玉米不同杂种优势近等基因系及其杂交种的基因表达分析

以自发突变产生的籽粒大小和穗位叶片不同的 3对近等基因系及其杂交种为材料 ,研究了 6个基因在其叶片组织中的表达情况 ,以评价这些基因在杂种优势形成中可否作为“限率性基因”模式存在。结果表明 ,有些基因通过增强或减弱表达在杂种优势现象中起一定作用。将被测基因视为整体时会发现它们依次增强表达趋势为 :近等基因系 4 88 1>4 88 2 >4 78;相应杂交种 4 88 1× 340 >4 88 2× 340 >4 78× 340。对近等基因系及其杂交种个体千粒重和穗粒重的相关分析表明 ,Sus1、Sh1和ZmSps34 3′基因亦可作为适合的限率性候选基因以研究杂种优势分子机理。     

水稻白叶枯病新抗源Y238的鉴定及其近等基因系培育

从269份普通野生稻中鉴定出一个高抗白叶枯病的新抗源,编号为Y238.通过多茵系鉴定、抗谱分析及与目前国际上已知基因比较,证明该新抗源含有一个新基因,暂命名为WBB2.对JG30/Y238杂交后代成株期接种鉴定、遗传分析表明,WBB2为完全显性基因.通过杂交和回交,已将WBB2导入栽培稻中构建近等基因系.     

水稻抗稻瘟病近等基因系的cDNA微阵列分析

应用cDNA微阵列对来源于中156／谷梅2号重组自交系的水稻抗稻瘟病近等基因系G205和G71的稻瘟病菌胁迫基因表达谱进行了分析,发现有3个cDNA克隆的表达仅在抗病基因系G205接种病原菌12h后受到诱导,其中两个为功能已知基因,另一个为功能未知的新基因。另有35个差异表达克隆在两个近等基因系中均检测到,其中17个克隆的表达在G205和G71均受到病原菌的诱导,另外18个克隆的表达则在G205和G71均受到病原菌原抑制。序列分析表明,这些稻瘟病菌应答基因分别与防卫反应,信号传递,逆境胁迫和光合作用及糖代谢等功能相关,为植物抗病机制提供了相关信息。另外,Northern还证实了编码富含甘氨酸蛋白基因（Glycinerich protein Grp)的表达受稻瘟病病原菌的诱导,是一个稻瘟病诱导相关基因。     

甜菜夜蛾抗高效氯氟氰菊酯近等基因系的构建

将甜菜夜蛾Spodoptera exigua抗性种群雄成虫与敏感种群雌成虫杂交,杂交后代再自交后,用高效氯氟氰菊酯杀死敏感和部分杂合个体的剂量(50 μg/mL)处理4龄幼虫,对1天后存活的幼虫再用250 μg/mL汰选,将存活幼虫发育成的雄成虫与敏感种群的雌成虫回交,再经自交和选育,如此循环6次,获得了甜菜夜蛾抗高效氯氟氰菊酯近等基因系。对该基因系构建过程中各代幼虫的抗性水平和相关生物学特性进行了监测和比较,结果表明获得的抗高效氯氟氰菊酯近等基因系抗性种群的抗药性仍保持较高水平;酯酶同工酶的电泳分析表明抗高效氯氟氰菊酯近等基因系抗性种群与敏感种群图谱相近,而与亲本抗性种群之间存在明显差异。     

甜菜夜蛾抗高效氯氟氰菊酯近等基因系的RAPD标记

用随机引物扩增的方法对近等基因系抗高效氯氟氰菊酯种群和敏感种群甜菜夜蛾进行比较分析,从160条引物中筛选,引物S42(GGACCCAACC)和OPH13(GACGCCACAC)在抗性种群中扩增分别得到了1条特异扩增带,带型清晰、重复性较好,所以S42和OPH13产生的两条特异扩增带是分别与甜菜夜蛾对高效氯氟氰菊酯抗性相关的RAPD分子标记。     

