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Genome-Wide Identification and Analysis of Dof Gene Family in Upland Cotton
Ju Longzhen, Zhao Ting, Fang Lei, Hu Yan, Zhang Tianzhen
Cotton Science    2020, 32 (4): 279-291.   DOI: 10.11963/1002-7807.jlzhy.20200630
Abstract678)      PDF(pc) (14548KB)(272)       Save
[Objective] The DNA binding with one finger proteins (Dof) are plant-specific transcription factors that play a very important role in plant growth, development and in response to abiotic stress. The aim of this study was to provide a reference for further functional study of the Dof genes associated with plant growth and development by genome-wide analysis of Dof transcription factor family in Gossypium hirsutum. [Method] Using the recently released upland cotton genomic data, the Dof genes in G. hirsutum were identified by bioinformatic method. The physico-chemical properties, sequence characteristics, gene duplication, phylogeny and expression pattern of Dof genes were analyzed. [Result] A total of 118 Dof genes were identified in G. hirsutum. Based on phylogenetic analysis, these Dof genes were divided into nine subfamilies, and the genes within the same subfamily had similar exon-intron organization and motif distribution. Gene duplication analysis revealed that whole genome duplication was the main contributor to the expansion of Dof genes in G. hirsutum. Cis-acting element analysis showed the promoters of Dof genes had cis-acting regulatory elements involved in stresses responses to different plant hormones. The results of RNA-seq data analysis revealed that Dof genes expression in G. hirsutum was classified into three types which had different expression pattern in different tissues, developmental stages and stresses, indicating gene functional differentiation. [Conclusion] The genome-wide identification and analysis of Dof gene family in G. hirsutum is helpful to understand the evolution and function, and provides a support for future studies.
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Identification of MADS-box Family and Analysis of Tissue Specific Expression in Gossypium hirsutum L.
Zhang Ai, Wang Caixiang, Su Junji, Zhang Xianliang, Shi Chunhui, Liu Juanjuan, Peng Yunling, Ma Xiongfeng
Cotton Science    2020, 32 (5): 404-417.   DOI: 10.11963/1002-7807.zamxf.20200831
Abstract261)      PDF(pc) (6829KB)(133)       Save
[Objective] MADS-box is an important transcription factor regulating the growth and development processes in cotton. Identification and screening upland cotton MADS-box genes is indispensible for accelerating their studying of biological functions. [Method] This article based on the upland cotton TM-1 reference genome assembled in 2019, all members of upland cotton MADS-box family were identified through HMMER 3.0 software. MapInspect, MEGA 7.0, MEME, TBtools and omicshare website were utilized to analyze chromosome location, phylogenetic relationship, motif prediction, gene structure, and tissue-specific expression of MADS-box genes. [Result] One hundred and eighty one upland cotton MADS-box genes (66 Type Ⅰ and 115 MIKC genes) were identified in total, which located on 26 chromosomes. The Type I proteins were scattered on 3 subfamilies and the MIKC proteins were divided into MIKC* and MIKCC which contain 10 subfamilies. The prediction results of motif showed that all proteins of MADS-box family contain MADS domain and gene structure analysis revealed that the structure and length of exons and introns are similar in one subfamily. Tissue-specific expression analysis found that 33 MADS-box genes were mainly expressed in fibrous tissue, 103 genes were primarily expressed in floral organs, and 41 genes were strongly expressed in roots and stems. [Conclusion] The MADS-box genes related to flowering regulation and fiber development of upland cotton were obtained in the study, which has theoretical significance for revealing the genetic regulation mechanism of cotton fiber quality and has application value for molecular breeding.
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Genome-Wide Identification and Functional Analysis of the NF-YA Gene Family in Gossypium hirsutum L.
Pan Ao, Wang Jingjing, Sun Fulai, Zhang Jingxia, Gao Yang, Du Zhaohai, Jiao Mengjia, Zhang Jun, Wang Furong, Liu Zhi
Cotton Science    2020, 32 (4): 316-328.   DOI: 10.11963/1002-7807.palz.20200611
Abstract244)      PDF(pc) (7726KB)(114)       Save
[Objective] The NF-YA gene family was identified in upland cotton (Gossypium hirsutum L.), and their expression characteristics were analyzed to discover the genes related to cotton flowering regulation. [Method] Using bioinformatic methods, the NF-YA genes were identified and their physical and chemical properties, gene structure, cis-element, collinearity, Ka/K swere investigated. The expression pattern was performed using quantitative real-time polymerase chain reaction (qRT-PCR). The gene function was verified by virus-induced gene silencing (VIGS). [Result] A total of 29 GhNF-YA genes were identified from upland cotton genome, these genes were divided into five clades and distributed on 18 chromosomes. The expansion of the NF-YA genes in upland cotton was mainly driven by whole genome duplication and segmental duplication. A lot of light responding elements were identified in the promoter regions of GhNF-YA genes. According to the expression profiling, the GhNF-YA genes were highly expressed in stems and leaves. Meanwhile, analysis of qRT-PCR showed that 12 GhNF-YA genes had a relatively high expression in the leaf of both early-maturing cotton cultivar Lumianyan 19 and late-maturing cotton cultivar Lumianyan 37 from three- to six-leaf stage, and expression of the most genes was significantly different between the two cultivars. The function of GhNF-YA18 in Lumianyan 37 was verified by VIGS, and the results indicated that the bud emergence in silenced plants was 11 d earlier than that of unsilenced control plants. In addition, the expression of GhNF-YA18 was lower in silenced plants than unsilenced plants at albino and budding stages. [Conclusion] In this study, we identified and characterized the 29 GhNF-YA genes in upland cotton. The results will provide a foundation for future studies in molecular regulatory mechanisms of cotton early fiowering.
