中国蓼属分叉蓼组和冰岛蓼属植物叶下表皮微形态研究

采用光学显微镜对中国蓼属分叉蓼组(Polygonum section Aconogonon)和冰岛蓼属(Koenigia)的21种植物的叶下表皮微形态进行了观察研究。结果表明,其叶下表皮微形态特征可分为三种类型：（1）气孔器类型为无规则型,表皮细胞多边形,垂周壁为波状;（2）气孔器类型为无规则型兼有非典型不等型,表皮细胞为多边形或不规则形,垂周壁为平直弓形或波状;（3）气孔器类型为不等细胞型,表皮细胞多边形,垂周壁为平直弓形。分叉蓼组（西伯利亚蓼除外）植物的叶表皮气孔器类型具有高度一致性,说明该组是一个自然类群。本文研究结果不支持将大铜钱叶蓼(Polygonum forrestii Diels)、铜钱叶蓼(P. nummularifolium Meissner)划归于冰岛蓼属的处理意见;支持将西伯利亚蓼(P. sibiricum Laxm.)独立成属,即西伯利亚蓼属(Knorringia Tzvel.)的观点;同时也支持仍将多穗蓼(P. polystachyum Wall.ex Meissner)和松林蓼(P. pinetorum Hemsl.)归在分叉蓼组的处理意见。     

中国蓼属头状蓼组植物叶表皮微形态及其分类学意义

采用光学显微镜对中国蓼属头状蓼组17种7变种植物的叶片下表皮微形态进行了观察研究,结果表明,其叶片下表皮微形态特征分为4种类型:(1)气孔器类型为无规则型,表皮细胞不规则形,垂周壁波状或深波状;(2)非典型不等型,偶有无规则型,表皮细胞多边形或不规则形,垂周壁弓形、波状或深波状;(3)平列型,表皮细胞不规则形,垂周壁深波状;(4)平列型兼有非典型不等型,表皮细胞不规则形,垂周壁波状。根据其叶片下表皮气孔器类型,结合该组植物形态、习性等特征,将中国蓼属头状蓼组植物划分为4个系,即掌裂叶系、多年生系、蓼子草系以及一年生直立系。     

中国蓼属萹蓄组植物叶下表皮微形态研究

在光学显微镜下对中国蓼属萹蓄组15种植物的叶片下表皮微形态进行了观察研究。结果表明,萹蓄组15种植物气孔器类型为不等型,气孔长宽比为1.19~1.72,气孔密度为122.07~376.38个/mm2,气孔指数为17.22%~25.17%,具有较高的一致性,支持将萹蓄组section Avicularia升为萹蓄属Polygonums. str.的观点。但表皮细胞形状及垂周壁式样变异较大,可将其分为2种类型:A.表皮细胞为多边形,垂周壁平直弓形;B.表皮细胞为不规则形,垂周壁浅波纹状。基于叶下表皮微形态并结合植物外部形态特征,将直立萹蓄P.aviculareL.f.erectum Y.M.Zhang et J.X.Li合并到展枝蓼P.patulum Bieb.,作为其异名处理。     

