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A SNP mutation in Clbl gene encoding TERMINAL FLOWER 1 protein is responsible for watermelon branchless. Lateral branching is one of the most important traits, which directly determines plant architecture and crop productivity. Commercial watermelon has the characteristics of multiple lateral branches, and it is time-consuming and labor-costing to manually remove the lateral branches in traditional watermelon cultivation. In our present study, a lateral branchless trait was identified in watermelon material WCZ, and genetic analysis revealed that it was controlled by a single recessive gene, which named as Clbl (Citrullus lanatus branchless). A bulked segregant sequencing (BSA-seq) and linkage analysis was conducted to primarily map Clbl on watermelon chromosome 4. Next-generation sequencing-aided marker discovery and a large mapping population consisting of 1406 F2 plants were used to further map Clbl locus into a 9011-bp candidate region, which harbored only one candidate gene Cla018392 encoding a TERMINAL FLOWER 1 protein. Sequence comparison of Cla018392 between two parental lines revealed that there was a SNP detected from C to A in the coding region in the branchless inbred line WCZ, which resulted in a mutation from alanine (GCA) to glutamate (GAA) at the fourth exon. A dCAPS marker was developed from the SNP locus, which was co-segregated with the branchless phenotype in both BC1 and F2 population, and it was further validated in 152 natural watermelon accessions. qRT-PCR and in situ hybridization showed that the expression level of Cla018392 was significantly reduced in the axillary bud and apical bud in branchless line WCZ. Ectopic expression of ClTFL1 in Arabidopsis showed an increased number of lateral branches. The results of this study will be helpful for better understanding the molecular mechanism of lateral branch development in watermelon and for the development of marker-assisted selection (MAS) for new branchless watermelon cultivars.

Fruit spine is an important trait in cucumber, affecting not only commercial quality, but also fruit smoothness, transportation and storage. Spine size is determined by a multi-cellular base. However, the molecular mechanism underlying the regulation of cucumber spine base remains largely unknown. Here, we report map-based cloning and characterization of a spine base size 1 (SBS1) gene, encoding a C2H2 zinc-finger transcription factor., Near-isogenic lines of cucumber were used to map, identify and quantify cucumber spine base size 1 (CsSBS1). Yeast-hybrid, bimolecular fluorescence complementation (BiFC), co-immunoprecipitation (Co-IP) and RNA-sequencing assays were used to explore the molecular mechanism of CsSBS1 in regulating spine base size development., CsSBS1 was specifically expressed in cucumber ovaries with particularly high expression in fruit spines. Overexpression of CsSBS1 resulted in large fruit spine base, while RNA-interference silencing of CsSBS1 inhibited the expansion of fruit spine base. Sequence analysis of natural cucumber accessions revealed that CsSBS1 was lost in small spine base accessions, resulting from a 4895 bp fragment deletion in CsSBS1 locus. CsSBS1 can form a trimeric complex with two positive regulators CsTTG1 and CsGL1 to regulate spine base development through ethylene signaling., A novel regulator network is proposed that the CsGL1/CsSBS1/CsTTG1 complex plays a significant role in regulating spine base formation and size, which offers a strategy for cucumber breeders to develop smooth fruit., This work was supported by the National Natural Science Foundation of China (31902020, 31972427), the Zhongyuan Youth Talent Program (ZYQR201912161), the Key Research Project of Henan institutions of higher learning (20A210015), the Program for Science & Technology Innovation Talents of Henan Province (21HASTIT038), and the Major Science and Technology Projects of Henan Province (201300111300).

3D cell culture technologies have recently shown very valuable promise for applications in regenerative medicine, but the most common 3D culture methods for mesenchymal stem cells still have limitations for clinical application, mainly due to the slowdown of inner cell proliferation and increase in the cell death rate. We previously developed a new 3D culture of adipose-derived mesenchymal stem cells (ASCs) based on its self-feeder layer, which to a certain extent simultaneously solves the two issues of ASC-3D cell culture. In this study, we compared the 3D spheroids formed on the self-feeder layer (SLF-3D ASCs) with the spheroids formed by using ultralow attachment plates (ULA-3D ASCs). We discovered that the cells of SLF-3D spheroids still have a greater proliferation ability than ULA-3D ASCs, and the volume of these spheroids increases rather than shrinks, with more viable cells inside the 3D spheroids compared with the ULA-3D ASCs. Furthermore, it was discovered that the SLF-3D ASCs are likely to exhibit the above-mentioned unique properties due to change in the expression level of ECM-related genes, like as Col3a, Mmp3, Has1, and Fn1. These results indicate that the SLF-3D spheroid is a promising way forward for clinical application.

