MicroRNA‐145 regulates immune cytokines via targeting FSCN1 in Staphylococcus aureus‐induced mastitis in dairy cows

Dairy cow mastitis is a detrimental factor in milk quality and food safety. Mastitis generally refers to inflammation caused by infection by pathogenic microorganisms. Our studies in recent years have revealed the role of miRNA regulation in Staphylococcus aureus‐induced mastitis. In the present study, we overexpressed and suppressed miR‐145 to investigate the function of miR‐145 in Mac‐T cells. Flow cytometry, ELISA and EdU staining were used to detect changes in the secretion of several Mac‐T cytokines and in cell proliferation. We found that overexpression of miR‐145 in Mac‐T cells significantly reduced the secretion of IL‐12 and TNF‐α, but increased the secretion of IFN‐γ; the proliferation of bovine mammary epithelial cells was also inhibited. Using quantitative real‐time PCR (qRT‐PCR), Western blotting and luciferase multiplex verification techniques, we found that miR‐145 targeted and regulated FSCN1. Knock‐down of FSCN1 significantly increased the secretion of IL‐12, while the secretion of TNF‐α was significantly downregulated in Mac‐T cells. Upon S. aureus infection of mammary gland tissue, the body initiated inflammatory responses; Bta‐miR‐145 expression was downregulated, which reduced the inhibitory effect on the FSCN1 gene; and upregulation of FSCN1 expression promoted mammary epithelial cell proliferation to allow the recovery of damaged tissue. The results of the present study will aid in understanding the immune mechanism opposing S. aureus infection in dairy cows and will provide a laboratory research basis for the prevention and treatment of mastitis.     

Rno‐miR‐425‐5p targets the DLST and SLC16A1 genes to reduce liver damage caused by excessive energy mobilization under cold stress

MicroRNAs (miRNAs) are a class of single‐stranded non‐coding small RNA molecules, which participate in the regulation of many physiological processes, and play a crucial role in cancer, metabolism and other processes. Rno‐miR‐425‐5p has been shown to play a role in the response to cold stress. To explore the mechanism by which rno‐miR‐425‐5p regulates the response to cold stress, we analysed the candidate target genes of rno‐miR‐425‐5p. After verification in rat hepatocyte BRL cells and in rat liver tissue, we identified several target genes that were altered in expression in response to cold stress. In rat liver tissue, the expression of rno‐miR‐425‐5p was significantly increased and the expression levels of target genes DLST and SLC16A1 were decreased under cold stress. The miRNA and mRNA levels were analysed by quantitative real‐time PCR and the protein levels were detected by Western blot analysis. Combined with the results of bioinformatic analysis, we concluded that rno‐miR‐425‐5p reduced the expression of DLST and SLC16A1, inhibiting energy release from the tricarboxylic acid cycle and preventing the liver from being injured by excessive energy mobilization.     

植物miR156及靶基因SPL家族的研究进展

SPL(SQUAMOSA promoter binding protein-like)是绿色植物所特有的一类转录因子,其部分家族成员含有miR156结合位点,在转录水平上受miR156负调控.在拟南芥(Arabidopsis thaliana)、水稻(Oryza sativa)等模式植物中,miR156-SPL不仅参与植物的开花调控,同时具有介导植物生育阶段转变、调节植物次生代谢、参与植物光信号转导和胁迫响应等作用.随着生物技术的发展,关于植物中miR156-SPL的研究越来越多,本研究重点阐述miR156-SPL模块在调控植物开花、株型结构、果实发育、胁迫响应等方面的作用及其机制,为进一步探索miR156-SPL模块在草类植物中的功能奠定基础.     

西瓜miR164b靶基因鉴定及其对黄瓜绿斑驳花叶病毒侵染应答的分析

将miR164b序列与西瓜基因组进行比对,获得miR164b前体Pre-miR164b基因并克隆。Pre-miR164b长度为97 bp,含有完整的茎环结构。Pre-miR164b启动子区含有光响应元件、赤霉素响应元件、乙烯响应元件、水杨酸响应元件、真菌诱导响应元件等多个顺式作用元件。降解组测序获得miR164b的4个靶基因,均注释为NAC转录因子基因,剪切位点位于miR164b序列5′ 端第10位碱基。miR164b及靶基因在黄瓜绿斑驳花叶病毒（CGMMV）侵染不同时间的表达模式不同,靶基因Cla018596的表达模式与miR164b呈负相关,而Cla019099、Cla023219和Cla023357的表达模式与miR164b没有相关性。     

