应用植物形态学和AFLP分子标记鉴别陕西漆树品种

应用植物形态学和AFLP分子标记,对陕西8个漆树栽培品种和6个野生居群进行了鉴别.结果表明:从26个形态性状中筛选出3个主成分,其中:第1主成分指标7个(第5小叶长/宽、花瓣长、花瓣宽、花丝长、花丝直径、花药长和宽)贡献率为30.383%;第2主成分指标5个(小叶数、复叶长、复叶柄长、第5小叶叶柄长、顶端小叶叶柄长)贡献率为19.321%;第3主成分指标2个(第5小叶顶角角度、顶端小叶顶角角度)贡献率为13.777%.筛选的8对AFLP引物组合,均可将漆树14个样品完全区分开.植物形态学和AFLP分子标记相结合,不仅可用于漆树不同品种及野生居群间的鉴别,而且可反映其相互间的亲缘关系.     

漆树科植物漆酚的多样性及演化关系研究

漆酚是漆树科植物独有的化学成分,具有良好的抗氧化、抗肿瘤、抗病毒等生物活性,极具药物和工业生产开发价值。该研究通过对国内外有关漆树科各类群有关漆酚数据的搜集和整理,开展漆树科漆酚的多样性与分布情况的综合统计;同时在利用核基因ETS和叶绿体基因rps16和trnL-F重建漆树科系统发育树的基础上,探讨漆酚在漆树科及各属间的起源和演化关系。结果表明:(1)漆树科漆酚具有较高的多样性,在其苯环和-R链上都有很大的变异。(2)发现漆酚仅存在于Anacardioideae亚科中,是该亚科的一个重要界定特征。(3)基于漆酚苯环结构和-R链特征的祖先性状重建分析表明,漆树科的漆酚是由邻苯二酚向间苯二酚演化,而漆酚-R链的十五烃(烯)基则是向十七烃(烯)基平行演化。     

带叶兜兰5个野生居群表型多样性分析

以云南、广西、贵州的5个带叶兜兰主要野生居群分布区为研究对象,采用巢式方差分析、表型分化系数、主成分分析、相关分析、聚类分析等分析方法,对带叶兜兰的18个性状的表型多样性进行分析。结果显示:带叶兜兰表型性状在居群间表现出显著或极显著差异,居群内变异较少。带叶兜兰平均表型分化系数为75.18%,居群间的变异是居群多样性的主要来源;18个表型性状变异系数平均值的变化范围为6.36%～18.51%,离散程度较高。主成分分析表明,造成带叶兜兰表型变异的主要来源是株高、子房长、花茎高、最大叶长、花横径和合萼片长6个性状。Pearson相关分析表明,株高、合萼片宽和中萼片宽等与大部分性状均呈显著或极显著正相关;18个性状中只有合萼片长与地理因子极显著相关。聚类分析结果表明,5个居群大致依据地理距离聚为3个类群。本研究发现,带叶兜兰居群遗传多样性较高,居群间的变异远高于居群内的变异,因此,应尽可能保护更多的野生居群,在原地保护的基础上建立种质资源保存圃、库,进行人工繁殖和育种,以利用促保护。     

8个栽培水杉居群遗传多样性的等位酶分析

水杉(Metasequoia glyptostroboides)是我国特有的活化石植物,有50多年的栽培历史。栽培水杉在多大程度上涵盖了野生水杉的遗传多样性,是评价水杉保育策略成功与否的关键所在。为了评估水杉栽培居群的遗传变异水平,作者采用超薄平板微型聚丙烯酰胺等电聚焦电泳方法对水杉的8个人工栽培居群的等位酶变异和遗传多样性进行了研究。对9个酶系统23个酶位点的检测结果表明: 有8个酶位点的等位基因频率分布差异非常显著, 在2个居群中存在稀有等位基因;平均等位基因数Ａ=1.375,多态位点百分率PPL=33.15,平均预期杂合度H_e=0.161。与孑遗居群相比,　栽培水杉的遗传变异水平偏低,表明栽培居群的遗传多样性不能完全涵盖孑遗居群。将采于潜江的9个丛枝水杉（Metasequoia glyptostroboides var.caespitosa）样品与分别采自潜江、武汉和上海的各9个水杉样品的等位酶资料进行了UPGMA单株聚类分析,并将丛枝水杉作为一个居群与其他8个水杉居群作UPGMA聚类分析,其结果不支持将丛枝水杉作为水杉的一个变种的分类处理。     

