诸葛菜种子的油脂分析与利用评价

一、概述诸葛菜 Orychophragmus Bunge 属十字花科 Cruciferae,又名二月兰、翠紫花。该属共有2种约5变种,广泛分布于东亚、南亚和地中海地区。我国发现有1种及3变种,分别为原种诸葛菜 O.violaceus(L.)O.E.Schulz;缺刻叶诸葛菜 var.intermedius(Pamp.)O.E.Schulz;毛果诸葛菜 var.lasiocarpus Migo 以及湖北诸葛菜 var.hupohansis 等,产于华北、     

埃塞俄比亚芥与诸葛菜属间杂种的基因组原位杂交分析

用幼胚培养方法重新获得的埃塞俄比亚芥(Brassica carinata A.Braun,2n=34)与诸葛菜(Orychophragmus violaceus (L.)O.E.Schulz,2n=24)的属间杂种仍为混倍体(2n=14～34),2n=34细胞的频率最高;绝大多数花粉母细胞(PMCs)表现正常的17个二价体配对和17：17的分离。基因组原位杂交分析结果表明,在所有体细胞和PMCs中不含有整条的诸葛菜染色体,2n=34的体细胞和PMCs中包含了来自黑芥(B．nigra (L．)Koch,2n=16)的16条染色体。这些具有完全或部分埃塞俄比亚芥染色体组成的细胞,可能来源于以前提出的杂种细胞在有丝分裂中完全或部分亲本染色体组分开和染色体复制,并伴随诸葛菜染色体的消除。     

中国杜英属的一些分类问题

本文根据最新研究结果,对国产杜英属植物进行了清理。这里报道的是对《中国植物志》 49(1)杜英属的修订和补充。它包括：(1)纠正3个错误鉴定：Elaeocarpus rugosus Poxb．=E． apiculatus sensu Merr.;E.sikkimensis Mast.=E.fleuryi sensu H.T.Chang;E.decandrus Merr.=E.chinensis sensu H.T.Chang pro parte。 (2) 发表1个新种和1个新变种：E． limitaneioides Y.Tang;E.glabripetalus Merr．var．grandifructusy．Tang． (3)归并了4种2 变种：E．boreali-yunnanensis H．T．Chang归并为E．lacunosus Wall. ex Kurz,E.floribundioides H．T．Chang归并到E．austro-yunnanensis Hu,E.fengjieensis P.C.Tuan归并至E．duclouxii Gagnep．,E．kwangsiensis H．T．Chang归并为E.glabripetalus Merr. var．alatus(Knuth) H. T. Chang,E. glabripetalus Merr. var．teres H.T.H.T.Chang归并为E.glabripetalus var．glabripetalus,E.prunifolioides Hu var．rectinervis H.T.Chang归并至E.prunifolioidesHu。 (4)报道了一些省级分布新记录;并简单讨论了属下系统。     

10种披碱草属植物的RAPD分析及其系统学意义

利用随机扩增多态性DNA(RAPD)技术分析了10种披碱草属植物,即：Elymus sibiricus L.,E． caninus(L．)L¨E．lanceolatus(Scribner et Smith)Gould,E．nutans Griseb,E．dahuricus Turcz,, E．tangutorum(Nevski)Hand．-Mazz., E．brachyaristatus ALöve,E.submuticus(Keng)Keng f.,E． cylidricus(Franch)Honda和E．excelsus Turcz. 。对34个OPRON公司十聚体随机引物进行多态性筛 选,25个(75．53％)能产生多态性。16个引物产生的136个DNA片断,用于计算种间Nei氏遗传相似 性系数分析,在NTSYS程序中利用UPGMA构建系统发育树状图。分析结果表明：(1)四倍体和六倍体 物种各自聚为一支,它们之间的亲缘关系较远;(2)E．sibiricus和E. caninus;亲缘关系较近,支持把 Roegneria caninus (L．)Nevski归入Elymus;(3)E．nutans和E．dahuricus亲缘关系密切;(4)形态相 似、地理分布一致的E．brachyaristatus和E．submuticus存在着一定程度的核苷酸序列的差异,它们与 E．nutans和E．dahuricus有一定的亲缘关系;(5)E．excelsus 与 E．cylindricus的亲缘关系较近,它们 又与E．tangutorum有亲缘关系;(6)RAPD结果与形态学和细胞学等分析结果基本一致,RAPD分析方法将为Elymus系统分类提供DNA水平上丰富的资料。     

