短命植物东方旱麦草Rubisco大亚基基因(rbcL)的克隆及表达分析

利用PCR、DNA重组、原核与真核生物表达等技术从新疆短命植物东方旱麦草(Eremopyrum orientale)基因组中扩增出Rubisco大亚基基因rbcL(GenBank登录号为FJ346562),并构建了原核与真核表达载体pGEX4T-1-rbcL和pcDNA3-rbcL,随后分别在原核宿主BL21(DE3)和小鼠中进行了表达,并利用真核表达载体和纯化蛋白制备的抗体,对表达产物的特异性进行了Western印迹检测。结果表明:东方旱麦草的rbcL基因可读区包含1431 bp,与旱麦草rbcL基因的同源性达99.86%,推测编码476个氨基酸,分子量56 kD。该基因在BL21中以融合蛋白GST-RBCL形式表达并存在于包含体中,融合蛋白大小约为82 kD。RT-PCR检测到该基因在小鼠肝脏中的表达。利用真核表达载体与纯化融合蛋白进行联合免疫制备的RBCL抗体可与RBCL特异性地结合,这为短命植物东方旱麦草基因后续的免疫定位检测奠定了基础。     

Morphological and genetic evidence of contemporary intersectional hybridisation in Mediterranean Helichrysum (Asteraceae,Gnaphalieae)

Hybridisation is considered an important evolutionary phenomenon in Gnaphalieae, but contemporary hybridisation has been little explored within the tribe. Here, hybridisation between Helichrysum orientale and Helichrysum stoechas is studied at two different localities in the islands of Crete and Rhodes (Greece). Using three different types of molecular data (AFLP, nrDNA ITS sequences and cpDNA ndhF sequences) and morphological data, the aim is to provide simultaneous and direct comparisons between molecular and morphological variation among the parental species and the studied hybrid populations. AFLP profiles, ITS sequences and morphological data support the existence of hybrids at the two localities studied, shown as morphological and genetic intermediates between the parental species. Chloroplast DNA sequences show that both parental species can act either as pollen donor or as maternal parent. Fertility of hybrids is demonstrated by the viability of seeds produced by hybrids from both localities, and the detection of a backcross specimen to H. orientale. Although there is general congruence of morphological and molecular data, the analysis of morphology and ITS sequences can fail to detect backcross hybrids.     

A Supplement to the Chloranthaceae of China

In the present paper the longstanding confusion of three uncertain species of Chloranthus, C. monostachys R. Br., C. pernyanus Solms-Laub. and C. kiangsiensis Metcalf is clarified. In addition to the new record of C. nervosus Coll. et Hemsl. to the flora of China, the distributional ranges of Sarcandra hainanensis (Pei) Swamy et Bailey and Hedyosmum orientale Merr. et Chun are also found to be much bigger than previously known.     

Effort to reconstruct past population history in the fern <Emphasis Type="Italic">Blechnum spicant</Emphasis>

Our aim was to evaluate the potential of existing herbarium collections in reconstructing the past population history of the rediscovered clone of the regionally extinct Blechnum spicant by comparing it with herbarium material from previous sites of occurrences in Finland and elsewhere in northern Europe, as well as to reveal the genetic and geographic relationship among the samples. We detected a total of nine polymorphic sites within the three sequenced regions, two SCAR markers developed now for B. spicant and the chloroplast trnL-trnF spacer, totalling 763 bp. Despite low variability, the phylogeographic analysis revealed the presence of some geographic pattern among the samples, of which all except one represented herbarium samples of up to 100 years of age. The rediscovered clone of B. spicant proved to represent the prevalent genotype occurring in northern Europe. The studied sequences were neutral in terms of evolution. It is apparent that existing herbarium collections are useful resources for a range of evolutionary and population studies.     

A new Miocene fauna of snakes from eastern Siberia,Russia.: Was the snake fauna largely homogenous in Eurasia during the Miocene?