用新的分子标记方法（RAPD）分析小麦抗白粉病基因Pm4α的近等基因系

ＲＡＰＤ是一种新发展的分子标记技术,本实验利用这种技术对小麦抗白粉病基因Ｐｍ４ａ的近等基因系进行分析。在１００个随机引物中找到了３个引物在这对抗白粉病的近等基因系中所扩增出的带型出现差异。并根据理论计算所找到的差异与抗生基因Ｐｍ４ａ连锁的概率是０．７,即３个差异中应有２个标记与抗性基因连锁。     

在遗传连锁研究中应用作物近等基因系的探讨

通过理论计算得知,在随机抽取的BC₅S₁近等基因系中,回交亲本的分子位点约有96%得到了恢复,供体亲本的分子位点将只保留4%,且其中约2/3因与转育位点相连锁而位于标记连锁区。这表明在遗传连锁研究中,可以应用作物近等基因系把分子位点和常规位点定位到同一连锁群上。     

水稻广亲和基因（WCG）近等基因系的随机引物PCR分析

应用随机引物ＰＣＲ（ＲａｎｄｏｍＰｒｉｍｅｒＰＣＲ）技术分别在水稻广亲和基因（ＷＣＧ）的近等基因系和色素原基因（Ｃ）的分离群体库中寻找与ＷＣＧ和Ｃ基因连锁的分子标记。对于ＷＣＧ近等基因系,在２２６个随机引物中初筛到２２个显示多态性片段的引物。根据理论值计算,在２２个多态性片段中预期有２０个与ＷＣＧ连锁。在这些连锁标记中距ＷＣＧ最近的可达０．５ｃＭ。同样在分离群体库的筛选中有１０个扩增产物与Ｃ基因连锁。     

爪哇根结线虫Mi-毒性和无毒近等基因系的制备及其遗传变异的初步分析

爪哇根结线虫是以孤雌生殖方式繁殖的农作物重要病原物,通过将爪哇根结线虫-无毒群体在抗病番茄品种Momotaro上连续选择19代,获得了一对针对抗性基因Mi的毒性和无毒近等基因系,用102种引物对该近等基因系进行随机扩增多态性DNA（RAPD）分析显示,除一种引物外两者的RAPD带谱基本一致,证实了该毒性和无毒群体的基因组组成十分近似,对一种引物扩增出的一个无毒群体特异性的RAPD片段进行了克隆,Southern杂交结果提示该多态性片段及其同源序列在毒性选择过程中被从毒性线虫的基因组中删除,但DNA序列分析和数据库检索表明,该片段与迄今发表的核酸序列均不存在显著的同源性,因此尚无法预测其潜在的功能性,爪哇根结线虫Mi-毒性和无毒近等基因系的制备为进一步鉴定和分离该线虫毒性相关基因打下了坚实的基础。     

水分亏缺冬小麦近等基因系冠气温差与群体总耗水量的关系

研究品种之间群体耗水特性的差异及其关键影响因素,为品种耗水特性评价与低耗水品种鉴选方法提供依据。选用水分亏缺条件下产量差异不显著,但耗水量差异极显著的冬小麦品种晋麦47和京411及其15个近等基因系为实验材料。利用防雨池和防雨棚开展实验,模拟水分亏缺条件。监测全生育期土壤水分含量,计算总耗水量,收获后测定籽粒产量,计算水分利用效率(WUE)。同时,分别在拔节-孕穗期、抽穗-开花期和灌浆期3个不同生育期监测冠层-大气温度差值(CTD)、叶片蒸腾速率和气孔导度。结果表明,3个不同生育期,15个近等基因系及其亲本之间,CTD均达到显著差异。CTD的方差分析表明,基因型和年份均对不同生育期的CTD有显著影响,但是二者之间仅在抽穗-开花期存在互作(P=0.0002)。15个近等基因系及其亲本之间耗水量存在显著差异,产量没有显著差异。源于耗水量的差异,部分品种/系之间WUE达显著差异。3个不同生育期,15个近等基因系及其亲本之间,CTD与总耗水量均呈极显著负相关关系。抽穗-开花期最高,2012—2013年度和2016—2017年度分别达到0.7042和0.6095。叶片蒸腾速率和气孔导度与群体总耗水量之间相关性很弱,3个生育期均未达到显著水平。对该组近等基因系材料,影响群体总耗水量的关键因素不是叶片蒸腾生理特性,而是群体冠层生长特性。表明构建合理的群体冠层结构不仅是获得高产的途经,而且是调控群体总耗水量,提高品种水分利用效率的重要途径。     