封面说明

正封面图片由中国农业科学院农业资源与农业区划研究所曹卫东研究员于2013年4月23日拍摄于中国农业科学院国际农业高新技术产业园(河北廊坊)的公益性行业(农业)科研专项绿肥项目核心试验区.二月兰(Orychophragmus violaceus),又名诸葛菜,为十字花科越年生草本植物,耐寒、耐旱、耐瘠薄,多作为野菜食用及观赏.近年来,二月兰被引入农田及果园,用作绿肥及裸露地覆盖,     

铁线莲属研究随记(Ⅴ)

(1)描述了9新种,4新变种;做出了2新等级,1新组合和1新名称。(2)归并了以下拉丁学名： Clematis dioica L．ssp．virginiana(L．)Kuntze var．bahamica Kuntze,C．bahamica (Kuntze) Britton,C．orbic- ulata Correll,C．brasiliana DC．var．laxa St．Hilaire,C．perulata Kuntze,C．barranacae Jones,C．discolor Gardn., C．laxiflora Baker,C．bathiei Lévl．和C．mauritiana Lam．var．sulfurea Viguier &; Perrier。(3)对威灵仙C．chinensis Osbeck的5个变种进行了分类;特产日本的C．fujisanensis Hisauti &; Hara与C.chinensis 极为相近,主要区别在于具较大的花,但有时花与C.chinensis的花同样大,由于区别不大,在本文中被 降级作变种处理;与其近缘、具强烈退化花序、特产华东的C．anhweiensis M．C．Chang也随之被处理为 变种。(4)瑞典学者Johnson在最近出版的铁线莲属专著中将特产西印度群岛东部的C．Flukenetii DC． 归并于特产美国东南部的C．catesbyana Pursh;本文不同意他做出的归并,并列出了这两个种的明显区 别特征,确认后者是一个应该得到承认的独立的种。(5)根据墨西哥标本描述的C．acapulcensis Hook． &; Arn．原知分布于中美一带,而其在南美的居群和一小叶多毛的新变种(var．puberula)过去长期间被误 定为其近缘种C．affinis St．Hilaire;这个混乱在本文中得到澄清,同时,本文给出了这二近缘种的区别 特征。(6)Viguier和Perrier两位学者在上世纪四十年代末研究马达加斯加一带的黄花铁线莲组-怀特铁 线莲亚组(sect．Meclatis subsect．Wightianae)植物时做出了不少错误鉴定：(a)将特产马达加斯加的具三 出复叶的C．mauritiana Lam. var．mauritiana,var．coriacea和C．microcuspis Baker,以及具单叶的C．ac- tinostemmatifolia W．T．Wang均鉴定为分布于非洲大陆的C．simensis Fresen., 并将后者作为分布于亚洲和欧洲的C．orientalis L．的亚种处理;(b)将特产马达加斯加具三出复叶的C．laxiflora Baker和具一回羽状复叶的C．ibarensis Baker鉴定为特产印度南部的C．wightiana Wall.,也将后者作为C．orientalis的亚种处理;(c)将特产科摩罗(Comoros)具二回羽状复叶的C．comoresensis W．T. Wang鉴定为C．brachiata Thunb.,也将后者作为C．orientalis L．的亚种处理;(d)将C．ibarensis Baker(具一回羽状复叶和两性花)这一拉丁学名用在了另一特产马达加斯加具2～3回羽状复叶和单性花、应属于单性铁线莲组sect． Aspidanthera的种(C．rutoides W．T．Wang)之上;(e)将另一特产马达加斯加具一回羽状复叶和单性花、 也属于单性铁线莲组的C．edentata Baker(＝C．insidiosa Baill．)降级作为这两位学者曲解的C．ibarensis Baker的亚种处理;(f)将特产马达加斯加的C．dissecta Baker归并到属于茴芹叶铁线莲组(sect． Pseudanemone)的C．pimpindllifolia Hook．中。此次本文第五部分继去年第一、二部分之后对上述混乱情况做出了完全澄清,并给出了分布于马达加斯加及其邻近岛屿的怀特铁线莲亚组8种植物的检索表;由于C．ibarensis Baker的定义被上述两位学者严重误解,本文根据较多具花、果的标本写出了此种完整、正确的形态描述。     