对翼蓼(蓼科)属级位置的重新探讨

通过对翼蓼Pteroxygonum giraldii Damm. &; Diels及相关属(蓼属Polygonum、何首乌属Fallopia、虎杖属Reynoutria、荞麦属Fagopyrum和金线草属Antenoron)的形态观察、果实解剖学观察、花被片脉序观察、花粉形态、核型分析, 以及ITS序列的分析确定了翼蓼和荞麦F. esculentum Moench较远的亲缘关系。其中我们发现翼蓼果实基部有三个角状物明显不同于其他属果实的形态特征。翼蓼外果皮明显加厚, 并有零星散布的波状内腔, 而荞麦的外果皮很薄, 细胞不等径, 中果皮极厚。以上证据证明了翼蓼与荞麦属亲缘关系较远。在观察荞麦属和翼蓼的花被片脉络时发现了两种不同的脉序类型, 符合将荞麦属分为两个组的划分。翼蓼花被片脉序为三出状, 支持将翼蓼归为Persicarieae族。对翼蓼及荞麦属植物的花粉进行比较后, 发现荞麦属植物的花粉网孔有明显的内凹穿孔而翼蓼却没有, 结果表明二者亲缘关系较远。通过对nrDNA ITS区域序列分析得出翼蓼及相关属为一个单系类群, 含有两个稳定的分支: 第一个分支由蓼属(萹蓄组sect. Avicularia)、何首乌属、虎杖属的植物组成, 第二个分支由蓼属(刺蓼组sect. Echinocaulon、蓼组sect. Polygonum、分叉蓼组sect. Aconogonon、拳参组sect. Bistorta、翼蓼和荞麦属植物组成。同时第二个分支又分成了两个亚分支, 蓼属(刺蓼组、蓼组、分叉蓼组、拳参组)和翼蓼属Pteroxygonum植物属于第一个亚支而荞麦属植物属于第二个亚支。结果支持翼蓼不属于荞麦属的范畴。实验结果显示翼蓼是个单型属, 属于Persicarieae族。     

中国蓼属萹蓄组植物果实形态的研究

应用解剖镜和电子扫描显微镜对中国产蓼属萹蓄组Polygonum sect.Polygonum 16种1变种及1变型植物的果实形态进行了研究。发现该组5种一年生植物的果实具有两型性,即有长、短两种果实类型。短果类型为卵形,具三棱,包于宿存花被内或与花被近等长,呈黑褐色。在解剖镜下观察到其表面具小点、无光泽,而在电子扫描显微镜下观察到其表面具有散乱分布或排列成行的瘤状颗粒;长果类型为狭卵形,具三棱,长于宿存花被,黄褐色或淡黄绿色,在解剖镜下观察到其表面平滑、有光泽,而在电子扫描显微镜下观察到其表面上部具有散乱分布的瘤状颗粒,中、下部则具不规则洼点、星花状褶皱或浅波纹状。果实具两型性的植物有萹蓄P.aviculare、乌鲁木齐萹蓄P.urumqiense、展枝萹蓄P.patulum、直立萹蓄P.avicularef.erectum和尖果萹蓄P.rigidum。果实微形态可分为两种类型：1.果实表面无瘤状颗粒或疣状突起、有光泽,此类型又可以分为2种亚类型：（1）果实表面平滑、浅波纹状,如岩萹蓄P.cognatum、松叶萹蓄P.acerosum和帚萹蓄P.argyrocoleum;（2）果实表面具浅洼点或洼点,如铁马鞭P.plebeium和针叶萹蓄P.polycnemoides。2.果实表面有瘤状颗粒或疣状突起,无或具微光泽,此类型又可分为4种亚类型：（1）果实表面具疣状突起,如圆叶萹蓄P.intramongolicum.;（2）果实表面具散乱分布的瘤状颗粒,如普通萹蓄P.humifusum和展枝萹蓄的短果;（3）果实表面具有纵行排列的瘤状颗粒,如萹蓄和乌鲁木齐萹蓄的短果;（4）果实表面上部具散乱分布的瘤状颗粒,中、下部具不规则洼点、星花状褶皱或浅波纹状,如萹蓄、展枝萹蓄、直立萹蓄和尖果萹蓄的长果。因此果实两型性和果实微形态特征对蓼属萹蓄组种、变种的鉴定具有较大的分类学价值。基于果实微形态特征以及其他形态特征,确定了新种晖春萹蓄Polygonum huichunense F.Z.Li,Y.T.Hou＆C.Y.Qu,同时恢复褐鞘萹蓄Polygonum aviculareL.var.fusco-ochreatum（Kom.）A.J.Li为原种,即Polygonum fusco-ochreatum Kom.。     

