新疆枸杞种质资源遗传多样性分析及DNA指纹图谱构建

利用SCoT分子标记对新疆枸杞种质资源进行遗传多样性分析和DNA指纹图谱构建,为杂交育种和种质鉴定提供理论依据。结果显示:9条SCoT引物扩增出条带256条,其中219条为多态性条带,多态性比率达85.62%,多态性信息含量(PIC)值变化范围在0.77~0.91之间,平均值为0.85,观测等位基因数(Na)、有效等位基因数(Ne)、Nei's基因多样性指数(H)和Shannon信息指数(I)的平均值分别为1.8562、1.4350、0.2611、0.3989,聚类分析表明,遗传相似系数变化范围在0.5938~0.8398之间,在遗传相似系数为0.66和0.71处,可将30份材料分别分为2大类和4个亚类,主坐标分析结果和聚类结果基本一致,同时利用5条多态性SCoT引物构建了30份材料的DNA指纹图谱。新疆枸杞种质资源遗传多样性水平较高,且SCoT分子标记适于新疆枸杞种质资源遗传多样性分析和DNA指纹图谱构建,该研究结果为新疆枸杞种质资源评价、鉴定和新品种选育奠定了基础。     

猕猴桃SCoT遗传多样性分析及指纹图谱的构建

利用SCoT标记对32个猕猴桃品种进行了遗传多样性分析。从47个SCoT引物中筛选了11个引物进行PCR扩增,共扩增出185个条带,其中多态性条带180个,多态性比率为97.30%。各引物Nei's基因多样性指数（H）平均为0.238 4,Shannon's信息指数（I）平均为0.377 8。利用UPGMA构建32份猕猴桃种质资源的聚类树状图。在遗传相似系数为0.78处可将32个猕猴桃品种分为5组,聚类结果与形态学分类基本一致。利用4条引物扩增的16个多态性位点构建了32个猕猴桃品种的DNA指纹图谱,可以将32个猕猴桃品种区分并准确鉴定。     

32个柿主栽品种SSR图谱构建及遗传变异分析

以32份柿主栽品种为试验材料,利用SSR标记技术构建其指纹图谱并进行遗传多样性分析。从78对候选引物中筛选出20对多态性高、稳定性好的引物作为核心引物,构建柿主栽品种SSR指纹图谱。结果显示:(1)20对引物在32份材料中共扩增出183条多态性条带,每对引物扩增出3～20条不等,平均每对引物扩增出9.15条,多态性比率为81.3%。各个位点的杂合度在0.410 3～0.914 3之间,平均为0.702 7。(2)8对引物在12个品种上具有特征带型,采用5对引物进行组合鉴定即可将32个柿品种完全区分开。(3)依据SSR带型特征,对每对引物生成的不同带型直接编号,简化柿SSR带型记录方法,并利用每个品种的带型编号,建立其DNA指纹图谱,结果表明,核心引物组合法比特征谱带法更适用于构建中国柿主栽品种DNA指纹图谱。(4)根据系统聚类分析将32个柿主栽品种分为两大类,品种间的亲缘关系与地理来源具有一定的相关性。     

基于引物“随机组合”构建观赏桃SSR指纹图谱

近年来,我国观赏桃新品种日渐繁多、名称混乱、市场难以监管,同时用以区分品种的SSR指纹图谱的构建方法在研究界无统一的科学标准,尤其是构成最终引物组合的核心引物的确定,具体操作流程层出不穷、五花八门。为探索筛选SSR指纹图谱核心引物的科学方法,同时构建观赏桃SSR指纹图谱,该研究选用35对已报道的SSR引物对22份观赏桃种质进行试验。结果表明:通过PCR扩增与分析,多态性较高的8对引物——候选引物总共扩增出31个多态性条带,变幅为3~5个,PIC值变幅为0.458~0.668,MI值变幅为1.374~3.340。采用"随机组合"法对8对引物进行C_8~1、C_8~2、C_8~3…依次分析,得到区分能力最强的3种不同的最少引物组合方式——候选组合,并能区分出18份种质,从中发现区分能力最强的3种引物组合方式并不都是由引物PIC值、alleles数量或MI值等多态性指标最高的引物组成,而是由互补性最强的引物组成。选用组合内各引物多态性条带总数最多的组合方式"4-3"(BPPCT001+BPPCT015a+BPPCT017+BPPCT025)为22份观赏桃种质构建了指纹图谱。基于此,通过常规多态性指标筛选候选引物可以确定出单对引物鉴别能力最强的少量引物;通过"随机组合"筛选候选组合可以进一步确定出引物之间互补性最强的几种组合方式;根据组合内各引物的多态性条带总数确定最终核心引物可以确定出可扩容性最大的引物组合。该研究最终建立了候选引物——候选组合——核心引物组合"三步法"确定SSR指纹图谱核心引物组合的科学方法,不仅为22份供试观赏桃种质构建了SSR指纹图谱,也为其它作物SSR指纹图谱的构建提供了新的思路。     