油菜品质改良的重要天然遗传资源——诸葛菜(ORYCHOPHRAGMUS VIOLACEUS)利用前景展望

Orychophragmus Violaceus L.O.E.Schulz属,中国种,诸葛菜,其种子含有特高含油率和特高亚油酸,且具有十分理想的脂肪酸组成的天然植物资源。从现资料看,属世界上的首次发现,也是我国第一个低芥酸源,就油苯脂肪酸组成改良育种的前景看,本文已论证,这里是一个极其宝贵的天然资源。     

Nuclear reconstitution of demembranated Orychophragmus violaceus sperm in Xenopus laevis egg extracts

The cell-free extracts from animal Xenopus laevis egg could induce chromatin decon-densation and pronuclear formation from demembranated plant (Orychophragmus violaceus) sperm. The demembranated Orychophragmus violaceus sperm began to swell in 30 min incubation, and then were gradually decondensed. The reassembly of nuclear envelope in the reconstituted nuclei had been visualized by means of electron microscopy and fluorescent microscopy. Membrane vesicles fused to form the double envelope around the periphery of the decondensed chromatin. The morphology of the newly formed nucleus, with a double membrane, was similar to those nuclei after fertilization. Transmission electron microscope micrograph of the whole mount prepared nuclear matrix-lamina showed the reconstituted nucleus to be filled with a dense network.     

诸葛菜组织培养成苗

植物名称:诸葛菜(Orychophragmus viola-ceus)别名“二月兰”。材料类别:花梗、花蕾培养条件: 一、诱导培养基:(1)花梗,MS附加IBA1mg/l(单位下同)、KT4、BA80。(2)花蕾,MS附加2,4-D2.KT1。二、分化培养基:MS附加NAA0.2、6-BA4.     

Elymus sibiricus, E. nutans 和 E. burchan-buddae 的形态学鉴定及其染色体组亲缘关系的研究

为了进一步研究Elymus sibiricus L.、E．nutans Griseb．和E．burchan-buddae(Neuski)Tzelev [＝Roegneria nutans(Keng)Keng]的外部形态差异及其系统学关系,本文对这三种植物的6个穗部形 态性状进行了观测和比较,并对这三个Elymus种进行了种间杂交及杂种F1的减数分裂染色体配对行 为的分析研究。结果表明：这三个Elymus种的穗长及颖长等性状均变异很大,而内稃的长、宽则变异 不大并具有明显的种间差异。E. nutans×E．barchan-buddae及E.nutans×E．sibiricus的杂种F1均完 全不育,减数分裂不规则。E．nutans×E．burchan-buddae杂种F1的减数分裂构型为：7.70I+13.40 Ⅱ+0.06Ⅲ+O.08 Ⅳ,而E．sibiricus×E．nutans杂种F1的构型为11.98 Ⅰ+9.61Ⅱ+O.64Ⅲ+0.39 Ⅳ+0.01V。由本实验的形态学和细胞学的研究结果得出以下结论：1．利用内稃形态性状结合穗部 其它性状的差异能对这三个物种进行较准确的鉴定;2．E．nutans与E.burchan-buddae的亲缘关系较 近,而E．nutans与E．sibiricus的亲缘关系则较远;3. E,burchan-buddae×E．nutans的杂种Fl中存在着染色体配对控制因子。     

