Analysis of quinolizidine alkaloids in Sophora flavescens Ait. by capillary electrophoresis with tris(2,2 '-bipyridyl) ruthenium (II)-based electrochemiluminescence detection

A capillary electrophoresis method coupled with electrochemiluminescence detection for the analysis of quinolizidine alkaloids was established, especially, oxymatrine (OMT) which could not be measured by previous electrochemiluminescence methods was detected sensitively herein. Complete separation of sophoridine (SR), matrine (MT) and OMT was achieved within 13 min using a background electrolyte of 50mM phosphate buffer at pH 8.4 and a separation voltage of 15 kV. The calibration curves showed a linear range from 2.8 x 10(-8) to 4.4 x 10(-7) M for SR, 2.7 x 10(-8) to 4.4 x 10(-7) M for MT.

Isolation and characterization of microsatellites in yellow perch (Perca flavescens)

A total of 45 microsatellite loci from yellow perch, Perca flavescens, were isolated and characterized. Among the 45 microsatellite loci, 32 had more than two alleles. A wild population of P. flavescens (n = 48) was used to examine the allele range of the microsatellite loci. Mendelian inheritance of alleles was confirmed by examining the amplified products in pair-mated families. The number of alleles for the 32 polymorphic loci varied from two to 16, and observed heterozygosity ranged between 0.024 (YP79) and 0.979 (YP60). Cross-species polymorphic amplification in four other Percidae species was successful for 22 loci.