Background MicroRNA-101 (miR-101) is a tumor suppressor microRNA (miRNA) and its loss is associated with the occurrence and progression of various diseases. However, the biological function and target of miR-101 in the pathogenesis of hypertrophic scars (HS) remains unknown. Methods We harvested HS and paired normal skin (NS) tissue samples from patients and cultured their fibroblasts (HSF and NSF, respectively). We used quantitative reverse transcriptase polymerase chain reaction (qRT-PCR), fluorescence in situ hybridization (FISH), enzyme-linked immunosorbent assays (ELISA) and Western blot analyses to measure mRNA levels and protein expression of miR-101, enhancer of zeste homolog 2 (EZH2), collagen 1 and 3 (Col1 and Col3) and α-smooth muscle actin (α-SMA) in different in vitro conditions. We also used RNA sequencing to evaluate the relevant signaling pathways and bioinformatics analysis and dual-luciferase reporter assays to predict miR-101 targets. We utilized a bleomycin-induced fibrosis mouse model in which we injected miR-101 mimics to evaluate collagen deposition in vivo. Results We found low expression of miR-101 in HS and HSF compared to NS and NSF. Overexpressing miR-101 decreased Col1, Col3 and α-SMA expression in HSF. We detected high expression of EZH2 in HS and HSF. Knockdown of EZH2 decreased Col1, Col3 and α-SMA in HSF. Mechanistically, miR-101 targeted the 3′-untranslated region (3′UTR) of EZH2, as indicated by the decreased expression of EZH2. Overexpressing EZH2 rescued miR-101-induced collagen repression. MiR-101 mimics effectively suppressed collagen deposition in the bleomycin-induced fibrosis mouse model. Conclusions Our data reveal that miR-101 targets EZH2 in HS collagen production, providing new insight into the pathological mechanisms underlying HS formation.

Abnormal activation of Wnt signaling has been demonstrated in the wound healing process and the pathogenesis of fibrotic disorders, with Wnt4 specifically identified as having a key role in the pathogenesis of renal, pulmonary and liver fibrosis. Wnt4 also was found to be upregulated by transforming growth factor-β1 (TGF-β1) in fetal and postnatal murine fibroblasts and bone marrow mesenchymal cells, suggesting an underlying cooperation between Wnt4 and TGF-β1 in fibrosis. However, the specific roles of Wnt4 in TGF-β1-induced skin myofibroblast transition and hypertrophic scar formation remain unclear. In the present study, we first observed reduced Wnt4 expression in hypertrophic scar tissue compared with that in normal skin tissue. Following upregulation by TGF-β1, Wnt4 inhibited the TGF-β1-induced transdifferentiation of fibroblasts into myofibroblasts. Using fibroblast-populated collagen lattice contraction assays, we showed that the increased contractility induced by TGF-β1 was significantly blocked by exogenous Wnt4 and the α-smooth muscle actin (α-SMA) expression was decreased in fibroblasts in the collagen lattices. In addition, knockdown of Wnt4 resulted in further increases in α-SMA and collagen I expressions. Further investigation showed that Wnt4 could inhibit the autocrine effect of TGF-β1 as well as block the phosphorylation of Smad3 and ERK but not of AKT or JNK. Lastly, using hypertrophic scar-derived fibroblasts, we showed that the elevated α-SMA and collagen I levels were markedly reduced after treatment with Wnt4. Taken together, our results suggest that Wnt4 negatively regulates TGF-β1-induced fibroblast activation, which may represent a novel therapeutic strategy for the treatment and prevention of hypertrophic scars.

Melon, Cucumis melo L., is an important horticultural crop with abundant morphological variability, but the genetic diversity and relationships within wild and cultivated melons remain unclear to date. In this study, thick-skinned (TC) (cultivated subspecies melo), thin-skinned (TN) (cultivated subspecies agrestis), and wild accessions were analyzed for genetic diversity and relationships using 36 microsatellite markers. A total of 314 alleles were detected with a mean allelic number of 8.72 and polymorphism information content of 0.67. Cluster analysis of the accessions resulted in four distinct clusters (I, II, III, and IV) broadly matching with the TC, TN, and wild groups. Cluster I contained only two Indian wild accessions. Cluster II was consisted of 49 South Asian accessions, 34 wild accessions, and 15 TN accessions. Cluster III was a typical TC group including 51 multiorigin TC accessions and one wild accession. The remaining 88 accessions, including 75 TN accessions, 6 wild accessions, and 7 TC accessions, formed the cluster IV, and all the TN and wild accessions in this cluster were from China. These findings were also confirmed by Principal component analysis and STRUCTURE analysis. The South Asian subspecies agrestis accessions, wild and cultivated, had close genetic relationships with a distinctive genetic background. Chinese wild melons showed closeness to cultivated subspecies agrestis landraces and could be a return from the indigenous cultivated melons. The AMOVA and pairwise F statistics (FST) presented genetic differentiation among the three groups, with the strongest differentiation (FST = 0.380) between TC and TN melons. These results offer overall information on genetic diversity and affiliations within a variety of melon germplasms and favor efficient organization and utilization of these resources for the current breeding purpose.

