Changwei Zhang

Southwest University

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Xilin Hou

Nanjing Agricultural University

Ying Li

Lanzhou Jiaotong University

Xilin Hou

Nanjing Agricultural University

Tongkun Liu

Nanjing Agricultural University

Dong Xiao

Nanjing Agricultural University

Di Cai

Beijing University of Chemical Technology

Shanwu Lyu

South China Botanical Garden

Xianchun Sang

Southwest University

Guanghua He

Key Laboratory of Guangdong Province

Xu Zhang

University of Shanghai for Science and Technology

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Nanjing Agricultural University

100

Ministry of Agriculture and Rural Affairs

Chinese Academy of Sciences

Beijing University of Chemical Technology

Southwest University

Rice Research Institute

Chinese Academy of Agricultural Sciences

Sun Yat-sen University

Peking University

Chinese Academy of Medical Sciences & Peking Union Medical College

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Genetic Diversity Analysis of Non-Heading Chinese Cabbage of Resistance to Clubroot Disease Based on SNP Molecular Markers

Agronomy (2023)

Liming Miao Dandan Xi Xiaofeng Li Changwei Zhang Lu Gao

Clubroot disease is a kind of soil-borne disease that seriously infects Brassica species. In this study, we collected 121 varieties of non-heading Chinese cabbages. In order to better understand the genetic variation and to screen suitable clubroot disease-resistant parental material, we re-sequenced them to examine the population genetic structure, population genetic diversity, population differentiation index, and selective sweep based on SNPs. The mapping rate with the reference genome was high, and data quality analysis revealed that the sequencing quality was good. The annotated data indicated that intronic and intergenic areas held the majority of SNPs and indels. Four subgroups of 121 non-heading Chinese cabbages were identified using principal component analysis, phylogenetic tree, and genetic structure analysis. An examination of genetic diversity revealed that while selfing may happen in subgroups C and D, heterozygosity may exist in subgroups A and B. In subgroup B, self-fertilization is not possible. There was a moderate degree of genetic differentiation between subgroups B and C (Fst = 0.0744347). For genes in certain sweep regions, we also ran GO enrichment and KEGG enrichment analysis. Two disease resistance-related genes, BraA01g042910.3.5C and BraA06g019360.3.5C, were examined. These findings will serve as a theoretical foundation for developing novel, clubroot disease-resistant types of non-heading Chinese cabbages.

Clubroot

Indel

10.3390/agronomy13010245

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Citations (3)

Non-Heading Chinese Cabbage Database: An Open-Access Platform for the Genomics of Brassica campestris (syn. Brassica rapa) ssp. chinensis

Plants (2022)

Zhidong Li Ying Li Tongkun Liu Changwei Zhang Dong Xiao

The availability of a high-quality genome sequence of Brassica campestris ssp. chinensis NHCC001 has paved the way for deep mining of genome data. We used the B. campestris NHCC001 draft genome to develop a comprehensive database, known as the non-heading Chinese cabbage database, which provides access to the B. campestris NHCC001 genome data. The database provides 127,347 SSR, from which 382,041 pairs of primers were designed. NHCCDB contains information on 105,360 genes, which were further classified into 63 transcription factor families. Furthermore, NHCCDB provides eight kinds of tools for biological or sequencing data analyses, including sequence alignment tools, functional genomics tools, comparative genomics tools, motif analysis tools, genome browser, primer design, and SSR analysis tools. In addition, eight kinds of graphs, including a box plot, Venn diagram, corrplot, Q-Q plot, Manhattan plot, seqLogo, volcano plot, and a heatmap, can be generated rapidly using NHCCDB. We have incorporated a search system for efficient mining of transcription factors and genes, along with an embedded data submit function in NHCCDB. We believe that the NHCCDB database will be a useful platform for non-heading Chinese cabbage research and breeding.

