北美鹅掌楸LtuFPPS1基因克隆与组织表达分析

法尼基焦磷酸合酶(farnesyl diphosphate synthase,FPPS)是植物萜类物质合成前体法尼基焦磷酸(farnesyl pyrophosphate,FPP)的关键合成酶。为探究北美鹅掌楸(Liriodendron tulipifera)萜类合成途径的分子机制,该文利用北美鹅掌楸转录组数据,通过设计特异性引物,采用RACE技术克隆得到LtuFPPS1基因的全长序列并进行生物信息学分析。结果表明:(1) LtuFPPS1基因全长1 460 bp,开放阅读框(ORF)长1 056 bp,编码351个氨基酸,蛋白质分子量为40 589.45 D,等电点为5.19,不稳定系数为43.50,归类为不稳定蛋白; LtuFPPS1基因所编码的蛋白不含信号肽,定位于线粒体,是一种不稳定亲水性蛋白,具有类异戊二烯类化合物合酶的特征结构域,α-螺旋为其氨基酸序列的主要二级结构。(2)进化树和同源序列分析表明,北美鹅掌楸LtuFPPS1蛋白与乐昌含笑(Michelia chapensis)的FPPS蛋白的亲缘关系更近。(3)组织表达分析结果发现,LtuFPPS1基因在北美鹅掌楸雌蕊中的表达量最高,其高低顺序为雌蕊花芽茎雄蕊萼片叶片花瓣根,据此推测萜类代谢物在花器官中的合成相对较多。综上所述,LtuFPPS1基因为萜类合成酶基因,可为从分子生物学层面探究北美鹅掌楸萜类物质的合成提供一定的理论帮助。     

Geographic variation in leaf monoterpenes of sequoia sempervirens

Sequoia sempervirens leaf monoterpenes were analyzed from 13 wild native populations (sampling the geographic range of the species) and eight population samples (composed of 18 open-pollinated familles) growing at Russell Reservation, Contra Costa Co., California. Statistically significant variation was found among the wild populations for quantitative composition of all major monoterpenes. Wild populations also displayed high infrapopulation variation. Trees at Russell Reservation also showed significant variation in quantitative composition among population samples, as well as among open-pollinated familles within populations. Patterns among the wild populations were similar to those among the respective planted populations; the greatest disjunction was between those north compared to those south of San Francisco Bay. Additionally, open-pollinated cones of a single tree appear capable of producing progeny that differ greatly in their monoterpene profiles.     

Geographic differentiation of monoterpenes from Abies procera and Abies magnifica

Cortical oleoresins from 352 Abies magnifica and A. procera trees, collected in 35 localities, were analysed for composition of their monoterpene fractions. The monoterpenes were composed primarily of -pinene, β-pinene, limonene and β-phellandrene, with camphene and 3-carene appearing only sporadically in larger amounts. Limonene, β-phellandrene, -pinene and 3-cerene were used to study the transitional populations in northern California and southern and central Oregon and the transitional-like populations in southern Sierra Nevada. The populations segregated latitudinally into three related clusters—above 44° (A.procera), between 44° and 40° (transitional), and below 40° (A.magnifica). A.magnifica from southern Sierra Nevada and A.magnifica from Mtn. Shasta differed in a number of parameters, with southern Sierra Nevada populations being chemically much closer to typical A. magnifica from central and northern Sierra Nevada. While monoterpene and morphological data did not allow a clear Interpretation of the status of the transitional and southern populations, the paleobotanical evidence favored the recent evolution over introgression. The desirability of additional studies was indicated.     

Geographical variation in the monoterpenes of Pinus contorta oleoresin

The monoterpene composition of the shoot cortical oleoresin allowed the natural range of Lodgepole pine (Pinus contorta) to be divided into chemically distinct regions. The trees examined were all grown in Britain from seed originally collected from 150 localities in north-west America. Those from the central area of the natural range contained a wide variety of monoterpene pattern-types, while those from more peripheral areas tended to be more exclusively characterized by their own specific pattern-types. Chemical evidence for introgression with Jack pine (P. banksiana) was detected in trees originating from central British Columbia, and there was also a highly localized and pronounced region of introgression chemotypes in coastal populations near the border of Oregon and California. Variations between trees grown in replicate plots and at different sites in Britain were small, and the results indicated that resin analysis of a sample of 25 trees was usually sufficient to determine the region of origin of an unknown population.     