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Identification and Analysis of Fiber Quality and Yield Related Traits of Interspecific  (Gossypium hirsutum L. × G. barbadense L.) Hybrids
Li Tengyu, Xu Chao, Li Yaoming, Gou Chengfei, Hong Zhu, Ding Mingquan, Sun Chendong
Cotton Science    2020, 32 (4): 348-359.   DOI: 10.11963/1002-7807.ltyscd.20200612
Abstract238)      PDF(pc) (1238KB)(95)       Save
[Objective] The aim of this study is to study the hereditary of heterosis of fiber quality and yield-related traits in the upland-island interspecific hybrids, and breed new interspecific hybrid varieties with high yield and fine fiber quality. [Method] In this study, 12 upland cotton materials and 5 sea-island cotton materials were selected to determine the fiber quality and yield traits of their parents and F1 in Lin’an, Zhejiang and Sanya, Hainan. [Result] It was found that fiber length and fiber strength of F1 (Gossypium hirsutum × G. barbadense) generally had significant mid-parent heterosis (MPH), some hybrid combinations showed strong over-parent heterosis (OPH), fiber length had a small coefficient of variation between the two places and could be stably inherited. And in terms of yield, seed cotton weight, lint weight, and lint percentage of some upland-island hybrids had MPH, but they were still significantly lower than those of upland cotton parents. [Conclusion] Two long-staple cotton hybrid combinations T035 and T044 with 5A grade high-quality were obtained, and an excellent material of G. barbadense Ta10-280 was screened. This study provides valuable data for the genetic law of fiber quality heterosis of upland-island hybrid cotton.
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Biocontrol Effect of Bacillus velezensis Strain SZAD1 on Verticillium dahliae
Zhang Qiong, Zabihullah Sherzad, Tang Canming
Cotton Science    2020, 32 (4): 329-338.   DOI: 10.11963/1002-7807.zqtcm.20200609
Abstract235)      PDF(pc) (5811KB)(64)       Save
[Objective] Verticillium wilt is the main fungal disease of cotton, lacking high resistant varieties and safe and effective chemical fungicides. Biological control of Verticillium wilt is of great significance to cotton production. The purpose of this study was to analyze the antagonistic activity and biocontrol potential of endophytic bacteria in cotton against Verticillium dahliae. [Method] SZAD1 was identified by 16S rDNA and gyrB sequence analysis. The ability of SZAD1 to inhibit the mycelial growth of V.dahliae strain VD080 was tested by the agar plate test, and the control effect on Verticillium wilt was tested by the seed soaking and root filling method. The agar plate containing carboxy methyl cellulose (CMC) or chitin was prepared to test whether the strain can secrete cellulase and chitinase, and the enzyme activity was determined by the dinitrosalicylic acid reagent method. [Result] SZAD1 belongs to Bacillus velezensis. SZAD1 can significantly inhibit the growth of VD080 mycelium. The control effect of SZAD1 on Verticillium wilt was 60.10% and 56.00% in the seed soaking and root filling  experiments respectively. SZAD1 strain can produce cellulase and chitinase with the maximum activity at 72 h after culture. The antibacterial effect of fermented supernatant was the strongest at 72 h after culture. The content of VD080 spores in liquid potato dextrose agar medium decreased by 30.20% and 96.53% compared with the control respectively after adding supernatant with concentrations of 25 mg·L-1 and 50 mg·L-1. [Conclusion] The supernatant of bacteria SZAD1 can reduce the colonization of VD080 in the stem of cotton plant by root filling and decreased the withering degree of cotton leaves. SZAD1 has the potential of controlling cotton Verticillium wilt as a biocontrol bacteria.