Brassica rapa

Genome browser

Comparative Genomics

Synteny

10.3390/plants11081005

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Citations (7)

The SNARE Protein Syp71 Is Essential for Turnip Mosaic Virus Infection by Mediating Fusion of Virus-Induced Vesicles with Chloroplasts

PLoS Pathogens (2013)

Tàiyún Wèi Changwei Zhang Xilin Hou Hélène Sanfaçon Aiming Wang

All positive-strand RNA viruses induce the biogenesis of cytoplasmic membrane-bound virus factories for viral genome multiplication. We have previously demonstrated that upon plant potyvirus infection, the potyviral 6K2 integral membrane protein induces the formation of ER-derived replication vesicles that subsequently target chloroplasts for robust genome replication. Here, we report that following the trafficking of the Turnip mosaic potyvirus (TuMV) 6K2 vesicles to chloroplasts, 6K2 vesicles accumulate at the chloroplasts to form chloroplast-bound elongated tubular structures followed by chloroplast aggregation. A functional actomyosin motility system is required for this process. As vesicle trafficking and fusion in planta are facilitated by a superfamily of proteins known as SNAREs (soluble N-ethylmaleimide-sensitive-factor attachment protein receptors), we screened ER-localized SNARES or SNARE-like proteins for their possible involvement in TuMV infection. We identified Syp71 and Vap27-1 that colocalize with the chloroplast-bound 6K2 complex. Knockdown of their expression using a Tobacco rattle virus (TRV)-based virus-induced gene silencing vector showed that Syp71 but not Vap27-1 is essential for TuMV infection. In Syp71-downregulated plant cells, the formation of 6K2-induced chloroplast-bound elongated tubular structures and chloroplast aggregates is inhibited and virus accumulation is significantly reduced, but the trafficking of the 6K2 vesicles from the ER to chloroplast is not affected. Taken together, these data suggest that Syp71 is a host factor essential for successful virus infection by mediating the fusion of the virus-induced vesicles with chloroplasts during TuMV infection.

Turnip mosaic virus

10.1371/journal.ppat.1003378

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Citations (130)

BcTFIIIA Negatively Regulates Turnip Mosaic Virus Infection through Interaction with Viral CP and VPg Proteins in Pak Choi (Brassica campestris ssp. chinensis)

Genes (2022)

Rujia Zhang Changwei Zhang Shanwu Lyu Huiyuan Wu Mengguo Yuan

TFIIIA is a zinc-finger transcription factor that is involved in post-transcriptional regulation during development. Here, the BcTFIIIA gene was isolated from pak choi. Sequence analysis showed that BcTFIIIA encodes 383 amino acids (aa) with an open reading frame (ORF) of 1152 base pairs (bp). We investigated the subcellular location of BcTFIIIA and found the localized protein in the nucleus. BcTFIIIA was suppressed when the pak choi was infected by the turnip mosaic virus (TuMV). The BcTFIIIA mRNA expression level in a resistant variety was higher than that in a sensitive variety, as determined by qRT-PCR analysis. Yeast two hybrid (Y2H) assay and bimolecular fluorescence complementation (BiFC) suggested that BcTFIIIA interacts with TuMV CP and VPg in vivo, respectively, and in vitro. A virus-induced gene silencing (VIGS) experiment showed that the silencing of BcTFIIIA gene expression in pak choi promoted the accumulation of TuMV. These results suggest that BcTFIIIA negatively regulates viral infection through the interaction with TuMV CP and VPg.