Geographical variation in the monoterpenes of Valeriana officinalis Leaf

The variation in terpene composition of the leaf essential oil was studied in three populations of Valeriana officinalis L. subsp, collina. These populations grow in Italy in environmentally very different areas. Our examination of geographically and ecologically distant populations has provided evidence for substantial Iocal intrasubspecific differentiation. Statistical treatment of chemical data revealed two distinct chemical varieties of subspecies collina.     

Geographic variability of monoterpenes from cortex of Abies concolor

Cortical oleoresin from 351 Abies concolor trees collected in 43 localities was analyzed for composition of its monoterpenoid fraction. The populations could be segregated into three geographic groups: (1) Northern and Central California (equivalent to var. Iowiana), producing camphene and 3-carene in trace to small amounts; (2) Southern California (the Cuyamaca race), producing camphene in trace to small and 3-carene in larger amounts; and (3) states to the east of California (equivalent to var. concolor) producing camphene and 3-carene in larger amounts. California populations from the southernmost Sierra Nevada and Northern Transversal Ranges were found to be chemically intermediate between var. Iowiana and var. concolor. Statistical analysis indicated the presence of additional second-order segregations in each of the four population groups, The results are explained on the basis of existing paleobotanical records.     

Within-tree spatial variation in the leaf monoterpenes of Sequoia sempervirens

Location within a tree was analyzed as a source of variation in Sequoia sempervirens leaf monoterpenes. No differences were found for quantitative composition or total yield/dry wt among lower, middle and upper canopy positions. The awlshaped, spirally arranged leaves of vigorous upper shoots showed small quantitative compositional differences, but not differences in total yield. The intermediate leaf form of young sprouts had the most different monoterpene quantitative composition and about three times the total yield of the above two leaf forms. Analysis of a clonal ring of 17 adult trees resulted in coefficients of variation similar to those for samples collected from different canopy levels of the same shoot. Results revealed the sources and magnitudes of experimental error in comparative studies of this species' leaf monoterpenes, and did not support the concept that somatic mutation provides an important source of variation in a large, long-lived organism such as coast redwood.     

鹅掌楸LcUGE基因的克隆和组织表达分析

为了解鹅掌楸(Liriodendron chinense)的UGE基因功能,采用RACE和EPIC-PCR技术克隆到2个UGE基因,命名为LcUGE1和LcUGE2。结果表明,LcUGE1基因的c DNA全长为1 531 bp,包含1 050 bp的开放阅读框,编码349个氨基酸, gDNA长度为11 920 bp;LcUGE2基因的c DNA长度为1 378 bp,包含1 056 bp的开放阅读框,编码351个氨基酸,g DNA长度为6544 bp。LcUGE1和LcUGE2基因均含有9个外显子和8个内含子,且外显子长度和内含子剪切位点序列几乎一致,但内含子片段长度存在显著差异。编码的LcUGE1和LcUGE2蛋白高度保守,保守性达到82%。LcUGE1基因在雄蕊中表达量最高,而LcUGE2基因则在花萼中表达量最高。这表明LcUGEs基因可能参与鹅掌楸的生殖发育过程。     

Geographic variation in field body temperature of sceloporus lizards

1. Using data from the literature, I assessed how broad climatic patterns affected field body temperatures (T_b’s) of lizards in the genus Sceloporus.

2. Sceloporus at temperate latitudes had mean T_b’s of 35°C throughout their elevational range. This pattern is associated with “tropical” temperatures that extend into high north latitudes during the summer and the relatively low elevations occupied by the lizards.