鹰嘴豆miR164基因家族的生物信息学分析及靶基因预测

miR164是植物特有的一类miRNA,参与调控植物的生长发育和对逆境胁迫的响应.为深入理解Car-miR164的功能和调控机制,本研究通过全基因组手段鉴定鹰嘴豆Car-mmiR164家族成员并预测靶基因功能,在PmiREN中获得Car-miR164家族5个成员的序列,通过染色体定位技术表明其位于鹰嘴豆Ca3、Ca4和...     

MiR156 and its Target Gene TBCC in Flower Development of Gossypium harknessii Cytoplasmic Male Sterile Line and its Maintainer

This study aimed to analyze the expression level and relationship between miR156 and its target gene TBCC (Tubulin binding Cofactor C) in buds of Gossypium at different developmental stage in a cytoplasmic male sterile(CMS) line and its maintainer line, to clone the full-length cDNA of the TBCC gene, and to determine the possible role of the TBCC gene in pollen development. Real-time RT-PCR showed that the expression levels of miR156 gradually increased with the development of pollen between the CMS line and the maintainer line. The expression of GhTBCC was strikingly complementary with the expression of miR156 in the maintainer line, but showed very low expression levels in the CMS line, which demonstrated that TBCC is related to male sterility. The full-length cDNA sequence of TBCC was cloned from the buds of the maintainer line, and the gene was named as GhTBCC. The CDS of GhTBCC is 1713 bp and encodes a predicted protein of 570 amino acids with a predicted molecular weight of 62.79 kDa. Conserved TBCC and CARP domains were observed in the predicted protein sequence. GhTBCC showed 81 %, 80 %, and 77 % sequence similarity to the TBCC protein sequences of Populous trichocarpa, Ricinus communis, and Arabidopsis thaliana, respectively.     

vvi-miR172s及其靶基因响应赤霉素调控葡萄果实发育的作用分析

[目的]miR172是植物生长发育的重要调节因子,阐释vvi-miR172s及其靶基因应答赤霉素在葡萄果实不同组织发育过程中的作用,从miRNA角度认识GA调控葡萄果实发育的作用机制.[方法]以'白罗莎里奥'葡萄为材料,以miR-RACE和PCR技术克隆vvi-miR172a/b/c/d的成熟体和前体序列,由psRNA...     

植物Cu,Zn-SOD分子调控机理研究进展

铜/锌超氧化物歧化酶(Cu,Zn-SOD)能清除植物体内有害的活性氧(ROS),参与植株遭受逆境胁迫时的应激反应等过程。铜分子伴侣(CCS)可以传递铜离子到Cu,Zn-SOD当中,并将其激活生成有活性的酶分子,依赖CCS协助是Cu,Zn-SOD主要的激活途径。植物在铜缺乏的环境下会诱导启动子结合蛋白(SPL7)和小RNA398(miR398)的表达,miR398通过降解编码Cu,Zn-SOD的mRNA抑制Cu,Zn-SOD的生成,从而调控植物体内铜平衡。本文主要对植物Cu,Zn-SOD激活和调控途径进行综述。     

Screening of Brown Planthopper Resistant miRNAs in Rice and Their Roles in Regulation of Brown Planthopper Fecundity

MicroRNAs (miRNAs) can participate in plant-insect interactions, which regulate plant defense networks. In this study, we analyzed the miRNA expression profiles of six rice varieties before and after brown planthopper (BPH)-feeding. We identified 45 differentially expressed miRNAs between BPH- susceptible and BPH-resistant rice varieties and 144 miRNAs that responded to BPH-feeding. Thus, miRNAs may be involved in multiple pathways regulating rice defense response against BPH. In addition, we found that the genetic history of rice varieties determined the regulation mode of the miRNA and affected the amounts, types, changing trends and response periods of miRNAs in response to BPH- feeding. To conclude, we scanned seven potential cross-kingdom miRNAs, of which miR5795 may target the vitellogenin gene in BPH, causing a 16.07% reduction in BPH oviposition. The results provide new miRNA information of rice-BPH interactions and BPH-resistant rice variety breeding.