茶树荧光性绿斑病叶黄绿色自发荧光的显微分析     

姜淑媛  ZHANG Li-xia  张丽霞  郭延奎  赵友全  贾明  翟衡 《茶叶科学》2008,28(2):135-141

茶树荧光性绿斑病是一种茶树成叶生理性病害,病叶上的绿色病斑可见自发的黄绿色荧光。为了以黄、绿色自发荧光为探针了解病害的发生规律,同时为黄色、绿色荧光物质的分离鉴定提供光谱学依据,应用荧光显微技术、显微荧光光谱成像技术、激光扫描共聚焦显微技术、流式细胞术研究了茶树荧光性绿斑病叶中黄、绿色自发荧光的显微观察条件、发射光谱和发光部位。结果表明:在不同激发光照射下,茶树荧光性绿斑病叶能发射多种自发荧光,其中黄色、绿色荧光显微观察的最佳条件为:蓝光激发,彩色和绿色单色光模式记录图像;在绿光、黄光范围有三个荧光峰,其波长分别为515nm、535nm和585nm;最初发出绿色、黄色荧光的病变部位是维管束鞘细胞,随后出现在部分海绵细胞、栅栏细胞和表皮细胞中;黄色、绿色荧光在细胞中的发光位置定位于液泡。  相似文献   

Milling characteristics and distribution of phytic acid and zinc in long-, medium- and short-grain rice   总被引：2，自引：0，他引：2  

Jianfen Liang  Zaigui Li  Kouichi Tsuji  Kazuhiko Nakano  M.J. Robert Nout  Robert J. Hamer 《Journal of Cereal Science》2008,48(1):83-91

Milling and polishing are important operations during the production of white rice. The degree of milling and polishing has a significant effect on the nutritional aspects of white rice, especially on minerals, due to a non-uniform distribution of nutrients in the kernel. Information on the distribution of nutrients in rice will greatly help in understanding the effect of milling and aid in designing procedures that improve technological and sensory properties of rice while retaining its essential nutrients as much as possible. In this study, three kernel shapes (short-, medium- and long-grain) of rice were selected for the study of milling characteristics and distribution of zinc (Zn) and phytic acid using abrasive milling and X-ray fluorescent microscope imaging approaches.Milling characteristics differed with kernel shapes and cultivars. Mass loss (y, %) correlated well with milling duration (x, s) and was fitted using a polynomial equation of y=ax²+bx+c (R²=0.99). Different kernel shapes of rice resulted in different patterns. Breakage in milling increased with longer duration of milling. The relation between breakage (y, %) and milling duration (x, s) fitted the exponential equation y=ae^bx. Levels of phytic acid, as well as Zn, decreased with prolonged milling. Phytic acid decreased at a higher rate than Zn. The analysis of different milling runs showed that the concentration of phytic acid decreased from the surface region inward, whereas X-ray fluorescent images indicated that the highest concentration of phosphorus was at the interface of the embryo and perisperm.Our results help in understanding the milling characteristics of different rice cultivars. Understanding these characteristics offers opportunities to optimize milling procedures for maximum phytate removal at minimum mineral losses and yield loss.  相似文献   

液相色谱-安培法检测养殖水体中的几种染料类抗菌剂     

钱疆  刘晓东  龚晖 《福建农业学报》2008,23(1):86-91

利用液相色谱—安培法检测分析养殖水体中3种染料—孔雀石绿、结晶紫、亚甲基蓝及其代谢物,无需衍生,养殖水样品采用阳离子反相混合模式MCX小柱富集,色谱柱为Waters Xettra C18(250×4.6 mm,5μm)流动相采用70%甲醇[甲醇∶0.1 mol.L-1乙酸钠=7∶3(V/V),pH4.5],流速为1 mL.min-1。采用直流安培检测,氧化电位1.4 V,工作电极为玻碳电极。3种染料及其代谢物在0.05~100μg.mL-1范围内具有良好的线性关系,检测限在0.2~0.5μg.L-1范围内,回收率为66%~101%,方法的精密度在0.5%~4.8%。该方法具有灵敏度高、选择性好的优点。  相似文献   