3. At tropical latitudes, mean T_b declined from 35°C at low elevations to 31°C at high elevations. This pattern is associated with low seasonal variation in temperature at tropical latitudes and the relatively high elevations occupied by the lizards.     

Geographic pattern of genetic variation in the fox tapeworm Echinococcus multilocularis

Intraspecific genetic variation of Echinococcus multilocularis, the etiologic agent of human alveolar echinococcosis, has been evaluated among 76 geographic isolates from Europe, Asia and North America by using sequence data of mitochondrial and nuclear DNA. Relatively low genetic variation was found only in the mitochondrial DNA sequence consisting of 3 protein-coding genes. Pairwise divergence among the resultant 18 haplotypes ranged from 0.03 to 1.91%. Phylogenetic trees and parsimony network of these haplotypes depicted a geographic division into European, Asian and North American clades, but 1 haplotype from Inner Mongolia was unrelated to other haplotypes. The coexistence of the Asian and North American haplotypes could be seen, particularly on the St. Lawrence Island in the Bering Sea. These data suggest an evolutionary scenario in which distinct parasite populations derived from glacial refugia have been maintained by indigenous host mammals. The nuclear DNA sequence for the immunodominant B cell epitope region of ezrin/radixin/moesin-like protein (elp) was extremely conservative, indicating that the elp antigen is available for immunodiagnosis in any endemic areas.     

Terpenes from the essential oil of Cymbopogon distans

A GC/MS study of the hydrocarbon fraction and the fraction containing oxygenated compounds showed the presence of 12 monoterpene hydrocarbons (28.4%), 13 sesquiterpene hydrocarbons (32.8%), 3 sesquiterpene alcohols (27.2%), 2 esters (7.2%) and 3 carbonyl compounds (4.4%) in the essential oil of Cymbopogon distans. Of these, 27 compounds have been identified.     

北美鹅掌楸原生质体的分离与培养

以鹅掌楸属植物北美鹅掌楸的悬浮细胞和组培苗叶片为材料,对北美鹅掌楸原生质体分离、纯化与培养条件进行研究.结果表明:叶片和悬浮细胞用含有0.1％2-吗啉乙磺酸(MES)和0.6 mol/L甘露醇的Cell ProtoplastWash(60M-CPW)溶液25℃预处理lh效果最好;悬浮细胞最佳酶解液为60M-CPW+ 1％纤维素酶+1％半纤维素酶+0.2％果胶酶Y-23+0.1％ MES,每克材料25℃酶解6h有效原生质体产量可以达到3×106个;叶片最佳酶解液为60M-CPW+2％纤维素酶+1％半纤维素酶+0.2％果胶酶Y-23+0.1％ MES,每克材料25℃酶解10 h有效原生质体产量可以达到11×106个;悬浮细胞原生质体易于培养,在KM8p+1.0 mg/L 2,4-D+0.5 mg/L 6-BA培养基中培养25 d可形成肉眼可见的愈伤组织.     

Compositional characteristics of the essential oil of Artemisia campestris var. glutinosa

The chemical variations of the essential oil from the aerial parts of Artemisia campestris var. glutinosa have been studied. Plant material has been harvested at each phenological status (vegetative, before anthesis, full flowering and seed-bearing). Analysis by GC and GC/MS of the essential oils have allowed to identify 51 components. The main components were γ-terpinene, capillene, 1-phenyl-2,4-pentadiyne, spathulenol, methyleugenol, p-cymene and β-pinene. Aromatic polyacetylens have been characterized for the first time in this species. Flowering involves an increase of these compounds in the essential oil.     