Lateral branching is one of the most important traits, which directly determines plant 27 architecture and crop productivity. Commercial watermelon has the characteristics of multiple 28 lateral branches, and it is time-consuming and labor costing to manually remove the lateral 29 branches in traditional watermelon cultivation. In our present study, a lateral branchless trait was 30 identified in watermelon material W CZ, and genetic analysis revealed that it was controlled by a 31 single recessive gene, which named as Clbl . A bulked segregant sequencing (BSA seq) and 32 linkage analysis was conducted to primarily mapping of Clbl on watermelon chromosome 4 33 Next-generation sequencing aided marker discovery and a large mapping population consisting of 34 1406 F 2 plants was used to further mapped the Clbl locus into a 9011 bp candidate region which 35 harbored only one candidate gene Cla018392 encoding a TERMINAL FLOWER 1 gene. Sequence 36 comparison of Cla018392 between two parental lines revealed that there was a SNP detected from 37 C to A in the coding region in the branchless inbred line WCZ , which resulted in a mutation of 38 Alanine (GCA) to Glutamate (GAA) at the fourth exon A dCAPS marker was developed from the 39 SNP locus, which was co-segregated with the branchless phenotype in both BC 1 and F 2 population, 40 and it was also further validated in 152 natural watermelon accessions. qRT PCR and in situ 41 hybridization showed that the expression levels of Cla0 18392 was significantly reduced in the 42 axillary bud and apical bud in the branchless line WCZ Ectopic expression of ClTFL1 in 43 Arabidopsis showed an increased number of lateral branches. The results of this study will be 44 useful for better understanding the molecular mechanism of lateral branch development in 45 watermelon and for the development of marker-assisted selection (MAS) for new branchless 46 watermelon cultivars.

Glucocorticoid counteracts cellular mechanoresponses by a novel lncRNA-dependent mRNA decay machinery., Mechanical stimuli on cells and mechanotransduction are essential in many biological and pathological processes. Glucocorticoid is an important hormone, roles, and mechanisms of which in cellular mechanotransduction remain unknown. Here, we report that glucocorticoid counteracted cellular mechanoresponses dependently on a novel long noncoding RNA (lncRNA), LINC01569. Further, LINC01569 mediated glucocorticoid effects on mechanotransduction by destabilizing messenger RNA (mRNA) of mechanosensors including early growth response protein 1 (EGR1), Cbp/P300-interacting transactivator 2 (CITED2), and bone morphogenic protein 7 (BMP7) in glucocorticoid receptor–mediated mRNA decay (GMD) manner. Mechanistically, LINC01569 directly bound to the GMD factor Y-box–binding protein 1 (YBX1). Then, the LINC01569-YBX1 complex was guided to the mRNAs of EGR1, CITED2, and BMP7 through specific LINC01569-mRNA interaction, thereby contributing to the successful assembly of GMD complex and triggering GMD. Our results uncovered roles of glucocorticoid in cellular mechanotransduction and novel lncRNA-dependent GMD machinery and provided potential strategy for early intervention in mechanical disorder–associated diseases.

Partial photooxidation of micropollutants may lead to various degradation intermediates, obviously affecting disinfection byproducts (DBPs) formation during the post-chlorination process. The photooxidation of atrazine (ATZ) in aqueous solutions with low-pressure mercury UV lamps in UV, UV/H2O2 and UV/TiO2 treatment system and the formation of chlorinated disinfection byproducts (DBPs) during subsequent chlorination processes including dichloroacetic acid (DCAA), trichloroacetic acid (TCAA), 1,1,1-trichloro-2-propanone (TCP), trichloromethane (TCM) and chloropicrin (CHP) were investigated in this study. The effect of solution pH on the oxidation pathway of ATZ in three UV photooxidation treatment process and the impact of photooxidation on the DBPs formations were assessed. Based on UPLC-ESI–MS/MS analyses, identification of main oxidation intermediates was performed and the plausible degradation pathways of ATZ in photooxidation system were proposed, indicating that photooxidation of ATZ in UV/H2O2 and UV/TiO2 process system was significantly pH-dependent processes. Dichloroacetic acid (DCAA), trichloroacetic acid (TCAA), 1,1,1-trichloro-2-propanone (TCP), trichloromethane (TCM) and chloropicrin (CHP) were detected in photooxidized ATZ solutions. Compared to the other three DBPs, TCM and TCP were the main DBPs formed. The DBPs formations were greatly promoted in oxidized ATZ solutions. Solution pH and UV irradiation time exhibited obvious impact on the DBPs formation on the basis of DBP species. The variation tendency of DBPs observed relates to the combustion of ATZ in photooxidation system and the production oxidation intermediates.

Although it is well-known that human skin aging is accompanied by an alteration in the skin microbiota, we know little about how the composition of these changes during the course of aging and the effects of age-related skin microbes on aging. Using 16S ribosomal DNA and internal transcribed spacer ribosomal DNA sequencing to profile the microbiomes of 160 skin samples from two anatomical sites, the cheek and the abdomen, on 80 individuals of varying ages, we developed age-related microbiota profiles for both intrinsic skin aging and photoaging to provide an improved understanding of the age-dependent variation in skin microbial composition. According to the landscape, the microbial composition in the Children group was significantly different from that in the other age groups. Further correlation analysis with clinical parameters and functional prediction in each group revealed that high enrichment of nine microbial communities (i.e., Cyanobacteria, Staphylococcus, Cutibacterium, Lactobacillus, Corynebacterium, Streptococcus, Neisseria, Candida, and Malassezia) and 18 pathways (such as biosynthesis of antibiotics) potentially affected skin aging, implying that skin microbiomes may perform key functions in skin aging by regulating the immune response, resistance to ultraviolet light, and biosynthesis and metabolism of age-related substances. Our work re-establishes that skin microbiomes play an important regulatory role in the aging process and opens a new approach for targeted microbial therapy for skin aging.

