum cultivar Junmian 1 (J1) and resistant cotton G. hirsutum cultivar Liaomian 38 (L38) upon Vd991 inoculation at two time points compared with the mock inoculated control plants. The potential function of DEGs uniquely expressed in J1 and L38 was also analyzed by GO enrichment and KEGG pathway associations. Most DEGs were assigned to resistance-related functions. In addition, resistance gene analogues (RGAs) were identified and analyzed for their role in the heightened resistance of the L38 cultivar against the devastating Vd991. In summary, we analyzed the regulatory network of genes in the resistant cotton cultivar L38 during V. dahliae infection, providing a novel and comprehensive insight into VW resistance in cotton.

SEVEN IN ABSENTIA Ubiquitin Ligases Positively Regulate Defense Against Verticillium dahliae in Gossypium hirsutum.

Ubiquitination is a post-translational regulatory mechanism that controls a variety of biological processes in plants. The E3 ligases confer specificity by recognizing target proteins for ubiquitination. Here, we identified SEVEN IN ABSENTIA (SINA) ubiquitin ligases, which belong to the RING-type E3 ligase family, in upland cotton (Gossypium hirsutum). Twenty-four GhSINAs were characterized, and the expression levels of GhSINA7, GhSINA8, and GhSINA9 were upregulated at 24 h after inoculation with Verticillium dahliae. In vitro ubiquitination assays indicated that the three GhSINAs possessed E3 ubiquitin ligase activities. Transient expression in Nicotiana benthamiana leaves showed that they localized to the nucleus. And yeast two-hybrid (Y2H) screening revealed that they could interact with each other. The ectopic overexpression of GhSINA7, GhSINA8, and GhSINA9 independently in Arabidopsis thaliana resulted in increased tolerance to V. dahliae, while individual knockdowns of GhSINA7, GhSINA8, and GhSINA9 compromised cotton resistance to the pathogen. Thus, GhSINA7, GhSINA8, and GhSINA9 act as positive regulators of defense responses against V. dahliae in cotton plants.

Universal fabrication of superhydrophobic and UV resistant cotton fabric with polyphenols

Universal fabrication of superhydrophobic and UV resistant cotton fabric with polyphenols

Rapid Mining of Candidate Genes for Verticillium Wilt Resistance in Cotton Based on BSA-Seq Analysis.

Cotton is a globally important cash crop. Verticillium wilt (VW) is commonly known as “cancer” of cotton and causes serious loss of yield and fiber quality in cotton production around the world. Here, we performed a BSA-seq analysis using an F2:3 segregation population to identify the candidate loci involved in VW resistance. Two QTLs (qvw-D05-1 and qvw-D05-2) related to VW resistance in cotton were identified using two resistant/susceptible bulks from the F2 segregation population constructed by crossing the resistant cultivar ZZM2 with the susceptible cultivar J11. A total of 30stop-lost SNPs and 42 stop-gained SNPs, which included 17 genes, were screened in the qvw-D05-2 region by SnpEff analysis. Further analysis of the transcriptome data and qRT-PCR revealed that the expression level of Ghir_D05G037630 (designated as GhDRP) varied significantly at certain time points after infection with V. dahliae. The virus-induced gene silencing of GhDRP resulted in higher susceptibility of the plants to V. dahliae than the control, suggesting that GhDRP is involved in the resistance to V. dahlia infection. This study provides a method for rapid mining of quantitative trait loci and screening of candidate genes, as well as enriches the genomic information and gene resources for the molecular breeding of disease resistance in cotton.