中国蒿属植物比较形态和解剖学研究：（1）叶表皮结构

本文通过光学显微镜和扫描电镜,观察国产菊科蒿属２亚属７组３０种代表植物成熟叶片的表皮细胞,气孔器,表皮毛的特征和气孔形状,气孔外拱盖及拱盖内缘,角质膜和蜡质纹饰。其中表皮细胞多角（边）形,大小不等,垂周壁平直,弓形,或者表皮细胞形状不规则,大小不等,垂周壁波浪状或波纹状。下表皮均有气孔,上表皮有或无气孔器,气孔器类型有无规则型,无规则四细胞型,不等细胞型,十字型,横列型和平列型。大多数种类具多细胞     

中国广义冰岛蓼属果实形态特征

应用扫描电镜对冰岛蓼属（Koenigia L.）8个物种的果实形态与果皮微形态进行了观察。结果表明,这些果实可分为3类：类型Ⅰ：瘦果长卵形至宽卵形,中部或中上部最宽,双凸镜状或3棱状,棱脊有或无;表面粗糙或光滑,具纵向断棱状纹饰,或具波状纹饰,这种类型有冰岛蓼、铜钱叶蓼、青藏蓼、大铜钱叶蓼4种;类型Ⅱ：瘦果卵形,中部膨大,具有3棱脊,但棱脊不显著。这种类型仅小叶蓼1种;类型Ⅲ：瘦果卵形,中间最宽但不膨大,具有3棱脊,且棱脊显著;表面光滑,具多皱纹饰,或凹凸不平纹饰,或具疣状纹饰,这种类型包括细茎蓼,蓝药蓼和柔毛蓼3种。研究结果支持将柔毛蓼与蓝药蓼作独立分种处理、而冰岛蓼与青藏蓼关系很近。     

中国蒿属植物比较形态和解剖学研究——(Ⅰ)叶表皮结构

本文通过光学显微镜和扫描电镜,观察国产菊科蒿属2亚属7组30种代表植物成熟叶片的表皮细胞、气孔器、表皮毛的特征和气孔形状、气孔外拱盖及拱盖内缘、角质膜和蜡质纹饰。其中表皮细胞多角(边)形,大小不等,垂周壁平直、弓形,或者表皮细胞形状不规则,大小不等,垂周壁波浪状或波纹状。下表皮均有气孔器,上表皮有或无气孔器,气孔器类型有无规则型。无规则四细胞型、不等细胞型、十字型、横列型和平列型。大多数种类具多细胞单列毛。多细胞双列毛和腺毛仅发现在腺毛蒿组。这些微形态特征在组间存在着某些差异,具有一定的分类学和生态学意义。     

黑龙江悬钩子属植物叶表皮形态结构的研究

利用扫描电子显微镜对黑龙江悬钩子属植物的叶表皮形态结构进行比较研究。结果显示:(1)悬钩子属植物叶的上表皮细胞呈多边形,垂周壁平直,或无规则形,垂周壁浅波纹;下表皮细胞无规则形,垂周壁浅波纹或深波纹。(2)表皮毛类型有单细胞直立不分支、卷曲不分支,头状腺毛和盾状腺毛四种类型。(3)气孔器均分布于下表皮,且气孔器类型为无规则形;气孔外拱盖单层、内缘平滑或不规则波状。研究表明,黑龙江悬钩子属植物的叶表皮微形态学特征表现出一定差异性,对种间的划分和鉴定具有一定的分类学意义。     

香辣蓼,中国春蓼属(蓼科)一新归化植物

报道了中国春蓼属一新归化植物：香辣蓼[Persicaria odorata（Lour.） Soják],并对其形态学特征进行了详细描述,提供了形态照片,同时分析了其用途。该种原产于中南半岛,被作为香料植物在云南、广西、广东及江西等地的傣族或客家人聚居区广泛栽培,现在云南西双版纳傣族自治州及德宏傣族景颇族自治州发现大量归化居群。香辣蓼与特产于中国江西的武功山春蓼（P.wugongshanensis B.Li）最为近似,二者叶片均具有强烈的气味,叶片及花被片具腺点,二型花柱,花序基部间断,花梗长于苞片,区别在于本种为多年生草本植物,具发达的根状茎,叶片披针形至狭披针形,果实表面光滑有光泽。     

