采后黄瓜响应低温胁迫的转录组学分析

采后黄瓜对低温敏感,在低温贮藏期间很容易发生冷害,会造成较大的采后损失。本研究采用转录组测序结合生物信息学分析的方法,分析了采后黄瓜在短时冷害温度处理后的转录组变化。结果显示,采后黄瓜在5℃贮藏期间,冷害指数和相对电导率逐渐增加,叶绿素荧光逐渐降低,表明采后黄瓜在5℃贮藏期间产生了明显的冷害。与处理前相比（0 h）,处理12 h导致1500个基因差异表达,其中706个基因表达上调,794个基因表达下调。与0 h相比,处理72 h导致7799个基因差异表达,其中3995个基因表达上调,3804个基因表达下调。KEGG富集结果显示,低温处理导致的差异表达基因显著富集在植物激素信号转导、苯丙氨酸代谢、植物与病原菌互作和苯丙烷素合成途径中。GO富集分析的结果显示,在生物过程分类下,差异表达基因主要参与以DNA为模板的转录调控、蛋白磷酸化和跨膜转运等过程;在细胞组份分类下,差异表达基因主要富集在细胞膜组份和细胞核;在分子功能分类下,差异表达基因主要与ATP结合、蛋白苏氨酸/丝氨酸激酶活性、DNA结合和转录因子活性等功能有关。在低温处理12 h时,与激素信号有关的基因显著表达。但在低温处理72 h...     

睡莲叶片胎生发育转录组分析

睡莲叶片胎生现象是繁育途径的重要补充,对种群的传播、扩散和生态环境的适应性有重要作用。通过转录组测序技术筛选和分析睡莲叶片胎生现象相关的代谢路径和调控基因,为深入认识睡莲叶片胎生发育的分子机制提供参考。以叶片具有胎生现象的‘小花睡莲’（X）和叶片无胎生现象的‘蓝星睡莲’（L）为材料,利用RNA-Seq技术对叶片4个发育阶段的叶脐部分进行生物信息学分析。分析对照（L）和样品（X）叶片不同发育阶段测序结果：筛选出的差异表达基因（DEGs）中,34 909个基因（48.65%）表达上调,36 850个基因（51.35%）表达下调。DEGs分析显示,随着叶片的发育,X和L上调基因和下调基因数均呈增加趋势。对L1-vs-X1、L4-vs-X4阶段的GO和KEGG功能富集分析表明,DEGs主要富集在质膜和膜相关成分、胞外区域、细胞壁等相关的细胞组分中,涉及到代谢过程、生物合成和应急响应等;Pathway代谢通路表明,DEGs主要参与到植物激素信号转导、苯丙烷类生物合成、氨基酸类代谢、类黄酮生物合成、甘油磷脂类代谢以及细胞周期相关等过程。对DEGs进一步分析,克隆出了4个可能参与睡莲叶片胎生发育的转录因子。     

The Emerging Evidence for a Protective Role of Fucoidan from Laminaria japonica in Chronic Kidney Disease-Triggered Cognitive Dysfunction

This study aimed to explore the mechanism of fucoidan in chronic kidney disease (CKD)-triggered cognitive dysfunction. The adenine-induced ICR strain CKD mice model was applied, and RNA-Seq was performed for differential gene analysis between aged-CKD and normal mice. As a result, fucoidan (100 and 200 mg kg⁻¹) significantly reversed adenine-induced high expression of urea, uric acid in urine, and creatinine in serum, as well as the novel object recognition memory and spatial memory deficits. RNA sequencing analysis indicated that oxidative and inflammatory signaling were involved in adenine-induced kidney injury and cognitive dysfunction; furthermore, fucoidan inhibited oxidative stress via GSK3β-Nrf2-HO-1 signaling and ameliorated inflammatory response through regulation of microglia/macrophage polarization in the kidney and hippocampus of CKD mice. Additionally, we clarified six hallmarks in the hippocampus and four in the kidney, which were correlated with CKD-triggered cognitive dysfunction. This study provides a theoretical basis for the application of fucoidan in the treatment of CKD-triggered memory deficits.     