Oligosaccharide Composition, Localization, and Developmental Changes of a CNS-Specific (F3-87-8) Glycoprotein

The F3-87-8 glycoprotein was isolated from rat brain by immunoaffinity chromatography after biosynthetic labeling by intracerebral administration of [3H]glucosamine, and the oligosaccharide composition of pronase-derived glycopeptides was determined by sequential lectin affinity chromatography and alkali treatment. Triantennary complex oligosaccharides (65%) and O-glycosidic oligosaccharides (18%) were the predominant types present, accompanied by 7-10% each of biantennary and high-mannose oligosaccharides. Twenty-two percent of the complex oligosaccharides had a fucose residue linked to the proximal N-acetylglucosamine of the chitobiose units. No poly(N-acetyllactosaminyl) or hybrid oligosaccharides were detected. Immunocytochemical studies on the localization of this glycoprotein in developing rat brain demonstrated that in 1-week postnatal cerebellum, there is light staining of the internal granule cell layer and surrounding the Purkinje cells. By 2 weeks, an intense staining of myelinating fiber tracts appears, accompanied by much lighter staining in the granule cell layer and at the base of the molecular layer. Staining of the white matter remains strong at 3 weeks postnatal, together with significant staining throughout the molecular layer, and then decreases in both areas by 1 month. In adult brain there is relatively uniform staining of approximately equal intensity in the white matter, granule cell layer, and molecular layer, whereas the Purkinje cell bodies appear unstained throughout development. In agreement with a previously reported immunochemical analysis, no staining was seen in other tissues, confirming the CNS-specific localization of this glycoprotein.     

Oligosaccharide Portion of GM1 Enhances Process Formation by S20Y Neuroblastoma Cells

The oligosaccharide portion of ganglioside GM1 was found to enhance neuritogenesis by S20Y murine neuroblastoma cells grown in vitro. The average length of the neurites produced by cells grown in the presence of the oligosaccharide portion of GM1 was comparable to that of cells grown in the presence of intact GM1. The processes of these cells were significantly longer (p less than 0.005, pooled t test) than those of cells grown in the presence of comparable concentrations of sialic acid, lactose, sialyllactose, GD1a, or the oligosaccharide moiety of GD1a. These results suggest that it is the oligosaccharide portion of GM1 that is responsible for the ability of GM1 to enhance process outgrowth by S20Y neuroblastoma cells.     

Oligosaccharide structure of a functional unit RvH1-b of Rapana venosa hemocyanin using HPLC/electrospray ionization mass spectrometry

In the present study the structures of two glycopeptides (G1 and G1'), isolated from FU RvH(1)-b and two glycopeptides (G2 and G3), isolated from the structural subunit RvH(1) of Rapana venosa hemocyanin, were determined. To structurally characterize the site-specific carbohydrate heterogeneity and binding site of the N-linked glycopeptide(s), a combination of capillary reversed-phase chromatography and ion trap mass spectrometry was used. The amino acid sequences of glycopeptides G1 and G1' determined by Edman degradation and MS/MS sequencing demonstrated that the oligosaccharides are linked to N-glycosylation sites. Two peptides (a glycosylated (G1) and non-glycosylated one) were identified in this fraction and no linkage sites were observed in the latter one. Based on the sequencing of the glycosylated fractions G1, G1', G2 and G3, the carbohydrate structure Man(alpha1-6)Man(alpha1-3)Man(beta1-4)GlcNAc(beta1-4)[Fuc(alpha1-6)]GlcNAc-R could be identified for glycopeptides G1 and G3, and only the typical core structure Man(alpha1-6)Man(alpha1-3)Man(beta1-4)GlcNAc(beta1-4)GlcNAc-R was found for G1' and G2. The Fuc residue found in glycopeptides G1 and G3 is attached to N-acetyl-glucosamine of the carbohydrate core, as often found in other glycoproteins.     

