银杏叶中叶绿素衍生物的反相高效液相色谱分析

以检测植物色素降解过程中叶绿素衍生物的变化为主要目的,采用反相高效液相色谱,结合光电二极管阵列和荧光检测技术,分析了银杏叶中叶绿素衍生物的组成和含量变化,发现在衰老的银杏叶中主要存在叶绿素a、叶绿素b和脱镁叶绿素a,而其他降解产物并未大量累积。     

反相高效液相色谱法测定厌氧菌有机酸代谢产物

建立反相高效液相色谱测定厌氧菌代谢发酵有机酸产物(乙酸、乳酸)的方法并用于测定乳酸菌代谢发酵产物中的含量。反相高效液相方法是一种简单、准确、灵敏的方法,可用于同时定量测定厌氧菌的有机酸代谢产物。     

反相高效液相色谱法测定不同品种花椒中芦丁和槲皮素的含量

目的:建立RP-HPLC测定花椒中芦丁与槲皮素含量的方法,并对不同种花椒的中芦丁与槲皮素含量进行测定与比较。方法:Zorbax Eclipse C18色谱柱(150 mm×4.6 mm,5μm),流动相∶甲醇-0.4%磷酸(50∶50);流速1 mL/min;检测波长:360 nm;柱温25℃。结果:芦丁在0.25~5.0μg,r=0.999 9峰面积与质量浓度呈良好的线性关系;平均回收率为99.1%,RSD为4.3%(n=3)。槲皮素在0.25~0.5μg,r=0.999 9峰面积与质量浓度呈良好的线性关系;平均回收率为111.2%,RSD为5.1%(n=3)。结论:该方法可用于花椒中芦丁和槲皮素的测定。测定结果表明,韩城红花椒中芦丁含量最高,茂汉红花椒次之,四川青花椒较少,云南青花椒最低。槲皮素在韩城红花椒中含量较高,在其他三种花椒中差别不大。     

反相高效液相色谱法测定烟叶中的游离氨基酸

用不同浓度的乙醇溶液提取烟样中的游离氨基酸 ,结果显示 ,存在最佳的乙醇溶液浓度 ,使烟样中被提取的游离氨基酸总量最大 ;对比了活性炭、乙醚、5 %磺基水杨酸、阳离子交换柱的纯化效果 ,发现阳离子交换柱的纯化效果较其它三种试剂要好。在提取和纯化之后 ,采用OPA、FMOC联合在线衍生反相高效液相色谱法测定了烟样中的游离氨基酸 ,该方法使烟样中的氨基酸和亚氨基酸能被同时测定 ,并且分析方法的重现性和回收率均令人满意。最后用该方法对云南B2 F98(上部、橘黄、二等烟叶 ,98年产 )烟叶中的游离氨基酸进行了测定 ,有 15种氨基酸被测出 ,其中Pro含量最高 ,约占总量的 2 5 % ,Thr含量最低 ,约占总量的 1%。     

蚕豆种子贮藏蛋白质组分的比较研究

以10份蚕豆品种为材料,利用SDS-PAGE方法,对其种子贮藏蛋白亚基组成进行了分析,结果表明:不同品种间的种子贮藏蛋白具有一定的差异,表现出一定的多态性;共分离出23条迁移率不同的亚基条带,13条具有多态性;利用贮藏蛋白亚基条带的信息,分析了品种间的蛋白相似度,相似度指数0.619～0.947,平均0.744;并通过聚类分析,在遗传相似系数0.8260水平上,将供试材料分为了3类,其中第2类具有较丰富的多样性,多样性指数达3.9296;共筛选出7对亲缘关系较远的杂交组合,可为蚕豆品种选育提供一定的科学依据。     

反相高效液相色谱法分析聚乙二醇修饰的水蛭素

目的：采用反相高效液相色谱法（RP-HPLC）对聚乙二醇（PEG）修饰的水蛭素进行分析分离,用以分析修饰产物中不同修饰度水蛭素的组成和比例。方法：色谱柱为Hypersil C18,5μm,4.6mm×150mm;流动相A为H2O＋0.01％的三氟乙酸,流动相B为乙腈＋0.01％的三氟乙酸。40min内由10％-50％流动相B进行梯度洗脱,洗脱流速1mL／min,上样量50μL,检测波长为214nm。结果：在单甲基化PEG-丙酸琥珀酰亚胺和水蛭素摩尔比不同的的反应产物中,PEG1-水蛭素、PEG2-水蛭素均可以达到基线分离,且不同批次的反应产物进行RP-HPLC的重复性良好。结论：RP-HPLC可以有效地对PEG修饰的水蛭素产物进行分析分离,为PEG化水蛭素的长效、缓释剂型的开发提供技术支持。     