Influence of Fucoidan Extracts from Different Fucus Species on Adult Stem Cells and Molecular Mediators in In Vitro Models for Bone Formation and Vascularization

Fucoidans, sulfated polysaccharides extracted from brown algae, are marine products with the potential to modulate bone formation and vascularization processes. The bioactivity and safety of fucoidans are highly associated with their chemical structure, which may vary with algae species and extraction method. Thus, in depth evaluation of fucoidan extracts in terms of endotoxin content, cytotoxicity, and their detailed molecular biological impact on the individual cell types in bone is needed. In this study, we characterized fucoidan extracts from three different Fucus species including Fucus vesiculosus (Fv), Fucus serratus (Fs), and Fucus distichus subsp. evanescens (Fe) for their chemical features, endotoxin content, cytotoxicity, and bioactive effects on human outgrowth endothelial cells (OEC) and human mesenchymal stem cells (MSC) as in vitro models for bone function and vascularization. Extracts contained mainly high molecular weight (HMW) fucoidans and were free of endotoxins that may cause inflammation or influence vascularization. OEC tolerated fucoidan concentrations up to 200 µg/mL, and no indication of cytotoxicity was observed. The inflammatory response, however, investigated by real-time PCR (RT-PCR) and enzyme-linked immunosorbent assay (ELISA) and endothelial barrier assessed by impedance measurement differed for the individual extracts. MSC in comparison with endothelial cells were more sensitive to fucoidans and showed partly reduced metabolic activity and proliferation at higher doses of fucoidans. Further results for MSC indicated impaired osteogenic functions in alkaline phosphatase and calcification assays. All tested extracts consistently lowered important molecular mediators involved in angiogenesis, such a VEGF (vascular endothelial growth factor), ANG-1 (angiopoietin 1), and ANG-2 (angiopoietin 2), as indicated by RT-PCR and ELISA. This was associated with antiangiogenic effects at the functional level using selected extracts in co-culture models to mimic bone vascularization processes during bone regeneration or osteosarcoma.     

不同品种茶树新梢响应"倒春寒"的转录组分析

为探究“倒春寒”冻害对不同品种茶树新梢转录水平的影响,本研究以茶园“倒春寒”发生时受冻害和未受冻害的龙井43和中茶126新梢为研究材料,对两组样品进行转录组分析,分别鉴定到1 012个和1 079个差异基因,以及284个共同差异基因。从中选取18个差异基因进行实时荧光定量PCR验证,结果与转录组测序结果基本一致,证明转录组数据可靠。利用GO和KEGG数据库,对差异基因进行代谢通路富集分析,发现两个品种的差异基因主要集中在光合作用、碳代谢和细胞色素P450等代谢进程,说明“倒春寒”对新梢生长发育相关的基础代谢造成了严重损伤,抑制了相关基因的正常表达;随后对284个共同差异基因进行表达聚类分析,结果表明,其中99个差异基因在两个茶树品种中的表达模式完全相反,涉及的生物过程包括MAPK信号通路、谷胱甘肽和苯丙烷代谢等,推测这些差异基因与龙井43和中茶126在受到低温胁迫后产生的不同信号传导模式有关。     

A 3D-Printed Polycaprolactone/Marine Collagen Scaffold Reinforced with Carbonated Hydroxyapatite from Fish Bones for Bone Regeneration

In bone tissue regeneration, extracellular matrix (ECM) and bioceramics are important factors, because of their osteogenic potential and cell–matrix interactions. Surface modifications with hydrophilic material including proteins show significant potential in tissue engineering applications, because scaffolds are generally fabricated using synthetic polymers and bioceramics. In the present study, carbonated hydroxyapatite (CHA) and marine atelocollagen (MC) were extracted from the bones and skins, respectively, of Paralichthys olivaceus. The extracted CHA was characterized using Fourier transform infrared (FTIR) spectroscopy and X-ray diffraction (XRD) analysis, while MC was characterized using FTIR spectroscopy and sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE). The scaffolds consisting of polycaprolactone (PCL), and different compositions of CHA (2.5%, 5%, and 10%) were fabricated using a three-axis plotting system and coated with 2% MC. Then, the MC3T3-E1 cells were seeded on the scaffolds to evaluate the osteogenic differentiation in vitro, and in vivo calvarial implantation of the scaffolds was performed to study bone tissue regeneration. The results of mineralization confirmed that the MC/PCL, 2.5% CHA/MC/PCL, 5% CHA/MC/PCL, and 10% CHA/MC/PCL scaffolds increased osteogenic differentiation by 302%, 858%, 970%, and 1044%, respectively, compared with pure PCL scaffolds. Consequently, these results suggest that CHA and MC obtained from byproducts of P. olivaceus are superior alternatives for land animal-derived substances.     