Geographic variation among Iberian communities of the exotic halophyte Cotula coronopifolia

The exotic annual forb Cotula coronopifolia has locally invaded salt marsh communities naturally dominated by Spergularia marina along large parts of the Iberian coastline. While the floristic composition of C. coronopifolia dominated communities on the Atlantic coast of northern Portugal has already been described, this information was missing for the southern part of the country. We carried out a phytosociological study at 21 sites along the south-western and southern coast of mainland Portugal to determine the ecology and syntaxonomical status of these communities, and to compare them to the Atlantic ones. Plant communities co-dominated by C. coronopifolia and S. marina were generally found in the sub-halophilous inner part of salt marshes. They occupied gaps within taller perennial vegetation (Juncus maritimus formations), such as small pools temporarily inundated with brackish waters. Southern communities differed from the northern ones by the high abundance of Triglochin barrelieri and the absence of salt-intolerant species that are present in the Atlantic communities. This difference may result from the drier summer climate in southern regions, leading to intense water evaporation and thus, more saline conditions. We propose to classify the Mediterranean communities in a new association, the Cotulo coronopifoliae–Triglochinetum barrelieri. High abundance of Limonium diffusum further allowed the distinction of a new subassociation limonietosum diffusi for the southernmost part of Portugal. Results show that invasive exotic plants can form new phytosociological associations with a characteristic species composition, but that these community types, despite being dominated by the same invader, still show ecological and geographical differentiation.     

GC-MS分析九头狮子草挥发油的化学成分

采用水蒸气蒸馏法提取九头狮子草挥发油,利用 GC-MS 联用技术,结合化学计量方法对其化学成分进行定性和定量分析,用面积归一法测定各组分的相对质量分数.结果表明：九头狮子草中挥发油主要由植酮(19．82％)、丁香油酚甲醚(3．96％)、β-石竹烯(3．75％)、肉豆蔻醚(3．08％)、3-甲基-2-(3,7,11-三甲基十二烷基)呋喃(3．64％)、2-戊基呋喃(2．73％)和氧化石竹烯(2．69％)等51种成分组成,占总量的98．82％.     

Laurus novocanariensis essential oil: Seasonal variation and valorization

We investigated the qualitative and quantitative seasonal variation of the leaf and fruit oils of the Macaronesian endemism Laurus novocanariensis and their plant defensive potential. The monoterpene fraction dominated the leaf (74%) and berry essential oils (73–44%, ripe–unripe). The insect antifeedant effects of these oils were species- and season-dependent against the aphids (Myzus persicae and Rhopalosiphum padi). Overall, the biological effects of these oils correlated with the oxygenated terpene fraction. Among the pure components tested, β-caryophyllene and its oxide were strong antifeedants to Leptinotarsa decemlineata and Spodoptera littoralis. The aphids responded to β-ocimene, β-pinene, 1,8-cineole, linalool (antifeedants) and linalool oxide (attractive to M. persicae). The antifungal effects of the leaf oils on Fusarium spp. were season-dependent. β-Caryophyllene oxide proved to be a strong antifungal. L. novocanariensis oils inhibited Lactuca sativa germination and radicle elongation, the leaves being more effective. Linalool also inhibited seed germination.     

北美鹅掌楸LtAGO1基因的克隆、表达及其启动子分析

为深入探究叶原基分化成叶器官的形态建成机制,该研究以北美鹅掌楸为材料,采用RT-PCR和RACE克隆技术获得LtAGO1的cDNA全长和启动子序列并预测其功能,通过RT-qPCR分析LtAGO1在鹅掌楸属中的组织表达模式。同时,经抗性筛选和DNA鉴定获得ProAGO1 ∷ GUS的转基因拟南芥株系,并进一步对T2代阳性植株进行表型和GUS组织化学染色分析。结果表明:(1) LtAGO1基因包含3 300 bp的开放阅读框,编码1 100个氨基酸,分子量为122.14 kD,理论等电点(pI)为9.36。(2)氨基酸序列分析显示LtAGO1含Gly-rich-AGO1和Piwi两个典型的AGO基因结构域,同源性分析显示LtAGO1蛋白与沉水樟AGO1蛋白(RWR84608.1)亲缘关系最近。(3)组织表达特异性分析显示LtAGO1在北美鹅掌楸不同组织间的相对表达量为雄蕊>花芽>花瓣>花萼>叶片>雌蕊>叶芽>茎,LtAGO1在北美鹅掌楸叶片不同发育阶段的相对表达量为叶芽萌动期>幼叶期>衰老期>成熟期,AGO1在鹅掌楸属叶缘的表达量高于叶片的其他部位且北美鹅掌楸叶凹陷部位的表达量高于叶尖部位。(4)获得叶中-侧轴向和基-顶轴向的极性缺失、叶缘锯齿、重瓣花型的转化株系,GUS组织染色显示ProAGO1启动GUS基因在叶芽顶端稳定表达且在新分化的叶柄上表达较强,在成熟期的茎、叶、花和果的维管束中均特异表达。LtAGO1启动子的GUS活性强度为叶顶芽>花>维管束,这与实时定量PCR结果相一致。综上认为,LtAGO1基因在顶端分生组织特异表达且受到多种途径的调控而参与到叶和花器官的发育进程中。该研究结果为进一步了解北美鹅掌楸LtAGO1基因的基本功能及其调控叶形发育机制提供了理论基础。     