A simple method using β-globin polymerase chain reaction for the species identification of animal cell lines—A progress report

Continuous cell lines are widely used in cell biology and serve as model systems in basic and applied research. Fundamental requirements for the use of cell lines are a well-identified origin and the exclusion of cross-contamination by prokaryotic or eukaryotic cells. Because the cross-contamination of one cell line with another cell line may occur in a concealed manner, special emphasis must be taken to (1) prevent such an "accident" and (2) monitor regularly the identity of the cell line(s) in use. Apart from human cell lines, mouse-, rat-, and hamster-derived cell lines are used in basic cell culture and biotechnology. We established a polymerase chain reaction (PCR) assay to detect and confirm the species origin for these species and to detect interspecies cross-contamination. Our PCR method is based on oligonucleotide primers annealing to specific sequences in the beta-globin gene, which were designed to amplify one deoxyribonucleic acid (DNA) segment only per analyzed sample. We confirmed the species identity of 82 cell lines as human, mouse, rat, and Syrian hamster by beta-globin PCR. The DNAs from eight additional cell lines of less frequently used species were not amplified with the primers chosen. Cross-contamination of 5-10% of either mouse or rat DNA was detectable. One species-specific primer pair was sufficient for confirmation of the expected species, and for identification of an unknown cell line the combination of two or more primer pairs is suggested. Our PCR assay represents a powerful, fast, easy, robust, and inexpensive method for speciation and does not need any elaborate sequencing or computer-based analysis system.     

Analysis of the proteoglycans synthesized by human bone cells in vitro

Proteoglycans were isolated by ion-exchange chromatography from the extracted cell layer and culture medium of human bone cell cultures following incubation in the presence of [35S]sulfate and [3H]leucine. On sodium dodecyl sulfate-polyacrylamide gel electrophoresis (SDS-PAGE), the synthesized proteoglycans consisted of at least five polydisperse species having median apparent Mr = 600,000, 400,000, 270,000, 135,000 and 40,000. When chromatographed further on octyl-Sepharose CL-4B, the proteoglycans of the cell layer resolved into three peaks. The unbound fraction (peak cell layer-I) contained a 40,000 species consisting of a single glycosaminoglycan chain with or without peptide. Peak cell layer-II contained three sulfated species on electrophoresis: a 600,000 species uniformly distributed across the peak, a 135,000 species enriched in the ascending limb (similar to bone PG-I as described previously), and a 270,000 species (similar to bone PG-I) enriched in the descending limb. Peak cell layer-III, eluting at 0.2% Triton X-100, was highly enriched in a 400,000 proteoglycan component. When media proteoglycans were chromatographed on octyl-Sepharose, two labeled peaks were found. Peak medium-I (unbound) contained a species exhibiting electrophoretic mobility similar to that of the 400,000 species present in peak cell layer-III. Peak II of the culture medium (medium-II) was apparently identical to that of peak cell layer-II, containing the 600,000, 270,000 and 135,000 species. No appreciable 40,000 species was observed in the medium. Treatment of the 600,000 species with either chondroitinase ABC or ACII generated a core protein preparation with bands of 390,000 and 340,000 on SDS gels. Neither the intact nor the chondroitinase ABC-treated 600,000 species was immunoprecipitated by a purified, polyclonal antiserum raised against the core protein of the large chondroitin sulfate proteoglycan of human articular cartilage. Treatment of the 270,000 and 135,000 proteoglycans with chondroitinase ABC, but not ACII, generated a core protein preparation with bands of 52,000 and 49,000 on SDS gels, indicating that they were dermatan sulfate-containing species. The 400,000 species contained both heparan sulfate and chondroitin sulfate, in approximately a 3:1 labeling ratio. This species changed in electrophoretic mobility following treatment with chondroitinase ABC, heparatinase, or both enzymes in combination, which suggested that it may be a hybrid proteoglycan (i.e. both types of glycosaminoglycan chain on the same core protein).(ABSTRACT TRUNCATED AT 400 WORDS)     