BackgroundAs an important agronomic trait affecting breeding and consumption choice, rind color of watermelon [Citrullus lanatus (Thunb.) Matsum. and Nakai] receives considerable attention from breeders. Although rind color occupies an important place in watermelon breeding work, the underlying molecular mechanism governing this complex trait has still not been determined. ResultsIn the present study, chlorophyll and carotenoid contents were hypothesized to be the main factors determining rind color. Clearly different structures and numbers of chloroplasts and grana lamellae were found among materials with different rind colors. To study the genes involved in the coloration process of the yellow, dark green, light green, and light green-gray rinds, 84,516 unigenes were obtained from the rind transcriptome. Through DEGs screening and pathway analysis of chlorophyll and carotenoid mechanism, the genes with abnormal expression levels were identified. Then, the nucleotide sequences and expression levels of the putative genes responsible for different rind colors were studied.ConclusionsThis study provides a comprehensive insight on the factors affecting rind coloration and investigated expression levels of the critical genes concerning pigment metabolism in watermelon. These results would help to analyze the molecular mechanisms of rind coloration regulation and development of the related organelles.

SummaryPlant height is one of the most important agronomic traits that directly determines plant architecture, and compact or dwarf plants can allow for increased planting density and land utilization as well as increased lodging resistance and economic yield. At least four dwarf/semidwarf genes have been identified in different melon varieties, but none of them have been cloned, and little is known about the molecular mechanisms underlying internode elongation in melon. Here, we report map-based cloning and functional characterization of the first semidwarf gene short internode (Cmsi) in melon, which encodes an ERECTA-like receptor kinase regulating internode elongation. Spatial-temporal expression analyses revealed that CmSI exhibited high expression in the vascular bundle of the main stem during internode elongation. The expression level of CmSI was positively correlated with stem length in the different melon varieties examined. Ectopic expression of CmSI in Arabidopsis and cucumber suggested CmSI as a positive regulator of internode elongation in both species. Phytohormone quantitation and transcriptome analysis showed that the auxin content and the expression levels of a number of genes involved in the auxin signaling pathway were altered in the semidwarf mutant, including several well-known auxin transporters, such as members of the ABCB family and PIN-FORMED genes. A melon polar auxin transport protein CmPIN2 was identified by protein–protein interaction assay as physically interacting with CmSI to modulate auxin signaling. Thus, CmSI functions in an auxin-dependent regulatory pathway to control internode elongation in melon. Our findings revealed that the ERECTA family gene CmSI regulates stem elongation in melon through auxin signaling, which can directly affect polar auxin transport.

The stripe pattern is an important agronomic trait in watermelon, which determines the fruit rind pattern and consumer choice. However, the genes controlling these traits are still largely unknown. In the present study, a dark-green stripe inbred line WT2 and a netted stripe inbred line WM204 were used for genetic analysis, which revealed that the dark-green stripe is controlled by a single dominant gene ClGS. By bulked-segregant analysis (BSA), the ClGS was primarily mapped on watermelon chromosome 6 by using F2 plants developed from a cross between WT2 and WM204. Next-generation sequencing-aided marker discovery and a large mapping population consisting of 1206 F2 plants was used for fine mapping of the ClGS gene, and it was further mapped into a 107 kb candidate region. There were 11 genes predicated in this candidate region and 10 of them were differentially expressed in the 1-DAP fruit rind between two parental lines. Furthermore, 64 SNPs and 3 Indels were detected in the CDS region of these candidate genes. To further confirm the candidate gene of ClGS, we investigated the sequence variations among 74 re-sequenced natural watermelon accessions by in silico bulk segregant analysis. A 3-bp insertion was identified in all the non-dark green stripe watermelon accession group, which was located on the 8th exon of Cla019205. A Indel marker developed harboring the insertion showed co-segregation with the phenotype in the F2 mapping population, and it was also in completely agreement with another 25 watermelon accessions by electrophoretic analysis. These evidences suggested Cla019205 is probably the candidate gene controlling dark-green stripe in watermelon. The results of this study will be helpful for better understanding of the stripe formation and marker-assisted selection in watermelon.

The genetic diversity and population structure of oriental thin-skinned melons, Cucumis melo subsp. agrestis, remain incompletely clear to date. In the present study, 163 melon accessions (153 subsp. agrestis and 10 subsp. melo accessions) mainly from India and China were examined for genetic diversity and population structure using a core set of 37 simple sequence repeat (SSR) markers evenly distributed across the 12 melon chromosomes. A total of 274 alleles were detected with the allelic number per marker ranging from 3 to 16 (mean = 7.40) and polymorphism information content (PIC) from 0.342 to 0.770 (mean = 0.524). With the SSR genotype data, two main groups (I and II), mainly represented by Indian and Chinese subsp. agrestis accessions, respectively, were obtained by the unweighted pair group method with arithmetic mean (UPGMA) and the principal component analysis (PCA). Group I contained 24 Indian subsp. agrestis landraces and four accessions from America and Maldives while group II was consisted of 122 Chinese accessions and 13 accessions from other seven countries. There existed a clear divergence between Indian and Chinese subsp. agrestis melons, resulting in two independent germplasm pools. Group II mainly represented by East Asian accessions could be further divided into three subgroups (II-1, II-2, and II-3). The analysis of molecular variance (AMOVA) and pairwise F statistics (FST) were performed to quantify the genetic differentiation among the whole panel, displaying a strong differentiation between Indian and Chinese subsp. agrestis accessions but a comparatively weak differentiation within East Asian pool. From these findings, Indian and Chinese subsp. agrestis accessions have a distant relationship with each other and probably undergo different domestication pathways in their domestication history. Our results provide useful information for pursuing the genetic diversity and structure of oriental thin-skinned melon resources, which will favour efficient utilization of these valuable germplasms in modern agriculture and breeding.