皂荚种质资源SCoT遗传多样性分析及指纹图谱的构建

采用SCoT标记分析了18个皂荚种质的遗传多样性,并采用UPGMA法对18个皂荚种质进行了聚类分析。在此基础上,通过筛选出的多态性条带构建了18个皂荚种质的SCoT指纹图谱。扩增结果表明:从51个SCoT引物中筛选了15个引物进行PCR扩增,共扩增出226条带,其中多态性条带216条,多态性比率为96.61%。各引物多态性信息含量(PIC)、观测等位基因数(Na)、有效等位基因数(Ne)、Nei’s基因多样性指数(H)和Shannon’s信息指数(I)的平均值分别为0.875 9、1.964 9、1.440 1、0.272 6、0.426 1。18个皂荚种质的遗传相似系数在0.491 4~0.938 1之间,表明供试材料之间具有较丰富的遗传多样性。聚类分析结果表明:在遗传相似系数为0.60处可将18个皂荚种质分为3组,其中野皂荚单独为一组,山皂荚和皂荚-T聚为一组,其它皂荚材料聚为一组。利用3个引物扩增的8个多态性位点构建了18份皂荚种质资源的DNA指纹图谱,可以将其区分并精准鉴定。该研究结果为皂荚种质的鉴定和新品种选育提供了一定的理论依据。     

应用ISSR指纹图谱鉴定上饶早梨3个主栽品种及其不同继代次数试管苗

为鉴定上饶早梨3个主栽品种("花厅六月雪","花厅黄皮消"和"田墩六月雪")的亲缘关系以及其不同继代次数试管苗的遗传变异,本研究应用筛选出的9条ISSR引物对上饶早梨3个主栽品种20份样本进行分子标记分析。结果显示,9条引物共获得47条清晰可辨条带,其中多态性条带24条,平均多态性比率为51.06%,平均每对引物扩增出4.7条带,其中2.4条具有多态性。其中,引物CW32438多态性最高,为100.00%,引物CW32466多态性最低,只有20.00%。20个样品的GS值范围为0.617 3~1.000 0,平均值为0.806 6,变幅为0.382 7。UPGMA法聚类分析结果表明,在GS值为0.817 7处,可将上饶早梨主栽品种分为两类,"花厅六月雪"和"花厅黄皮消"为第Ⅰ类,"田墩六月雪"为第Ⅱ类。根据引物扩增位点的特异性,挑选了5条核心引物,并依据其扩增模式图进行编码组合,构建了上饶早梨3个主栽品种的DNA分子指纹图谱。利用这些指纹图谱的差异,可鉴定上饶早梨主栽品种的归属地。上饶早梨3个主栽品种继代8~13次的试管苗虽然存在一定程度的变异,但这种遗传差异无显著性,说明通过带芽茎段对上饶早梨3个主栽品种进行离体快繁可保证其遗传稳定性。     

莲藕品种DNA指纹图谱的绘制

采用RAPD技术对14个莲藕品种进行遗传多态性分析,用5个Operon引物和80个SBS的RAPD引物进行筛选,从中选出来自SBS的RAPD-C13和RAPD-D15扩增出的8条多态性条带,绘制了14个品种的DNA指纹图谱,在该图谱中每个品种均有各自特异的DNA指纹。     

地黄SCoT分子标记体系的建立和指纹图谱的构建

该研究采用L_(25)(5~6)正交设计和单因素两种方法,对影响地黄SCoT-PCR反应的5个因素(模板DNA浓度,引物浓度,ddH_2O和Mix的用量以及退火温度)进行了优化。结果表明:优化后的反应体系总体积为25μL,含有8μL ddH_2O,1μL模板DNA(80 ng·μL~(-1)),1μL引物(8μmol·L~(-1))和15μL Mix,退火温度为45℃。运用30份地黄种质材料,对优化的SCoT-PCR正交体系进行多次重复验证,获得了多态性丰富、条带清晰的扩增图谱,证明该反应体系稳定可靠。利用该体系对32条SCoT引物进行两次筛选,得到14个扩增产物清晰、重复性好且多态性条带相对较高的引物。利用SCoT_4等5条引物构建了上述地黄2个种共30份种质的SCoT指纹图谱。利用这5个SCoT引物指纹图谱可将7个地黄常用栽培品种区分开。这表明SCoT分子标记体系适用于地黄主要品种亲缘关系及遗传多样性的研究,所构建的指纹图谱也为地黄常见的7个栽培品种的区分提供参考依据。     

