一个特殊的顺式发夹核酶对感染烟草原生质体的影响

研究一个烟草花叶病毒(TMV)cDNA3′末端被特异切割的发夹核酶对感染原生质体的作用。选用一个外源基因绿色水母荧光蛋白(GFPuv)作为报道基因，取代了大部分的CP编码区域，以研究在烟草原生质体中发夹核酶对TMV载体的复制，通过GFP的表达来确定表达量。顺式发夹核酶对TMV载体感染效率的影响通过体外转录来观察。TMV GFPRIB体外转录感染原生质体的效率比与其不含该核酶的对应物的感染效率高3～5倍。Northern杂交结果表明，包括核酶在内的3’末端非病毒序列在体外转录反应开始后就被顺式发夹核酶切除了。当直接用载体DNA pSTMVGFPRIB感染原生质体时，感染原生质体的数量比与之相对应的pSTMVGFP NOS感染原生质体数多50％～80％。     

氢化物发生-原子荧光法测定烟叶中的铅

建立了一种测定烟叶中微量铅的方法--氢化物发生-原子荧光光谱法.研究了光电管负高压、灯电流、屏蔽气和载气流量、载流的选择、酸度等对实验的影响.在选定的工作参数下,检出限为0.08 μg/L.用标准物质桃叶(GBW08501)对该法进行了验证,验证结果与推荐值一致,相对标准偏差为0.34%.该方法简便、快速、灵敏,对于实际样品的测定得到了满意的结果.     

Induction of nicotine in tobacco by herbivory and its relation to glucose oxidase activity in the labial gland of three noctuid caterpillars

Tobacco Nicotiana tabacum L. is a host plant ofHelicoverpa armigera (Huibner), Helicoverpa assulta Guenee and Spodoptera litura (Fabricius) (Lepidoptera, Noctuidae). The difference in leaf nicotine response to the feeding by these three larvae and the mechanical simulation of their feeding was examined by HPLC. Results indicated that nicotine induction was suppressed by H. armigera and H. assulta larvae feeding or by simulated damage treated with their labial glands extracts. The production of nicotine was also suppressed by the glucose oxidase from Aspergillus niger when it was treated on mechanically wounded leaf area. On the contrary, the nicotine production was stimulated by S. litura larva feeding or by simulated damage treated with its labial gland extract. Heat denature can not counteract the stimulation effect of the S. litura labial gland extracts to tobacco nicotine production. The glucose oxidase activity was detected in labial gland extracts of both H. armigera and H. assulta, but the activity in H. armigera was significantly higher than that in H. assulta. No glucose oxidase activity was detected in labial gland extracts of S. litura. It is shown that the glucose oxidase activity in labial glands of caterpillars plays an important role in the nicotine response to herbivory. The glucose oxidase was mainly contained in the labial gland of H. armigera larva, and had the highest activity at pH 7.0. D-Glucose was the optimal substrate of the glucose oxidase. Labial gland glucose oxidase activities varied daily during larval development with high activities found when larvae were actively feeding.     

不同供钾水平下烟草钾积累特性研究

通过水培试验,研究了不同供钾水平下烟草上、中、下部叶以及根、茎中钾的积累特点,结果表明:供钾水平的提高能显著的增加叶片以及根、茎的含钾量.在供钾水平为0.1—50mmol/L范围内,烟草各器官干重没有显著的变化,烟叶中钾积累呈现出快速-缓慢-快速-缓慢的交替变化,其中在外界钾离子浓度0.1-0.2mmol/L和0.4-10mmol/L范围内,随钾浓度提高而积累量快速提高,这两个积累阶段与植物钾吸收的两个不同机制相一致.试验还证明:随着培养液中钾浓度的提高,中、下部叶钾含量增加较快,上、中、下三部位叶片钾含量的高低顺序发生改变.     

样本筛选与操作优化的可视化方法实现

可视化方法通过将多维空间数据映射到二维平面上，依据“物以聚类”属性对二维平面中各样本点进行分类、识别，找出离群样本并将其剔除掉．与此同时，产生目标函数等值线．沿着等值线值增大或减小的趋势方向寻优，就很容易地确定出最优点或最优化操作区域．以某卷烟厂生产烟丝为例进行分析和计算，结果表明，文中筛选样本数据方法中，离群点的剔除提高优化结果的准确度。最佳操作条件的确定可为工业生产或科学实验提供决策支持。     

Simultaneous quantification of ten Amadori compounds in tobacco using liquid chromatography with tandem mass spectrometry

Amadori compounds are aroma precursors formed in the initial phase of the Maillard reaction. Based on their similar structures, simultaneous quantification of more than six Amadori compounds in tobacco has not been reported yet. In this study, a simple and rapid method was developed to simultaneously quantify ten Amadori compounds including the isomers of Fructose‐isoleucine and Fructose‐leucine in tobacco. The separation was performed on an Atlantis T₃ column (2.1 × 250 mm, 5 μm) by gradient elution using acetonitrile and water as the mobile phases. The quantification method was systematically evaluated and proven to be sensitive and accurate. The linearity was good, with correlation coefficients of 0.9977–0.9999. The limits of detection and quantitation were 1.354–2.532 and 4.516–8.444 ng/mL, respectively. The recoveries were 84.0–119.6%, and the relative standard deviations were 1.33–5.40%. The method was used to analyze the changes in the amounts of ten Amadori compounds in tobacco before and after tobacco primary processing. The analysis shows that the Maillard reaction occurs during the short processing period.     