Breast cancer resistance to the monoclonal erbB2/HER2 antibody trastuzumab (or herceptin) has become a significant obstacle in clinical targeted therapy of HER2-positive breast cancer. Previous research demonstrated that such drug resistance may be related to dysregulation of miRNA expression. Here, we found that knockdown of the long non-coding RNA, urothelial cancer associated 1 (UCA1), can promote the sensitivity of human breast cancer cells to trastuzumab. Mechanistically, UCA1 knockdown upregulated miR-18a and promoted miR-18a repression of Yes-associated protein 1 (YAP1). A luciferase reporter assay confirmed the association of miR-18a with wild-type UCA1 but not with UCA1 mutated at the predicted miR-18a-binding site. The direct targeting of YAP1 by miR-18a was verified by the observation that miR-18a mimic suppressed luciferase expression from a construct containing the YAP1 3' untranslated region. Meanwhile, reciprocal repression of UCA1 and miR-18a were found to be Argonaute 2-dependent. Knockdown of YAP1 recapitulated the effect of UCA1 silencing by reducing the viability of trastuzumab-treated breast cancer cells, whereas inhibition of miR-18a abrogated UCA1 knockdown-induced improvement of trastuzumab sensitivity in breast cancer cells. These findings demonstrate that the UCA1/miR-18a/YAP1 axis plays an important role in regulating the sensitivity of breast cancer cells to trastuzumab, which has implications for the development of novel approaches to improving breast cancer responses to targeted therapy.

Plant appearance is one of the most important characteristics affecting consumption and breeding choices. In watermelon, except for the normal phenotypes such as the basic foreground pattern (solid/stripe), background pattern (yellow/green rind), and fruit shape (round/oblong), there was a novel and unique appearance called ‘moon and star’ trait with big and tiny yellow spots on the fruit rind. Following a dominant inheritance pattern, the attracting appearance is convenient for variety breeding. The result of this study revealed that the chlorophyll content dramatically decreased and chloroplast structure significantly changed for the yellow spot, which were supposed to be the direct reason for fruit color change. Contrary to a previous study, genetic analysis showed that there were two interacting genes responsible for this novel trait. Using bulk segregant analysis, two candidate regions located on chromosome 1 and 8 were identified to contain these two genes regulating the ‘moon and star’ trait.

Loss of key components that form cell-cell adherens junctions, such as α-catenin, triggers severe epidermal hyperproliferation. However, the underlying molecular mechanisms remain largely unknown. We report here that neuroblastoma breakpoint family (NBPF) genes are upregulated and that NBPF7 specifically promotes cellular proliferation of α-catenin-silenced HaCaT cells through functional linkage with the NF-κB pathway. Genome-wide profiling of HaCaT cells shows that NBPF genes are upregulated following α-catenin knockdown. Data from western blot analyses are consistent with the activation of the NF-κB pathway as well as increased expression of NBPF7 by α-catenin knockdown. Co-immunoprecipitation assays indicate that NBPF7 could be detected in endogenous activated NF-κB immunoprecipitates. Immunoflurence analyses demonstrate that NBPF7 co-localizes with activated NF-κB in the nucleus after α-catenin silencing. Moreover, inhibition of NBPF7 decreases the proliferation of HaCaT cells and abolishes the enhanced proliferation associated with α-catenin knockdown in HaCaT cells. These results indicate that NBPF7 plays a key role in the α-catenin signaling pathway that regulates cell proliferation of keratinocytes. Our findings suggest that the classical NF-κB pathway plays a critical role in cellular proliferation and that NBPF7 is a functional mediator for α-catenin in the regulation of keratinocyte growth.