肉苁蓉水提液对大鼠成骨细胞骨形态发生蛋白2基因表达的影响     

邢晓旭  刘钟杰  韩博 《中国畜牧兽医》2013,40(1):17-21

本试验采用酶消化法分离得到大鼠颅骨成骨细胞,取第3代成骨细胞,通过加入不同浓度的肉苁蓉水提液(5×10^-5~5×10^-1 mg/mL)来提取细胞总RNA,再采用实时荧光定量PCR方法,来观察不同浓度的肉苁蓉水提液对大鼠成骨细胞骨形态发生蛋白2(bone morphogenetic protein 2,BMP2)基因表达的变化。从试验结果可以得出,肉苁蓉水提液对大鼠成骨细胞BMP2基因表达具有促进作用。  相似文献   

橡胶树原生质体的分离及红色荧光蛋白的瞬时表达研究   总被引：1，自引：1，他引：0  

于晓玲阮孟斌  王树昌 《中国农学通报》2013,29(28):18-22

目的：分离巴西橡胶树松散愈伤组织原生质体,建立目标基因在橡胶原生质体的瞬时表达系统。方法：以巴西橡胶树松散愈伤组织为材料,酶解分离原生质体,通过电击介导的转化方法,将编码红色荧光蛋白（RFP）的植物表达载体转入原生质体中,用激光扫描共聚焦显微镜检测原生质体中RFP的表达情况。结果：分离出大量高纯度的橡胶原生质体,成功获得转基因原生质体细胞,RFP在原生质体中得到成功表达。初步建立了橡胶原生质体瞬时表达系统。  相似文献   

猪圆环病毒2型TaqMan实时荧光定量PCR检测方法的建立   总被引：1，自引：1，他引：0  

曾思遥  张淑琼  余绍华  唐志玲  陈瑞爱  罗满林 《中国畜牧兽医》2012,39(6):41-46

本研究以猪圆环病毒2型(PCV2)核衣壳蛋白ORF2基因为目的基因,设计合成特异性引物和探针。PCR扩增得到目的基因,并克隆到pMD18-T载体,筛选得到标准阳性质粒。通过对荧光定量PCR反应条件的优化,建立了PCV2的TaqMan实时荧光定量PCR检测方法。试验结果表明,该方法特异性强,对猪伪狂犬病病毒(PRV)、猪繁殖与呼吸综合征病毒(PRRSV)、猪瘟病毒(CSFV)等猪常见病原的检测结果均为阴性;与普通PCR方法相比,其敏感性高出100倍,可达100拷贝/μL;对临床样品的检测证明,该方法可有效检测出淋巴结、肺脏等组织中的PCV2。  相似文献   

The Chemically Synthesized Ageladine A-Derivative LysoGlow84 Stains Lysosomes in Viable Mammalian Brain Cells and Specific Structures in the Marine Flatworm Macrostomum lignano     

Thorsten Mordhorst  Sushil Awal  Sebastian Jordan  Charlotte Petters  Linda Sartoris  Ralf Dringen  Ulf Bickmeyer 《Marine drugs》2015,13(2):920-935

Based on the chemical structure and the known chemical synthesis of the marine sponge alkaloid ageladine A, we synthesized the ageladine A-derivative 4-(naphthalene-2-yl)-1H-imidazo[4,5-c]pyridine trifluoroacetate (LysoGlow84). The two-step synthesis started with the Pictet-Spengler reaction of histamine and naphthalene-2-carbaldehyde to a tetrahydropyridine intermediate, which was dehydrogenated with activated manganese (IV) oxide to LysoGlow84. Structure and purity of the synthesized LysoGlow84 were confirmed by NMR spectroscopy and mass spectrometry. The fluorescence intensity emitted by LysoGlow84 depended strongly on the pH of the solvent with highest fluorescence intensity recorded at pH 4. The fluorescence maximum (at 315 nm excitation) was observed at 440 nm. Biocompatibility of LysoGlow84 was investigated using cultured rat brain astrocytes and the marine flatworm Macrostomum lignano. Exposure of the astrocytes for up to 6 h to micromolar concentrations of LysoGlow84 did not compromise cell viability, as demonstrated by several viability assays, but revealed a promising property of this compound for staining of cellular vesicles. Conventional fluorescence microscopy as well as confocal scanning microscopy of LysoGlow84-treated astrocytes revealed co-localization of LysoGlow84 fluorescence with that of LysoTracker^® Red DND-99. LysoGlow84 stained unclear structures in Macrostomum lignano, which were identified as lysosomes by co-staining with LysoTracker. Strong fluorescence staining by LysoGlow84 was further observed around the worms’ anterior gut and the female genital pore which were not counterstained by LysoTracker Red. Thus, LysoGlow84 is a new promising dye that stains lysosomes and other acidic compartments in cultured cells and in worms.  相似文献   