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Cloning and Expression Profiling of Cuticular Protein Genes of Spodoptera litura
Zhao Peng, Zhang Shuai, Zhao Chenchen, Hu Fangmei, Cui Jinjie, Li Shaoqin
Cotton Science    2020, 32 (4): 292-304.   DOI: 10.11963/1002-7807.zplsq.20200521
Abstract233)      PDF(pc) (5483KB)(68)       Save
[Objective] The expression pattern of fourteen cuticular protein (CP) genes (ICPG) in Spodoptera litura were analyzed in this study, which provided reference for further study on the function of cuticular proteins in the growth and development of S. litura. [Method] Polymerase chain reaction (PCR) was used to amplify the cDNAs of the genes encoding CPs of S. litura. Their nucleotide sequences and corresponding amino acid sequences were analyzed, and lepidopteran phylogenetic tree was constructed using MEGA5.2 software. The expression of CP genes was analyzed in the third to the sixth instar larvae and 2-day-old adult genital organs by the reverse transcription PCR (RT-PCR) technique. [Result] Fourteen cDNAs coding 14 CPs of S. litura were obtained. The sequence analysis showed 14 CPs harbor the conserved motif of 35-36 amino acids that is a typical feature of chitin-binding proteins and share high sequence homology with CPs of Helicoverpa armigera. RT-PCR results showed that these CP genes were expressed in the third to the sixth instar larvae and 2-day-old adults. In particular, their relative expression levels are higher in the cuticulars of older larvae and lower in the fat bodies of younger larvae. In addition, the relative expression of 14 CP genes was low in testes and ovaries. The highest relative expression of SlICPG-11 and SlICPG-14 in the pupal stage. [Conclusion] In this study, the cDNA of 14 S. litura CP genes were cloned. Their expression exhibited tissue and developmental stage specificity indicating their putative roles in S. litura development. It lays the foundation for target site selection for the control of S. litura.
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Cloning and Functional Analysis of Low Temperature Response Gene, GhZAT10, in  Upland Cotton at Seedling Stage
Yang Yongfei , Ge Changwei, Shen Qian, Zhang Siping, Liu Shaodong, Ma Huijuan, Chen Jing, Liu Ruihua, Li Shicong, Zhao Xinhua, Li Zhikun, Pang Chaoyou
Cotton Science    2020, 32 (4): 305-315.   DOI: 10.11963/1002-7807.yyfpcy.20200703
Abstract233)      PDF(pc) (6553KB)(108)       Save
[Objective] The aim of this study is to identify chilling tolerant genes which can provide a basis for breeding upland cotton cultivars with tolerance to chilling. [Method] The GhZAT10 (Zinc finger of Arabidopsis thaliana 10) gene was cloned, and its expression in roots, stems, and leaves was analyzed with cold treatment by quantitative real-time polymerase chain reaction (qRT-PCR). Then we analyzed the structural characteristics of GhZAT10, protein properties and constructed the phylogenetic tree by bioinformatics methods. Next, we constructed subcellular localization vector 35S::GhZAT10-GFP by Gateway technology. Finally, we studied the effect of GhZAT10 in chilling response by virus induced gene silencing of cotton. [Result] The Open reading frame(ORF) length of GhZAT10 gene is 813 bp (base pair), encoding 270 amino acid residues. The expression of GhZAT10 in roots was higher than that in stems and leaves, and up-regulated after chilling stress in these tissues. GhZAT10 does not possess signal peptides or transmembrane helices; its activity is closely related to the phosphorylation regulation. GhZAT10 has homology with the ZAT10 of Theobroma cacao, Arabidopsis thaliana and Citrus sinensis. GhZAT10 protein is located in the nucleus. The GhZAT10-silenced cotton plants were more sensitive to low temperature than wild type. [Conclusion] GhZAT10 belongs to C2H2 zinc finger protein, and plays a positive role to response the chilling stress in upland cotton.
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Effect of Coronatine on AsA-GSH Cycle of Cotton Seedling under Low Temperature Stress
Li Jin, Zhai Menghua, Yu Chunxin, Wang Li, Zhang Jungao, Zhou Xiaoyun, Liang Jing, Duan Liusheng, Lei Bin
Cotton Science    2020, 32 (5): 381-391.   DOI: 10.11963/1002-7807.ljlb.20200729
Abstract218)      PDF(pc) (3693KB)(88)       Save
[Objective] In order to explore the stress-resistant mechanism of coronatine, an indoor experiment was conducted to study the effect of coronatine on ascorbate-glutathione circulatory system of the vegetative organs of cotton seedling under low temperature stress. [Method] Xinluzao 57 was used as the experimental material, when the seedlings reached the two-leaf stage, they were treated at 25 ℃ and 4 ℃ after spraying water, marked as CK and LT; the same treatment was conducted after spraying 0.01 μmol·L-1 coronatine, marked as COR and (LT+COR). After 1 d of treatment, the roots, stems and leaves of cotton seedling were collected to determine the antioxidant contents and antioxidant enzyme activities. [Result] Compared with the control treatment (CK), the APX, MDHAR activities and DHA, GSH, glutathione contents decreased, the GPX activity, AsA and ascorbate contents increased, and no significant effect on DHAR and GR activities were observed in roots, stems and leaves of cotton seedling after spraying water at low temperature (LT), while in leaves, the GSSG content decreased, AsA-DHA ratio and GSH-GSSG ratio showed the maximum increase. Compared with the LT treatment, spraying COR at low temperature (LT+COR) showed increased APX, MDHAR, DHAR, GPX, GR activities and AsA, DHA, GSH, ascorbate contents; glutathione content in roots, stems and leaves of cotton seedling, and GSSG content, AsA-DHA ratio and GSH-GSSG ratio obviously changed in leaves. [Conclusion] Spraying COR at low temperature could regulate the AsA-GSH metabolism and alleviate the damage caused by low temperature to cotton seedling. COR had the strongest relieving effect on seedling leaves.