细菌脂多糖及其寡糖链结构分析技术研究进展

脂多糖是大多数革兰氏阴性细菌细胞壁的主要成分,能诱发宿主细胞固有免疫反应,在细菌的识别、黏附、转移、致病等过程中发挥非常重要的作用.其结构组成与细菌的血清型和致病能力息息相关,因而对其结构进行精准分析,有助于研究其结构与生物效应间的关系,便于鉴别菌种和研发相关的抗生素及疫苗.由于脂多糖具有两亲性、多电荷且结构复杂等特点,为研究分析带来很多难题.本文全面归纳了细菌脂多糖分析技术的相关研究成果,阐述脂多糖及其寡糖链的提取、分离纯化及鉴定方法.     

Versatile High Resolution Oligosaccharide Microarrays for Plant Glycobiology and Cell Wall Research

Microarrays are powerful tools for high throughput analysis, and hundreds or thousands of molecular interactions can be assessed simultaneously using very small amounts of analytes. Nucleotide microarrays are well established in plant research, but carbohydrate microarrays are much less established, and one reason for this is a lack of suitable glycans with which to populate arrays. Polysaccharide microarrays are relatively easy to produce because of the ease of immobilizing large polymers noncovalently onto a variety of microarray surfaces, but they lack analytical resolution because polysaccharides often contain multiple distinct carbohydrate substructures. Microarrays of defined oligosaccharides potentially overcome this problem but are harder to produce because oligosaccharides usually require coupling prior to immobilization. We have assembled a library of well characterized plant oligosaccharides produced either by partial hydrolysis from polysaccharides or by de novo chemical synthesis. Once coupled to protein, these neoglycoconjugates are versatile reagents that can be printed as microarrays onto a variety of slide types and membranes. We show that these microarrays are suitable for the high throughput characterization of the recognition capabilities of monoclonal antibodies, carbohydrate-binding modules, and other oligosaccharide-binding proteins of biological significance and also that they have potential for the characterization of carbohydrate-active enzymes.     

Effects of Inhibitors of Oligosaccharide Processing on P0Protein Synthesis and Incorporation into PNS Myelin

Four inhibitors of oligosaccharide processing were used to investigate their effects on the transport of PNS myelin glycoproteins through the secretory pathway, as well as to gain further insight into the structure of the oligosaccharide chains of the P0 and 19-kDa glycoproteins. Several different inhibitors of oligosaccharide processing were incubated with chopped peripheral nerves from young rats (21-24 days of age) and the uptake of 14C-amino acid and [3H]fucose or [3H]mannose was measured in P0 and the 19-kDa glycoprotein after separation of homogenate and myelin proteins on polyacrylamide gels. [3H]Mannose was not found as suitable as [3H]fucose as an oligosaccharide precursor because glucose used as an energy source profoundly inhibited the uptake of [3H]mannose. The substitution of pyruvate as an energy source, however, resulted in incomplete glycosylation, poor amino acid uptake, and truncated oligosaccharide chains. Endoglycosidase H cleaved approximately 50% of the P0 labeled with [3H]fucose and 14C-amino acid. The lower molecular weight protein resulting from endoglycosidase H cleavage contained approximately one-half the [3H]fucose label on the protein, whereas one-half remained on the oligosaccharide chain of the undegraded P0, indicating that at least one-half the P0 has a hybrid structure. Deoxynojirimycin, deoxymannojirimycin, and castanospermine inhibited incorporation of [3H]fucose into the oligosaccharide chains of P0 and the 19-kDa glycoprotein as predicted from their action in blocking various stages of trimming of high mannose structures before the addition of fucose. P0 synthesized in the presence of these inhibitors was cleaved to a greater extent by endoglycosidase H than the normal protein, indicating increased vulnerability to this enzyme with arrest of normal processing. Similar results were obtained for the 19-kDa glycoprotein. Both the incompletely processed P0 and the 19-kDa glycoprotein formed in the presence of these inhibitors appeared to be transported normally into myelin.     