Intraspecific variation of Chiliadenus iphionoides essential oil in Israel

Chiliadenus iphionoides (Asteraceae), a shrub endemic to the Mediterranean region and widespread throughout Israel, is used in the traditional eastern Mediterranean medicine. Although recent research confirmed its pharmacological potential, C. iphionoides essential oil has not been adequately characterized chemically. Essential-oil samples were collected from representative wild populations throughout Israel and characterized by GC/MS analysis. Considerable interpopulation variation was found for the composition of the essential oils. Multivariate analysis showed a significant correlation between the chemical composition and the geographic location, with three main chemotypes identified.     

有机栽培龙脑百里香精油对白色念珠菌体外抑菌活性的研究

【背景】白色念珠菌(Candidaalbicans)属于条件致病性真菌,可引起严重的黏膜真菌感染及全身系统性真菌感染,是导致患者高发病率和高死亡率的主要菌群之一。【目的】探究百里香精油对白色念珠菌的抑菌活性及抑制机理。【方法】测定5种百里香精油对白色念珠菌的抑菌圈直径,分析具有高抑菌活性的精油成分。在此基础上,通过扫描电子显微镜(scanning electron microscope, SEM)观察精油对白色念珠菌菌体细胞形态的影响。测定碱性磷酸酶(alkaline phosphatase, AKP)含量、胞外溶液电导率并进行碘化丙啶(propidium iodide, PI)染色分析,探究精油对白色念珠菌生物膜的形成与黏附及磷脂酶活性的影响,并通过实时荧光定量PCR法分析与白色念珠菌生物膜形成相关基因(凝集素样序列基因ALS4,从酵母型向菌丝型细胞的形态转变基因HWP1、磷脂酶基因PLB1)的表达水平,探究该精油对白色念珠菌的抑菌机制。【结果】筛选出了对白色念珠菌高度敏感的有机栽培龙脑百里香精油(Thymus vulgaris CT borneol essential oil, T...     

Geographic variation in clutch and brood size of the Eagle OwlBubo bubo in the Western Palearctic

Summary In the western Palearctic, the clutch size of the Eagle Owl (Bubo bubo) is constant with latitude and longitude whereas the brood size decreases from south to north, a reverse trend compared to other Strigiforms. This tendency seems independent of the population density of Eagle Owls and the availability of prey with high food value and may be related to climatic constraints which could determine lower viability of eggs and young at high latitudes.

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Zusammenfassung In der Westpaläarktis ist die Gelegegröße des Uhus in Gebieten unterschiedlicher geographischer Länge und Breite konstant. Im Unterschied dazu nimmt die Brutgröße von Süden nach Norden ab, umgekehrt zum Trend bei anderen Eulen. Die Tendenz der Abnahme der Brutgröße scheint unabhängig von der Siedlungsdichte und der Erreichbarkeit von Beute mit hohem Nährwert. Sie kann mit klimatischen Bedingungen zusammenhängen, die eine höhere Mortalität der Eier bzw. Nestlinge in hohen Breiten verursachen.