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Study Crystallinity of the Developing Cotton Fibers by Micro-Fourier Transform Infrared Spectroscopy (FTIR) and X-Ray Diffraction (XRD)
Zhang Lipeng, Zhang Shiding, Xu Peng, Li Xianchang, Zhang Zhen, Fan Senmiao, Gong Juwu, Yuan Youlu, Shang Haihong, Zou Huawen
Cotton Science    2020, 32 (4): 370-380.   DOI: 10.11963/1002-7807.zlpzhw.20200610
Abstract211)      PDF(pc) (5428KB)(47)       Save
[Objective] The change of crystalline index (CI) of two different cultivated cotton fibers at the development stage was studied by micro-Fourier transform infrared spectroscopy (micro-FTIR) and X-ray diffraction (XRD). The feasibility was verified by measuring CI in developing cotton fibers with micro-FTIR method. And it was utilized to estimate the CI of mature cotton fibers. [Method] Upland cotton 0-153 and sea island cotton S-6 were selected as examined materials. The cotton fibers of 2 varieties were obtained at 5-30 d post anthesis respectively, with a sampling interval of 5 d. After obtaining the fiber, it was washed several times with distilled water and then placed in an oven at 40℃ for 48 hours. After drying, the FTIR and XRD spectra of each sample were obtained. The CI is calculated according to 4 different FTIR-CI calculation methods, the crystallinity changes of different cotton varieties in the developing period were compared, and the correlation between the FTIR-CI and XRD-CI was studied. [Result] Only FTIR-CI calculated by the Carrillo-Colom index(FTIR-CCI) had a good correlation with XRD-CI method, with higher R2 than 0.9 of both varieties. The fitting model (IR-CI) between the FTIR-CCI and XRD-CI was used to calculate the CI of the twenty-three randomly selected mature fibers. The results showed that the accuracy of IR-CI was good, and the XRD-CI results were within the error range of the calculated results according to the IR-CI model, while the precision of IR-CI could not reach anticipation. [Conclusion] Micro-FTIR can be used to study the change of cellulose in cotton fiber during the developing period. The model of IR-CI established by the correlation between the FTIR-CCI and XRD-CI, can be used to evaluate the crystallinity of developing period in cotton fiber. However, for the study of the crystallinity of mature fibers, it is necessary to use a large number of samples in the later experiment to establish an optimized model.
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The Effects of Increased-Density on Canopy Apparent Photosynthesis, Dry Matter Accumulation and Distribution of Cotton under Late-Sown Condition
Li Hui, Wan Hualong, Tian Liwen, Liu Liantao, Zhang Yongjiang, Bai Zhiying, Zhang Ke, Wang Guoping, Sun Hongchun, Li Cundong
Cotton Science    2020, 32 (4): 339-347.   DOI: 10.11963/1002-7807.lhlcd.20200622
Abstract204)      PDF(pc) (3579KB)(104)       Save
[Objective] The effects of high planting densities on canopy apparent photosynthesis, leaf area index, dry matter accumulation and distribution features of cotton were studied, with purpose to elucidate the suitable density for late cotton-sown system in the Yellow River cultivation region. [Method] Field experiments including two planting density treatments (D1 of 9.0×104 plant·hm-2 and D2 of 12.0×104 plant·hm-2) were conducted during the 2017 and 2018 growth seasons, using cultivars Nongda 601(ND-601) and Guoxin Cotton 9(GX-9) in 2017 and Nongda 601 in 2018 as the materials. The effects of densities on photosynthetic characteristics and yield components were investigated. [Result] At fast growth stage, the leaf area index under D2 was significantly higher than that of D1, which sustained longer peak duration under the former condition. These results suggested that higher density can promote the generation of enlarged canopy structure at vigorous growth stage. However, much intensified leaf area index resulted in population shading and reduced canopy apparent photosynthesis. Although D2 treatment was shown to benefit the biomass accumulation of the population plants and the vegetative organs. But the enhanced vegetative growth led to restriction on the reproductive tissue, which resulted in lowered nutrient distribution to reproductive organ; the increase of density was no significant difference on boll weight and lint percentage. Perhaps because of the interannual climate variation, the results of other yield components obtained across the two growth seasons were inconsistent with each other. [Conclusion] Under the suitable conditions, D1 treatment is beneficial to establish the reasonable population structure and to achieve stable yield. In contrast, the D2 treatment has the potential to achieve higher yield. Our results suggested that D2 together with adoption of chemical control technology can help high-yielding cultivation, through increasing population dry matter amount, improving nutrient distribution across organs, and elevating harvest index of the cotton plants.