利用反相高效液相色谱测定香椿叶中槲皮素含量

用乙醇对香椿叶粉末进行提取,树脂柱浓缩,真空冷冻干燥,甲醇溶解,制备香椿黄酮。用7.6 mol.L-1盐酸85℃水浴回流2 h进行水解,以反相ODS柱甲醇-水(体积比为50:50,磷酸调pH至2.53)为流动相,在波长368 nm处对香椿叶中的槲皮素进行分离、测定。结果表明:香椿叶中含槲皮素平均质量分数为1.28%,平均加样回收率为98.9%,RSD为1.75%,反相高效液相色谱法测定香椿叶中槲皮素含量操作简便易行、结果准确可靠。     

大麦花药离体诱导的基因型稳定性和培养基的环境指数分析

本研究以湖北啤酒大麦优良新品种（系）为材料,分析了基因型的稳定性、培养基的环境指数以及基因型与培养基的互作效应.各基因型花药愈伤组织诱导率的稳定性分析表明,E2具有较强的普遍适应性,但稳定性好的材料愈伤组织诱导率并不一定最高,各种配方培养基的环境指数值大小不同,方向也有正有负,基因型效应以及基因型和培养基的互作效应均达极显著水平。     

Screening for Barley yellow dwarf virus-Resistant Barley Genotypes by Assessment of Virus Content in Inoculated Seedlings

The content of Barley yellow dwarf virus (BYDV) in roots and leaves of barley seedling plants differing in their level of resistance was assessed by quantitative ELISA 1–42 days after inoculation with the strain of BYDV (PAV). High virus accumulation in roots and low concentration in leaves was characteristic of the period 9–15 days after inoculation. In leaves, the differences in virus content between resistant and susceptible genotypes became significant after 15 days and resistance to virus accumulation was better expressed 30–39 days after inoculation. Roots of resistant materials exhibited evident retardation of virus accumulation and the greatest difference in virus content between resistant and susceptible plants was detected 9 days after inoculation. By these criteria, the selected winter and spring barley cultivars and lines (in total 44 materials) fell in to five groups according to field reactions and the presence or absence of the Yd2 resistance gene. There were highly significant and positive relations between ELISA values and 5‐year field data on symptomatic reactions and grain‐yield reductions due to infection. Using the described method, resistant and moderately resistant genotypes (both Yd2 and non‐Yd2) were significantly differentiated from susceptible genotypes. The possible use of this method in screening for BYDV resistance is discussed.     

Identification of Phthalate Ester-assimilating Bacteria

Seventeen bacterial capable of utilizing phthalate esters isolated from natural sources were identified. Based on morphological and biochemical characteristics, type of cell division, GC content in DNA, principal amino acid in the cell wall and cellular fatty acid composition, 10 isolates were identified as Nocardia erythropolis, one isolate as Pseudomonas acidovorans, another as Pseudomonas cepacia and four as members of the genus of Corynebacterium.     

Simultaneous Measurement of Nicotinic Acid and Its Major Metabolite,Nicotinuric Acid in Blood and Urine by a Reversed-phase High-performance Liquid Chromatography

The linear chromosome of Streptomyces griseus IFO13350 contains not only atypical telomere sequences but also probable pseudogenes for two typical telomeric proteins. Two identical operons (SGR98t-SGR97t near one telomere and SGR7041t-SGR7042t near the other telomere) in the terminal inverted repeat sequence were predicted to encode a novel pair of telomeric proteins. SGR97t, a 185-amino-acid protein showing only 18% amino acid sequence identity to typical terminal proteins of Streptomyces, was found to be attached to the chromosomal ends, as determined by immunological analysis. On the other hand, electrophoretic mobility shift assays showed that SGR98t, an 837-amino-acid protein having a DnaB-like helicase C-terminal domain, was capable of binding specifically to the single-stranded terminal DNA corresponding to the 3′ overhang of the replication intermediate. These results indicate that SGR97t (and SGR7042t) and SGR98t (and SGR7041t) were the functional telomeric proteins in the replication of the linear chromosome of S. griseus IFO13350.     

Analysis of Some Barley Chlorophyll Mutants and Their Response to Temperature Stress

Six barley chlorophyll (Chl) mutants, viridis, flavoviridis, chlorina, xanhta, lutea, and albina, differed in the contents of Chl (a+b) and carotenoids (Cars). In accordance with their Chl-deficient phenotype, the Chl a and b and Car contents of mutants decreased from viridis to albina, only xantha had the same or even higher concentration of Cars as the wild type plant. The albina mutant completely lacked and xantha had a significantly reduced photosynthetic activity. We found quantitative differences in protein contents between wild type and mutant plants, with the lowest concentration per fresh mass in the albina mutant. Chl fluorescence analysis revealed that heat-treated barley leaves of both the wild type and Chl mutants had a lower photosystem 2 efficiency than the untreated ones. With ³⁵S-methionine labelling and SDS-PAGE we found that six to nine de novo synthetized proteins appeared after heat shock (2 h, 42 °C) in the wild type and Chl mutants. In albina the expression of heat shock proteins (HSPs) was reduced to 50 % of that in the wild type. Hence mainly albina mutants, with a completely destroyed proteosynthetic apparatus of the chloroplasts, are able to synthesize a small set of HSPs. The albina mutant is a very useful tool for the study of different gene expression of chloroplast and nuclear DNA.     