Turnip mosaic virus

Bimolecular fluorescence complementation

Two-hybrid screening

Protein-fragment complementation assay

10.3390/genes13071209

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Citations (2)

Host Factors Genes BcCLC1 and BcCLC2 Confer Turnip Mosaic Virus Resistance in Non-Heading Chinese Cabbage (Brassica campestris ssp. chinensis)

Plants (2023)

Mengguo Yuan Shanwu Lyu Yaolong Wang Liu E Tongkun Liu

Clathrin is an evolutionarily highly conserved evolutionary protein consisting of clathrin light chains (CLC) and clathrin heavy chains (CHC), and these form its basic structure. Clathrin is an important host factor in the process of viral infection. In this study, we cloned the BcCLC1 gene and the BcCLC2 gene from the '49CX' variety of non-heading Chinese cabbage (NHCC, Brassica campestris L. ssp. chinensis Makino) and verified their functions. The results showed that BcCLC1 was mainly localized in the cytomembrane and cytoplasm, and only a small amount entered the nucleus. BcCLC2 encoded a protein comprising 265 amino acids that were distributed in the cytomembrane, nucleus, and cytoplasm. A BiFC assay and yeast two-hybrid (Y2H) analysis showed that BcCLCs (BcCLC1 and BcCLC2) could interact with several TuMV proteins. We further investigated the mechanism of BcCLCs in regulating TuMV virus infections in NHCC, and observed that BcCLCs gene silencing inhibited TuMV infections and overexpression of BcCLCs in Arabidopsis promoted TuMV infections in NHCC. Finally, mutants of Arabidopsis homologs of BcCLCs were also screened and subjected to TuMV inoculation tests. In conclusion, we speculate that BcCLCs confer Turnip mosaic virus (TuMV) resistance in NHCC by interacting with TuMV proteins to promote the intracellular transport of the virus.

Turnip mosaic virus

Bimolecular fluorescence complementation

Two-hybrid screening

10.3390/plants12122269

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Citations (1)

Depolymerization of post-consumer PET bottles with engineered cutinase 1 fromThermobifida cellulosilytica

Green Chemistry (2022)

Zixuan Zhang Shiming Huang Di Cai Chaofeng Shao Changwei Zhang

The enhanced enzymatic activity and thermal stability of cutinase 1 from Thermobifida cellulosilytica by enzyme engineering were utilized to achieve efficient degradation of post-consumer polyethylene terephthalate (PET) bottle particles.

Depolymerization

Cutinase

10.1039/d2gc01834a

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Citations (20)

Genome-wide identification of turnip mosaic virus-responsive microRNAs in non-heading Chinese cabbage by high-throughput sequencing

Gene (2015)

Zhen Wang Dahua Jiang Changwei Zhang Huawei Tan Yanxiao Li

Identification

Turnip mosaic virus

10.1016/j.gene.2015.06.047

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Citations (21)

Microtubule and male sterility in a gene‐cytoplasmic male sterile line of non‐heading Chinese cabbage

Journal of the Science of Food and Agriculture (2012)

Jingyi Zhang Changwei Zhang Yan Cheng Qi Li Shumin Wang

Abstract BACKGROUND: Microtubules are the basic components of the cytoskeleton in eukaryotic cells and are made up of 13 parallel protofilaments, each composed of α‐ and β‐tubulin unit molecules aligned along the longitudinal axis of the microtubule. RESULTS: α‐Tubulin gene TUBA2 from non‐heading Chinese cabbage ( Brassica campestris ssp. chinensis Makino) was expressed at the highest level in stamens and at lower levels in other organs. In addition, it was expressed at a much lower level in the cytoplasmic male sterile (CMS) line than in the maintainer line. Furthermore, at the microsporocyte stage of development in the CMS line the microtubule bundles were knitted together in random organisation, which differed significantly from the radiate microtubule bundles running circumferentially around the nucleus in the maintainer line. Also, large vacuoles appeared within the cytoplasm in the CMS line with no dyed microtubules. CONCLUSION: TUBA2 was very important to pollen development, which might be closely related to male sterility. Large vacuoles might replace the nuclei close to the cell walls and lead to a lack of microtubules when the cells abort. Abnormalities and defects in the organisation and composition of microtubules in the male sterile line highlighted the complex interaction between microtubules and cytoplasmic male sterility. Copyright © 2012 Society of Chemical Industry

Cytoplasmic male sterility

10.1002/jsfa.5722

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Citations (5)

