用柱色谱法提制替告皂甙元和海柯皂甙元标准品

本文叙述了用硅胶为吸附剂,分别用乙酸乙酯—苯,丙酮—苯为洗脱剂的柱色谱法,有效地提制了替告皂甙元和海柯皂甙元标准品。提制试验是用Ⅱa级活性硅胶进行的,样品与硅胶之比为1:30。在两种洗脱剂系统中,乙酸乙酯或丙酮对苯的比例根据试验的洗脱顺序而定。     

《中国药典》中山茱萸含量测定方法的改进

按2000年版药典一部山茱萸项下的薄层条件进行操作,存在斑点不集中、发散甚至拖尾、显色不易控制、扫描背景噪声大等问题,影响积分测定,使含量计算易产生误差。通过反复试验采用环己烷-氯仿-醋酸乙酯-甲酸(20：5：8：0．1)为展开剂,用λs=545nm,λk=430nm进行扫描取得了理想结果。     

双波长——薄层扫描法测定舒心素注射液中人参皂甙的含量

本文采用双波长薄层扫描法,以精制西洋参总皂甙,人参单体皂甙—Rb_1—Rb_2—Rb_3—Rd—Re—Rg_1及假人参皂甙—F_(11)为标准品,测定舒心素注射液中西洋参茎叶总皂甙及单体皂甙的含量,此法简便准确.     

薄层扫描法测定薏苡仁人参中人参皂苷Re的含量

采用薄层扫描法测定人参皂苷Re含量,测定波长λs=530nm,参比波长λR=650nm.展开剂为（正丁醇∶乙酸乙酯∶水=4∶1∶5）放置后的下层液,显色剂为10％的硫酸乙醇溶液,参考了有关文献[1,2].经测量,人参经过薏苡仁为辅料的加工炮制后的人参皂苷Re含量为0.76mg/g.     

金白龙茶中γ-氨基丁酸的双波长薄层扫描测定法研究

本研究对展开剂和扫描波长进行了系统的研究,点样硅胶板在正丁醇"冰醋酸"水=4"1"1溶剂系统中展开,采用双波长(λs=515nm;λR=680nm)锯齿扫描法进行检测,研究表明,该方法快速准确,操作简单,精密度,线性范0.5μL#15μL,重复性好,适用于γ-氨基丁酸的痕量检测。     

西洋参中掺入中国人参的半定量鉴别研究

人参皂甙在展开剂系统:氯仿:甲醇:水(65:35:10)的薄层展开时,在单体 Re与Rg1之间有一特征斑点R(x),而西洋参没有;在梯度点样测试中,该斑点与点样量呈线性关系,相关系数r= 0.996;通过西洋参总皂甙对照品与人参皂甙提取液叠加试验,Rx叠加回收率101%;因此经叠加后,测试掺有人参的西洋参样品皂甙中Rx,经CS- 930 薄层扫描,测得Rx峰面积,可估算出酉洋参中掺入人参的量,同时可补偿因取样不均,而Rx斑点不明显,可能发生的误检。本研究同时给出该系统条件下,CS- 930 薄层扫描的特征鉴别图谱。     

薄层扫描法比较红参及葛根人参中人参皂苷Re的含量

目的探讨新型辅料人参———葛根人参的加工工艺,并建立葛根人参及红参中人参皂苷Re的含量比较方法。方法采用70%乙醇超声提取葛根中的有效成分,旋转蒸发仪适当浓缩,浸入人参煎煮,待葛根提取液完全浸入参体后烘箱35℃低温干燥得葛根人参;以甲醇超声提取红参及葛根人参,点板(硅胶G板),进行薄层扫描(λS=550 nm,λR=650nm)。结果人参皂苷Re点样量在2～18ul范围内,点样量与峰面积呈良好的线性关系,得标准曲线Y=161.2X-166.84,r=0.9994;红参及葛根人参中人参皂苷Re的含量分别为0.19%和0.21%。结论薄层扫描法操作简便,结果准确,可以用作比较红参及葛根人参中人参皂苷Re的含量。     

不同参龄西洋参中皂甙的含量变化

应用CS—930型双波长薄层扫描仪分析不同产地、不同参龄、不同采收期的西洋参中的皂甙含量,结果表明:(1)不同产地的西洋参中皂甙含量有差异;(2)西洋参中的皂甙含量随着参龄的增长而增加;(3)西洋参中的皂甙含量随采收期的不同而变化;(4)8月31日采收的西洋参(柏同参龄),其单体皂甙和总皂甙含量最高。(5)8月31日采收的西洋参(相同参龄),其Rb_1含量较高,9月23日采收的西洋参,基Re含量较高。     

西洋参茎叶总皂甙的品质评价(一)——总皂甙、单体皂甙、分组皂甙的定性定量分析

以人参皂甙-Rb_1、-Rb_2、-Rb_3、-Rd、-Re、-Rg_1、-Rg_2、-Rh_1、-Rh_2、拟人参皂甙-F_(11)及-RT_5为标准品,利用薄层层析对西洋参茎叶总皂甙进行了定性分析,采用比色法和双波长薄层扫描法测定了总皂甙、单体皂甙、分组皂甙含量。结果表明,总皂甙中含有16种以上的单体皂甙,其含量为83％以上;单体皂甙中拟人参皂甙-F_(11)含量最高;分组皂甙中原人参二醇皂甙高于原人参三醇皂甙含量,奥科提罗醇(Ocotillol)皂甙含量最低。此文属首次报道。     