The kinetics, degradation mechanism and degradation pathways of atrazine (ATZ) during sole-UV and UV/H

Dwarf habit is one of the most important traits in crop plant architecture, as it can increase plant density and improved land utilization, especially for protected cultivation, as well as increasing lodging resistance and economic yield. At least four dwarf genes have been identified in watermelon, but none of them has been cloned. In the current study, the Cldw-1 gene was primary-mapped onto watermelon chromosome 9 by next-generation sequencing-aided bulked-segregant analysis (BSA-seq) of F2 plants derived from a cross between a normal-height line, WT4, and a dwarf line, WM102, in watermelon. The candidate region identified by BSA-seq was subsequently validated and confirmed by linkage analysis using 30 simple sequence repeat (SSR) markers in an F2 population of 124 plants. The Cldw-1 gene was further fine-mapped by chromosome walking in a large F2 population of 1,053 plants and was delimited into a candidate region of 107.00 kb. Six genes were predicted to be in the candidate region, and only one gene, Cla010337, was identified to have two single nucleotide polymorphisms (SNPs) and a single nucleotide deletion in the exons in the dwarf line, WM102. A derived cleaved amplified polymorphic sequence (dCAPS) marker was developed from the single nucleotide deletion, co-segregated with the dwarf trait in both the F2 population and a germplasm collection of 165 accessions. Cla010337 encoded an ATP-binding cassette transporter (ABC transporter) protein, and the expression levels of Cla010337 were significantly reduced in all the tissues tested in the dwarf line, WM102. The results of this study will be useful in achieving a better understanding of the molecular mechanism of the dwarf plant trait in watermelon and for the development of marker-assisted selection (MAS) for new dwarf cultivars.

As an important appearance trait, the rind color of watermelon fruit affects the commodity value and further determines consumption choices. In this study, a comparative transcriptome analysis was conducted to elucidate the genes and pathways involved in the formation of yellow rind fruit in watermelon using a yellow rind inbred line WT4 and a green rind inbred line WM102. A total of 2,362 differentially expressed genes (DEGs) between WT4 and WM102 at three different stages (0, 7, and 14 DAP) were identified and 9,770 DEGs were obtained by comparing the expression level at 7 DAP and 14 DAP with the former stages of WT4. The function enrichment of DEGs revealed a number of pathways and terms in biological processes, cellular components, and molecular functions that were related to plant pigment metabolism, suggesting that there may be a group of common core genes regulating rind color formation. In addition, next-generation sequencing aided bulked-segregant analysis (BSA-seq) of the yellow rind pool and green rind pool selected from an F2 population revealed that the yellow rind gene (Clyr) was mapped on the top end of chromosome 4. Based on the BSA-seq analysis result, Clyr was further confined to a region of 91.42 kb by linkage analysis using 1,106 F2 plants. These results will aid in identifying the key genes and pathways associated with yellow rind formation and elucidating the molecular mechanism of rind color formation in watermelon.

We investigated the mechanisms mediating hepatic metabolic responses to an acute lipopolysaccharide (LPS) challenge in goats. Guanzhong dairy goats (15) were randomly divided into three groups: control (CTL, saline, 0.2 ml/kg BW), lower dose LPS (LPS-L, 20 μg/kg BW) and higher dose LPS (LPS-H, 40 μg/kg BW). All injections were administered intraperitoneally twice with a 24-h interval. Forty-eight hours after the first injection, blood samples were collected to extract plasma for biochemical analysis, and liver tissues were biopsied and stored in liquid nitrogen for metabonomics analysis. We found that plasma levels of alanine aminotransferase, aspartate aminotransferase and total bilirubin increased (p

Cucumber fruit wart composed of tubercule and spine (trichome on fruit) is not only an important fruit quality trait in cucumber production, but also a well-studied model for plant cell-fate determination. The development of spine is closely related to the initiation and formation of tubercule. The spine differentiation regulator CsGL1 has been proved to be epistatic to the tubercule initiation factor CsTu, which is the only connection to be identified between spine and tubercule formations. Our previous studies found that the MIXTA-LIKE transcription factor CsMYB6 can suppress fruit spine initiation, which is independent of CsGL1. How the formation of spine and tubercule is regulated at the molecular level by CsMYB6 remains poorly understood. In this study, we characterized cucumber 35S:CsMYB6 transgenic plants, which displayed an obvious reduction in the number and size of fruit spines and tubecules. Molecular analyses showed that CsMYB6 directly interacted with the key spine formation factor CsTTG1 in regulating the formation of fruit spine, and CsTu in regulating the initiation of fruit tubercule, respectively. Based on these evidences, a novel regulatory network is proposed by which CsMYB6/CsTTG1 and CsMYB6/CsTu complexes play an important role in regulating epidermal development, including spine formation and tubercule initiation in cucumber.

Melon (Cucumis melo L.) is an economically important fruit crop with high phenotypic diversity, but the high-density genetic map suitable for QTL analysis of multiple phenotypes is unavailable to date. In this study, we adopted double-digest restriction-site associated DNA sequencing (ddRAD-seq) for large-scale discovery of single nucleotide polymorphisms (SNPs) and construction of a high-density genetic map in melon. A F2 population containing 140 individuals was developed from two parental lines, H581 and H906, which respectively belonged to flexuosus and reticulatus botanical groups and showed a significant phenotypic difference. Approximately 441 Gb of data containing 3.4 × 109 paired-end reads were obtained following the ddRAD-seq. Mining this dataset, 11,073 SNPs were detected between the two parents, of which 4654 valid SNPs were finally retained in the population for genetic map construction. In total, 1090 bin markers, consisting of all the 4654 SNPs, were sorted into 12 linkage groups (LGs). The total length of the bin map was 1462.30 cM, with an average marker density of 1.39 cM and a recombination rate of 3.90 cM/Mb. Haplotype map and linkage relationship heat map revealed that most of the recombination blocks were well defined in the segregating population, confirming a high accuracy of the bin map. This map could be used for discovery of QTLs/genes related to the diverse phenotypes between the two botanical groups, further facilitating molecular breeding in melon.

