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Epigenomics ; 11(14): 1613-1625, 2019 Nov.
Artigo em Inglês | MEDLINE | ID: mdl-31701765


Aim: To understand whether the anatomical location of origin plays a role in shaping the DNA methylation (DNAm) landscape of psoriatic skins. Patients & methods: A number of 108 psoriatic and 57 control skin samples were grouped based on their anatomical locations. Two group t-tests were used to identify those differentially methylated sites and regions. Target region methylation loci were validated by bisulfate conversion sequencing. The correlations of DNAm with pathological features, DNAm and gene expression were also interrogated. Results: Our analysis revealed 315 location-specific differentially methylated sites for back, 291 for the extremities and 801 for abdomen. Moreover, we observed that the extremity-specific loci cg21942490 located on HOXA9 is associated with hyperkeratosis. We further observed that HOXA5 and KIAA1949 are differential methylation regions. Conclusion: Our study shown evidence of anatomical location-dependent DNAm pattern in psoriasis skins, and thus provided new insights into the pathogenesis of this disease.

Theor Appl Genet ; 132(1): 113-123, 2019 Jan.
Artigo em Inglês | MEDLINE | ID: mdl-30334067


KEY MESSAGE: We identified two curly-leaf (cul) mutants in cucumber. Map-based cloning revealed that both mutants are due to allelic mutations in the CsPHB gene, a homolog of the Arabidopsis PHABULOSA which encodes a class III homeodomain-leucine zipper (HD-ZIP III) transcription factor. Leaf rolling is an important agronomic trait in crop breeding. Moderate leaf rolling minimizes shadowing between leaves, leading to improved photosynthetic efficiency. Although a number of genes controlling rolled leaf have been identified from rice and other plant species, none have been mapped or cloned in cucurbit crops. In this study, we identified and characterized two curly leaf (cul) mutants, cul-1 and cul-2 in cucumber. With map-based cloning, we show that cul-1 and cul-2 are allelic mutations and CsPHB (Csa6G525430) was the candidate gene for both mutants. The CsPHB gene encoded a class III homeodomain-leucine zipper (HD-ZIP III) transcription factor. A single non-synonymous mutation in the fourth and fifth exons of the CsPHB was responsible for the cul-1 and cul-2 mutant phenotypes, respectively. The single-nucleotide substitutions in cul-1 and cul-2 were both located in cs-miRNA165/166 complementary sites of CsPHB. The expression level of CsPHB gene in multiple organs of cul-1 and cul-2 mutants was higher than that in the wild type, while the expression of cs-miRNA165/166 in the two genotypes showed the opposite trend. We speculate that disruption of the binding between the mutant allele of CsPHB and cs-miRNA165/166 leads to the curly-leaf phenotype. This is the first report to clone and characterize the CsPHB gene in the family Cucurbitaceae. Taken together, these results support CsPHB as an important player in the modulation of leaf shape development in cucumber.

Cucumis sativus/genética , Proteínas de Homeodomínio/genética , Zíper de Leucina , Folhas de Planta/crescimento & desenvolvimento , Proteínas de Plantas/genética , Fatores de Transcrição/genética , Alelos , Mapeamento Cromossômico , Clonagem Molecular , Genótipo , Fenótipo , Mutação Puntual
Theor Appl Genet ; 131(6): 1379-1389, 2018 Jun.
Artigo em Inglês | MEDLINE | ID: mdl-29541828


KEY MESSAGE: Two round-leaf mutants, rl-1 and rl-2, were identified from EMS-induced mutagenesis. High throughput sequencing and map-based cloning suggested CsPID encoding a Ser/Thr protein kinase as the most possible candidate for rl-1. Rl-2 was allelic to Rl-1. Leaf shape is an important plant architecture trait that is affected by plant hormones, especially auxin. In Arabidopsis, PINOID (PID), a regulator for the auxin polar transporter PIN (PIN-FORMED) affects leaf shape formation, but this function of PID in crop plants has not been well studied. From an EMS mutagenesis population, we identified two round-leaf (rl) mutants, C356 and C949. Segregation analysis suggested that both mutations were controlled by single recessive genes, rl-1 and rl-2, respectively. With map-based cloning, we show that CsPID as the candidate gene of rl-1; a non-synonymous SNP in the second exon of CsPID resulted in an amino acid substitution and the round leaf phenotype. As compared in the wild type plant, CsPID had significantly lower expression in the root, leaf and female flowers in C356, which may result in the less developed roots, round leaves and abnormal female flowers, respectively in the rl-1 mutant. Among the three copies of PID genes, CsPID, CsPID2 and CSPID2L (CsPID2-like) in the cucumber genome, CsPID was the only one with significantly differential expression in adult leaves between WT and C356 suggesting CsPID plays a main role in leaf shape formation. The rl-2 mutation in C949 was also cloned, which was due to another SNP in a nearby location of rl-1 in the same CsPID gene. The two round leaf mutants and the work presented herein provide a good foundation for understanding the molecular mechanisms of CsPID in cucumber leaf development.