Roles of cell walls and intracellular contents in supercooling capability of xylem parenchyma cells of boreal trees

The supercooling capability of xylem parenchyma cells (XPCs) in boreal hardwood species differs depending not only on species, but also season. In this study, the roles of cell walls and intracellular contents in supercooling capability of XPCs were examined in three boreal hardwood species, Japanese beech, katsura tree and mulberry, whose supercooling capability differs largely depending on species and season. XPCs in these species harvested in winter and summer were treated by rapid freezing and thawing (RFT samples) or by RFT with further washing (RFTW samples) to remove intracellular contents from XPCs in order to examine the roles of cell walls in supercooling. RFT samples were also treated with glucose solution (RFTG samples) to examine roles of intracellular contents in supercooling. The supercooling capabilities of these samples were examined by differential thermal analysis after ultrastructural observation of XPCs by a cryo‐scanning electron microscope to confirm effects of the above treatments. XPCs in RFTW samples showed a large reduction in supercooling capability to similar temperatures regardless of species or season. On the other hand, XPCs in RFTG samples showed a large increase in supercooling capability to similar temperatures regardless of species or season. These results indicate that although cell walls have an important role in maintenance of supercooling, change in supercooling capability of XPCs is induced by change in intracellular contents, but not by change in cell wall properties.     

Species variability in boron requirement is correlated with cell wall pectin

Fourteen species of crop plants which differ in their reportedtissue boron requirements were grown in B-replete or B-deficientmedium. Leaf samples were collected and analysed for B and cellwall components. There was a significant positive correlationamong the species between B concentration in the leaf or thecell wall and uronic acid, rhamnose and galactose (indicativeof pectin) in the cell wall. The concentration of cell wallpectin was also positivety related with reported tissue-B requirementsand observed sensitivity to B deficiency. Boron deficiency didnot alter the amount of uronic acid present in cell walls, suggestingthat there was no effect of B deficiency on pectin metabolism.Under B-deficient conditions the amount of ‘soluble’B (i.e. B not associated with the cell wall) declined dramaticallywhile the proportion of cellular B that was ‘insoluble’(i.e. B associated with the cell wall) increased. The positiverelationship between pectin content, insoluble B and tissue-Brequirement of diverse species suggests that the amount of cellwall pectin may be significant in determining the relative tissue-Brequirements of the species. These results indicate that either(1) species with high cell wall pectin contents require greateramounts of B for the construction of the cell wall, or (2) pectinin cell walls forms an insoluble complex with B, thereby reducingits availability for other putative B-requiring metabolic functions.Thus, species with a high pectin content would have a highertissue-B requirement. Key words: Boron, deficiency, uronic acid, pectin, cell wall     

Identification and authentication of animal cell culture by polymerase chain reaction amplification and DNA sequencing

Polymerase chain reaction (PCR) amplification and deoxyribonucleic acid (DNA) sequence analysis were used to identify the species origin of cell lines used in a cell culture facility where various cell lines of different species are routinely propagated. The aldolase gene family was selected for PCR amplification because the DNA sequences of this gene are highly conserved over a wide range of animals and humans. A total of 36 cell lines representing 13 different species were selected for this study. The DNA from each cell line was amplified, and PCR products were analyzed by agarose gel electrophoresis. The results showed unique profiles of amplified bands on agarose gels that allowed differentiation among non-closely related species. However, DNA amplification of closely related species, including rat and mouse or human and primate, resulted in similar and indistinguishable banding patterns that could be further differentiated by DNA sequence analysis. These results suggested that aldolase gene amplification coupled with DNA sequence analysis is a useful tool for identification of cell lines and has potential application for use in identification of interspecies cross-contamination.     