Temporal shedding patterns and virulence factors of Escherichia coli serogroups O26, O103, O111, O145, and O157 in a cohort of beef calves and their dams

This study investigated the shedding of Escherichia coli O26, O103, O111, O145, and O157 in a cohort of beef calves from birth over a 5-month period and assessed the relationship between shedding in calves and shedding in their dams, the relationship between shedding and scouring in calves, and the effect of housing on shedding in calves. Fecal samples were tested by immunomagnetic separation and by PCR and DNA hybridization assays. E. coli O26 was shed by 94% of calves. Over 90% of E. coli O26 isolates carried the vtx(1), eae, and ehl genes, 6.5% carried vtx(1) and vtx(2), and one isolate carried vtx(2) only. Serogroup O26 isolates comprised seven pulsed-field gel electrophoresis (PFGE) patterns but were dominated by one pattern which represented 85.7% of isolates. E. coli O103 was shed by 51% of calves. Forty-eight percent of E. coli O103 isolates carried eae and ehl, 2% carried vtx(2), and none carried vtx(1). Serogroup O103 isolates comprised 10 PFGE patterns and were dominated by two patterns representing 62.5% of isolates. Shedding of E. coli O145 and O157 was rare. All serogroup O145 isolates carried eae, but none carried vtx(1) or vtx(2). All but one serogroup O157 isolate carried vtx(2), eae, and ehl. E. coli O111 was not detected. In most calves, the temporal pattern of E. coli O26 and O103 shedding was random. E. coli O26 was detected in three times as many samples as E. coli O103, and the rate at which calves began shedding E. coli O26 for the first time was five times greater than that for E. coli O103. For E. coli O26, O103, and O157, there was no association between shedding by calves and shedding by dams within 1 week of birth. For E. coli O26 and O103, there was no association between shedding and scouring, and there was no significant change in shedding following housing.     

Expression of the cloned Escherichia coli O9 rfb gene in various mutant strains of Salmonella typhimurium.

To investigate the effect of chromosomal mutation on the synthesis of rfe-dependent Escherichia coli O9 lipopolysaccharide (LPS), the cloned E. coli O9 rfb gene was introduced into Salmonella typhimurium strains defective in various genes involved in the synthesis of LPS. When E. coli O9 rfb was introduced into S. typhimurium strains possessing defects in rfb or rfc, they synthesized E. coli O9 LPS on their cell surfaces. The rfe-defective mutant of S. typhimurium synthesized only very small amounts of E. coli O9 LPS after the introduction of E. coli O9 rfb. These results confirmed the widely accepted idea that the biosynthesis of E. coli O9-specific polysaccharide does not require rfc but requires rfe. By using an rfbT mutant of the E. coli O9 rfb gene, the mechanism of transfer of the synthesized E. coli O9-specific polysaccharide from antigen carrier lipid to the R-core of S. typhimurium was investigated. The rfbT mutant of the E. coli O9 rfb gene failed to direct the synthesis of E. coli O9 LPS in the rfc mutant strain of S. typhimurium, in which rfaL and rfbT functions are intact, but directed the synthesis of the precursor. Because the intact E. coli O9 rfb gene directed the synthesis of E. coli O9 LPS in the same strain, it was suggested that the rfaL product of S. typhimurium and rfbT product of E. coli O9 cooperate to synthesize E. coli O9 LPS in S. typhimurium.     