Simultaneous determination of residues of metalaxyl,cyazofamid and a cyazofamid metabolite in tobacco leaves and soil by liquid chromatography with tandem mass spectrometry

A simple method was developed and validated for the simultaneous determination of metalaxyl, cyazofamid and the cyazofamid metabolite 4‐chloro‐5‐p‐tolylimidazole‐2‐carbonitrile (CCIM) by liquid chromatography with tandem mass spectrometry. The three target compounds were extracted from tobacco and soil with acetonitrile containing 0.1% acetic acid, and the extracts were purified using octadecylsilane. The proposed method showed satisfactory linearity (R² ≥ 0.9985) for the target compounds. The limits of detection for metalaxyl, cyazofamid and CCIM were 0.006, 0.06 and 0.06 mg/kg in soil and green tobacco leaves and 0.03, 0.3 and 0.3 mg/kg in cured tobacco leaves, respectively. The limits of quantification for metalaxyl, cyazofamid and CCIM were 0.02, 0.2 and 0.2 mg/kg in soil and green tobacco leaves and 0.1, 1 and 1 mg/kg in cured tobacco leaves, respectively. The average recoveries from soil and tobacco were 72.91–98.40% for metalaxyl, 76.73–105.80% for cyazofamid and 74.48–106.45% for CCIM. The relative standard deviation range was 1.23–6.99%. The developed method was successfully applied to analysis of residues of metalaxyl, cyazofamid and CCIM in real soil and tobacco samples. The results indicated that the established method could meet the requirement for the analysis of trace amounts of all three analytes in soil and tobacco.     

天然脂肪醇抑芽剂对烤烟抑芽效果研究

为解决烟草抑芽剂农药残留潜在风险,将一种由天然脂肪醇配置的抑芽剂作为材料,以氟节胺为对照,通过田间试验的方法,研究该抑芽剂不同稀释倍数及不同施药方法的抑芽效果,结果表明:天然脂肪醇抑芽剂6个处理的抑芽率均显著高于氟节胺处理的,第40 d时,其抑芽率达到87.38%以上。其中,天然脂肪醇抑芽剂15倍、20倍稀释处理的抑芽效果达到97.94%以上,与氟节胺处理间差异不显著;同时,这两个处理可以增大顶部两片叶面积,提高上桔二烟叶的还原糖、总糖含量。     

Two New 4‐Hydroxyisoflavanes from the Root of Codonopsis cordifolioidea and Their Anti‐virus Activities

Two new 4‐hydroxyisoflavanes, cordifoliflavanes A and B (1 and 2), were isolated from the roots of Codonopsis cordifolioidea. Their structures were elucidated by spectroscopic methods, including extensive 1D‐ and 2D‐NMR techniques. Compounds 1 and 2 were tested for their anti‐HIV‐1 activities and anti‐tobacco mosaic virus activities. The results showed that compounds 1 and 2 have modest anti‐HIV‐1 activities and anti‐tobacco mosaic virus activities, respectively.     

Quantitation of perfluoroalkyl acids,parabens and cotinine from single low volume serum sample by validated analytical method

ABSTRACT

In this paper, we present the results of an analytical method that has been recently developed, validated and successfully applied in a biomonitoring approach. In the environmental pollutant studies it is desirable that the analytical method can determine multiple classes of compounds from a single, small volume sample. The presented analytical method with a simple sample pre-treatment allows the quantitation of 13 perfluoroalkyl acids (PFAAs), 6 parabens and cotinine (used as nicotine biomarker) from a single, small volume of 100 µL serum sample by liquid chromatography-triple quadrupole mass spectrometer (LC-MS/MS). The limits of quantitation (LOQ) for PFAAs, parabens and cotinine were 0.10–0.50, 0.20–0.80 and 0.10 ng/mL, respectively. Besides sensitivity the method has excellent trueness/accuracy and repeatability. The trueness of the method for the determination of PFAAs ranged from 95% to 106% and the repeatability (as RSD %) from 0.6% to 5.6%. The accuracy and RSD for parabens were 73–120% and 1.3–9.7%, respectively, and 100–106% and 1.3–3.5 % for cotinine. Biomonitoring data reveals the presence of several PFAAs and parabens in serum samples of Finnish population. The total concentrations for PFAAs and parabens were from 2.0 to 33 ng/mL and from <LOQ to 1100 ng/mL, respectively. Nearly all non-smokers had the serum cotinine concentration below 1.0 ng/mL, which can be suggested as the cut point for cotinine concentration to identify smoking.