兰科槽舌兰属的系统学研究

槽舌兰属为兰科树兰亚科万代兰族指甲兰亚族植物，大部分种类为中国特有种，部分种类分布到越南、泰国、缅甸等国家和地区。长期以来，本属植物由于与近缘属间的界线模糊不清，而属内性状变异大，是兰科植物系统学研究的疑难属之一。本文通过对槽舌兰属及部分近缘属植物的形态地理学、细胞学、传粉生物学、分子系统学等研究，重新界定了槽舌兰属，确认该属植物的种类，提出一个新的属下分类系统，讨论了槽舌兰属属内和近缘属间的系统学关系，同时对槽舌兰属高山组植物的演化进行了探讨。 1．槽舌兰属及近缘属形态学性状 　　我们通过标本研究和野外观察，对槽舌兰属和近缘属植物的形态学性状及其变异式样进行了详细比较分析，重新评价各性状的分类学价值，认为蕊喙、蕊柱翅、花粉块类型等对于槽舌兰属与近缘属的划分具有重要意义;胼胝体类型、粘盘柄形状、距的类型、唇瓣中裂片边缘状况等对属下分类具有重要的意义;而中裂片、侧裂片和距的形态对槽舌兰属的种的划分具有重要的意义。传统上作为划分种依据的胼胝体的数目、花序上花的数目、茎的长短等在种内变异较大，不宜用作种划分的依据。 2. 槽舌兰属植物核型研究 　　我们报道了8种槽舌兰属植物的体细胞的染色体数目，其中6种为首次报道，并对其中7个种的核型进行初步分析。目前已进行研究的槽舌兰属植物的染色体数为2n=38，体细胞分裂中期的染色体长小于2 um, 染色体类型以中部和近中部着丝粒染色体为主，部分染色体成点状。槽舌兰属不同染色体数目相等、染色体形态和大小相似，从一个方面反映了槽舌兰属植物是一个单系类群。 3. 滇西槽舌兰（Holcoglossum rupestre）的传粉生物学研究 　　我们选择滇西槽舌兰的进行传粉生物学研究以探讨本属植物系统学和进化，发现滇西槽舌兰的传粉系统为兰科植物中少见的甲虫专化传粉系统，而且滇西槽舌兰的传粉系统处于一种不稳定的状态。弯腿金龟（Hybovalus bioculatus）是滇西槽舌兰唯一的传粉者，以两个不同的部位携带花粉团，即前腿和臀部，并存在着两种不同的传粉机制。滇西槽舌兰这种不稳定的传粉系统似乎表明该系统可能正处于转变阶段。繁殖系统的实验表明自然状况下滇西槽舌兰结实率由传粉率决定，而自然状况下低的结实率可能由两方面的原因导致：很低的传粉效率和有效的防止自花传粉机制。 4. 槽舌兰属植物的分子系统学研究 　　我们对9种槽舌兰属植物和2种万代兰属植物的叶绿体基因组trnL-F序列进行了测定，并从基因库下载了14种万代兰族植物和2种用作外类群的美洲附生兰亚科植物的trnL-F序列。测序结果发现槽舌兰属的trnL-F序列存在着很大的差异：H. lingulatum的trnL-F存在大片段的缺失，而H. kimballianum的trnL-F存在许多小片段的插入。分子系统学分析表明槽舌兰属为一单系类群，支持将Vanda subulifolia和V. amesiana转置入槽舌兰属中，强烈支持由H. sinicum、H. flavescens、H. rupestre和 H. weixiense构成槽舌兰属属内分支。 5．槽舌兰属的分类学修订 　　在广泛查阅国内外馆藏标本和开展多次野外调查的基础上，并结合细胞学和分子系统学研究结果，我们重新界定了槽舌兰属。该属植物的主要特征：茎短;叶肉质，细长半圆柱形，部分种叶略扁而成“V”形，其近轴面具一纵槽，先端渐尖，不裂;总状花序侧生，花疏生;花苞片比花梗和子房短;侧萼片或多或少呈镰形;花瓣通常具爪，与中萼片相似;唇瓣3裂;侧裂片直立;中裂片较大，基部常有附属物;蕊柱粗短，具翅，无蕊柱足或具很短的足;蕊喙2裂;花粉团蜡质，2个，球形，具裂隙。确定槽舌兰属目前包括11种（包括2个新种）：H. subulifolium、H. amesianum、H. kimballianum、H. lingulatum、H. omeiense、H. quasipinifolium、H. sinicum、H. rupestre、H. flavescens、H. wangii和H. weixiense。 H. tangii和Vanda saprophytica被分别处理为H. lingulatum与H. kimballianum的异名，而H. tsii的分类学地位尚有待进一步研究。同时在我们研究的基础上，提出槽舌兰属一个新属下的分类系统： 槽舌兰属 Holcoglossum 　　短距亚属 Holcoglossum subgen. Amesianum 　　H. subulifolium，H. amesianum; 槽舌兰亚属Holcoglossum subgen. Holcoglossum 　槽舌兰组Holcoglossum sect. Holcoglossum 　　H. kimballianum，H. lingulatum，H. wangii，H. omeiense，H. quasipinfolium; 高山组Holcoglossum sect. Sinicum 　H. sinicum，H. rupestre，H. flavescens，H. weixiense。 6．槽舌兰属属内和近缘属间系统学关系探讨 　　通过广泛的形态地理研究并结合传粉生物学和分子系统学研究结果，我们对槽舌兰属属内和属间关系进行的探讨。槽舌兰属短距亚属与万代兰属和指甲兰属比较接近，是槽舌兰属中比较原始的类群; 槽舌兰组植物的分布中心介于短距亚属和高山组分布中心之间，而该组植物的许多性状也表现出过渡性质。在‘Aerides-Vanda alliance’中，我们认为钻喙兰属和万代兰属是该类群中比较原始的属，而鸟舌兰属（Ascocentrum）可能是介于指甲兰属和举喙兰属（Seidenfadenia）之间，支持传统的观点认为Papilionanthe位于Vanda和Aerides之间但更接近于Aerides，但同时我们认为举喙兰属可能是‘Aerides-Vanda alliance’中最为特化的类群。 7． 高山组所属物种形成探讨 　　槽舌兰属高山组的四个种均为中国特有植物，主要分布云南西北部地区，其中H. weixiense、 H. rupestre、H. sinicum3个种只局限分布在云南的西北部地区。尽管高山组各个种比较相似，但形态上已经分化，生境也不同而且存在着对应关系。细胞学实验结果发现槽舌兰属高山组的四种植物均为二倍体（2n=38），与槽舌兰属的其他种类一致，这表明该组植物在横断山区的分化时染色体的数目并没有变化，高山组植物的三种横断地区特有植物可能为裂生特有。高山组4个种的1300bp的trnL-F总共只有10个碱基的差异，在trnL-F分支图中，高山组分支作为单系类群得到强有力的支持（BP=94%）。高山组植物trnL-F序列微小的差别反映了该组植物可能是在近期通过快速物种形成进行分化的。这些资料说明槽舌兰属高山组植物是近期通过适应辐射进行分化以适应不同生境，而且可能是生境和传粉系统共同驱动下形成的。