人参皂甙化学研究的回顾(续)

(接上期)二、人参中皂甙的化学分析(一)人参皂甙定量方法的建立人参中皂甙的定量方法有重量法、比色法、薄层扫描法、气相色谱法及高效色谱法等。为了适应国内各厂家的现有条件,建立一个简而易行,而相对准确的人参皂甙的定量方法是非常必要的。邵春杰和笔者以人参皂甙 Re(从人参地上部位分离易得)为标准物质进行比色测定,方法简便,结果准确,易于在国内推广使用,目前国内有关单位几乎皆采用此法测定人参中的总皂甙含量。此外,我们也利用薄层扫描法对人参总皂甙中各种单体皂甙进行了定量分析。     

Synthesis and characteristic of a novel green fluorescent protein eYGFPuv

Recently, a novel green fluorescent protein eYGFPuv has been identified in the marine organism Chiridius poppei which displays high fluorescence intensity and can be visible by eyes in dark. Although strong green fluorescence was achieved in transgenic petunia, 3 expression cassettes (about 8 kb) complicate its application. In this study, to confirm whether 1 expression cassette could be used as a transgenic marker in prokaryotes and eukaryotes, eYGFPuv was cloned into prokaryotic expression vector pET28α-eYGFPuv- His and plant binary expression vector 35S::eYGFPuv. Compared to EGFP, eYGFPuv protein exhibited stronger dazzling green fluorescence in E. coli under excited light at 365 nm and maintains steadily over a long period of time without degradation. When transiently expressed in tobacco leaves, eYGFPuv protein displayed strong green fluorescence. Moreover, the fluorescence of eYGFPuv protein also could be directly observed in living plant, and thus can be used easily as a marker to screen transformed lines in transgenic research. Overall, compared to previous studies on eYGFPuv tandem repeats, our data confirmed that single eYGFPuv sequence still possesses high fluorescence intensity and quenching resistance. Furthermore, because of small size of expression cassette,it is suitable for efficient transformation in both prokaryotic and eukaryotic organisms.     

解淀粉芽胞杆菌的GFP标记及定殖能力

采用改良的海藻糖电转化法获得GFP标记菌株X5-GFP、BQA2-GFP,显微镜观察标记菌株在土壤和水培中的番茄根际的定殖情况。结果表明:标记菌株单独或与青枯菌先后加入进行处理,番茄根际标记菌株的种群数量变化趋势都是一致的。在土壤番茄根际中,X5-GFP单独处理后当天的菌量为1.05×106CFU/g,随之菌量逐渐下降;处理30 d后,为1.50×10~3CFU/g;而BQA2-GFP单独处理后当天菌量为1.68×106 CFU/g,随之菌量也逐渐下降;处理30 d后,为2.50×10~3CFU/g;在水培番茄根际中,X5-GFP单独处理后当天的菌量为2.70×10~5CFU/g,随之菌量逐渐下降;处理30 d后,为1.00×10~3 CFU/g;而BQA2-GFP单独处理后当天菌量为3.60×10~5CFU/g,随之菌量同样逐渐下降;处理30 d后,为1.50×10~3 CFU/g。这表明解淀粉芽胞杆菌X5、BQA2均在番茄根际有较强的定殖能力。     

Cell and nuclear degradation in root meristems following exposure of potatoes (Solanum tuberosum L.) to salinity

Summary In many plant species, it has been demonstrated at the whole plant level that supplemental Ca²⁺ alleviates the effects of salinity stress. These effects have been attributed to physiological processes, but there are no reports of the effects of supplemental Ca²⁺ on preventing nuclear damage to the root meristematic cells following exposure to NaCl salinity. Two in vitro cultured potato (Solanum tuberosum L.) cultivars, analysed by fluorescent microscopy and flow cytometry, showed a similar pattern of salinity-induced changes to the nuclei of root meristematic cells. Damage occurring after only a few hours was followed by nuclear degradation at 24 h. Flow cytometry histograms showed a reduction in G1 and G2 nuclei and an increase in degraded nuclei, in NaCl-stressed roots. Salinity-induced nuclear degradation was alleviated by the addition of CaCl₂.     