锥栗种质资源遗传多样性的SRAP分析

应用SRAP分子标记法,采用筛选出的15个引物组合,对采自湖南、浙江、四川、贵州、福建5省的17个野生锥栗居群资源及23份锥栗栽培品种进行了遗传多样性分析,旨在为锥栗遗传背景分析和种质创新提供依据。17个野生锥栗居群共扩增出221个位点,平均多态性位点数为155.06,多态性位点百分率为70.16%。总遗传多样性指数(Ht)、居群内遗传多样性指数(Hs)、Nei's基因多样性指数、Shannon's信息指数分别为0.3636、0.2466、0.2460、0.3677,说明野生锥栗居群具有较高的遗传多样性和变异度。其中,衡山(HS)居群的多态性位点百分率最高,为85.07%,吉首小溪村(JS)居群次之,为83.26%,表明衡山、湘西一带为湖南省野生锥栗居群遗传多样性最丰富的区域。UPGMA聚类分析表明,浏阳溪江居群和衡山居群间的遗传相似性最大,亲缘关系最近;贵州黎平居群和岳阳月田镇居群间的遗传相似性最小,遗传差异较大。23个锥栗品种多态性位点百分率占比为89.14%,遗传相似性系数在0.66~0.85之间,且在遗传相似性系数0.66处,大尖嘴单独聚为一类,反映出它的遗传基础与其他品种具有较大的差异;在遗传相似性系数0.85处,华栗2号和铁锥被聚为一类,表明这2个品种亲缘关系最近,遗传差异最小。此外,还构建了Em1-Me2、Em2-Me1、Em2-Me2和Em2-Me3共4个引物组合的SRAP数字指纹图谱,为锥栗品种的快速鉴定提供了参考依据。     

浙江扇脉杓兰野生居群的表型性状变异及其与地理-土壤养分因子的相关性

为揭示扇脉杓兰(Cypripedium japonicum Thunb.)表型性状的变异规律,以浙江临安6个野生居群(老殿、宝剑石、红蛇洞、清凉峰、大明山和里横塘)为研究对象,分析了叶片、萼片、子房及唇瓣等18个表型性状的变异幅度,并讨论了表型性状变异与地理-土壤养分因子间的相关性以及居群间和居群内的表型多样性.结果显示:6个居群18个表型性状(地径、下叶长、下叶宽、下叶长/宽比、上叶长、上叶宽、上叶长/宽比、中萼片长、中萼片宽、苞片长、苞片宽、子房长、花梗直径、花梗长、唇瓣长、唇瓣宽、植株展度和株高)测量值的F值为1.240 ～ 18.666,除花梗直径和株高外,其他16个性状均有极显著差异.各性状的变异系数为5.937％ ～ 26.349％,其中,唇瓣长的平均变异系数最小(6.119％)、苞片宽的平均变异系数最大(24.124％);居群间各性状的变异系数为8.139％～ 13.663％,平均变异系数为11.143％,其中,里横塘居群的平均变异系数最大、宝剑石居群的最小.各性状的多样性指数平均为1.977,其中地径的多样性指数最大(2.045)、苞片宽的最小(1.663).通过UPGMA聚类分析,可将6个居群划分为3组:清凉峰、大明山、老殿和宝剑石居群为1组;红蛇洞和里横塘居群分别各自成组,其中清凉峰和大明山居群的遗传距离最近.各表型性状间存在显著或极显著的相关关系;但仅地径与速效钾含量显著正相关(r=0.854),子房长、唇瓣长和宽与土壤全磷含量显著正相关(r值分别为0.916、0.854和0.914),植株展度与土壤pH显著正相关(r=0.810),其余性状与地理因子(包括经度、纬度和海拔)及土壤养分因子均无显著相关性.研究结果表明:扇脉杓兰野生居群表型性状存在丰富的变异和多样性,且与生境土壤中速效钾及全磷含量及土壤pH有关.     

