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Comparative phosphoproteomic analysis of BR-defective mutant reveals a key role of GhSK13 in regulating cotton fiber development

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Science China(Life Sciences) 2020年第12期63卷 1905-1917页

作者： Lingling Wang Han Cheng Fangjie Xiong Shuya Ma Lei Zheng Yun Song Kexuan Deng Huanhuan Wu Fuguang Li Zuoren Yang Zhengzhou Research Base State Key Laboratory of Cotton BiologyZhengzhou UniversityZhengzhou 450001China State key laboratory of cotton Biology Institute of Cotton ResearchChinese Academy of Agricultural SciencesAnyang 455000China key laboratory for Ecology of Tropical Islands Ministry of EducationCollege of Life SciencesHainan Normal UniversityHaikou 571158China School of Life Sciences Chongqing UniversityChongqing 401331China

Brassinosteroid(BR), a steroid phytohormone, whose signaling transduction pathways include a series of phosphorylation and dephosphorylation events, and GSK3 s are the main negative regulator kinases. BRs have been shown to play vital roles in cotton fiber elongation. However, the underlying mechanism is still elusive. In this study, fibers of a BR-defective mutant Pagoda 1(pag1), and its corresponding wild-type(ZM24) were selected for a comparative global phosphoproteome analysis at critical developmental time points: fast-growing stage(10 days after pollination(DPA)) and secondary cell wall synthesis stage(20DPA). Based on the substrate characteristics of GSK3, 900 potential substrates were identified. Their GO and KEGG annotation results suggest that BR functions in fiber development by regulating GhSKs(GSK3s of Gossypium hirsutum L.) involved microtubule cytoskeleton organization, and pathways of glucose, sucrose and lipid metabolism. Further experimental results revealed that among the GhSK members identified, GhSK13 not only plays a role in BR signaling pathway, but also functions in developing fiber by respectively interacting with an AP2-like ethylene-responsive factor GhAP2L, a nuclear transcription factor GhDNFYB19, and a homeodomain zipper member GhHDZ5. Overall, our phosphoproteomic research advances the understanding of fiber development controlled by BR signal pathways especially through GhSKs, and also offers numbers of target proteins for improving cotton fiber quality.

关键词： phosphoproteome brassinosteroid GSK3 GhSK13 cotton fiber development

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Identification and characterization of cell cultures with various embryogenic/regenerative potential in cotton based on morphological,cytochemical,and cytogenetical assessment

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Journal of Integrative Agriculture 2019年第1期18卷 1-8页

作者： GUO Hui-hui WU Jian-fei CHEN Cui-xia WANG Hong-mei ZHAO Yun-lei ZHANG Chao-jun JIA Yin-hua LIU Fang NING Tang-yuan CHU Zhao-hui ZENG Fan-chang State key laboratory of Crop Biology Ministry of Science and Technology/College of AgronomyShandong Agricultural University State key laboratory of cotton Biology Ministry of Science and Technology/Cotton Research InstituteChinese Academy of Agricultural Sciences key laboratory of Water Resources and Drought Resistant germplasm improvement Ministry of Agriculture

Somatic embryogenesis(SE) plays a vital role in genetic transformation and massive propagation of important agronomical and economical ***,we conducted a systematic assessment of the morphological,cytochemical,and cytogenetical characteristics of six culture strains with various embryogenic/regenerative potential during SE process in *** indicated that the six cell culture strains had stable ploidy levels,and did not reveal any relationship between the cytogenetic state and their morphogenetic ***,the six culture strains were compared via double staining with Evans blue and Acetocarmine to efficiently distinguish embryogenic and non-embryogenic cells and determine the embryogenic nature of the *** addition,the kind of auxins added in medium affected not only growth property,color,size of cell clumps but also ploidy level and regeneration *** combining analysis of morphological,cytochemical,and cytogenetical characteristics of the cell cultures,we are able to obtain and maintain homogeneous cell population with high morphogenic and regeneration ability and establish efficient somatic embryogenesis and regeneration system from short-term cell cultures in upland cotton,which highlight the application of biotechnological approaches in crop breeding,and above all,to better understand totipotency of cells in higher plants.