Production of Monoclonal Antibody Discriminating Serological Difference in Escherichia coli O9 and O9a Polysaccharides

A monoclonal antibody (mAb) with a unique antigenic specificity against Escherichia coli O9 was produced. The O9a mAb was reactive with a part of the strains in E. coli O9. The O9a mAb did not react with LPS from the E. coli O9 test strain Bi316-42. The distribution of the antigen defined by the O9a mAb in E. coli O9 was consistent with that of E. coli O9a present in E. coli O9 strains. The chemical structure of the repeating unit of the O-specific polysaccharide detected by the mAb was demonstrated to be a mannotetraose by two-dimensional nuclear magnetic resonance spectroscopy. It was confirmed that the mAb recognized E. coli O9a serotype in E. coli O9 serotype strains, suggesting that E. coli O9a serotype might be a dominant strain in E. coli O9.     

QCM immunosensor detection of Escherichia coli O157:H7 based on beacon immunomagnetic nanoparticles and catalytic growth of colloidal gold

In this paper, we describe a novel method for detecting Escherichia coli (E. coli) O157:H7 by using a quartz crystal microbalance (QCM) immunosensor based on beacon immunomagnetic nanoparticles (BIMPs), streptavidin-gold, and growth solution. E. coli O157-BIMPs were magnetic nanoparticles loaded with polyclonal anti-E. coli O157:H7 antibody (target antibody, T-Ab) and biotin-IgG (beacon antibody, B-Ab) at an optimized ratio of 1:60 (T-Ab:B-Ab). E. coli O157:H7 was captured and separated by E. coli O157-BIMPs in a sample, and the streptavidin-gold was subsequently conjugated to E. coli O157-BIMPs by using a biotin-avidin system. Finally, the gold particles on E. coli O157-BIMPs were enlarged in growth solution, and the compounds containing E. coli O157:H7, E. coli O157-BIMPs, and enlarged gold particles were collected using a magnetic plate. The QCM immunosensor was fabricated with protein A from Staphylococcus aureus and monoclonal anti-E. coli O157:H7 antibody. The compounds decreased the immunosensor's resonant frequency. E. coli O157-BIMPs and enlarged gold particles were used as "mass enhancers" to amplify the frequency change. The frequency shift was correlated to the bacterial concentration. The detection limit was 23 CFU/ml in phosphate-buffered saline and 53 CFU/ml in milk. This method could successfully detect E. coli O157:H7 with high specificity and stability. The entire procedure for the detection of E. coli O157:H7 took only 4 h.     

Low level of cross-resistance between triclosan and antibiotics in Escherichia coli K-12 and E. coli O55 compared to E. coli O157

Misuse of biocides has encouraged the emergence of resistance and cross-resistance in certain strains. This study investigated resistance of triclosan-adapted Escherichia coli K-12 and E. coli O55 to antimicrobial agents and compared these to E. coli O157:H7. Cross-resistance in E. coli K-12 and E. coli O55 was observed however to a lesser extent than in E. coli O157:H7. Triclosan-adapted E. coli K-12 demonstrated cross-resistance to chloramphenicol, whereas triclosan-adapted E. coli O55 exhibited resistance to trimethoprim. In comparison, E. coli O157:H7 was resistant to chloramphenicol, tetracycline, amoxicillin, amoxicillin/clavulanic acid, trimethoprim, benzalkonium chloride and chlorohexidine suggesting strain specific rather than general resistance mechanisms.     

乳酸杆菌体外对大肠埃希菌O-78拮抗作用试验

目的：研究乳酸杆菌体外对大肠埃希菌O-78的拮抗作用。方法：将乳酸杆菌与大肠埃希菌O-78体外共培养观察拮抗情况及用电镜观察形态变化。结果：等量同时间接种,24h内乳杆菌就将大肠埃氏菌O-78全部抑杀;乳酸杆菌早24h接种,4-8h内将大肠埃希菌O-78全部抑杀;大肠埃希菌O-78早24h接种,在12-24h内全部被抑杀。电镜观察发现,经乳酸杆菌培养液处理,大肠埃希菌O-78细胞膜形态发生变化。结论：本试验中所用乳酸杆菌体外具有抑制杀死大肠埃希菌O-78的作用。     