川西亚高山人工针叶林下11种植物对两种光生境的适应性研究

青藏高原东缘的亚高山针叶林是长江上游重要的生态屏障，经过近六十年的采伐后，取而代之的是大量人工种植的云杉纯林。目前，这些人工林已经表现出树种单一，结构层次简单等生态问题，其物种多样性及生态效益与同地带天然林相比差距较明显。如何丰富该地区物种多样性，完善人工林生态系统的生态功能是一个十分重要的课题。林下植物是人工林群落的重要组成部分，对维持群落的生物多样性及完善生态系统功能具有明显的作用。因此，研究该地区人工针叶林的林下植被对不同生境的适应性对于理解人工林生态系统物种多样性的形成和维持机制都具有重要的意义。 本文以青藏高原东部亚高山针叶林的主要森林类型----云杉人工林为研究对象，选择林下11种具有不同喜光特性的常见植物，分别设置人工林林冠下及成熟林窗为研究样地，通过对各种植物叶片形态与物质分配特征、叶片解剖学特征、叶片光合生理特性、植物自然分布特征等方面的比较分析，研究林下植物对不同光生境的适应策略及其适应能力，揭示不同物种对人工林生境的适应共性，为西南亚高山地区植被恢复及人工林的经营管理提供科学依据。具体研究结果如下： 在叶片形态和物质分配特征方面：在林窗光生境中，11种林下植物叶片比叶重(LMA)显著高于林下光生境的同种植物。同时，林窗下生长的植物叶片叶片厚度及栅栏细胞长度显著增加，这是影响叶片比叶重变化的直接原因。而多数植物叶重比在两种生境中无明显变化。说明在长期适应自然生境之后，植物可能更多地采取调节叶片组织细胞水平（即叶片功能细胞形态）及叶片器官水平（即单个叶片形态）特征的策略来适应各类生境，而非整株水平上的叶片总比重的增减。 在叶片解剖结构特征方面：多数阔叶物种栅栏组织厚度(PT)、栅栏组织厚度/海绵组织厚度(PT/ST)、栅栏细胞层数及近半数种的气孔密度(SD)在林窗生境中更大或更多，而叶片表皮细胞厚度(UET、LET)气孔长径(SL)及海绵组织厚度(ST)受两种生境影响不大。喜光特性相似的物种在生境适应策略上具有一定的趋同性。 在光合生理特征方面：在林窗生境中多数种植物的最大光合速率(Amax)、暗呼吸速率(Rd)及喜光植物光补偿点(LCP)显著或极显著高于林内生境同种植物。且在同一生境条件下，多数深度耐荫植物比喜光及轻度喜光植物有稍低的Rd和LCP。各植物在林内低光生境中具有更大的内禀光能转化效率，并在中午12:00～14:00之间光强最大的时刻发生了的最深程度的光抑制。多数种能通过调节自身某种光合素含量或色素之间的比例来适应不同的光生境，即通过增加叶绿素含量或降低Chla/b值来适应林内弱光生境，通过提高类胡萝卜素含量或单位叶绿素的类胡萝卜素含量降低强光带来的伤害。绝大多数物种并不采取调节叶片C、N含量的策略来适应不同的光生境。总之，植物部分光合参数(Amax、Rd、LCP)受生境的影响与其自身喜光特性有关，但另一些参数（Fv/Fm日变化、色素含量及比例、叶氮相对含量）受生境影响与其自身喜光特性无明显关联。 在表型可塑性方面：在叶片各表型参数中，器官水平及细胞水平的形态特征参数平均可塑性大于整株水平形态和物质分配特征参数可塑性；叶片光合组织的可塑性大于非光合组织可塑性；反映植物光合能力的参数可塑性大于叶片色素含量参数可塑性。植物叶片形态和物质分配、解剖学特征参数平均可塑性大小与其自身喜光特性基本吻合，即喜光种及轻度耐荫种各参数可塑性最高，深度耐荫种可塑性最小，而这种规律并未在光合生理参数的可塑性大小上体现出来。但是叶片形态和物质分配参数、光合生理参数的平均可塑性水平却大于叶片解剖学参数。 在植物自然分布特征方面：喜光物种云杉幼苗及歪头菜在林内生境中分布密度明显降低，深度耐荫种疏花槭却恰恰相反，更多数物种（7种植物）在两种生境中密度变化趋势不明显。从分布格局来看，7种植物在两种生境中均为聚集分布，但聚集强度为林窗>林内；少数物种桦叶荚迷、直穗小檗、冰川茶藨、黄背勾儿茶在林窗中为聚集型，在林内生境中的分布型发生改变而成为随机型，说明光生境的差异能影响到植物种群的分布特征。但这种影响程度与植物自身的喜光特性无关，同时与各物种叶片表型平均可塑性的大小也无明显关联。 The subalpine coniferous forest area in eastern Qinghai-Tibet Plateau is important ecology-barrier of upriver Yangtze. In past sixty years, those forests had been cut down and replaced with a lot of spruce plantations. At now, there are many ecology problems presenting to us such as singleness species, simple configuration, lower species diversity and ecological benefit than natural forests at the same belt. How to restore the species diversity and enhance the eco-function of the plantations is a very important issue. The understory plants are important part of plantation community, which improved the bio-diversity and eco-function distinctly of forests. So, it is very significance to study the adaptation of understory plants to different environment in plantation, and this study would helping us to understand how plantations to develop and remain their biodiversity. This study was conducted in a 60a spruce plantation in Miyaluo located in western Sichuan, China, and spruce plantation is major types of subalpine coniferous forest in eastern Qinghai-Tibet Plateau. In this paper, the leaf morphological and biomass-distributed characteristics, the anatomical characteristics, the photosynthetic characteristics and the distribution patterns characteristics of eleven different light-requirement understory species grown in two different environments (forest gaps and underneath close canopy) were studied and compared. The purpose of this study was to analyze the adaptation of this forest understory plants, to show up the commonness of these different light-requirement understory species in light acclimation, and to provide some scientific reference to manage and restore the vegetation of subalpine plantation of southwest China. The results were as follows: The leaf morphological and biomass-distributed characteristics: These eleven species in forest gaps had significantly higher dry weight per leaf area (LMA) than those under close canopy. The palisade parenchyma cells of the broad-leaved species in gaps were significantly longer than those grown under the canopy, which been a directed factor for the change of leaf mass per unit area (LMA) in different environment. But the leaf weight ratio (LWR) of most plants species were not evidently changed by the contrasted environments in our study. It was shown the morphological characteristics