不同产地无患子果皮皂苷含量的地理变异研究

通过对不同产地无患子果皮皂苷含量的测定分析,结果表明,14个产地间果皮皂苷含量差异显著,平均含量为8.85%。果皮皂苷含量与产地经度间存在显著性正相关,东西部产地果皮皂苷含量差异明显,西部产地平均含量高于东部产地。聚类分析结果亦表明,东西部产地间果皮皂苷含量出现明显分化,西南部产地的江西宜丰、贵州榕江、广西龙州分化尤为明显,表现出较高的皂苷含量,为无患子高皂苷含量筛选的优良产区。     

Preparation of Oligosaccharide Units Library and Its Utilization

There is high current interest in developing synthetic routes to oligosaccharides involved in glycoconjugates. Significant attention has been focused on the application of glycosidase-catalyzed transglycosylation for practical synthesis of oligosaccharides. The enzymatic synthesis has become more practical by the use of several glycosidases available in sufficient quantities. This review describes convenient syntheses of di- and trisaccharide units, Which are related to molecular recognition, by using regioselective trans-galactosylation, trans-N-acetylglucosaminylation, transfucosylation, and transmannosylation. The region-selectivity could be controlled to some extent by using the following techniques: (1) varying enzymes, (2) organic co-solvent system, (3) the configuration of the existing glycosidic linkage of the acceptor and (4) inclusion complex of acceptor glycoside with cyclodextrin. Furthermore, glycopolymers carrying a series of disacchariues containing β-D-galactosyl residues were synthesized and used as a model in oligosaccharide-lectin interaction analysis. These water-soluble glycopolymers were shown to be useful as probes of carbohydrate recognition.     

外源DNA导入大豆后代种皮颜色变异的过氧化物酶分析

本实验用聚丙烯酰胺凝胶电泳的方法,对外源DNA导入大豆引起种皮颜色变异的后代进行了过氧化物酶同工酶的酶谱分析.结果表明:不同种皮颜色的后代含有供体的谱带.这说明了外源DNA片断已整合到受体基因组中并得到表达.     

大豆蛋白质含量的遗传变异及其与主要农艺性状的相关性分析

摘要：以综合性状优良的黄淮海区主栽大豆品种中黄13为轮回亲本,从大豆微核心种质中选择蛋白质含量显著低于或高于轮回亲本的中黄20、东山69、迟黄豆-1和泰兴牛毛黄乙等4个品种作为供体亲本,比较分析了4个组合的RP、DP、F2、BC1F2和BC2F2蛋白质含量的遗传变异及其与主要农艺性状的相关性,结果表明,双亲蛋白质含量高有利于提高其杂交、回交后代的蛋白质平均含量及超轮回亲本个体比例;F2、BC1F2和BC2F2群体蛋白质含量的变异系数依次降低,BC2F2的蛋白质平均含量及其变异系数接近于轮回亲本;蛋白质含量在F2群体内呈正态分布,在双亲蛋白质含量高的组合中,其BC1F2群体呈偏态分布,但在BC2F2群体恢复了正态分布,稳定较快;供体亲本与其杂交2代、回交1代和回交2代在蛋白质含量、脂肪含量、株高、单株荚数、单株粒数、百粒重等性状上呈显著或极显著相关。