草鱼肠系膜脂肪组织高质量总RNA提取方法的研究     

姜鹏  骆丽婷  李胜杰  樊佳佳  马冬梅 《水产科学》2021,(2):166-171

基于传统的酸性酚—异硫氰酸胍—氯仿一步提取法,比较分析多种优化操作步骤,摸索出一种高质量提取体质量为80~150 g草鱼肠系膜脂肪组织总RNA的改良方法。试验结果显示,相较于肝脏、脾脏、肠道等脏器组织,草鱼肠系膜脂肪组织RNA丰度低,且极易在样品前处理阶段出现顽固性降解问题。探索发现,将取样量增至约30 mg,可提升RNA产量以满足常规试验需求。针对降解难题,改良常规的样品前处理技术流程,采用鲜样液氮速冻,冻样直接放入TRIzol试剂中裂解,并即刻进行长时间机械匀浆,时长约3 min等核心操作步骤,可显著降低脂肪组织样品RNA的降解。Agilent生物分析仪检测结果显示,改良方法提取的草鱼肠系膜脂肪组织RNA完整度高,关键RIN值为8.7~9.0。研究推测,长时间机械匀浆所形成的持续剪切冲击力或许有助于TRIzol试剂中的异硫氰酸胍等成分突破油滴阻碍而有效抑制内源性RNA酶。本方法提取的草鱼脂肪组织总RNA质量可满足高通量转录组测序要求。  相似文献   

Expression Analysis of Five Genes in the Gonadal Axis of Small Tail Han Sheep and Sunite Sheep     

ZHANG ZhuangBiao  DI Ran  LIU QiuYue  HU WenPing  WANG XiangYu  TIAN ZhiLong  ZHANG XiaoSheng  ZHANG JinLong  CHU MingXing 《中国农业科学》2018,51(24):4710-4719

  相似文献   

Relationships of Spectral Reflectance with Plant Tissue Mineral Elements of Common Bean (Phaseolus Vulgaris L.) Under Drought and Salinity Stresses     

Blagoja Boshkovski  Constantinos Tzerakis  Georgios Doupis  Anhelina Zapolska  Chariton Kalaitzidis 《Communications in Soil Science and Plant Analysis》2020,51(5):675-686

ABSTRACT

Salinity and drought stresses are critical for Phaseolus vulgaris L. growth and development. They affect plants in various ways, including tissue mineral element content. Micro- and macro-elements in leaves of Phaseolus vulgaris L. (cv. ‘Blue lake’ and cv. ‘Terli’) subjected to deficit irrigation and salinity treatments were investigated, both analytically and with regards to their effect on the leaves’ spectral reflectance. B (boron), K (potassium), Mn (manganese), Na (sodium), Si (silicon) and Zn (zinc) appeared to be influenced by stress factors, mainly responding to salinity increase. The leaf spectral reflectance of the plants appeared to be significantly correlated with most of the elements under investigation. Multivariate regression identified a relationship of the reflectance at particular regions of the spectrum with phosphorus and NDVI (normalized difference vegetation index) and indicated a significant correlation with B, Fe (iron), K, Mn, P (phosphorus) and Zn. Moreover, customized spectral indices, exhibiting significantly high correlation with B, Fe, K, Mg (magnesium), Mn, Na, P, Zn and N (nitrogen), were developed.  相似文献   

茶树蛋白激酶基因CsCIPK的克隆与表达特性分析     

刘昊  王文丽  滕瑞敏  李辉  王瑜  王永鑫  庄静 《园艺学报》2019,46(1):96

以茶树品种‘龙井43’作为材料,利用RT-PCR方法,从茶树的cDNA中克隆得到1个编码蛋白激酶的基因,命名为CsCIPK。序列分析表明,CsCIPK开放阅读框长度为1 341 bp,编码446个氨基酸,蛋白质分子量为414234。蛋白功能域预测和多重对比显示,CsCIPK蛋白含有1个保守的N端激酶结构域和1个相对不保守的C端调节结构域,即丝氨酸/苏氨酸激酶结构域和NAF结构域。理化性质、亲/疏水性、无序化分析显示,CsCIPK属于疏水性蛋白,理论等电点为7.04,有4段无序化区域,其二级结构分析显示主要由α螺旋、不规则卷曲组成。通过实时荧光定量PCR对‘龙井43’和‘安吉白茶’中的CsCIPK表达特性进行分析。结果显示‘龙井43’中CsCIPK的相对表达量在高温、干旱及盐处理4 h、低温处理24 h时达到最高。‘安吉白茶’中CsCIPK的相对表达量在高温及盐处理4 h、低温及干旱处理1h时达到最高。CsCIPK在‘龙井43’的根中,‘安吉白茶’茎中表达量最高。不同浓度的GA和IBA处理‘龙井43’茶苗,结果显示0.2 mmol·L-1 GA处理后,CsCIPK表达量先升高后下降,6 d时处理组为对照组的62倍;0.6 mmol·L-1 IBA处理后,CsCIPK的表达量在3 d时显著高于对照组;不同浓度GA和IBA处理后,9 d时CsCIPK表达量均显著低于对照。  相似文献   

大蒜不定芽的诱导及其增殖系数的调节     

汤青林  王志敏  宋明  王小佳  王平利  向巧彦 《勤云标准版测试》2006,(6)