玉米苗期根部比较转录组分析揭示耐盐性差异机制

以耐盐自交系郑58和盐敏感自交系昌7-2为材料,分析供试材料在无胁迫(CK)和200 mmol/L NaCl溶液胁迫下5 h的转录组数据。与无胁迫(CK)相比较,在郑58和昌7-2分别鉴定出390和421个上调、390和421个下调差异基因。对耐感材料间共同上调和郑58特有上调DEGs进行GO富集、REVIGO分析和KEGG富集分析以及重点通路分析结果表明：郑58和昌7-2间共同上调DEGs显著富集的条目包括对活性氧代谢过程的调节、内质网应激反应等GO条目;郑58特有上调显著富集含氧化合物的反应、对化学物质的反应等GO条目。共同上调DEGs仅在共单萜生物合成的代谢通路上显著富集;郑58特有上调DEGs在植物激素信号转导、苯丙烷生物合成等6个通路上显著富集。在植物激素信号转导、苯丙烷生物合成两个通路上分别有9和7个相关基因,耐盐自交系郑58上调的转录因子在MYB和NAC家族分布数量较多。植物激素信号转导、苯丙烷生物合成通路相关的基因以及MYB和NAC家族基因可能与玉米耐盐性密切相关。     

小麦品种西农979抗赤霉病基因的转录组测序鉴定

小麦品种西农979对赤霉病具有较好的抗性。为了发掘西农979的抗赤霉病基因,本研究采用禾谷镰刀菌菌株BN-8分别接种抗病品种西农979与感病品种周麦18,取接种前西农979颖壳及接种后12、24和96 h的西农979与周麦18颖壳进行转录组分析,筛选抗感赤霉病差异表达基因,并对其进行GO功能注释及KEGG富集分析。结果表明,接种后12、24和96 h各出现14、1 978和142个差异表达基因,共计2 122个基因。这些基因经GO功能注释分类分成3大类43个亚类,主要涉及代谢过程、细胞过程、应激反应、生物调控、细胞部件、细胞、催化活性、结合、转运活性等。KEGG通路分析结果显示,内质网中的蛋白质加工通路中差异基因最多,有48个;其次是光合作用—天线蛋白通路及淀粉和蔗糖代谢通路,各含41和30个差异基因;显著富集的通路有光合作用—天线蛋白、内质网中的蛋白质加工、萜骨架的生物合成等。植物激素信号转导通路中发现27个差异基因,参与脱落酸和茉莉酸等路径;植物-病原体互作通路中发现19个差异基因,编码钙结合蛋白、WRKY转录因子、抗病蛋白等。此外,还筛选到UDP-葡萄糖基转移酶基因等脱毒相关蛋白基因。     