云南含笑天然居群的表型多样性分析

为揭示云南含笑天然居群表型变异程度和变异规律,以云南昆明地区天然分布的云南含笑为研究对象,调查了6个居群180个单株的14个表型性状,采用巢式方差分析、变异系数、相关分析、Shannon-Weaver多样性指数分析和聚类分析等方法,分析了居群间和居群内表型多样性.结果表明:(1)云南含笑表型性状在居群间和居群内存在极其丰富的多样性,14个表型性状平均表型分化系数(24.38％)小于居群内变异(75.62％),居群内变异是表型变异的主要来源;14个表型性状的变异系数(CV)在16.20％～60.11％之间,表明云南含笑居群内表型性状离散程度较高.(2)对云南含笑各居群的Shannon-Weaver指数分析表明,云南含笑各居群具有丰富的多样性,总体表型多样性指数为1.772.(3)利用居群间欧氏距离进行的UPGMA聚类分析结果表明,云南含笑6个天然居群可以聚为3类,而且表型性状并没有严格依地理距离而聚类.     

同质园栽培下的湖南鱼腥草居群叶表型性状多样性分析

将采自湖南的20个鱼腥草(Houttuynia cordata Thunb.)居群栽培于同质园内,通过测定9个叶片表型性状和6个叶片相对性状,采用方差分析、变异系数、相关性分析、主成分分析以及聚类分析等方法,对鱼腥草居群叶片形态的变异特征进行研究。结果表明:(1)同质园栽培的20个鱼腥草居群叶片15个表型性状均存在显著差异,居群间差异大于居群内差异;(2)居群内叶性状的平均变异系数变化幅度为5.63%~9.08%,说明居群内多样性程度较低,其中株洲攸县居群(P16)变异系数最高(9.08%),怀化溆浦居群(P3)变异系数最小(5.63%);(3)15个叶性状的平均变异系数变化幅度为3.71%~10.28%,说明各性状的多样性程度也较低,其中叶面积变异系数最大(10.28%),叶宽与叶中部宽之比(W_2/W_4)的变异系数最小(3.71%);(4)相关性分析显示,叶面积与叶宽、叶中部宽、叶长、叶基至叶宽处的距离、叶柄长都呈现出显著正相关,与叶尖夹角呈显著负相关,而与叶基夹角关系不大;(5)叶片15个性状中前3个主成分的贡献率为83.66%,聚类分析可将20个居群划分为心形、小心形、狭心形和大心形4类,呈现出间断分布的地理格局;(6)同质园实验说明叶表型性状多样性主要是由遗传物质决定的,对其多样性分析能为优良种质资源的选育提供理论依据。     

云南滇龙胆居群表型多样性及其与环境关系研究

为揭示滇龙胆天然居群表型变异程度和变异规律,以云南省5个区域天然分布的20个野生居群400个单株20个表型性状指标为调查研究对象,并用变异系数、Shannon-Weiner多样性指数和巢式方差分析对滇龙胆居群间和居群内的表型变异进行分析,用相关性分析对滇龙胆表型性状与地理气象因子间的变异格局进行分析,用类平均聚类法对20个滇龙胆居群进行分类.结果表明:5个区域的滇龙胆以楚雄地区变异最大,变异系数为39.8%,变异最小的是昆明地区,为31.4%;不同居群的20个表型性状变异程度差异明显,变异系数在14.4%～91.8%之间,平均为40.4%,20个居群的平均变异为26.8%～37.0%;滇龙胆地理居群的表型分化较高,20个性状居群间的分化系数平均为73.14%,变化范围为36.03%～91.94%,居群间变异高于居群内;滇龙胆20个表型性状的总的多样性指数平均为2.547,5个不同分布区域多样性指数存在差异,最大为楚雄(1.271 4),最小为玉溪(1.266 7);表型性状变异受经度和降雨量影响较大,与纬度、海拔和温度相关性不显著;通过UPGMA聚类,滇龙胆被分成3个组,性状的表型特征并没有依地理距离而聚类....     