为获得白皮大蒜组培繁殖体系,以其茎尖为材料,通过BA和NAA不同激素配比试验研究,结果表明:诱导愈伤组织的最适培养基为:MS BA0.2mg/L NAA0.5mg/L,诱导率达60%;诱导不定芽的最适培养基为:MS BA1.0mg/L NAA0.1mg/L,丛生芽繁殖系数高达80%。同时,为建立扩繁最佳体系,分别研究了激素、pH、谷氨酸钠三因素对不定芽增殖系数的调节效果,研究表明:添加0.5mg/L的GA3使增值系数提高到13.8;pH5.8最利于不定芽增值;添加10mg/L谷氨酸钠能在多次继代培养中保持较高的增值系数。  相似文献   

柚木(Tectona grandis)无性系繁育及造林技术     

陈玉桥  梁坤南 《林业调查规划》2006,31(2):106-108

柚木为马鞭草科柚木属植物,是世界上最珍贵的用材树种之一。文章介绍了柚木的生物学特性、无性繁殖即组培技术要点以及造林技术,论述了柚木在我省的推广前景。  相似文献   

月季组织培养过程中的影响因素   总被引：12，自引：0，他引：12  

王正加  高云振  李军萍 《浙江林业科技》2003,23(4):51-56

综述了月季组织培养过程中的影响因素，重点介绍了植物生长调节剂、外植体、培养基和培养条件等对月季试管苗生长的影响。同时分析了月季组培影响因素研究中存在的不足，提出对多因素、复杂因子共同影响行为的研究和探讨是今后月季组培影响因素研究中的重点内容。  相似文献   

Expression and modulation of connective tissue growth factor in renal interstitial fibrosis     

LIU Yan  FAN Jun-ming 《园艺学报》2004,20(9):1725-1728

CTGF, a member of the CCN family of immediate early genes, is a recently discovered profibrotic growth factor, which is involved in many pathophysiologic procedures. CTGF acts as a downstream effector of TGF-β acting on interstitial cells to enhance the progression of fibrotic renal diseases. It has been shown that CTGF gene expression can be induced or blocked by some kinds of cytokine and drugs. It is an interesting candidate target for future intervention strategies of renal interstitial fibrosis.  相似文献   

贮藏中福桔果皮细胞结构的显微观察及果实理化成分测定     

陈文军  洪启征  郑伸坤 《福建农林大学学报(自然科学版)》1991,(2)

贮藏期间福桔果皮细胞始终具有分裂增殖能力,细胞生长的养分主要通过维管束向果肉吸取,因而促进果肉衰老,形成枯水.枯水果实在贮藏早期就发现果皮细胞层和油腔分泌细胞无丝分裂旺盛,双核细胞多,果皮增厚多.严重枯水时,果皮细胞还具细胞核、线粒体、有色体等超微结构,未枯水果实细胞分裂少见.经预贮后贮藏的果实,由于中断了果皮组织与维管束的联系,果皮细胞生长受抑制,果肉水分、养分消耗少,枯水率最低,  相似文献   

In vitro expression and inhibition of procoagulant activity produced by bovine alveolar macrophages and peripheral blood cells     

J. Rashid  D. J. Weiss  S. K. Maheswaran  M. P. Murtaugh 《Veterinary research communications》1996,20(6):519-531

Local and systemic activation of coagulation is frequently associated with bacterial sepsis. The coagulopathy is due, at least in part, to expression of tissue factor (TF) by monocytes and macrophages. The purpose of this study was to evaluate the expression of procoagulant activity by bovine alveolar macrophages, leukocytes and platelets, and to determine the relative potency of three chemical inhibitors of TF expression (pentoxifylline, retinoic acid, and cyclosporin A). Bovine alveolar macrophages were stimulated with lipopolysaccharide (LPS) derived from Pasteurella haemolytica or recombinant bovine tumour nervous factor (TNF) and dose- and time-dependent effects on TF expression were studied. LPS and TNF induced TF expression in alveolar macrophages and LPS treatment of whole blood induced TF expression in mononuclear cells. Neutrophils and platelets also expressed procoagulant activity, but this activity was not inhibited by anti-bovine TF monoclonal antibody. Pentoxifylline (40 mol/L), retinoic acid (0.01 mmol/L) and cyclosporin A (0.08 mol/L) inhibited TF expression when added concurrently with LPS or TNF, but not when added 4 h after stimulation. TF mRNA was not detected in unstimulated alveolar macrophages by Northern blot analysis. In contrast, exposure to LPS or TNF for 6 h induced marked expression of TF mRNA, which was inhibited by treatment with pentoxifylline, retinoic acid and cyclosporin A. Expression of TNF by alveolar macrophages stimulated with LPS was also inhibited by these compounds. Our results indicate that procoagulant activity expressed by alveolar macrophages and monocytes is associated with expression of TF, whereas procoagulant activity expressed by neutrophils and platelets is not. The concentrations of pentoxifylline and retinoic acid necessary for inhibition of TF expression in vitro may not be achievable in vivo owing to their toxic effects. However, the in vitro concentration of cyclosporin A that inhibited TF expression did not exceed the plasma concentration observed in humans, and therefore may be useful for inhibition of TF expression in vivo.Abbreviations BAL bronchoalveolar lavage - LPS lipopolysaccharide - cDNA cloned deoxyribonucleic acid - cAMP cyclic adenosine monophosphate - GAPDH glyceraldehyde phosphate dehydrogenase - mRNA messenger ribonucleic acid - TF tissue factor - TNF tumour necrosis factor - DPBS Dulbecco's phosphate-buffered saline  相似文献