猪Oct4-EGFP多能性报告载体的构建与验证     

朱蒙  付博  刘娣  杨秀芹 《中国畜牧兽医》2015,42(8):1993-1999

本研究旨在构建一套猪Oct4-EGFP多能性报告载体,可在不破坏细胞的前提下研究Oct4的表达规律,从而有利于早期胚胎发育研究及干细胞的研究。试验采用无缝克隆(In-Fusion PCR cloning)技术,将Oct4启动子序列直接重组到pEGFP-N1载体上,用Oct4代替质粒pEGFP-N1中增强型绿色荧光蛋白(EGFP)原有的CMV启动子构建出Oct4-EGFP报告载体,并用脂质体转染技术转染入大白猪胎儿成纤维细胞中,分析Oct4-EGFP报告载体,表达情况。结果发现,经PCR及测序验证,成功构建了Oct4-EGFP多能性报告载体,并在孤雌囊胚上初步验证了载体的有效性;经过脂质体转染,经筛选及PCR鉴定,获得了8株整合有Oct4-EGFP多能性报告载体的转基因细胞。研究结果表明,运用无缝克隆技术可高效率构建Oct4-EGFP多能性报告载体,且获得的转基因阳性细胞可为猪胚胎早期发育和胚胎干细胞研究奠定技术基础。  相似文献   

Development of a Fusarium oxysporum f. sp. melonis functional GFP fluorescence tool to assist melon resistance breeding programmes          下载免费PDF全文

B. Ramos  G. López  A. Molina 《Plant pathology》2015,64(6):1349-1357

Fusarium wilt, caused by Fusarium oxysporum f. sp. melonis (Fom), is one of the most widespread and devastating melon diseases. This vascular disease is caused by the colonization of melon xylem vessels by any of the four Fom races reported (r0, r1, r2 and r1,2, subdivided into r1,2w and r1,2y). The macroscopic evaluation of disease symptoms (disease rating, DR) at several days post‐inoculation (dpi) with Fom spores has been the traditional method to determine the resistance of melon accessions to this fungal pathogen. In this study, one isolate from each Fom race was transformed by Agrobacterium tumefaciens to constitutively express the green fluorescent protein (GFP). Fom‐GFP transformants, as virulent as the corresponding wildtype races, were selected to develop an inoculation assay based on the non‐invasive evaluation of the fluorescence emitted by Fom‐GFP. It was determined that melon root neck was the appropriate area to follow Fom‐GFP and a fluorescence signal rating (FSR) was established in parallel to DR determination. This method allowed the evaluation of GFP signal in the root neck of inoculated melon seedlings at 11–15 dpi. The GFP signal was scored in 62 melon accessions/breeding lines inoculated with different Fom‐GFP, followed by evaluation of the macroscopic DR in the aerial part of melon seedlings at 20–28 dpi. Correlation analysis demonstrated a direct and significant relationship between FSR and DR. This method has shown to be an effective and reliable tool that can assist Fom resistance breeding programmes in melon.  相似文献   

农杆菌介导的西瓜枯萎病菌遗传转化     

任俊杰  王丽霞  高洪波  吕桂云 《植物保护》2015,41(1):93-97

为获得带GFP标记的西瓜枯萎病菌转化株,用于后期观察病原菌侵染过程,采用农杆菌介导的方法,对西瓜枯萎病菌1号生理小种进行了遗传转化。结果表明:共培养时间为36h,枯萎病菌孢子和农杆菌AGL1比例为1∶1时该菌株的遗传转化效率最高,可以达到117.33个转化子/107个孢子。转化株的孢子、菌丝体及萌发的孢子均能发出稳定而强的绿色荧光。转化株的致病力检测显示其致病力与转化前的野生菌株致病力无明显差异。结果表明本研究获得的带GFP标记的西瓜枯萎病菌转化株可用于观察病菌在西瓜根系的侵染过程。  相似文献