适用于多重荧光SSR检测的玉米基因组DNA提取方法

通过比较植物DNA提取试剂盒法、SDS法、磁珠法和质粒DNA小提试剂盒法提取玉米叶片全基因组DNA的质量,确定最适用于多重荧光SSR检测的方法。结果表明,4种方法获得全基因组DNA的ODA260/ODA280平均值分别为1.92、2.16、2.22和2.00;DNA浓度平均值分别为19.16、2 050.58、69.12、53.56 ng/μL。比较发现,SDS法和植物DNA提取试剂盒法因为提取DNA杂质多和提取成本高,均不适用于多重SSR-PCR大量样本的检测;质粒DNA小提试剂盒法和磁珠法提取的全基因组DNA都适用于多重SSR-PCR检测,分别适合人工操作和机械自动化提取。     

茶尺蠖核型多角体病毒荧光定量PCR检测方法的建立

根据茶尺蠖核型多角体病毒（Ectropis oblique nucleopolyhedrovirus, EoNPV）基因序列设计一对引物,并从感染该病毒的虫体中提取EoNPV基因组DNA,进行PCR扩增,将该基因片段与载体pMD18-T连接,转化入大肠杆菌TG1中,经单克隆菌落筛选及测序鉴定后得到重组质粒,以该质粒为荧光定量PCR标准品模板稀释后建立标准曲线。构建的标准曲线线性关系良好,相关系数为0.989,检测范围为10³~10⁸拷贝/µL,特异性和重复性较好。该方法可以准确地判断出病毒拷贝数,为茶尺蠖核型多角体病毒在虫体内增殖动态的研究和病毒制剂的质量检测提供了方法依据。     

The use of citrus tristeza virus (CTV) containing a green fluorescent protein gene as a tool to evaluate resistance/tolerance of transgenic citrus plants

The T36 strain of citrus tristeza virus (CTV) expressing green fluorescent protein gene (CTV-GFP) was used to detect replication of CTV in wild-type (WT) ‘Hamlin’ sweet orange plants and those transformed with the use of Agrobacterium tumefaciens strain harboring a binary vector with a coat protein gene from either T36 or T30 isolate of CTV. Soil-adapted WT and transgenic plants were challenged with CTV by grafting shoots from CTV-GFP infected plants. None of the transgenic plants appeared to be able to inhibit CTV replication as CTV-associated GFP fluorescence in all of them was detected by fluorescent microscopy. For the purpose of comparison of two different methods, CTV multiplication in transgenic plants was also examined by enzyme-linked immunosorbent assay (ELISA) assays. GFP-labeled CTV represents a useful tool for estimation of susceptibility of citrus cultivars or transgenic lines to CTV infection. This method using CTV-GFP is simpler, cheaper and less time-consuming than ELISA.     

LED光源在马铃薯种质资源试管苗保存的应用

本研究分别以LED和日光灯为光源,以10份不同栽培区的马铃薯种质资源试管苗为试验材料,比较分别在两种光源下,不同栽培区马铃薯种质资源试管苗的生长差异,确定LED光源下马铃薯种质资源试管苗是否能正常生长。通过对马铃薯试管苗7个数量性状的统计分析,结果表明:两种光源下,10份不同栽培区的马铃薯种质资源试管苗生长趋势基本一致;两种光源下,不同栽培区马铃薯种质资源7个数量性状t测验结果,除南中552的根条数差异显著外,其他性状差异不显著。通过数量性状统计分析结果,说明在LED光源下保存马铃薯种质资源试管苗是可行的。     

Development of a formulation of Pseudomonas fluorescens PfALR2 for management of rice sheath blight

Pseudomonas fluorescens isolates that inhibited growth of Rhizoctonia solani, the rice sheath blight pathogen, were isolated from the rhizosphere of different crops. One of the most effective strains, PfALR2, was developed as an antibiotic resistant strain and a peat-based formulation was developed for this bacterium. The effective dose of a peat formulation was assessed for seed treatment, root treatment, soil application and foliar spraying. All individual treatments controlled the disease effectively. However, a combination of all four treatments resulted in the best sheath blight control in the greenhouse. In field trials, application of the peat-based formulation of PfALR2 effectively controlled the disease, increased yield, and efficacy was comparable to that of the commercially available fungicide, carbendazim.     

山东棕壤茶区茶树荧光性绿斑病因的营养诊断

茶树荧光性绿斑病是一种茶树成叶生理型病害,在叶片背面有深绿色隆起的颗粒或斑块,且在病变处可见自发的黄绿色荧光。为了探明该病害的发生原因,对山东不同茶区、品种和病害程度的荧光性绿斑病叶13种元素含量进行测定,并与对照叶含量和茶叶中元素正常范围进行综合比较分析,筛选出与病害发生相关的可能元素,在此基础上,利用砂培进行不同营养液诱导处理,观测各处理诱导病害发生情况,同时测定诱导病叶中相关元素含量,结果表明:⑴钙、锰、铝过量处理可以诱导茶树荧光性绿斑病发生;⑵山东棕壤茶区荧光性绿斑病的发生是由于成叶中钙、锰两种元素过量累积所致。此外,讨论了茶树钙、锰、铝中毒的营养诊断指标。     

一种可视化快速筛选转基因玉米的方法研究

利用可视化表型分析快速筛选转基因后代可以有效提高遗传转化和基因功能的验证效率.本文研究一种高效、低成本的筛选转基因玉米的新方法,该方法利用愈伤组织和糊粉层特异性表达启动子Ltp2和可视化红色荧光蛋白编码基因DsRed构建植物表达载体pCAMBIA3300-DsRed,通过农杆菌介导法侵染玉米HiⅡ幼胚,诱导出分子检测为...