关键词： cotton somatic embryogenesis cotton cell culture strains cytochemical EB-AC double staining cytogenetical ploidy stability embryogenic/regenerative potential, morphology

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A comparative analysis of small RNAs between two Upland cotton backcross inbred lines with different fiber length: Expression and distribution

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The Crop Journal 2019年第2期7卷 198-208页

作者： Guoyuan Liu Man Wu Wenfeng Pei Xihua Li Nuohan Wang Jianjiang Ma Xinshan Zang Shuxun Yu Jinfa Zhang Jiwen Yu State key laboratory of cotton Biology Cotton Institute of the Chinese Academy of Agricultural Sciences Key Laboratory of Cotton Genetic Improvement Ministry of Agriculture Department of Plant and Environmental Sciences New Mexico State University

The cotton fiber is the most important raw material for the textile industry and an ideal model system for studying cell elongation. However, the genetic variation of fiber elongation in relation to miRNA is poorly understood. A high-throughput comparative RNA-seq of two lines differing in fiber length(FL) from a backcross inbred line(BIL)population of G. hirsutum × G. barbadense revealed differentially expressed(DE) miRNAs and their targets in rapidly elongating fibers. A real-time quantitative PCR analysis was further performed to validate the results. A total of 463(including 47 DE) miRNAs were identified,and seven DE miRNAs were co-localized with seven FL quantitative trait loci(QTL)identified in the G. hirsutum × G. barbadense population. Of 82(including 21 DE) targets identified, nine(including one DE) were also co-localized with the seven FL QTL. The relationship between the allopolyploid and its diploid ancestral species with respect to miRNAs and their targets was also characterized. These results will facilitate the understanding of the molecular genetic mechanism of fiber elongation with regards to miRNAs in cotton.

关键词： Allopolyploid Gossypium Fiber elongation miRNA QTL hotspots

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Restructuring Tilth Layers Can Change the Microbial Community Structure and Affect the Occurrence of Verticillium Wilt in cotton Field

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Phyton-International Journal of Experimental Botany 2023年第10期92卷 2841-2860页

作者： Ming Dong Yan Wang Shulin Wang Guoyi Feng Qian Zhang Yongzeng Lin Qinglong Liang Yongqiang Wang Hong Qi Institute of cotton Key Laboratory of Biology and Genetic Improvement of Cotton in Huanghuaihai Semiarid AreaMinistry of AgricultureNational Cotton Improvement Center Hebei BranchShijiazhuang050051China

Restructuring tilth layers(RTL)is a tillage method that exchanges the 0–20 and 20–40 cm soil layers that can be applied during cotton cultivation to increase cotton yield,eliminate weeds and alleviate severe disease,including Verticillium ***,the mechanism by which RTL inhibits Verticillium wilt is ***,we investigated the distribution of microbial communities after rotary tillage(CK)and RTL treatments to identify the reasons for the reduction of Verticillium wilt in cotton fields subjected to *** high-throughput sequencing was used to sequence the bacterial and fungal *** disease incidence and severity of Verticillium wilt decreased by 28.57%and 42.64%,respectively,after ***,RTL significantly enhanced bacterial richness and evenness at 20–40 cm and-reduced the differences in fungal evenness and richness between soil depths of 0–20 and 20–40 *** number of Verticillium dahliae decreased,while the relative abundance of biocontrol bacteria such as Bacillus and Pseudoxanthomonas increased significantly following *** RTL improved bacterial diversity,decreased the number of Verticillium dahliae and increased the relative abundance of biocontrol bacteria,which may have suppressed the occurrence of Verticillium wilt in cotton fields.