changing been adopted as a strategy of light acclimation for plants wasn’t on whole plant level (leaf weight ratio) but cellular level (the function cells morphological characteristics) and organic level (the leaf morphological and biomass-distributed characteristics) mostly. The leaf anatomical characteristics: Most broad-leaved plants in gaps increased palisade parenchyma thickness (PT), the palisade parenchyma cell layers and the ratio of palisade to spongy parenchyma (PT/ST). So did as almost about half species in this study in stomatal density (SD). No significant differences in thickness of leaf epidermal cells (UET, LET), stomatal length (SL) and spongy parenchyma (ST) between two environments of most species were observed. The results suggested that species with light-requirement approximately had convergent evolution on adaptation to light condition. The leaf photosynthetic characteristics: The dark respiration rate (Rd) of most plants species, the light compensation point (LCP) of light-demanding plants species in gaps were significantly increased than under close canopy in this study. In a same habitat, most deep-shade-tolerant plants had lower Rd and LCP than those light-demanding plants and slight-shade-tolerant plants. Each species has bigger inherent electron transport rate under close canopy than in gaps, and the greatest photoinhibition happened during 12 to 14 in the daytime. Most species could adapt different light environment by the way of changing their photosynthetic pigments content or the ratio of pigments content. For example, some plants under close canopy increased chlorophyll (Chl) or reduced the values of the ratio Chla/b to adapted the low light condition, some plants in gaps increased carotenoid (Car) or reduced the weight ratio CarChl to avoid been hurt in high light. For most plants, changing the content of C and N in leaf wasn’t a strategy of light acclimation. In conclusion, the variation of some leaf photosynthetic parameters in different light environment such as Fv/Fm, pigments, C and N in leaf related with the light-requirmnet of species, but the others such as Amax, Rd, LCP did not. The leaf plasticity indexes: Among those leaf plasticity indexes, the leaf morphological and biomass-distributed parameters on cellular and organic level were greater than on whole plant level for same species, and the photosynthetic parenchyma parameters were greater than non-photosynthetic parenchyma parameters in same leaf, and photosynthetic capability parameters were greater than photosynthetic pigments content parameters for same species. The average plasticity indexes of leaf morphological and biomass-distributed and anatomical parameters were accordant with plants’ light-requirement approximately: those light-demanding plants and slight-shade-tolerant plants had bigger plasticity indexes than deep-shade-tolerant plants. But this regular wasn’t observed in physiological plasticity indexes for most plants, though the average leaf plasticity indexes of leaf morphological and biomass-distributed, photosynthetic characteristics parameters was greater than the anatomical characteristics parameters. The distribution patterns characteristics: Oppositely to the deep-shade-tolerant specie Acer laxiflorum Pax., the density of light-demanding species Picea asperata Mast. and Vicia unijuga A. Br. in gaps was bigger than under close canopy. Each of the other species has the approximately density in two different environment. The spatial patterns of seven species were aggregated distribution in two environments, but the trend of aggregation of population under close canopy was decrease from in gaps. A few species such as Viburnum betulifoium Batal., Berberis dasystachya Maxim., Ribes glaciale Wall. and Berchemia flavescens Brongn. were aggregated distribution in gaps while random distribution under close canopy. It was shown that the difference between two light environments could affect the distribution pattern of plant population, and the effect didn’t relate with the light-requirement or plasticity indexes of species.

