公牛(Bos taurus)去势和长期育肥能够促进内脏脂肪的沉积,过多的脂肪沉积易引发机体慢性炎症,然而有关牛网膜脂肪组织炎症的分子机理研究相对较少.本研究以牛网膜脂肪组织为研究对象,旨在探究性别和月龄对荷斯坦育肥牛网膜脂肪组织炎症相关基因表达的影响.选择16月龄荷斯坦公牛(BF16)、阉牛(SF16)及26月龄阉牛(SF26)各3头,测定生长和血液生化指标,屠宰后采集网膜脂肪组织,进行组织切片苏木精-伊红染色(hematoxylin-eosin staining,HE)染色,采用RNA-seq技术对3组牛网膜脂肪组织进行转录组学分析,筛选组间差异基因,并对差异基因进行GO功能注释和KEGG富集分析、qPCR验证、候选基因筛选及蛋白网络互作分析.结果显示,共得到1173个组间差异表达基因,富集在8个基因表达趋势模式中;功能分析显示差异基因富集多个与炎症相关的生物学过程和信号通路;随机选择6个差异表达的基因进行qPCR验证,表达趋势与转录组测序结果一致,表明测序结果可靠;筛选到与炎症相关的5个生物学过程条目和5个信号通路共50个非冗余基因,聚类分析发现,与BF16组相比,去势能够上调SF16组网膜脂肪组织中抗炎症和促脂肪细胞分化基因(如TNF-α诱导蛋白3(TNF alpha induced protein 3,TNFAIP3),骨形成蛋白2(bone morphogenetic protein 2,BMP2)等)的表达,下调促炎症基因(如白细胞介素1β(interleukin-1beta,IL1B))的表达;与BF16和SF16组相比,长期育肥能够上调SF26组网膜脂肪组织中衰老和慢性炎症相关的基因(如半乳糖凝集素3(galectin 3,LGALS3),C-C基序趋化因子受体7(C-C motif chemokine receptor 7,CCR7)等)的表达.蛋白网络互作分析显示IL1B为关键中心节点.本研究结果为优化利用荷斯坦阉牛生产大理石纹牛肉技术提供理论依据.

民猪具有良好的抗寒能力,研究表观遗传对民猪脂肪细胞分化的影响,揭示民猪脂肪细胞分化的分子机制,将为探索民猪抗寒性状形成原因提供依据.在体外培养的民猪前脂肪细胞培养液中分别添加不同浓度5-Aza-CdR和TSA处理一定时间,MTT法和流式细胞术检测前脂肪细胞活率,确定最适处理浓度;用最适浓度的5-Aza-CdR和TSA处理前脂肪细胞诱导形成成熟脂肪细胞后,分别检测细胞中表观遗传相关基因DNMT1、DNMT3B、ALDH1A3和ALDH1A9,以及脂肪细胞棕色化相关基因ZNF423、PPARα、NCOR1和EBF2的mRNA相对表达量,并用油红O染色观察脂滴形成情况.结果表明,在本实验研究条件下,500 nmol/L的5-Aza-CdR处理48 h和150 nmol/L的TSA处理24 h是对前脂肪细胞生长影响最小的最高浓度,作为后续实验的细胞处理浓度.5-Aza-CdR处理前脂肪细胞后能够降低成熟脂肪细胞中DNMT1、DNMT3B、ZNF423、PPARα、NCOR1和EBF2基因的mRNA相对表达量(P<0.05),并抑制脂滴生成.TSA处理后,成熟脂肪细胞中ALDH1A3和ALDH1A9基因的mRNA相对表达量略有降低,ZNF423、PPARα、NCOR1和EBF2基因的mRNA相对表达量上升(P<0.05).综上所述,猪脂肪细胞的分化受到表观遗传试剂5-Aza-CdR和TSA的影响,5-Aza-CdR处理会促进前脂肪细胞向棕色脂肪方向分化,同时抑制白色脂肪形成和沉积,而TSA的作用则正好相反.

【Objective】The objective of this study is to research into the early stage root gene expression profile of G.max induced by H.glycines and to explore the resistant mechanism to the pathogen at molecular level.【Method】SSH-cDNA libraries enriched with differentially expressed ESTs were constructed from SCN-challenged root tissues at pre-penetration and early infection stages from Xiaoli black bean(G.max) by suppression subtractive hybridization and reverse Northern blot.【Result】 One hundred and sixty-six unique ESTs were identified and analyzed with Blastx and Blastn by comparing sequences in the GenBank.One hundred and ninteen unigenes accounted for 83% of all the unigenes showed high homology with the function known genes or proteins.The function-known ESTs were annotated into functional categories including signal recognition and transduction,energy and material metabolism,stress responses,transcriptional regulation,protein synthesis and/or modification,transport functions,cellular architecture.【Conclusion】Discreet gene tag clusters primarily including catalase,ubiquitin,chitinase,lipoxygenase,aquaporin,ripening related protein,metallothionein,plasma membrane intrinsic protein,cytochrome P450,glyceraldehyde-3-phosphate dehydrogenase were abundant in the SCN-infected roots.It is speculated that those genes played an important function in the incompatible interaction between Xiaoli black bean and H.glycines.

Hsp90 had been identified to be highly conserved among different species.The Sl-hsp90 gene of tomato was amplified from Solanum lycopersicum L-402 cultivar genome DNA by DNA-PCR method.The plant expression vector pCAMBIA1302 containing Sl-hsp90 gene was constructed.Meanwhile,we quantitatively investigated the changes in the total mRNA levels of Hsp90 in tomato roots treated with Meloidogyne incognita.Expression levels of Sl-hsp90 were examined by fluorescent real-time PCR.The stability of mRNA was estimated.It was found that Hsp90 was more inducible after nematode infected.In addition,females and second-stage juveniles(J2) of M.incognita were exposed at 4℃ and 35℃ for different times in order to explore the impact of adverse temperature on Mi-hsp90 gene expression.Expression levels of Mi-hsp90 on 35℃ in females and J2 were more than that of Mi-hsp90 at 4℃.At 35℃ a burst of expression for Mi-hsp90 was observed in J2 after 2h of heat shock treatment,then expression dropped,although remaining still relatively high after 24h.Female and J2 are more suitable for high temperature.We established the Hsp90 high expression in nematode stressed by temperature and tomato roots infected by root knot nematode.