The wastewaters produced in chemical production contain large numbers of persistent organic pollutants, posing potential adverse impacts to aquatic ecosystem and human health. In this work, the electrochemical degradation of organic pollutants in chemical wastewater belonging to Sunny wealth chemicals (Linyi, Shandong Province) was conducted using anodic oxidation process. Three commercially available anodes (Pt, boron-doped diamond (BDD) and Ti-Ru-Sn ternary oxide (Ti/RuO2-SnO2)) were chosen to investigate the effect of several primary operating parameters, such as electrolyte composition, electrolyte concentration and applied current density. Total organic carbon (TOC) was detected as reference parameters to assess the performance of anodic oxidation. The experimental results show that BDD anode exhibited excellent oxidation capacity for degradation of organics in chemical wastewater, achieving over 30% removal efficiency in all cases within 4 h. TOC removal efficiency of 52% was obtained with BDD anode in 4 h of reaction time at applied current density 30 mA cm-2, NaCl concentration 0.05 mol L-1; nevertheless, less than 45 % of removal was achieved with Pt and Ti/RuO2-SnO2 anode under the same conditions.

Background Acute hyperglycemia is regarded as a risk factor for critically ill patients; however, insufficient understanding of its nature and underlying mechanisms hinders widespread adoption of glycemic control in critical care units. Methods A single center, prospective cohort study recruiting 107 burn patients and 62 controls was conducted to characterize the early phase of acute hyperglycemia in burn patients. A total of 1643 blood samples were collected and analyzed over the entire postburn 200 h. A mouse severe burn model was used to study the underlying mechanisms of acute hyperglycemia postburn. Results The dynamic change of postburn blood glucose represented a distinctive pattern in amplitude and duration that was in parallel with the degree of burn injury. Multiple linear regressions revealed that serum insulin, glucagon and glucocorticoid were the major factors affecting blood glucose postburn. Particularly, extensive burns impaired capacity and responsiveness of pancreatic insulin secretion, which was associated with increased serum IL-1β in burn patients. Mechanistically, acute IL-1β elevation specifically induced pancreatic beta cell apoptosis and dampened capacity of insulin secretion, leading to postburn hyperglycemia in burned mice. More importantly, inhibition of IL-1β not only alleviated pancreatic beta cell apoptosis, but also attenuated hyperglycemia and improved survival of burned mice. Conclusions Our findings reveal a novel mechanism of acute hyperglycemia postburn in which impaired insulin secretory capacity mediated by IL-1β leads to acute hyperglycemia. These data suggest that targeting IL-1β to restore endogenous insulin secretory function may be a novel glycemic control strategy to improve outcomes for burn patients.

Lipopolysaccharide (LPS) is a common pathogenic agent that causes many diseases and metabolic disorders. Hypoglycemia is often observed when animals are infected with LPS. To explore the influence of LPS on blood glucose and hepatic gluconeogenesis in goats, 12 goats were randomly assigned to 1 of 2 groups: the LPS-treated group (60 μg/kg BW of LPS; jugular vein injections) or the control group (saline vehicle; jugular vein injections). Blood samples were collected from jugular veins at 0, 1, 2, 4, 6, and 8 h, and liver tissue samples were biopsied 8 h after the injections. The dynamic changes in blood glucose levels as well as key hepatic gluconeogenic enzyme mRNA and protein expression, ATP and ADP levels, and glutathione reductase (GR) activity were determined. The results showed that blood glucose levels in the LPS group were dramatically reduced after an initial, short-term increase. In liver tissue, the mRNA of key gluconeogenic enzymes, phosphoenolpyruvate carboxykinase 1 (PEPCK1;P 0.05). The protein expression of PEPCK1 decreased (P < 0.01), whereas that of G6Pase-α increased (P < 0.05) significantly. The ratio of ADP to ATP ( < 0.05) and the activity of GR (P < 0.01) were markedly increased in the LPS group compared with those in controls. This research showed that LPS markedly affects and reduces blood glucose in dairy goats. The crucial reasons for the marked change in blood glucose are the altered expression of key gluconeogenic enzymes in different pathways and of essential factors associated with gluconeogenesis in the liver.

The transcription factor peroxisome proliferator-activated receptor-γ (PPAR-γ) functions to regulate cell differentiation and lipid metabolism. Recently, its agonist has been documented to regulate extracellular matrix production in human dermal fibroblasts. This study explored the underlying molecular mechanisms and gene interactions in hypertrophic scar fibroblasts (HSFBs) in vitro. HSFBs were cultured and treated with or without PPAR-γ agonist or antagonist for gene expression. Bioinformatical analysis predicted that miR-145 could target Smad3 expression. Luciferase assay was used to confirm such an interaction. The data showed that PPAR-γ agonist troglitazone suppressed expression of Smad3 and Col1 in HSFBs. PPAR-γ agonist induced miR-145 at the gene transcriptional level, which in turn inhibited Smad3 expression and Col1 level in HSFBs. Furthermore, ELISA data showed that Col1 level in HSFBs was controlled by a feedback regulation mechanism involved in PPAR-γ agonist and antagonist-regulated expression of miR-145 and Smad3 in HSFBs. These findings indicate that PPAR-γ-miR-145-Smad3 axis plays a role in regulation of collagen synthesis in HSFBs.