山罗花和天柱山罗花四个居群的数值分析

对山罗花和天柱山罗花的4个居群的15项形态特征进行了数值分析。结果表明:形态性状在4个居群间均存在一定程度的变异,居群间变异系数的平均值从小到大排列为:天柱山居群(天柱山罗花)、天柱山居群(山罗花)、黄山居群(山罗花)、鹞落坪居群(山罗花);天柱山罗花与山罗花种间的形态差异已达到极显著水平,山罗花的3个居群间部分性状也具有极显著的形态差异;以形态特征为基础的Q-聚类分析可以把天柱山罗花和山罗花聚为两类,但山罗花(天柱山居群)和其他2个居群在较远处聚在一起,R-聚类分析发现了强正相关关系、弱正相关关系、弱负相关关系的性状。     

Principal-Component Analysis of the Characteristics Desirable in Baker's Yeasts

Twenty-seven properties considered to be required for good bakery products were examined in 56 industrial and 2 laboratory yeast strains. The data obtained were applied to principal-component analysis, one of the multivariate statistical analyses. The first and second principal components together were extracted, and these accounted for 77.7% of the variance. The first principal component was interpreted as the glycolytic activity of yeast in dough, and the second one was interpreted as the balance of leavening abilities in sweet and flour doughs from the factor loadings. The scattergram on the two principal components was effective in grouping the 58 yeast strains used.     

Principal-Component Analysis of the Characteristics Desirable in Baker's Yeasts

Twenty-seven properties considered to be required for good bakery products were examined in 56 industrial and 2 laboratory yeast strains. The data obtained were applied to principal-component analysis, one of the multivariate statistical analyses. The first and second principal components together were extracted, and these accounted for 77.7% of the variance. The first principal component was interpreted as the glycolytic activity of yeast in dough, and the second one was interpreted as the balance of leavening abilities in sweet and flour doughs from the factor loadings. The scattergram on the two principal components was effective in grouping the 58 yeast strains used.     

两种标记方法对60mer寡核苷酸芯片背景信号影响的初步分析

研究两种不同的样本标记方法对人全基因组高密度60mer寡核苷酸芯片背景信号的影响。收集5对患病与健康人外周血单个核细胞,分别提取总RNA后,采用限制性显示技术（restriction display,RD）进行样本双色（Cy3/Cy5）荧光标记,与5张Agilent 60mer高密度（22K）Human 1B寡核苷酸芯片进行杂交。芯片全部杂交点分3组：基因探针组、阳性对照组和阴性对照组。阳性对照采用荧光标记寡核苷酸直接掺入法进行标记。对全部杂交信号点的Cy3和Cy5背景信号值,用SPSS软件进行数据转换、正态性检验、方差齐性检验、变异系数分析和重复数据的方差分析。数据分析结果显示,Cy3 标记的背景信号值均高于 Cy5标记的背景信号值。重复测量数据的方差分析表明,在Cy3 和Cy5标记中,两种不同标记方法间的背景信号值的差异极显著（PCy3＜0.01, PCy5＜0.01）,且RD标记点的背景信号平均值低于荧光标记寡核苷酸直接掺入标记法标记的阳性对照点。RD标记方法是一种有用的低背景信号的高密度长链寡核苷酸芯片样本标记方法。     

Study on Mineral Elements in Salvia roborwskii from the Qinghai-Tibet Plateau

The contents of trace elements, including aluminum, barium, beryllium, calcium, cobaltium, chromium, copper, iron, potassium, lanthanum, lithium, magnesium, manganese, sodium, nickel, phosphorus, lead, sulfur, silicon, titanium, vanadium, and zinc, were determined by ICP-AES. The mean concentrations of the 22 elements were as follows: K>Ca>Mg>Fe>Al>Na>S>P>Ba>Ti>Mn>Zn>Sr>Cu>Ni=Cr>Pb>V>Li=La>Co>Be. Principal components analysis of SPSS was applied to study the characteristic elements in Salvia roborwskii. Five principal components which accounted for 89.288% of the total variance were extracted from the original data. The first factor accounted for 56.401% of the total variance, which meant that aluminum, barium, beryllium, calcium, and cobaltium were the characteristic elements in S. roborwskii. This useful new method was used to evaluate the quality of S. roborwskii, and to provide the scientific foundation for its utilization and further research.     