番木瓜主栽品种SCoT指纹图谱构建及遗传变异分析

利用SCoT标记对中国22个番木瓜主要栽培品种进行遗传多样性分析和指纹图谱构建,为番木瓜品种资源的鉴定和杂交亲本的选配提供技术参考。结果显示:(1)SCoT标记有效等位基因数(Ne)平均为1.47,Nei’s基因多样性指数(H)平均为0.26,Shannon’s信息指数(I)平均为0.39,表明SCoT标记具有较高的多态性检测效率。(2)聚类分析和主坐标分析结果表明,大部分番木瓜品种间的遗传相似系数为0.79~0.90,品种间遗传多样性水平较低;在相似性系数为0.75时可将所有品种分为3个组群。(3)6个SCoT引物可以将22个番木瓜品种完全区分开,每个品种都各有独特的指纹图谱,置信概率达到99.976%。     

云南莲瓣兰主栽品种SSR指纹图谱的构建和遗传差异分析

为了解云南莲瓣兰(Cymbidium tortisepalum)的遗传多样性,利用SSR技术对32个莲瓣兰主栽品种进行遗传变异分析,并构建莲瓣兰栽培品种的指纹图谱。结果表明,筛选出的12对多态性高、稳定性好的引物共检测到95个等位基因,每对引物检测到4~18个等位基因,有效等位基因数(N E)为61.489,平均有效等位基因数(NA)为5.124,Shannon信息指数(I)和多态性信息含量(PIC)分别为0.806~2.624和0.789~0.953。12对引物中,以引物SSR03的等位基因数、NE、观测杂合度、I和PIC最高。32个品种在12对引物上都具有不同的特异性条带,可以彼此区别。从12对引物中筛选出3对核心引物SSR02、SSR03和SSR12构建了莲瓣兰主栽品种SSR分子指纹图谱,这3对核心引物组合即可鉴定32个莲瓣兰栽培品种。这为莲瓣兰的品种鉴定、遗传多样性分析和分子育种研究提供理论基础和技术支持。     

六种藓类植物茎的比较解剖学研究

通过对6种藓类植物,即褶叶青藓(Brachythecium salebrosum(Web.et Mohr.)B.S.G.)、湿地匐灯藓(Plagiomnium acutum(Lindb.)Kop.)、侧枝匐灯藓(Plagiomnium maximoviczii(Lindb.)Kop.)、大凤尾藓(Fissidensnobilis Griff.)、大羽藓(Thuidium cymbifolium(Doz.et Molk.)B.S.G.)和大灰藓(Hypnum plumaeforme Wils.)嫩茎和老茎的石蜡切片和显微观察发现,同一藓类植株的嫩茎和老茎,茎结构稳定,不同种藓类植物茎横切面具有不同特征.植物体茎横切面形状、表层细胞的层数、细胞大小和细胞壁厚薄、皮层细胞大小和形状、中轴的有无以及比例等特征可以作为藓类植物的分科分类依据之一.     

Photoregulation of seed germination of wild-type and of an aurea-mutant of tomato

Seed germination of an aurea mutant of tomato ( Lycopersicon esculentum Mill.) is promoted by continuous irradiation with red, far-red or long-wavelength far-red (758 nm) light as well as by cyclic irradiations (5 min red or 5 min far-red/25 min darkness). Far-red light applied immediately after each red does not change the germination behaviour. Seed germination of the isogenic wild-type, cv. UC-105, is promoted by continuous and cyclic red light while it is inhibited by continuous and cyclic far-red light and by continious 758 nm irradiation. Far-red irradiation reverses almost completely the promoting effect of red light. The promoting effect (in the aurea mutant) and the inhibitory effect (in the wild-type) of continuous far-red light do not show photon fluence rate dependency above 20 nmol m⁻² s⁻¹. It is concluded that phytochrome controls tomato seed germination throgh low energy responses in both the wild type and the au mutant. The promoting effect of continuous and cyclic far-red light in the au mutant can be attributed to a greater sensitivity to P_fr.     

真菌类遗传学分析的知识结构教学

本文以认知结构理论为指导,讨论了真菌类遗传分析与高等动植物遗传分析的内在联系,认为利用这种内在联系进行教学可收到好的效果并说明了作者的具体教学过程。 Abstract:In the paper, the relationship between genetic analysis of Fungi and genetic analysis of high animal and plant was discussed.A good results were obtained when we adopted this method in the teaching.     