Hydrophobic properties of gram-negative rods colonizing upper respiratory tract of healthy people

The cell surface hydrophobicity is one of the non specific factors of adhesion influencing the ability of microorganisms to colonize nasopharynx. The aim of this paper was to evaluate via salt aggregation test (SAT) the cell surface hydrophobicity of 150 strains of gram-negative rods isolated from the throat or/and nasal specimens of healthy people. It has been found that among the nonfermenting rods hydrophobic strains were predominant. In contrast, the isolates of Enterobacteriaceae family were characterized by the distinctive features of the cell surface within particular genera or even species. The obtained results show that, despite differences in cell surface hydrophobicity, numerous species of gram-negative rods have the ability to colonize the mucous membrane of upper respiratory tract. This suggests that the cell surface hydrophobicity is rather a feature of species or genus, but it is not related to the ecological niche of microorganisms in human body.     

Alginate cell encapsulation: new advances in reproduction and cartilage regenerative medicine

Cell lines represent valuable tools for basic research and diagnostic applications as well as for the production of biological products such as antibodies or vaccines. For all cell culturists, a well-identified origin of their cell lines as well as the periodic re-examination of their identity should be a basic requirement. We established a simple polymerase chain reaction (PCR) to verify or identify rodent and human cell lines. Since mouse-, rat-, Chinese hamster- and Syrian hamster-derived cell lines represent the most frequently used rodent cell lines, our investigations were focused on these species. Our assay used oligonucleotide primers annealing to sequences within the β-actin and the β-globin gene and to repetitive DNA. Primers were designed mostly from intron sequences of the genes aiming to amplify only one specific DNA segment and thus enabling to exclude easily false DNA. More than 130 cells lines originating from the five species were analyzed in that study. Our PCR revealed specific profiles for all species investigated. No further methods like DNA sequencing or fragment length polymorphism analysis were needed to differentiate these species. The results introduce our PCR-assay as a rapid, specific and routinely feasible tool in order to identify or distinguish rodent cell lines from each other and from human cell lines.     

Quantation by flow microfluorometry of total cellular DNA in Acanthamoeba.

The DNA content of five species of Acanthamoeba was determined by flow microfluorometry. Acanthamoeba castellanii (AC-30), acanthamoeba polyphaga (APG and P-23), acanthamoeba rhysodes, acanthamoeba culbertsoni (A-1), and acanthamoeba royreba were grown in a casitone based medium 24-48 HR. The trophozoites were harvested, and evaluated for DNA-bound fluorescence. All species tested has DNA values between 2.0-5.0 pg/cell. These results placed DNA/cell values of Acanthamoeba slightly lower than DNA/cell values of other eucaryotic cells and much lower than Amoeba proteus values. These results indicate that FMF may be a useful adjunct in distinguishing Acanthamoeba cells from either eucaryotic cells or some other amoeba. However, differences in DNA/cell between species of Acanthamoeba are small and would not be useful in identification of species.     

Callus induction and thallus regeneration in some species of red algae

Callus induction was obtained from axenic explants of 14 species of red algae. ASP12NTA solid medium (1.5% agar) supplemented with indole-3-acetic acid (IAA) and 6-benzylaminopurine (BAP) was used for callus induction. In most of the species, addition of IAA or BAP at 0.1 mg/L or 1.0 mg/L enhanced callus induction rate or callus size. The combination of IAA (0.1 mg/L) and BAP (0.1 mg/L) was more effective among eight species, while high concentrations of IAA (10 mg/L) showed an inhibitory effect. Great variation in callus form, source tissue, and color of the induced callus were observed. The callus mainly originated from medullary and cortical tissue of the explant. Callus with filamentous, oval and spherical cell chains or disorganized cell mass was observed. The excised calluses from the explants of six species showed sustained growth on subculture. On transfer of the subdivided callus mass of seven species to PES liquid medium, shoot formation and thallus regeneration were observed.