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Regulation of Free Amino Acid in Cotton at the Seedling Stage by Aphis gossypii Feeding
Ma Yan, Zhang Shuai, Luo Junyu, Zhu Xiangzhen, Gao Xueke, Cui Jinjie
Cotton Science    2020, 32 (4): 360-369.   DOI: 10.11963/1002-7807.mycjj.20200526
Abstract176)      PDF(pc) (962KB)(51)       Save
[Objective] Free amino acids are important nitrogen nutrients, and their changes reflect the interaction between insects and host plants. The purpose of this experiment was to study the effect of aphid feeding on the free amino acid content of host cotton in seedling stage. [Method] The seedling cotton infected by aphid with different damage grades was selected from the field and analyzed the content of free amino acid in euphylla, cotyledon, hypocotyl and root. [Result] The content of free amino acids in cotton tissues increased after feeding by Aphis gossypii, and the content of free amino acids in cotyledon, hypocotyl and root is more than 2 times of the level 0. The content of non-essential amino acids increased more significantly than that of essential amino acids. The changes of single free amino acid content in different tissues were not completely consistent, and the Asn and Glu increased significantly in all tissues. [Conclusion] The feeding behavior of cotton aphid affects the nutritional level of cotton tissue and the content of free amino acid increased significantly. 
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GhMYB43 Negatively Regulates Lignin Biosynthesis and Jasmonic Acid Signaling
Shen Jili, Xiao Shenghua, Xi Hui, Nurimanguli Aini, Hu Qin, Zhang Xiaojun, Yang Zhaoguang, Nie Xinhui , Zhu Longfu
Cotton Science    2020, 32 (6): 522-537.   DOI: 10.11963/1002-7807.sjlzlf.20200907
Abstract163)      PDF(pc) (10912KB)(289)       Save
[Objective] Verticillium wilt is a vascular fungal disease spread by soil, which causes serious loss of cotton yield and reduction in quality every year. This study provides a theoretical basis for the innovation of cotton multi-resistant germplasms by identifying disease resistance genes and studying disease resistance mechanisms. [Method] Yeast one hybrid was performed to screen the upstream regulatory factors of GhLac1, and the construction of phylogenetic trees, multiple alignment of amino acid sequences, reverse transcription-quantitative polymerase chain reaction (RT-qPCR), transient tobacco transformation, dual luciferase assay system, virus-induced gene silencing and cotton genetic transformation technology were performed to verify the function of this transcription factor. [Result] Gh_D12G0544 is the most homologous to AtMYB43 in Arabidopsis thaliana, so its encoding gene was named GhMYB43. GhMYB43 is located on chromosome 12 of the Dt subgroup of upland cotton and encodes a protein containing 376 amino acids that contains two MYB domains. RT-qPCR analysis showed that GhMYB43 was predominantly transcribed in the stem and was up-regulated by salicylic acid (SA) and H2O2, but down-regulated by methyl jasmonate (Me-JA); at the same time, the transcription level was induced by Verticillium dahliae. Subcellular localization results showed that GhMYB43 protein is localized in the nucleus. The dual luciferase assay system verified that the gene has transcriptional activation activity. Disease resistance identification found that the inhibition GhMYB43 transcription level enhanced the resistance of cotton to V. dahliae, and overexpression of GhMYB43 increased the susceptibility of plants to this pathogen. Histochemical staining and content determination of lignin showed that the lignin content of GhMYB43 overexpressing transgenic lines was significantly lower than that of control materials. RT-qPCR analysis showed that GhMYB43 negatively regulates the transcription level of enzyme genes involved in the lignin synthesis and JA signaling pathway. [Conclusion] GhMYB43 negatively regulates lignin synthesis and JA signal.
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Chemical Defoliation of Machine-Harvested Cotton was Accompanied by Intense Ethylene and Cytokinin Signal Responses
Gao Yu, Xu Jiao, Zhang Bing, Sun Weinan, Yang Xiyan
Cotton Science    2020, 32 (6): 491-500.   DOI: 10.11963/1002-7807.gyyxy.20201105
Abstract151)      PDF(pc) (4774KB)(55)       Save
[Objective] Chemical defoliation is a necessary technical measure for machine-harvested cotton, and the effect of chemical defoliation directly affects the efficiency of mechanical-picking and fiber quality. Study on the molecular mechanism of defoliant in regulating defoliation could effectively guide the screening and improvement of defoliant-sensitive cotton germplasms. [Method] In this study, 200 cotton accessions were collected and planted in greenhouse, and treated with defoliant at the early flowering stage. Different varieties with high or low sensitivity to defoliant were selected according to the defoliation rate on the 4th day after treatment. The selected cotton varieties were also treated with defoliant at the boll opening stage in the field, and the defoliation rate on the 7th day after treatment was calculated. Meanwhile, samples from the abscission zone of the  selected varieties were collected on the first and third day after treatment. And the expression of ethylene-, cytokinin- and hydrolase- related genes were analyzed by quantitative real-time polymerase chain reaction (qRT-PCR). [Result] Six defoliant sensitive and 6 insensitive varieties were selected from greenhouse and field tests. According to the qRT-PCR results, cytokinin-related genes, ethylene-related genes and hydrolase-related genes showed different expression patterns following defoliant treatment in different varieties, and most of these genes showed significant different expression in the defoliant sensitive and insensitive varieties. [Conclusion] This study indicates that different cotton germplasms respond to differently to defoliant, which might be the result of differential expression of cytokinin- and ethylene-related genes.