An automated SPE/LC/MS/MS method for the analysis of cocaine and metabolites in whole blood

As laboratories are called upon to develop novel, fast, and sensitive methods, here we present a completely automated method for the analysis of cocaine and its metabolites (benzoylecgonine, ecgonine methyl ester, ecgonine and cocaethylene) from whole blood. This method utilizes an online solid-phase extraction (SPE) with high performance liquid chromatographic separation and tandem mass spectrometric detection. Pretreatment of samples involve only protein precipitation and ultracentrifugation. An efficient online solid-phase extraction (SPE) procedure was developed using Hysphere MM anion sorbent. A gradient chromatography method with a Gemini C6-Phenyl (50mmx3.00mm i.d., 5microm) column was used for the complete separation of all components. Analysis was by positive ion mode electrospray ionization tandem mass spectrometry, using multiple reaction monitoring (MRM) to enhance the selectivity and sensitivity of the method. For the analysis, two MRM transitions are monitored for each analyte and one transition is monitored for each internal standard. With a 30-microL sample injection, linearity was analyte dependent but generally fell between 8 and 500ng/mL. The limits of detection (LODs) for the method ranged from 3 to 16ng/mL and the limits of quantitation (LOQs) ranged from 8 to 47ng/mL. The bias and precision were determined using a simple analysis of variance (ANOVA: single factor). The results demonstrate bias as <7%, and %precision as <9% for all components at each QC level.     

Full-scan high resolution accurate mass spectrometry (HRMS) in regulated bioanalysis: LC-HRMS for the quantitation of prednisone and prednisolone in human plasma

A liquid chromatography-full scan high resolution accurate mass spectrometry (LC-HRMS) method for quantifying prednisone and prednisolone in human plasma using a quadrupole time-of-flight mass spectrometer (Q-TOF) was developed. Plasma samples were extracted using a liquid-liquid extraction procedure. Full scan data were acquired in the TOF only mode and extracted ion chromatograms were generated post-acquisition with the exact masses of the analytes. The calibration range was 5-2500 ng/mL, with a Lower Limit of Quantitation (LLOQ) of 5 ng/mL. The assay accuracy was between 98.4% and 106.3%. The between-run (inter-day) and within-run (intra-day) precision were within 1.7% and 2.9%, respectively. The matrix effect was between 0.98 and 1.10 for the six different lots of human plasma evaluated. Pooled incurred samples were analyzed by the method and the results matched those obtained from an LC-MS/MS method. In addition, qualitative information on phospholipids, and other endogenous components were also extracted from the full-scan data acquired.     

Rapid solid-phase extraction method for automated gas chromatographic–mass spectrometric determination of nicotine in plasma

The aim of this study was to develop a simple, rapid and sensitive assay of nicotine in plasma for automated gas chromatographic–mass spectrometric analysis. Biological samples were extracted using pre-packed Extrelut-1 columns with 5 ml of ethyl acetate. Quantitative analysis was done using deuterium-labelled nicotine as internal standard. The limit of quantitation was 0.5 ng in 1-ml plasma samples. Precision was ranging from 13.3% to 1.64% (R.S.D.) depending on the concentration, while the deviation was 4.16%. This method has been used for determination of nicotine bioavailability from new, low-dosage, nicotine chewing gum strips.     

Effects of data imbalance on estimation of heritability

Summary Effects of data imbalance on bias, sampling variance and mean square error of heritability estimated with variance components were examined using a random two-way nested classification. Four designs, ranging from zero imbalance (balanced data) to low, medium and high imbalance, were considered for each of four combinations of heritability (h²=0.2 and 0.4) and sample size (N=120 and 600). Observations were simulated for each design by drawing independent pseudo-random deviates from normal distributions with zero means, and variances determined by heritability. There were 100 replicates of each simulation; the same design matrix was used in all replications. Variance components were estimated by analysis of variance (Henderson's Method 1) and by maximum likelihood (ML). For the design and model used in this study, bias in heritability based on Method 1 and ML estimates of variance components was negligible. Effect of imbalance on variance of heritability was smaller for ML than for Method 1 estimation, and was smaller for heritability based on estimates of sire-plus-dam variance components than for heritability based on estimates of sire or dam variance components. Mean square error for heritability based on estimates of sire-plus-dam variance components appears to be less sensitive to data imbalance than heritability based on estimates of sire or dam variance components, especially when using Method 1 estimation. Estimation of heritability from sire-plus-dam components was insensitive to differences in data imbalance, especially for the larger sample size.Supported by grants from the Illinois Agricultural Experiment Station and the University of Illinois Research Board. Charles Smith, H. W. Norton and D. Gianola contributed valuable suggestions     