铝诱导的茶树根系转录组变化分析

探讨茶树响应铝（Aluminum,Al）的基因调控网络和表达模式,确定一些关键候选基因,为茶树耐Al分子机制研究奠定基础。测定了0、0.2、1、2、4βmmol·L^-1 5个Al³⁺浓度处理7βd的福鼎大白茶根系抗氧化酶活性和Al含量变化,并提取0βmmol·L^-1（R0）、1βmmol·L^-1（R1）和4βmmol·L^-1（R4）3个浓度下的茶树根系总RNA,通过Illumina Hiseq Xten平台进行高通量转录组测序。结果表明,随着Al³⁺浓度的升高,根系POD（Peroxidase,过氧化物酶）活性逐渐下降,APX（Ascorbic acid peroxidase,抗坏血酸过氧化物酶）活性则逐渐升高。SOD（Superoxide dismutase,超氧化物歧化酶）活性在Al³⁺浓度为1βmmol·L^-1时最高,CAT（Catalase,过氧化氢酶）活性在各处理间无显著差异。根系中Al含量随着Al³⁺浓度的升高呈先上升后下降趋势,在Al³⁺浓度为1βmmol·L^-1时达到最高。经筛选得到R1 VS R0,R4 VS R0,R4 VS R1的DEGs（Differentially expressed genes）分别为1β894、2β439个和1β384个,显著上调（下调）的差异表达基因分别有733（1β161）、846（1β593）个和628（756）个。GO富集分析表明,3个处理组在生物学途径中富集最多的类别均为刺激响应。在分子功能和细胞组件方面,R1 VS R0和R4 VS R0富集最多的类别均为核酸结合转录因子活性和细胞外围,R4 VS R1富集最多的类别为氧化还原酶活性相关基因和膜区域。KEGG富集分析表明,R1 VS R0、R4 VS R0、R4 VS R1分别显著富集了29、41条和19条Pathway,它们包括转录因子、转运蛋白、植物-病原菌互作、苯丙烷生物合成途径等,鉴定到多个参与调控活性氧代谢、有机酸或金属转运蛋白、转录因子及细胞壁结构修饰等生理过程的基因在Al诱导后上调或抑制表达,显示这些基因与茶树耐Al分子机制密切相关。     

转Pi9抗稻瘟病基因水稻株系的比较转录组分析

【目的】在转录水平上解析Pi9基因介导的稻瘟病抗性调控机理,为培育抗病水稻品种提供理论依据。【方法】向水稻品种日本晴(NPB)及其转Pi9抗稻瘟病基因株系(NPB/Pi9)接种稻瘟菌。分别于接种后0 h、12 h、24 h、36 h提取叶组织样品,选取12503个水稻基因定制基因芯片,进行水稻基因转录组分析,并通过qRT-PCR对部分差异表达基因进行验证。【结果】NPB/Pi9在接种后12 h、24 h和36 h的基因表达量分别与其接种0 h表达量比较,共检测到7754个差异表达基因;相应地,感病水稻NPB在以上时间点共检测到7385个差异表达基因;在接种后36 h,NPB/Pi9的差异表达基因数目显著多于NPB。比较NPB/Pi9和NPB相同时间点的基因表达量,共获得4065个差异表达基因,其中接种后36 h的差异表达基因显著多于接种后0 h、12 h或24 h。因此,NPB/Pi9的稻瘟病防御反应更强烈。对NPB/Pi9与NPB相同时间点的差异表达基因进行GO和KEGG分析,细胞外区域、植物对刺激应答、转录调控、氧化还原、离子结合、次生代谢和植物激素相关的GO分类在接种后呈显著富集,苯丙氨酸代谢、类黄酮生物合成和植物激素信号途径的KEGG通路在接种后显著富集。与效应分子触发的免疫反应(ETI)相关的水杨酸信号途径、几丁质酶,以及与病原相关分子模式触发的免疫反应(PTI)相关的胞外区域、对刺激的应答、木质素合成等,均在抗感水稻之间差异表达。而且PTI/ETI共有的WRKY转录因子、MAPK激酶、茉莉酸和乙烯信号途径等发生差异表达。综上所述,NPB/Pi9和NPB的差异表达模式与ETI和PTI相关,两者相互联系并在Pi9介导的稻瘟病抗性中发挥作用。【结论】与日本晴比较,抗病基因型NPB/Pi9对稻瘟病防御反应更强烈。转录因子、激酶、NBS-LRR基因、几丁质酶、水杨酸、茉莉酸和乙烯信号途径,以及植物次生代谢在Pi9介导的稻瘟病抗病反应中发挥重要作用。     