关键词： Bacteria fungi restructuring tilth layers diversity disease reduction

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Mapping and Quantitative Trait Loci Analysis of Verticillium Wilt Resistance Genes in cotton

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Journal of Integrative Plant Biology 2008年第2期50卷 174-182页

作者： Hong-Mei Wang Zhong-Xu Lin Xian-Long Zhang Wei Chen Xiao-Ping Guo Yi-Chun Nie Yun-Hai Li National key laboratory of Crop Genetic improvement Huazhong Agricultural University Wuhan 430070 China key laboratory of cotton Genetic improvement of the Ministry of Agriculture Cotton Research Institute Chinese Academy of Agricultural Sciences Anyang 455112 China

Verticillium wilt is one of the most serious constraints to cotton production in almost all of the cotton-growing countries. In this study, ＂XinLuZaol＂（XLZl）, a susceptible cultivar Gossypium hirsutum L. and ＂Hai7124＂（H7124）, a resistant line G. barbadense, and their F2：3 families were used to map and study the disease index induced by verticillium wilt. A total of 430 SSR loci were mapped into 41 linkage groups; the map spanned 3 745.9 cM and the average distance between adjacent loci was 8.71 cM. Four and five quantitative trait loci （QTLs） were detected based on the disease index investigated on July 22 and August 24 in 2004, respectively. These nine QTLs explained 10.63-28.83% of the phenotypic variance, six of them were located on the D sub-genome. Two QTLs located in the same marker intervals may partly explain the significant correlation of the two traits. QTLs explaining large phenotypic variation were identified in this study, which may be quite useful in cotton anti-disease breeding.

关键词： cotton molecular marker quantitative trait locus Verticillium wilt.

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Genetic Effect on Yield and Fiber Quality Traits of 16 Chromosome Substitution Lines in Upland cotton

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Agricultural Sciences in China 2008年第11期7卷 1290-1297页

作者： LUAN Ming-bao GUO Xiang-mo ZHANG Yong-shan YAO Jin-bo key laboratory of cotton germplasm improvement Ministey of Agriculture/Cotton Research Institute Chinese Academy of Agricultural Sciences （ CAAS） A ny ang 455000 P.R. Ch ina

Evaluation of the genetic effect on yield and fiber quality can provide useful information on cotton breeding. Sixteen CSB lines and TM-1 introduced from USDA/ARS were used as male and top-crossed with three elite cultivars and the 51 F1 hybrids, 16 CSB lines, TM-1, and 3 elite cultivars were planted at the cotton Research Institute of CAAS, Anyang, Henan Province and Xiajin, Shandong Province, China. The yield traits and fiber quality data were obtained and additive and dominance effect on each trait were measured by AD model. Boll weight takes the largest additive proration, whereas boll number takes the least additive proration. The largest and the least dominant proration for lint yield and boll weight were measured, respectively. Fiber length has the additive and dominance effect, and dominance effect was slightly more than additive effect. Larger additive and no dominance effect on uniformity, micronaire, and fiber strength were measured. Significantly, positive additive effect on boll weight of CSB06 and CSB12Sh was observed. CSB14Sh and CSB01 have significantly positive additive effect on 4 and 3 traits of fiber quality, respectively. CSB01 has the greatest dominant effect on lint yield among CSB lines. The dominant effect on fiber length of CSB lines showed positive. It is beneficial to use CSB06 and CSB12Sh as parents to improve boll size, to use CSB14Sh and CSB01 as parents to improve fiber quality. As for hybrid cotton breeding, it is reasonable using CSB01 to improve lint yield traits, and using CSB01, CSB11Sh, and CSB06 to improve fiber length.