Cucumis sativus/genética , Genes de Plantas , Folhas de Planta/crescimento & desenvolvimento , Proteínas Serina-Treonina Quinases/genética , Alelos , Mapeamento Cromossômico , Clonagem Molecular , Cucumis sativus/enzimologia , Genes Recessivos , Ligação Genética , Mutação , Fenótipo , Filogenia , Folhas de Planta/genética , Polimorfismo de Nucleotídeo Único
Theor Appl Genet ; 130(8): 1549-1558, 2017 Aug.
Artigo em Inglês | MEDLINE | ID: mdl-28466109


KEY MESSAGE: Next-generation sequencing-aided map-based cloning delimited the cucumber tendril - less1 ( td - 1 ) locus into a 190.7-kb region in chromosome 6 harboring a putative, novel-function candidate gene encoding a histone acetyltransferase ( CsGCN5 ). The tendril initiated from the lateral meristem is an important and characteristic organ for the species in the Cucurbitaceae family including cucumber (Cucumis sativus L.). While the tendril has its evolutionary significance, it also poses a nuisance in cucumber cultivation under protected environments in which tendril-less cucumber has its advantages. From an EMS mutagenesis population, we identified a tendril-less mutant B007, which was controlled by a recessive gene td-1. Through next-generation sequencing-aided map-based cloning, we show CsGCN5 (Cucumis sativus GENERAL CONTROL NONDEREPRESSIBLE 5), a cucumber gene for a histone acetyltransferase as the most possible candidate for td-1. A non-synonymous SNP in the first exon of CsGCN5 resulted in an amino-acid substitution from Asp (D) in the wild type to Asn (N) in the tendril-less mutant. The candidacy of CsGCN5 was further confirmed by multiple lines of evidence in both biparental and natural cucumber populations. Non-significant expression of CsGCN5 in multiple organs was found between the wild type and the mutant. CsGCN5 exhibited strong expression in the tendril of wild-type plants suggesting its important roles in growth and development of plant tendrils. The identification and characterization of the td-1 mutant from the present study provided a useful tool in understanding the molecular mechanisms of tendril organogenesis and investigation of novel functions of the histone acetyltransferase in cucumber.

Cucumis sativus/genética , Genes Recessivos , Histona Acetiltransferases/genética , Mapeamento Cromossômico , Clonagem Molecular , Cucumis sativus/enzimologia , DNA de Plantas/genética , Genes de Plantas , Ligação Genética , Filogenia , Polimorfismo de Nucleotídeo Único , Análise de Sequência de DNA
J Biomater Sci Polym Ed ; 27(13): 1317-30, 2016 09.
Artigo em Inglês | MEDLINE | ID: mdl-27376548


Biomaterial surface topography significantly influences cellular form and function. Using poly(L-lactic acid) films with normal spherulites, banded spherulites, and amorphous surfaces as model substrates, we conducted a systematic assessment of the role for polymer crystallization induced surface morphologies on cell growth and contact guidance. Microscopy and image analysis showed that the MC3T3-E1 cells spread out in a random fashion on the amorphous substrate. At 24 h post-seeding, MC3T3-E1 cells on both types of spherulite surfaces were elongated and aligned along the spherulite radius direction. For the banded spherulite surface with radial stripes and coupling annular grooves, the cell orientation and cell nuclear localization were related to the grooves structure. With increasing time, this orientation preference was weaker. These results demonstrate that the patterning of polymer crystallization structure provide important signals for guiding cells to exhibit characteristic orientation and morphology especially in the early stages of regeneration.

Materiais Biocompatíveis/química , Osteoblastos/citologia , Poliésteres/química , Animais , Adesão Celular , Técnicas de Cultura de Células , Linhagem Celular , Proliferação de Células , Sobrevivência Celular , Cristalização , Camundongos , Propriedades de Superfície , Engenharia Tecidual
Carbohydr Polym ; 94(1): 444-8, 2013 Apr 15.
Artigo em Inglês | MEDLINE | ID: mdl-23544561


We report here the self-assembly of reversible pH intelligent aggregate from natural polysaccharide hyaluronic acid and functionalized polyhedral oligomeric silsesquioxane with tertiary amines and hydroxyls (POSS-AH). Various forms such as micelles, sacs and membranes were self-assembled through different procedures in this organic/inorganic hybrid system. The complex can be manipulated simply by changing pH to produce hydrogen-bond and ionic interaction. Furthermore the aggregates can form at a wide pH range by adjusting the proportion of the two constituents. These pH-responsive composites will offer potential opportunities in many fields, including cell immune barriers, drug and gene delivery systems and chemical sensors.

Ácido Hialurônico/química , Compostos de Organossilício/química , Concentração de Íons de Hidrogênio , Micelas , Nefelometria e Turbidimetria , Tamanho da Partícula