The Anticancer Effect of Fucoidan in PC-3 Prostate Cancer Cells

Fucoidan, a sulfated polysaccharide, has a variety of biological activities, such as anti-cancer, anti-angiogenic and anti-inflammatory. However, the mechanisms of action of fucoidan as an anti-cancer agent have not been fully elucidated. The present study examined the anti-cancer effect of fucoidan obtained from Undaria pinnatifida in PC-3 cells, human prostate cancer cells. Fucoidan induced the apoptosis of PC-3 cells by activating both intrinsic and extrinsic pathways. The induction of apoptosis was accompanied by the activation of extracellular signal-regulated kinase mitogen-activated protein kinase (ERK1/2 MAPK) and the inactivation of p38 MAPK and phosphatidylinositol 3-kinase (PI3K)/Akt. In addition, fucoidan also induced the up-regulation of p21^Cip1/Waf and down-regulation of E2F-1 cell-cycle-related proteins. Furthermore, in the Wnt/β-catenin pathway, fucoidan activated GSK-3β that resulted in the decrease of β-catenin level, followed by the decrease of c-myc and cyclin D1 expressions, target genes of β-catenin in PC-3 cells. These results suggested that fucoidan treatment could induce intrinsic and extrinsic apoptosis pathways via the activation of ERK1/2 MAPK, the inactivation of p38 MAPK and PI3K/Akt signaling pathway, and the down-regulation of Wnt/β-catenin signaling pathway in PC-3 prostate cancer cells. These data support that fucoidan might have potential for the treatment of prostate cancer.     

Injectable Thermosensitive Chitosan-Collagen Hydrogel as A Delivery System for Marine Polysaccharide Fucoidan

Fucoidans, sulfated polysaccharides from brown algae, possess multiple bioactivities in regard to osteogenesis, angiogenesis, and inflammation, all representing key molecular processes for successful bone regeneration. To utilize fucoidans in regenerative medicine, a delivery system is needed which temporarily immobilizes the polysaccharide at the injured site. Hydrogels have become increasingly interesting biomaterials for the support of bone regeneration. Their structural resemblance with the extracellular matrix, their flexible shape, and capacity to deliver bioactive compounds or stem cells into the affected tissue make them promising materials for the support of healing processes. Especially injectable hydrogels stand out due to their minimal invasive application. In the current study, we developed an injectable thermosensitive hydrogel for the delivery of fucoidan based on chitosan, collagen, and β-glycerophosphate (β-GP). Physicochemical parameters such as gelation time, gelation temperature, swelling capacity, pH, and internal microstructure were studied. Further, human bone-derived mesenchymal stem cells (MSC) and human outgrowth endothelial cells (OEC) were cultured on top (2D) or inside the hydrogels (3D) to assess the biocompatibility. We found that the sol-gel transition occurred after approximately 1 min at 37 °C. Fucoidan integration into the hydrogel had no or only a minor impact on the mentioned physicochemical parameters compared to hydrogels which did not contain fucoidan. Release assays showed that 60% and 80% of the fucoidan was released from the hydrogel after two and six days, respectively. The hydrogel was biocompatible with MSC and OEC with a limitation for OEC encapsulation. This study demonstrates the potential of thermosensitive chitosan-collagen hydrogels as a delivery system for fucoidan and MSC for the use in regenerative medicine.     

小麦近等基因系幼穗二棱期转录组分析

抽穗期是与小麦适应性直接相关的一个很重要的性状,直接或间接影响小麦的产量和抗性。为明确控制抽穗期的关键表达基因,以抽穗期分别为196d和184d的小麦近等基因系Y57和96-2为材料,对其二棱期幼穗分别构建转录组测序文库,利用Illumina HiSeq 2500平台进行RNA-seq测序;将获得的clean read与中国春参考序列比对,得到unique read,采用FPKM法计算基因表达量,对差异表达基因进行功能注释、GO和COG功能分类以及KEGG通路分析。结果表明,供试材料经过测序获得质量不低于30的碱基比例(Q30)均高于85.5%;Y57和96-2分别有60 246 194和60 963 580条clean read,其中63.11%和69.51%能与参考基因组序列匹配。共筛选到395个差异表达基因,有382个基因得到功能注释;有298个基因获得GO功能注释,主要涉及细胞组分、结合、细胞反应和代谢过程;COG注释的基因主要涉及一般功能、信号转导机制和转录;KEGG功能注释分析发现,显著富集在光合作用-天线蛋白通路上的基因均上调表达。编码丝氨酸/苏氨酸蛋白磷酸酶2A的基因在Y57中几乎不表达,而在96-2中表达量较高。进一步分析发现,多个与生长发育、抽穗开花相关的转录因子在两个材料中的表达差异显著。此结果可为深入研究抽穗期相关基因和穗发育调控机制奠定基础。     