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Analysis of Cotton Secondary Metabolites under the Interaction between Verticillium dahliae and Upland Cotton
Li Shezeng, Niu Luxin, Li Bochao, Chen Xiuye, Liu Chang, Lu Xiuyun, Guo Qinggang, Ma Ping, Ma Zhiying
Cotton Science    2020, 32 (6): 501-521.   DOI: 10.11963/1002-7807.lszmzy.20200807
Abstract148)      PDF(pc) (10107KB)(58)       Save
[Objective] The aim of this study is to investigate the cotton differential metabolites in the interaction between Verticillium dahliae and cotton plant, and to explore new clues for the further study on the defense mechanism of cotton against V. dahliae. [Method] To obtain the cotton root, stem and leaf samples of the pathogen treated and healthy controls, the upland cotton cultivar CCRI 24 was selected as tested cotton material in this paper, and inoculated with V. dahliae conidia or sterile water by injuring root at the 2-true-leaf stage. The 70% (volume fraction) methanol extracts of these samples were separated and detected by ultra-performance liquid chromatography-electrospray ionization-mass spectrometry (UPLC-ESI-MS), and the metabolite data were obtained by the online XCMS software. Through multivariate statistics and student’s t-test, the differential metabolites were investigated. The types of metabolites were putatively identified based on the comparison of the experimental molecular mass and the monoisotopic accurate molecular mass of cotton metabolites. [Result] The efficiency of negative ion mode was higher than that of positive ion detection mode in the cotton metabolites in UPLC-ESI-MS analysis. 576 ions mainly distributed in cotton root were found in the cotton seedling tissues including root, stem and leaf, which were the distinguished differential metabolites between V. dahliae treatment and healthy control. Among them, 77 ions were identified as sesquiterpenoids, diterpenoids, flavonoids, carbohydrates, aliphatics and phenols. In addition to sesquiterpenes, 17 compounds, including caffeic acid, astragalin, isoastragalin, dillenetin, ent-catechin, gossypetin 8-rhamnoside, gossypicyanin, herbacetin 7-glucoside, leucocyanidin, quercetin 3'-glucoside, quercetin 3-glucoside, quercetin 7-glycosides, α,2',3,3',4,4',6-heptahydroxychalcone 2'-glucoside, melibiose, sucrose, sucrose 6-phosphate and 1-tetratriacontanol, had not been reported in the literatures on the interaction between V. dahliae and cotton, which may be the novel pathogenesis-related metabolites on cotton Verticillium wilt. [Conclusion] The putative pathogenesis-related metabolites of cotton Verticillium wilt may play an important role in the interaction of cotton defense against V. dahliae, which provides an important clue for exploring the new resistant mechanism of cotton to Verticillium wilt.
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Genome-wide identification and expression analysis of REM gene family in Gossypium hirsutum
Shi Rongkang, Zhang Dongmei, Sun Zhengwen, Liu Zhengwen, Xie Meixia, Zhang Yan, Ma Zhiying, Wang Xingfen
Cotton Science    2021, 33 (2): 95-111.   DOI: 10.11963/1002-7807.srkwxf.20210113
Abstract137)      PDF(pc) (11066KB)(377)       Save
[Objective] The REM (Reproductive meristem) gene family encodes a class of transcription factors, which plays an important role in the growth and development of plants. However, it has not been reported in Gossypium hirsutum. [Methods] In this study, REM gene family were systematically identified via bioinformatics based on the third generation genomic data of G. hirsutum and the transcriptome data from public databases. The physical and chemical properties of REM protein, gene structure, gene expression patterns in different tissues and under different stresses for REM family were analyzed. [Results] REM gene family in G. hirsutum contained 79 members which distributed on 25 chromosomes and could be divided into five subfamilies. Each of REM proteins contained at least one B3 domain, and most of the members were located in the nucleus. Cis-acting elements responding to hormone and stress were located in the upstream 2 kb of genes. Gene expression analysis showed that most of the REM genes displayed higher expression in the ovules or fibers than in other tissues, and 29 REM genes responded to cold, hot, salt or drought. Expression patterns of six genes in different developmental stages detected by qRT-PCR(Quantitative real-time ploymerase chain reaction) were basically consistent with transcriptome data. [Conclusion] The distribution in the genome, gene structure, and phylogenetic characteristics of these genes were determined, and the roles of REM genes in growth, development and stress were preliminarily clarified according to transcription profile.