Competition for proline between indigenous Escherichia coli and E. coli O157:H7 in gnotobiotic mice associated with infant intestinal microbiota and its contribution to the colonization resistance against E. coli O157:H7

Previously, we produced two groups of gnotobiotic mice, GB-3 and GB-4, which showed different responses to Escherichia coli O157:H7 challenge. E. coli O157:H7 was eliminated from GB-3, whereas GB-4 mice became carriers. It has been reported that the lag time of E. coli O157:H7 growth in 50% GB-3 caecal suspension was extended when compared to GB-4 caecal suspension. In this study, competition for nutrients between intestinal microbiota of GB-3 and GB-4 mice and E. coli O157:H7 was examined. Amino acid concentrations in the caecal contents of GB-3 and GB-4 differed, especially the concentration of proline. The supplementation of proline into GB-3 caecal suspension decreased the lag time of E. coli O157:H7 growth in vitro. When E. coli O157:H7 was cultured with each of the strains used to produce GB-3 mice in vitro, 2 strains of E. coli (proline consumers) out of 5 enterobacteriaceae strains strongly suppressed E. coli O157:H7 growth and the suppression was attenuated by the addition of proline into the medium. These results indicate that competition for proline with indigenous E. coli affected the growth of E. coli O157:H7 in vivo and may contribute to E. coli O157:H7 elimination from the intestine.     

Characterization of Escherichia coli O3 and O21 O antigen gene clusters and development of serogroup-specific PCR assays

Escherichia coli O3 and O21 are associated with enteroaggregative E. coli (EAEC). EAEC strains are often non-typable using the routine agglutination method due to their aggregative phenotype. Typing of E. coli O3 and O21 may also be impeded by cross-reactions with O152 or O83. In this study, the O antigen gene clusters of E. coli O3 and O21 were characterized, and PCR assays based on O antigen specific genes wzx (encoding O unit flippase) and wzy (encoding O unit polymerase) from each strain were developed. By screening against all 186 known E. coli O serotypes, the PCR assays were shown to be highly specific to O3 and O21 respectively. The sensitivity of the assays was determined to be 1 pg per mul of chromosomal DNA and 2 CFU per 10 g of water samples. The PCR assays were also applied to 658 clinical E. coli isolates, and 100% of detection accuracy was obtained. The PCR assays developed here are suitable for the detection and identification of E. coli O3 and O21 strains in environmental and clinical samples.     

Experimental Escherichia coli O157:H7 carriage in calves.

Nine weaned calves (6 to 8 weeks of age) were given 10(10) CFU of a five-strain mixture of enterohemorrhagic Escherichia coli O157:H7 by oral-gastric intubation. After an initial brief period of pyrexia in three calves and transient mild diarrhea in five calves, calves were clinically normal throughout the 13- to 27-day study. The population of E. coli O157:H7 in the faces decreased dramatically in all calves during the first 2 weeks after inoculation. Thereafter, small populations of E. coli O157:H7 persisted in all calves, where they were detected intermittently in the feces and rumen contents. While withholding food increased fecal shedding of E. coli O157:H7 by 1 to 2 log10/g in three of four calves previously shedding small populations of E. coli O157:H7, the effect of fasting on fecal shedding of E. coli O157:H7 was variable in calves shedding larger populations. At necropsy, E. coli O157:H7 was not isolated from sites outside the alimentary tract. E. coli O157:H7 was isolated from the forestomach or colon of all calves at necropsy. Greater numbers of E. coli O157:H7 were present in the gastrointestinal contents than in the corresponding mucosal sections, and there was no histologic or immunohistochemical evidence of E. coli O157:H7 adhering to the mucosa. In conclusion, under these experimental conditions, E. coli O157:H7 is not pathogenic in weaned calves, and while it does not appear to colonize mucosal surfaces for extended periods, E. coli O157:H7 persists in the contents of the rumen and colon as a source for fecal shedding.