热胁迫下水仙花叶片的转录组分析

为研究水仙响应高温胁迫过程中相关基因的表达及响应热应激的分子机制,本研究以漳州水仙花为材料,使用BGISEQ-500测序技术,对高温（30、35 ℃）处理及正常生长条件（15 ℃）的水仙叶片进行转录组学的测序分析。使用Trinity软件对15（对照）、30、35 ℃（高温胁迫）处理24 h的水仙叶片测序数据进行从头组装,利用BLAST软件进行基因比对注释,通过DEGseq和PossionDis检测差异表达基因,并对差异表达基因进行GO和KEGG富集分析。结果表明：本次水仙花叶片测序共获得112 160条总长度为128 733 815 bp、平均长度为1147 bp的Unigene,其N50以及GC含量分别为1770 bp和42.33%。Nr比对注释匹配度最高的物种为石刁柏,利用KOG数据库进行比对,5560条Unigene序列分布于25个功能区域。差异表达基因GO富集显示与光合系统及膜系统相关的GO term被显著富集,KEGG分析显示次生代谢物的生物合成、苯丙烷类生物合成、代谢途径、光合作用、糖胺聚糖降解、乙醛酸和二羧酸代谢、光合作用-天线蛋白、单萜类生物合成等8条代谢途径在3个比较组中均显著性富集,硫胺素代谢途径是唯一在高温胁迫条件下均显著富集的代谢途径。本研究注释了大量相关基因序列,通过本研究为改善水仙花植物的耐热性及耐热性品种的培育提供了丰富的数据资源和分子基础。     

Effect of Oversulfation on the Composition,Structure, and In Vitro Anti-Lung Cancer Activity of Fucoidans Extracted from Sargassum aquifolium

Intensive efforts have been undertaken in the fields of prevention, diagnosis, and therapy of lung cancer. Fucoidans exhibit a wide range of biological activities, which are dependent on the degree of sulfation, sulfation pattern, glycosidic branches, and molecular weight of fucoidan. The determination of oversulfation of fucoidan and its effect on anti-lung cancer activity and related signaling cascades is challenging. In this investigation, we used a previously developed fucoidan (SCA), which served as a native fucoidan, to generate two oversulfated fucoidan derivatives (SCA-S1 and SCA-S2). SCA, SCA-S1, and SCA-S2 showed differences in compositions and had the characteristic structural features of fucoidan by Fourier transform infrared (FTIR) and nuclear magnetic resonance (NMR) analyses. The anticancer properties of SCA, SCA-S1, and SCA-S2 against human lung carcinoma A-549 cells were analyzed in terms of cytotoxicity, cell cycle, Bcl-2 expression, mitochondrial membrane potential (MMP), expression of caspase-3, cytochrome c release, Annexin V/propidium iodide (PI) staining, DNA fragmentation, and the underlying signaling cascades. Our findings indicate that the oversulfation of fucoidan promotes apoptosis of lung cancer cells and the mechanism may involve the Akt/mTOR/S6 pathway. Further in vivo research is needed to establish the precise mechanism whereby oversulfated fucoidan mitigates the progression of lung cancer.     

软腐病菌侵染下菜心叶片的转录组分析

为探讨软腐病菌侵染下菜心转录组功能基因信息,采用BGISEQ-500高通量测序技术对软腐病菌侵染下菜心叶片进行转录组测序,平均每个样获得47.27 M clean reads,Q20值均大于95%,共检测到表达的Unigene为36 760个,其中已知的有35 327个,预测新Unigene有1 433个;共检测出19 549个新转录本,其长度主要集中在300~2000 nt。功能注释结果显示,有32 047个Unigene在Nr数据库获得注释,其中注释到芸薹属白菜上的Unigene最多;GO功能共注释到12 588个Unigene;KEGG数据库注释到18 583个Unigene,涉及到137个代谢通路。共获得21 776个组间差异表达基因（differential expression genes, DEGs）,其中对照与发病前期组间的DEGs有15 007个,6 137个上调表达,8 870个下调表达;发病前期与发病中期组间DEGs有13 118个,10 278个上调表达,2 840个下调表达;发病中期与发病后期组间DEGs有11 293个,1 790个上调表达,9 503个下调表达。DEGs的Ven图分析显示5 110个基因在每组间均存在差异,DEGs功能分析显示DEGs参与泛素蛋白降解途径、过氧化物酶体途径、光合碳固定途径、糖酵解途径等多种生命活动。本研究结果为深入开展菜心抗软腐病基因组学和分子生物学研究奠定基础。     