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Responses and Underlying Mechanisms of Different Mainstem Leaves on Cotton to Defoliant Thidiazuron
Liao Baopeng, Wang Songman, Du Mingwei, Li Fangjun, Tian Xiaoli, Li Zhaohu
Cotton Science    2020, 32 (5): 418-424.   DOI: 10.11963/1002-7807.lbpdmw.20200817
Abstract127)      PDF(pc) (2346KB)(78)       Save
[Objective] This study elucidates the responses of different mainstem leaves on cotton to defoliant thidiazuron and reveal the underlying mechanisms. [Method] Cotton plants were raised hydroponically. At the ten-leaf stage, 4.54 mmol·L-1 thidiazuron was evenly applied to all mainstem leaves, the youngest full-expanded leaf (the fourth leaf from apex) and the youngest leaf (the first leaf from apex). Then, the progress of abscission zone formation and shedding of each leaf was observed, and the expression level of genes related to ethylene synthesis and signal transduction in the fourth and first leaf from apex was analysed. [Result] The abscission zone developed in the following sequence: the first leaf from the top, then the lowest leaves (the ninth or tenth leaf from the top, 35-40 days old), and the fourth leaf from the top. However, the leaf abscission was in a different order: the earliest was the first leaf from apex, then the fourth leaf from apex, and the latest was the lowest leaves which were prone to wither and stick to the stalk. The GhACS1 expression was up regulated in both the first leaf and the fourth leaf from apex within 24 h after thidiazuron treatment. However, the expression of this gene changed more rapidly and sharply in the first leaf from the top than that in the fourth leaf  from the top. In addition, the expression of GhACO1 (ethylene synthesis gene), GhEIN3 and GhERF23 (ethylene signal transduction genes) was significantly increased up to ten times in the first leaf from the top, whereas the expression of these genes did not change in the fourth leaf from the apex. [Conclusion] After exposed to defoliant thidiazuron, the time of the abscission zone formation of different leaves on cotton main stem is different, and the order of shedding is not consistent with that of abscission zone formation. The youngest ready to expand leaf abscises earlier than the youngest fully expanded leaf after thidiazuron treatment, which is possibly associated with the rapid expression change of genes related to ethylene synthesis and signal transduction in the former.
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QTL Mapping of Chlorophyll Content in Gossypium hirsutum and Gossypium barbadense Backcross Inbred Lines
Geng Yanhui, Bian Yingying, Pei Wenfeng, Liu Guoyuan, Wu Man, Zang Xinshan, Li Dan, Li Xingli, Zhang Jinfa, Yu Jiwen
Cotton Science    2020, 32 (5): 463-471.   DOI: 10.11963/1002-7807.gyhyjw.202009
Abstract126)      PDF(pc) (1408KB)(58)       Save
[Objective] The purpose of this study was to map quantitative trait loci (QTL) related to chlorophyll content based on Soil and Plant Analyzer Development (SPAD) readings in cotton. [Method] The 195 BILs (Backcross Inbred Lines) were produced by a cross between Gossypium barbadense Hai 7124 and G. hirsutum CRI 36, using CRI 36 as the recurrent parent for backcrossing with F1 to produce BC1F1, followed by seven generations of selfing. The genetic linkage map was constructed in a previous study. QTLs of chlorophyll SPAD value in the first flowering and boll development stages were identified with inclusive composite interval mapping (ICIM) method of the BIP and MET models in IciMapping 4.1 software, respectively. [Result] In total, nine chlorophyll SPAD reading QTLs were identified on 6 chromosomes. The q-SPAD-A11-1 detected at the first flowering stage overlapped with q-SPAD-A11-2 detected at the boll development stage, contributing 5.08% and 5.75% of the phenotypic variation, respectively. The q-SPAD-D08-2 physical position ranged from 48.71 to 53.65 Mb on chromosome D08, which overlapped with a chlorophyll content QTL detected in a previous study. [Conclusion] The novel stable QTLs, q-SPAD-A11-1 and q-SPAD-A11-2 detected in this study provide an important piece of information for fine mapping chlorophyll content in cotton.
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Recent Advances in the Enhancement of Agroecosystem Services and Functioning by Cotton-based Intercropping Systems
Li Xiaofei, Han Yingchun, Wang Guoping, Wang Zhanbiao, Feng Lu, Yang Beifang, Fan Zhengyi, Lei Yaping, Xiong Shiwu, Xing Fangfang, Li Yabing
Cotton Science    2020, 32 (5): 472-482.   DOI: 10.11963/1002-7807.lxflyb.20200826
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Cotton-based intercropping systems are one of modern agriculture farming systems aiming at improving overall economic profitability of cotton field, which not only release the competition for land between other crops and cotton and increase growing area and yield of both crops, but also represent a mechanistic approach to reconciling crop production and biodiversity conservation. Recently, cotton-based intercropping systems have been widely focused and applied. Here, we reviewed the potential of cotton-based intercropping systems to reinforce agroecosystem services and functioning, including promoted plant biodiversity, improved overall productivity and economic profits, increased light use efficiency, improved cotton quality, reduced pest and disease occurrence, and suppressed weed growth. Further, the underlying mechanisms behind the enhancement of agroecosystem services and functioning by cotton-based intercropping systems through niche complementarity, interspecific facilitation, and allelopathy between intercropped species were summarized in the paper. Finally, the research prospects were also pointed out.