关键词： chromosome substitution lines in upland cotton additive genetic effect dominance genetic effect AD model

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Mechanism of floral scent production in Osmanthus fragrans and the production and regulation of its key floral constituents,β-ionone and linalool

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Horticulture Research 2019年第1期6卷 432-443页

作者： Yuanji Han Hongyun Wang Xiaodan Wang Ke Li Meifang Dong Yong Li Qian Zhu Fude Shang School of Life Sciences State Key Laboratory of Cotton BiologyState Key Laboratory of Crop Stress Adaptation and ImprovementKey Laboratory of Plant Stress BiologyLaboratory of Plant Germplasm and Genetic EngineeringHenan UniversityKaifeng 475004 HenanChina

Sweet osmanthus(Osmanthus fragrans Lour.)is among the top ten most well-known flowers in China and is recognized as both an aromatic plant and ornamental ***,manual sectioning,scanning electron microscopy,and transmission electron microscopy of sweet osmanthus petals revealed that large amounts of lipids are present inside the petal cells and on the cell ***,no secretory structures were ***,the petal cells protrude slightly outward,and the surfaces of the cells are adorned with highly regular brush-shaped *** surfaces of the‘Yingui’petals possessed mostly curled and more numerous hairs,whereas the‘Dangui’petals possessed fewer brush-shaped and more sparsely arranged *** addition,many granular substances were attached to the brush-shaped hairs,and the granules were denser on the hairs of the‘Yingui’petals compared to the hairs on the‘Dangui’***,35 aromatic components in the‘Yingui’petals and 30 aromatic components in the‘Dangui’petals were detected via *** main aromatic component of the‘Yingui’petals wasβ-ionone,whereas that of the‘Dangui’petals was linalool and its *** sequencing and qRT-PCR indicated that the highβ-ionone content in the‘Yingui’petals was due to the overexpression of CCD1 and CCD4 and that the high linalool content in the‘Dangui’petals was due to the overexpression of MECS,HDR,IDI1,and LIS1,which function upstream of the linalool synthetic *** particular,the expression levels of CCD4 and LIS1 were upregulated by 5.5-and 5.1-fold in the‘Yingui’and‘Dangui’petals,*** transcription factor(ERF61)was cloned and named,and the expression pattern of ERF61 in sweet osmanthus petals was found to be generally consistent with that of *** transformation experiments,yeast one-hybrid experiments,and electrophoretic mobility shift assays indicated that ERF61 binds to the CCD4 promoter and stimulates CCD4 expression,thereby regulating the synthesis ofβ-iono

关键词： shaped flowers attached

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Histological and Ultrastructural Observation Reveals Significant Cellular Differences between Agrobacterium Transformed Embryogenic and Non-embryogenic Calli of cotton

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Journal of Integrative Plant Biology 2009年第5期51卷 456-465页

作者： Hai-Hong Shang Chuan-Liang Liu Chao-Jun Zhang Feng-Lian Li Wei-Dong Hong Fu-Guang Li key laboratory of cotton Genetic improvement Ministry of Agriculture Chinese Cotton Research Institute Chinese Agricultural Academy of Sciences Anyang 455004 China

Over the past few decades genetic engineering has been applied to improve cotton breeding. Agrobacterium medicated transformation is nowadays widely used as an efficient approach to introduce exogenous genes into cotton for genetically modified organisms. However, it still needs to be improved for better transformation efficiency and higher embryogenic callus induction ratios. To research further the difference of mechanisms for morphogenesis between embryogenic callus and non-embryogenic callus, we carried out a systematical study on the histological and cellular ultrastructure of Agrobacterium transformed calli. Results showed that the embryogenic callus developed nodule-like structures, which were formed by small, tightly packed, hemispherical cells. The surface of some embryogenic callus was covered with a fibrilar-like structure named extracellular matrix. The cells of embryogenic calli had similar morphological characteristics. Organelles of embryogenic callus cells were located near the nucleus, and chloroplasts degraded to proplastid-like structures with some starch grains. In contrast, the non-embryogenic calli were covered by oval or sphere cells or small clusters of cells. It was observed that cells had vacuolation of cytoplasm and plastids with a well organized endomembrane system. This study aims to understand the mechanisms of embryogenic callus morphogenesis and to improve the efficiency of cotton transformation in future.