Togay, a locality from Ol’khon Island, Baykal Lake, Russia, has yielded a fauna of snakes of Late Middle or early Late Miocene age. It is located in a broad area from which no Neogene snake has been reported; therefore, it represents an important landmark. The fauna includes a non-erycine boid, two or three colubrids, a viperid of the ‘oriental complex’ of Vipera, and perhaps another viperid. This assemblage is astonishingly reminiscent of the snake faunas from the late Early and early Middle Miocene from western and central Europe, it being understood that Miocene faunas are practically unknown in the geographically intermediary area. It may be entertained whether a homogenous snake fauna inhabited Eurasia (except the southern part of the continent) during the Miocene.     

基部被子植物雪香兰（金粟兰科）单性花的形态发生和发育

基部被子植物金粟兰科（Chloranthaceae）的单性花或两性花结构十分简单,雪香兰（Hedyosmum orientale）花单性、雌雄异株,花的形态及结构与其它属物种具有显著的差异,对于研究被子植物花特别是花被的起源和系统进化具有重要意义。该研究采用电子显微镜和光学显微镜观察了雪香兰单性花的器官发生及发育过程。结果表明,雌、雄花均为顶生和腋生,多个小花呈聚伞圆锥状排列。雄花外侧是苞片,每朵雄花上着生150–200个雄蕊,花轴基部着生少数退化的叶原体。苞片原基及其腋生的花原基最初呈圆丘状,随后伸长。在雄花发育过程中,苞片原基比雄蕊原基生长快,雄花原基纵向伸长,叶原体原基在基部发生,雄蕊原基自下而上发生。每2朵雌花底部合生形成小聚伞花序,每朵雌花被一苞叶包裹,由单心皮和三棱型子房构成,外覆三裂叶状花被。在雌花发育过程中,雌花原基比苞片原基生长快,花被原基首先于花顶端发生,随后花顶端中心凹陷,进一步发育成具有单心皮的子房原基。雪香兰的单性花发育不经过两性同体阶段,花分生组织只起始雄蕊器官或雌蕊器官的发育。研究结果支持雪香兰单性花是原始性状的观点,雄花叶原体与雌花三裂叶状花被同源,可能是花被（萼片与花瓣）的起源。     

乌毛蕨配子体发育的研究

采用混合土培养乌毛蕨（Blechnum orientale）孢子,显微镜下观察记录其孢子萌发及配子体发育过程。结果表明：孢子黑褐色,赤道而豆形,极而观椭圆形,单裂缝。播种1周左右孢子萌发,萌发类型为书带蕨型,配子体发育为叉蕨型。丝状体5—10细胞时开始发育为片状体。播种2周后发育形成幼原叶体,成熟原叶体呈心脏形。原叶体边缘及表面均可产生毛状体,数量丰富,为单细胞。播种后1个月左右开始有颈卵器出现,成熟颈卵器颈部由4列细胞组成,3—5层细胞高。精子器产,扛时间较颈卵器早10d左右,精子器近圆球形,由3细胞组成。精卵受精后2周左右即可观察到从原叶体上生成的幼胚。     

Chemical Diversity of Volatiles of Teucrium orientale L. var. orientale,var. puberulens,and var. glabrescens Determined by Simultaneous GC‐FID and GC/MS Techniques

In the present work, three varieties of Teucrium orientale, var. orientale, var. puberulens, and var. glabrescens, were collected and investigated for chemical composition of the oils. Subsequent gas chromatography (GC‐FID) and gas chromatography coupled to mass spectrometry (GC/MS) revealed high abundance of sesquiterpenes in the essential oils analyzed. All the oils contained β‐caryophyllene (22.6, 8.5, and 6.3%, resp.) and hexadecanoic acid (7.9, 12.8, and 13.1%). Germacrene D (24.6 and 33.4%) and bicyclogermacrene (6.7 and 8.5%) were found to be the main constituents of var. orientale and var. puberulens, respectively. The high percentages of β‐cubebene (26.9%), α‐cubebene (9.0%), and α‐copaene (7.2%) established the diversity of var. glabrescens. The qualitative difference between the essential oils allowed the differentiation between the varieties in agreement with the morphological observations described in Flora of Turkey for each variety studied. In addition, a cluster analysis of twelve Teucrium taxa based on the essential‐oil composition has been carried out. Hovewer, the analysis did not clearly reflect the infrageneric classification of the genus, it largely confirmed the relationships between the infraspecific taxa of Teucrium orientale and T. chamaedrys.