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Optimization and application of an artificial inoculation method for cotton boll blight
Lu Xiuyun, Shang Junyan, Shao Meiqi, Xie Xuejiao, Guo Qinggang, Li Shezeng, Ma Ping
Cotton Science    2021, 33 (1): 1-12.   DOI: 10.11963/1002-7807.lxymp.20201207
Abstract120)      PDF(pc) (1867KB)(66)       Save
[Objective] Cotton boll blight, which caused by Phytophthora boehmeriae, is one of the most important diseases of cotton boll rot. The aim of this study was to optimize the artificial inoculation method of cotton boll blight and to apply for further studies. [Method] The population of P. boehmeriae in adult bolls sampled from different parts of cotton plant was compared under the suitable culture conditions. The disease occurrences of surface sterilization and non-sterilization, as well as injury and non-injury cotton bolls inoculated with P. boehmeriae were compared. The artificial inoculation method of cotton boll blight was established through pathogenesis tests and used to screen 12 kind of chemicals against the disease, to identify the resistances of 16 cotton varieties and to test the pathogenicity of 10 strains of P. boehmeriae. [Result] The disease incidence of the boll from the 1-3rd fruit branches was more serious than that from the 4-6th fruit branches, the 7-9th fruit branches and the 10-12th fruit branches. P. boehmeriae and other microorganisms on cotton boll surface could be eliminated by soaking cotton bolls in 75% (volume fraction) alcohol for 2 min. Cotton bolls from the 4-9th fruit branches of cotton were soaked in 75% alcohol for 2 min, and then wounded with sewing needles on the middle and upper part of the cotton bolls. The wounded parts of cotton bolls were inoculated with the mycelium disks of P. boehmeriae. Cotton boll blight occurred completely in 3-7 d under moisture culture at 25 ℃. Among the 12 kind of chemical fungicides, 25% (mass fraction) Metalaxyl propamocarb hydrochloride wettable powder (WP), 70% Propineb WP and 52.5% Famoxadone cymoxanil wettable granule (WG) showed significant control effects against cotton boll blight with the artificial inoculation. For 16 cotton varieties, there were differences in resistance to the disease. Among the 10 strains of P. boehmeriae, JP18-4 showed the highest pathogenicity, while JP15-2 showed the lowest. [Conclusion] Artificial inoculation method of cotton boll blight was established and optimized in this study. The method can be used for screening control agents against cotton boll blight and the identification of resistant cotton varieties and pathogenicity difference of P. boehmeriae.
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Cotton Chemical Topping by Applying DPC in Different Cotton-Growing Regions 
Qi Haikun, Wang Sai, Xu Dongyong, Lu Zhengying, Zhao Wenchao, Hao Yanjie, Zhang Xiang, Li Wei, Han Huanyong, Wang Jiangtao, Wang Hongzhe, Chen Hongzhang, Wang Lin, Du Mingwei, Tian Xiaoli, Li Zhaohu
Cotton Science    2020, 32 (5): 425-437.   DOI: 10.11963/1002-7807.qhklzh.20200729
Abstract119)      PDF(pc) (2133KB)(70)       Save
[Objective] The objective of this study was to investigate the stability and universality of cotton chemical topping by applying mepiquat chloride (1,1-dimethyl-piperidinium chloride, DPC) in different cotton-growing regions. [Method] Field experiments were conducted in 2018 at 10 locations in the Yellow River basin (Hejian and Handan, Hebei province; Dezhou and Wudi, Shandong province), the Yangtze River basin (Dafeng, Jiangsu province; Huanggang, Hubei province), and Xinjiang area (Shihezi location I and loacation II, northern Xinjiang and Luntai and Shaya, southern Xinjiang). Local cultivars/lines were used, and the experiments were performed using a randomized complete block design with three or four replicates. Accompanied with typical DPC multi-application in each location, chemical topping was conducted at 10 days before manual topping (T1) or at the same time with manual topping (T2) by applying four dosages of DPC (0, 90, 180, 270 g·hm-2), manual topping was used as the first control and non-topping as the second control. [Result] The time of chemical topping significantly affected cotton plant height (except for the results in Handan, Dezhou and Wudi) and the number of fruit branches (except for the results in Dafeng and Huanggang). It was observed that earlier chemical topping would result in lower cotton plant height and a fewer fruit branches. In Hejian and Shihezi location I, the average plant height across DPC chemical topping at T1 stage was not only lower than that of T2 stage but also 3.3 cm and 4.6 cm lower than that of manual topping, respectively. In most locations, chemical topping at T1 stage increased around two fruit branches per plant compared with manual topping, while in T2 stage the increased fruit branches per plant ranged from 2.3 to 7.7. Also, we found that a higher dosage of DPC resulted in shorter plant height (except for that in Huanggang). In some locations, plant heights of chemical topping with 180 g·hm-2 or 270 g·hm-2 DPC were even shorter than that of manual topping. The number of fruit branches per plant of 0 g·hm-2 DPC increased by 2.4-8.3 compared with manual topping. However, chemical topping with 90-270 g·hm-2 DPC significantly reduced the number of fruit branches compared with 0 g·hm-2 DPC. There were no significant differences in the number of fruit branches among three DPC dosages (90, 180, and 270 g·hm-2). In Handan, seed cotton yield of chemical topping at T2 stage was significantly lower than that of manual topping due to the decreased boll number, which is possibly associated with the high temperature and drought weather after chemical topping. While at other locations, most treatments of chemical topping by using DPC did not produce significant effects on yield. In addition, chemical topping by using DPC did not delay cotton maturity, characterized by their similar boll-opening rate and the first harvest rate to those of manual topping before spraying harvest aids. [Conclusion] Cotton chemical topping with DPC is more stable and universal across different cotton-growing regions. We suggest that 90-180 g·hm-2 DPC could be used at the same time with manual topping for cotton chemical topping.
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