Cause of Senescence of Nine Sorts of Flowers

The levels of endogenous phytohormones and respiratory rate in nine sorts of flowers such as Cymbidium faberi Rolfe, Nopalxochia ackermannii Kunth and others were investigated both at full bloom and senescence and meanwhile the effect of exogenous phytohormones on prolonging the blossoms and promoting ethylene production were tested. There is a high content of endogenous ethylene in all the long-lived flowere, about 3–16 folds higer than the short-lived ones. There is a high level of ABA at full blooming flowers of short-lived flowers, in which there is no or only some cytokinins in it, but the ratio of CTK (6BA+zeatin)/ABA is smaller(l.7). The endogenous ABA reached a much higher level at senescence in all nine sorts of flowers, so it is reasonable to consider that it is ABA which plays an important role of regulation in controlling flower's senescence. There is a much higher level of GA3 and zeatin in the long-lived flowers which is not demonstrated in the shortlived ones. The respiratory rate is one of the factors controtling the longevity of flowers, but it does not play a decided role. Application of 6BA and zeatin prolongs distinctly orchid’s longevity, however exogenous IAA through the promotive action on ethylene production, evidently extends the longevity of the flowers of the Nopalxochia ackermannii Kunth.     

Interconnection of parameters of regulation system of lipid peroxidation and of morphophysiological parameters of mouse liver

A complex analysis of seasonal fluctuations of the mean group parameters of the system of regulation of lipid peroxidation has been performed in liver of Balb/c mice. Association of lipid characteristics and morphophysiological parameters is studied in the Balb/c mouse liver. An inter-connection is revealed between the liver index and the amount of lysoforms of phospholipids, the scale and character of the interconnection differing essentially depending on proportion of phos-phatidylcholine in mouse liver phospholipids.     

龙胆科药用植物化学成分的研究现状

龙胆科植物在我国的分布范围很广,且多数为药用植物,其多数种属的药用植物,至今其化学成分尚未被系统研究。综述了目前龙胆科药用植物的化学成分的研究现状及一般提取方法,对近年来发现的环烯醚萜及裂环烯醚萜类化合物进行了总结,为本科药用植物的更深入研究提供了参考。     

Physiological characteristics of various species of strains of carboxydobacteria

Seven strains of aerobic carbon monoxide-oxidizing bacteria (carboxydebacteria) when growing on CO as sole source of carbon and energy had doubling times which ranged from 12–42 h. The activity profiles obtained after discontinuous sucrose density gradient centrifugation indicated that the CO-oxidizing enzymes are soluble and the hydrogenases are membrane-bound in all strains examined. The CO-oxidizing enzymes of Pseudomonas carboxydohydrogena, Pseudomonas carboxydoflava, Comamonas compransoris, and the so far unidentified strains OM2, OM3, and OM4 had a molecular weight of 230,000; that of Achromobacter carboxydus amounted to 170,000. The molecular weights of the CO-oxidizing and H₂-oxidizing enzymes turned out to be identical. The cell sonicates were shown to catalyze the oxidation of both CO and H₂ with methylene blue, thionine, phenazine methosulfate, toluylene blue, dichlorophenolindophenol, cytochrome c or ferricyanide as electron acceptors. Methyl viologen, benzyl viologen, FAD⁺, FMN⁺, and NAD(P)⁺ were not reduced. The spectrum of electron acceptors was identical for all strains tested. Neither free formate, hydrogen nor oxygen gas were involved in the CO-oxidation reaction. Methylene blue was reduced by CO at a 1:1 molar ratio. The results indicate that CO-oxidation by carboxydobacteria is catalyzed by identical or similar enzymes and that the reaction obeys the equation CO+H₂OCO₂+2H⁺+2e^- as previously shown for Pseudomonas carboxydovorans.Dedicated to Otto Kandler remembering almost three decades of enjoyable cooperation     

Modulation of allostery of pyruvate kinase by shifting of an ensemble of microstates

Since the introduction of the concepts of allostery about four decades ago, much advancement has been made in elucidating the structure-function correlation in allostery. However, there are still a number of issues that remain unresolved. In this review we used mammalian pyruvate kinase (PK) as a model system to understand the role of protein dynamics in modulating cooperativity. PK has a triosephosphate isomerase (TIM)(α/β)₈ barrel structural motif. PK is an ideal system to address basic questions regarding regulatory mechanisms about this common (α/β)₈ structural motif. The simplest model accounting for all of the solution thermodynamic and kinetic data on ligand-enzyme interactions involves two conformational states, inactive E^T and active E^R. These conformational states are represented by domain movements. Further studies provide the first evidence for a differential effect of ligand binding on the dynamics of the structural elements, not major secondary structural changes. These data are consistent with our model that allosteric regulation of PK is the consequence of perturbation of the distribution of an ensemble of states in which the inactive E^T and active E^R represent the two extreme end states. Sequence differences and ligands can modulate the distribution of states leading to alterations of functions. The future work includes: defining the network of functionally connected residues; elucidating the chemical principles governing the sequence differences which affect functions; and probing the nature of mutations on the stability of the secondary structural elements, which in turn modulate allostery.