基于加权关联网络分析的辣椒（Capsicum annuum L.）果实颜色转变相关基因的鉴定

辣椒属于茄科（Solanaceae）辣椒属（Capsicum L.）一年生或多年生植物,其播种面积和产值在中国均居蔬菜首位。果色是辣椒众多经济性状中最直观的性状之一,直接影响人们的购买选择。辣椒果色受多种因素调控,相关色素生物合成的分子调控还需进一步解析。本研究以果实发育过程中呈现5种颜色的辣椒种质HNCA0076为实验材料,进行了生理检测、转录组测序以及WGCNA分析。在果实5个不同颜色时期,叶绿素含量变化较大,类胡萝卜素含量整体呈现递增趋势,类黄酮含量无明显变化,花色苷含量在紫色果期后逐渐降低,橙色果期后又逐渐升高。RNA测序数据质量满足进一步分析要求。4个转色期的上调（下调）表达基因数分别为826（276）、390（588）、1248（1800）和1528（2485）,随着果实的逐步发育,相邻比较组合中的差异基因的数量逐渐增加。差异表达基因经GO富集分析,主要集中在分子功能中的四吡咯结合、血红素结合、转移酶活性、氧化还原酶活性、铁离子结合和水解酶活性等过程;KEGG富集分析主要富集在类胡萝卜素生物合成、类黄酮生物合成和淀粉、蔗糖代谢等信号通路,其中类胡萝卜素代谢途径的3个基因在紫色时期向黄色时期的转变过程中表达上调,类黄酮代谢途径的11个基因在黄色时期向红色时期的转变过程中表达下调。4个对比组合差异基因分别注释到8877、8736、8689、8573个转录因子,共同注释到8049个转录因子,包括AP2、F-box、MYB和UDPGT等类型。利用非冗余的差异表达基因进行了加权关联网络分析,关联到4个与叶绿素、类胡萝卜素、花色苷、类黄酮含量相关的模块,进一步分析表明p450（LOC107859314）、Pkinase（LOC107839192、LOC107875415）、F-box（LOC107863894）和zf-MYND（LOC107852593）等29个转录因子可能参与辣椒果实转色的调控。本研究为进一步探索辣椒果实色素形成过程中的关键调控因子以及分析相关色素生物合成的调控提供了重要数据。     

Chondrogenic differentiation of rat mesenchymal stem cells on silk fibroin/chondroitin sulfate/hyaluronic acid ternary scaffolds

Cartilage repair is a challenge in bone tissue reconstruction. In this study, silk fibroin (SF), chondroitin sulfate (CS) and hyaluronic acid (HA) were employed to fabricate scaffolds for tissue engineered cartilage by freeze drying technique. The secondary pores were formed in the main pores of SF/CS/HA scaffold which improved the pore connectivity and equilibrium swelling of the scaffold. Furthermore, rat bone marrow mesenchymal stem cells were seeded on the scaffolds to evaluate the cell adhesion and proliferation. Results of hematoxylin/eosin staining and cell counting kit-8 assay showed that the cells migration and differentiation of SF/CS/HA (80/15/5) scaffold were better than that of SF/CS/HA scaffolds with different ratios after 7 days culture. Moreover, immunohistochemistry and scanning electron microscope demonstrated that large amounts of collagen II and proteoglycans of the cells were expressed in the SF/CS/HA 3D scaffold, while the expression of collagen I was barely visible by immunohistochemistry. Abound of extracellular matrix was formed to morphologically round and distributed uniformly throughout the scaffolds. The 3D ternary scaffold could promote the cells chondrogenic differentiation without using any inductive agent and offer potential for cartilage tissue regeneration.