Knowledge of boron and its isotope in plants is useful to better understand the transposition and translocation of boron within plant, the geochemical behavior in the interface between soil and plant, and the biogeochemical cycle of boron. It is critical to develop a useful method to separate boron from the plant for the geochemical application of boron and its isotope. A method was developed for the extraction of boron in plant sample, whose isotope was determined by thermal ionization mass spectrometry. The results indicated that this method of dry ashing coupled with two-step ion-exchange chromatography is powerful for the separation of boron in plant sample with large amounts of organic matters completely. The ratios of boron isotope composition in those plant tissue samples ranged from-19.45‰to+28.13‰(total range:47.58‰) with a mean value of2.61±11.76‰SD. The stem and root isotopic compositions were lower than those in flower and leaf. The molecular mechanism of boron isotope may be responsible for the observed variation of boron isotopic composition and are considered as a useful tool for the better understanding of boron cycling process in the environment and for the signature of living systems.

We have synthesized and characterized a series of functional binary graft copolymers PGMA-g-(PHFBMA-r-POEGMA)s(BGCs). First, PHFBMA–CCH, POEGMA–CCH and P(GMA-N3) were synthesized via sequential atom transfer radical polymerization (ATRP). BGCs were prepared by grafting of alkyne-end poly(hexafluorobutyl methacrylate) (PHFBMA–CCH) and poly(oligo (ethylene glycol) methyl ether methacrylate) (POEGMA–CCH) onto poly(3-azide-2-hydroxypropyl methacrylate) (P(GMA-N3)) via click chemistry. The self-assembly behaviors were investigated by combination of dynamic light scattering (DLS), transmission electron microscopy (TEM), and atomic force microscopy (AFM). Since POEGMA was soluble in water while PHFBMA was insoluble, BGCs self-assembled and produced stable PHFBMA-centered nano-micelles; then BGCs were used to fabricate hydrophobic cotton fabrics. While the PHFBMA block provided low surface free energy, the POEGMA block served as an anchor with cotton fibers, the modified cotton fabrics showed excellent superhydrophobic property. The results confirmed that fluorinated surface was formed onto substrate without changing the transparency and bulk composition of the cotton fabrics. Moreover, SEM and AFM analysis indicated that nano- and microscale roughness were created by combining BGC-based nano bumps onto surfaces of micro-sized cotton fabrics.

As a critical micronutrient, boron (B) plays an important role in plant growth and embryonic development. To further understand the effects of B uptake, transportation and isotopic fractionation, the contents and isotopic compositions of hydro-soluble B in the sap and structural B fixed in the cell within individual plant tissues were investigated. The B isotope ratio was determined by multi-collector inductively coupled plasma mass spectrometry. The δ11B values in hydro-soluble and structural B in the investigated plant samples ranged from −1.57‰ to +11.30‰ and from +6.57‰ to +16.64‰, respectively. Different fractionation factors of the B isotopes, in the range of 0.9954–1.0150, were observed in these samples, indicating that in most plant tissues, the heavy isotope (11B) was preferentially enriched in structural B, which was fixed into the cell. However, there was a reversal in the fractionation of B isotopic compositions in the fruit samples compared with the other plant tissue samples. It is more powerful to examine the molecular mechanisms of B transport, uptake and utilization than the use of limited plant organ samples containing a mixture of hydro-soluble and structural B within different intra-plant compartments and in inter-plant interactions. These isotopic shifts, which may be used as important isotopic indicators, contribute to the surface processes interactions in the plant–soil system and the knowledge of the molecular mechanisms of B in the uptake and absorption by different plant species in nature.

Map-based cloning identified CmGL that encodes a HD-ZIP type IV transcription factor that controls multicellular trichome initiation in melon. Trichomes are small hairs covering the aerial parts of plants that originate from the epidermal cells, which can protect plants against the damage by insects and pathogens. The regulatory pathway of unicellular trichomes has been well studied in the model plant Arabidopsis. Little is known about the genetic control and regulation of trichome development in melon (Cucumis melo L.) which has multicellular trichomes. In this study, we identified a melon mutant, cmgl, which showed completely glabrous on all aerial organs. A bulked segregant analysis was conducted to identify polymorphic markers for linkage analysis in a population with 256 F2 plants, which allowed to locate the cmgl locus in melon chromosome VIII. Next-generation sequencing-aided marker discovery and fine mapping in a large population with 1536 F2 plants narrowed the candidate gene region to 12 kb that harbored only one candidate gene for cmgl, which encoded a class IV homeodomain-associated leucine zipper transcription factor. Four SNPs in the coding region of the CmGL gene were identified between the two parental lines; a single base substitution from C to A resulted in a premature termination codon and a truncated protein in the cmgl. The SNP was converted into a dCAPS marker, which showed co-segregation in the F2 population and 564 melon accessions. Result of this study will be helpful for better understanding of genetic control of trichome development in melon and marker-assisted selection in developing new cultivars.