BcOPR3 Mediates Defense Responses to Biotrophic and Necrotrophic Pathogens in Arabidopsis and Non-heading Chinese Cabbage

Phytopathology (2022)

Yuan Wang Shuilin Yuan Cen Shao Weitong Zhu Dong Xiao

In plants, the salicylic acid (SA) and jasmonic acid (JA) signaling pathways usually mediate the defense response to biotrophic and necrotrophic pathogens, respectively. Our previous work showed that after non-heading Chinese cabbage (NHCC) was infected with the biotrophic pathogen Hyaloperonospora parasitica, expression of the JA biosynthetic gene BcOPR3 is induced; however, its molecular mechanism remains unclear. Here, we overexpressed BcOPR3 in Arabidopsis and silenced BcOPR3 in NHCC001 plants to study the defensive role of BcOPR3 in plants against pathogen invasion. The results showed that overexpression of BcOPR3 increased the susceptibility of Arabidopsis to Pseudomonas syringae pv. tomato DC3000 (Pst DC3000) but enhanced its resistance to Botrytis cinerea. BcOPR3-silenced NHCC001 plants with a 50% reduction in BcOPR3 expression increased their resistance to downy mildew by reducing the hyphal density and spores of H. parasitica. In addition, BcOPR3-partly silenced NHCC001 plants were also resistant to B. cinerea, which could be the result of a synergistic effect of JA and SA. These findings indicate a complicated role of BcOPR3 in the mediating defense responses to biotrophic and necrotrophic pathogens.

Jasmonic acid

Pseudomonas syringae

NPR1

Plant Immunity

Botrytis

10.1094/phyto-02-22-0049-r

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Citations (1)

<p>Characterization and Cytotoxicity of Polyprenol Lipid and Vitamin E-TPGS Hybrid Nanoparticles for Betulinic Acid and Low-Substituted Hydroxyl Fullerenol in MHCC97H and L02 Cells</p>

International Journal of Nanomedicine (2020)

Ran Tao Chengzhang Wang Yin Lu Changwei Zhang Hao Zhou

This study demonstrated an innovative formulation including the polyprenol (GBP) lipid and vitamin E-TPGS hybrid nanoparticles (NPs) which was aimed to control the transfer of betulinic acid (BA) and low-substituted hydroxyl fullerenol (C60(OH)n). Additionally, it developed BA-C60(OH)n-GBP-TPGS-NPs delivery system and researched the anti-hepatocellular carcinoma (HCC) effects.The NPs were prepared by nanoprecipitation with ultrasonic-assisted emulsification (UAE) method. It was characterized by scanning electronic microscopy (SEM), transmission electron microscopy (TEM), FTIR spectrum, size distribution and zeta potential. Physical and chemical properties were evaluated through measurement of drug release, stability studies, drug loading efficiency (DE) and encapsulation efficiency (EE). Biological activities were evaluated through measurement of MTT assay, lactate dehydrogenase leakage assay (LDH), cell proliferation assays, cell apoptosis analysis, comet assay, wound healing assay, cell invasion and Western blot analysis.The NPs exhibited clear distribution characteristics, improved solubility and stability. BA and C60(OH)n for the NPs displayed a biphasic release pattern with sustained drug release properties. The mixture of C60(OH)n with different hydroxyl groups may have a certain effect on the stability of the NPs system itself. The NPs could effectively inhibit MHCC97H cell proliferation, migration and invasion in vitro. Combined use of C60(OH)n and BA in GBP lipids may improve the inhibit effect of C60(OH)n or BA against HCC cells and reduce cytotoxicity and genotoxicity of C60(OH)n for normal cells. We concluded that one of the important mechanisms of BA-C60(OH)n-GBP-TPGS-NPs inhibiting MHCC97H cells is achieved by up-regulating the expression of Caspase-3, Caspase-8 and Caspase-9.

Betulinic acid

Zeta potential

MTT assay

10.2147/ijn.s249773

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Citations (17)