关键词： cell organization pattern chloroplasts embryogenic callus Gossypiurn hirsutum histological structure transformation ultrastructure

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Construction of a Full-Length cDNA Library of Gossypium hirsutum L. and Identification of Two MADS-Box Genes

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Agricultural Sciences in China 2011年第1期10卷 28-40页

作者： WANG Li-na WU Dong YU Shu-xun FAN Shu-li SONG Mei-zhen PANG Chao-you LIU Jun-jie cotton Research Institute Chinese Academy of Agricultural Sciences~Key Laboratory of Cotton Genetic Improvement Ministry of AgricultureAnyang 455000 P.R.China

A full-length normalized cDNA library for the flower development stages of short-season cotton （Gossypium hirsutum L.）（CCRI36） was constructed. A total of 3 421 clones were randomly selected for sequencing, with a total of 3 175 effective sequences obtained after removal of empty-carriers and low-quality sequences. Clustering the 3 175 high-quality expressed sequence tags （ESTs） resulted in a set of 2 906 non-redundant sequences comprised of 233 contigs and 2 673 singletons. Comparative analyses indicated that 913 （43.6%） of the unigenes had homologues with function-known genes or functionassumed genes in the National Center for Biotechnology Information. In addition, 763 （36.4%） of the unigenes were functionally classified using Gene Ontology hierarchy. Through EST alignment and the screening method, the full-length cDNA of two MADS-box genes viz., GhMADSll and GhMADS12 were acquired. These genes may play a role in flower development. Phylogenetie analysis indicated that GhMADS11 and GhMADS12 had high homology and close evolutionary relationship with AGL2/SEP-type and PI-type genes, respectively. The expression of both GhMADSll and GhMADS12, genes was high in reproductive organs. In floral organs, GhMADSll expression was high in petals （whor12） and ovules, while GhMADS12 expression was high in petals （whor12） and stamens （whor13）. Results show that the EST strategy based on a normalized cDNA library is an effective method for gene identification. The study provides more insights for future molecular research on the regulation mechanism of cotton flower development.

关键词： cotton normalized cDNA library EST MADS-box gene

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Sequence Divergence of Microsatellites and Phylogeny Analysis in Tetraploid cotton Species and Their Putative Diploid Ancestors

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Journal of Integrative Plant Biology 2005年第12期47卷 1418-1430页

作者： Wang-Zhen GUO Dong FANG Wen-Duo YU Tian-Zhen ZHANG State key laboratory of Crop Genetics and germplasm Enhancement Cotton Research Institute Nanjing Agricultural University Nanjing 210095 China

To determine the level of microsatellite sequence differences and to use the information to construct a phylogenetic relationship for cultivated tetraploid cotton （Gossypium spp.） species and their putative diploid ancestors, 10 genome-derived microsatellite primer pairs were used to amplify eight species, including two tetraploid and six diploid species, in Gossypium. A total of 92 unique amplicons were resolved using polyacrylamide gel electrophoresis. Each amplicon was cloned, sequenced, and analyzed using standard phylogenetic software. Allelic diversities were caused mostly by changes in the number of simple sequence repeat （SSR） motif repeats and only a small proportion resulted from interruption of the SSR motif within the locus for the same genome. The frequency of base substitutions was 0.5%-1.0% in different genomes, with only few indels found. Based on the combined 10 SSR flanking sequence data, the homology of A-genome diploid species averaged 98.9%, even though most of the amplicons were of the same size, and the sequence homology between G. gossypioides （Ulbr.） Standl. and three other D-genome species （G. raimondii Ulbr., G. davidsonii Kell., and G. thurberi Tod.） was 98.5%, 98.6%, and 98.5%, respectively. Phylogenetic trees of the two allotetraploid species and their putative diploid progenitors showed that homoelogous sequences from the A- and D-subgenome were still present in the polyploid subgenomes and they evolved independently. Meanwhile, homoelogous sequence interaction that duplicated loci in the polyploid subgenomes became phylogenetic sisters was also found in the evolutionary history of tetraploid cotton species. The results of the present study suggest that evaluation of SSR variation at the sequence level can be effective in exploring the evolutionary relationships among Gossypuim species.

关键词： diploid Gossypium microsatellite phylogenetic tree simple sequence repeat (SSR) tetraploid.

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