14个蝴蝶兰品种遗传关系的ISSR分析

采用ISSR分子标记技术对14个蝴蝶兰品种进行品种间遗传关系的研究。利用14个筛选的引物共扩增出179条带,其中多态性条带147条,多态性条带比率(PPB)为82%。品种之间的遗传相似系数范围在0.734~0.936间,说明部分蝴蝶兰品种间存在显著的遗传分化。14个引物组合可区分所有14个品种,并且检测到20条品种特异性条带,这些品种特异条带可用来鉴定供试蝴蝶兰中的10个品种。因此,ISSR分子标记能有效地进行蝴蝶兰品种鉴定。UPGMA聚类分析表明,14个品种可聚为2类,聚类情况与花色特征比较一致,但与花色的划分结果不完全相同,这可能是由于品种间杂交引起的。本文也论讨了ISSR分析结果对蝴蝶兰育种的指导意义。     

ISSR鉴定亲缘关系非常近的芒果栽培品种

用ISSR技术鉴定7个吕宋芒品种(系)和柳州吕宋芒。从30个引物中筛选出6个多态性好的ISSR引物建立DNA指纹图谱用于区分吕宋芒品种(系)。分析DNA指纹图谱,发现这6个引物中每个引物都能区分吕宋系列品种(系),表明ISSR-PCR技术对芒果品种(系)的鉴定非常有效,能区分亲缘关系很近的品种(系)。基于69条多态性条带的聚类分析结果,发现吕宋芒和其它供试的7个品种(系)同源性低,而这7个品种(系):高州吕宋芒、湛江吕宋芒、田阳香芒、金钱芒、柳州吕宋芒、粤西一号、攀西红吕宋同源性较高,可归为一类。     

Increase in Thermostability of Recombinant Barley β-Amylase by Random Mutagenesis

Random mutations were introduced into recombinant barley β-amylase by modified PCR to increase its thermostability. Two clones were obtained. One was found to have a change of Ser-351 to Pro and another, a change of Ala-376 to Ser, and 2.3°C and 1.0°C increases, respectively, in the thermostabilities compared with that of native recombinant β-amylase.     

不同来源大麦对条纹病抗性鉴定及遗传多样性分析

为了解不同来源大麦对条纹病的抗性及遗传多样性,本研究采用"三明治法"通过人工接种大麦条纹病菌对91份大麦材料进行抗性评价,并通过31对SSR标记对91份大麦材料进行遗传多样性和群体结构分析。结果表明,人工接种大麦条纹病菌后共鉴定出4份免疫、6份高抗、33份抗病、43份感病和5份高感大麦材料;31对SSR标记共检测出等位基因238个,平均每对标记可检测到7.677个等位基因,等位基因数的变幅为2～19;主基因频率变化范围为0.236～0.951,平均值为0.394;基因多样性指数的变幅为0.094～0.871,平均值为0.667;多态性信息含量变幅为0.091～0.860,平均值为0.613;遗传相似系数变异范围为0.103～1.000,平均值为0.522;在遗传相似系数为0.783水平上可将参试材料聚为3个大类群,各大类分别包含86份、2份和3份材料;群体遗传结构分析表明,供试大麦材料分为3个亚群,每个亚群分别包含47份、33份和11份材料,且在91份材料中,Q>0.6的材料占总数的97.80%。本研究经抗病鉴定及分子标记结果综合分析,可为挑选抗病亲本辅助抗大麦条纹病优良品种的选...     

取样策略对SSR标记鉴别紫云英品种能力的影响

紫云英属于异花授粉植物, 品种内个体基因型杂合, 品种鉴定难度大。本文以紫云英闽紫系列3个审定品种为材料, 采用SSR标记进行取样策略对3个品种鉴别能力影响的研究。结果表明：（1）固定4对引物组合,从5～50进行梯度取样时,品种内的扩增位点总数、观测等位基因数趋于增多,但有效等位基因数、Shanon信息指数、遗传多样性指数增大到最大值后趋于下降,其中总体样品的最大值是出现在取样量为30的时候;随着样品量的增加,品种间Nei氏遗传距离以及分子方差分析的品种间期望变异系数比例值（PhiPT）均呈减少趋势,但PhiPT值的置信度在增大;（2）固定品种的样品容量为30和50,再加入2对能扩增出在品种间形成频率差异的标记位点的SSR引物对,基于这6对SSR引物可以将参试的3个紫云英品种有效的区分,品种间期望变异系数比例值（PhiPT）提高且差异的置信度具有明显的统计学意义（P<0.001）。主成分分析进一步表明：3个参试品种30个样品与50个样品的散布状况基本一致。对紫云英取样策略的研究表明：为提高对参试品种的鉴别能力,样品取样量以30株为宜,即达到较佳鉴别效果又降低分析成本。