Mass spectrometric assay for analysis of haptoglobin fucosylation in pancreatic cancer

A mass spectrometric method was developed to elucidate the N-glycan structures of serum glycoproteins and utilize fucosylated glycans as potential markers for pancreatic cancer. This assay was applied to haptoglobin in human serum where N-glycans derived from the serum of 16 pancreatic cancer patients were compared with those from 15 individuals with benign conditions (5 normals, 5 chronic pancreatitis, and 5 type II diabetes). This assay used only 10 μL of serum where haptoglobin was extracted using a monoclonal antibody and quantitative permethylation was performed on desialylated N-glycans followed by MALDI-QIT-TOF MS analysis. Eight desialylated N-glycan structures of haptoglobin were identified where a bifucosylated triantennary structure was reported for the first time in pancreatic cancer samples. Both core and antennary fucosylation were elevated in pancreatic cancer samples compared to samples from benign conditions. Fucosylation degree indices were calculated and show a significant difference between pancreatic cancer patients of all stages and the benign conditions analyzed. This study demonstrates that a serum assay based on haptoglobin fucosylation patterns using mass spectrometric analysis may serve as a novel method for the diagnosis of pancreatic cancer.     

Evaluation of laboratory indexes of absorption of soil phosphorus by plants: I

Summary The yield of phosphorus in sorghum plants grown on samples of 22 different soils from the United States was determined in the greenhouse. Analyses of variance of the regressions of these values on the measurements of soil phosphorus by various laboratory methods were calculated as an aid in evaluating the methods.No general advantage was found from incubating the samples in moist condition for a week before analysis versus making the analysis on the initially-dry samples. In the order of decreasing precision of predicting the yields of phosphorus the methods are as follows: (1) anion-exchange resin method of Amer and coworkers, (2) 0.5M NaHCO₃ method of Olsen and coworkers, (3) phosphate potential method of Schofield and Aslyng, (4) phosphorus concentration in the 0.01M CaCl₂ extract of Schofield and Aslyng, and (5) 0.1N HCl, 0.03N NH₄F method of Bray and Kurtz.After taking into account the additional variables, little or no improvement in precision of prediction was obtained when the phosphorussorbedvs. time curves for the anion-exchange-resin method were (a) divided into four segments on the basis of time, the quantities of phosphorus in the four segments being used as independent variables in a multiple-regression equation, or (b) broken down into a maximum of four components on the assumption that the overall curve represents the summation of a group of simultaneous first-order reactions, the quantities of phosphorus in the several components being used as independent variables in a multiple-regression equation.The precision of prediction was improved by using as the soil-phosphorus measurement the sum of the products of the rate of phosphorus extraction by the anion-exchange-resin method and the quantities of phosphorus extracted within individual time intervals. A logarithmic expression was used to fit the relationship, however, and it appeared that the greater precision of prediction resulted from the logarithmic transformation rather than the superiority of the method as such.The precision of prediction was improved also by using the H₂PO ₄ ^– concentration instead of the total-inorganic-phosphorus concentration as the independent variable in the 0.01M CaCl₂ extracts of Schofield and Aslyng and by using the H₂PO ₄ ^– instead of the total inorganic phosphorus sorbed by the anion-exchange resin. This modification made the anion-exchange-resin method considerably better than the others tested.Journal Paper No. J-3452 of the Iowa Agricultural and Home Economics Experiment Station, Ames, Iowa. Project No. 1183. Contribution from the Department of Agronomy.Graduate Students and Professor of Soils, respectively.