长白山森林生态系统中的稀有和濒危多孔菌

In last 10 years,extensive field inventories were carried out to investigate Polypore species, the major wood decaying fungi in the Changbaishan Nature Reserve of Northeastern China. The following 27 species were treated as rare or threathened species: Amylocystis lapponica (Romell) Singer, Anomoporia albolutescens (Romell) Pouzar, Anomoporia bombycina (Fr.) Pouzar, Anomoporia vesiculosa Y.C. Dai & Niemel, Antrodia carbonica (Overh.) Ryvarden & Gilb., Antrodia crassa (P. Karst.) Ryvarden, Antrodiella citrinella Niemel & Ryvarden, Diplomitoporus flavescens (Bres.) Dománski, Donkioporia expansa (Desm.) Kotl. & Pouzar, Gloeophyllum carbonarium (Berk. & M.A. Curtis) Ryvarden, Haploporus odorus (Sommerf.) Bondartsev & Singer, Inonotopsis subiculosa (Peck) Parmasto, Nigroporus ussuriensis (Bondartsev & Ljub.) Y.C. Dai & Niemela, Oxyporus sinensis X.L. Zeng, Parmastomyces taxi (Bondartsev) Y.C. Dai & Niemela, Phellinidium sulphurascens (Pilat) Y.C. Dai, Phellinus vaninii Ljub., Polyporus vassilievae Thorn, Pycnoporellus fulgens (Fr.) Donk, Skeletocutis brevispora Niemela, Skeletocutis ochroalba Niemela, Skeletocutis perennis Ryvarden, Trechispora candidissima (Schwein.) Bondartsev & Singer, Wolfiporia dilatohypha Ryvarden & Gilb., Wolfiporia curvispora Y.C. Dai, Wrightoporia avellanea (Bres.) Pouzar and Wrightoporia lenta (Oveh. & J. Lowe) Pouzar. Polypores are richer in East Asia than in Europe and North America, not only because of destructive galciations and fewer hosts in the latters, but also because of the geography. NE Asia is a link between Europe and North America. Changbaishan Nature Reserve is very rich in polypores, and over 260 species were recorded in the reserve. Some rare species in North America and Europe, for instance, Anomoporia albolutescens, Antrodia crassa, Diplomitoporus flavescens, Inonotopsis subiculosa and Skeletocutis ochroalba etc. were found in Changbaishan Nature Reserve as well, and these species are in fact rare in the earth. Most of the 27 species occurred on fallen trunks or rotten wood in the reserve, but some of them grew on living trees. 18 species occured on substrate of gymnosperms, and 9 species grew on wood of angiosperm.Among the 27 species, 7 species caused a brown rot,and 20 species produced a white rot. The morphology, substrate and ecology of each species were briefly discussed. The most important tool for polypore conservation is the conservation of their habitats, and it is necessary to study the ecology of the rare and threathened species of polypores in the Changbaishan Nature Reserve. Because most of polypores live on the substrate of fallen trunks and rotten wood, it is very important to keep such substrate in the ecosystem.

Yellow perch strain evaluation I: Genetic variance of six broodstock populations

As a prelude to strain selection for domestication and future marker assisted selection, genetic variation revealed by microsatellite DNA was evaluated in yellow perch, Perca flavescens, from four wild North American populations collected in 2003-2004 (Maine, New York, North Carolina, and Pennsylvania,), and two captive populations (Michigan and Ohio). For the loci examined, levels of heterozygosity ranged from H-e=0.04 to 0.88, genetic differentiation was highly significant among all population pairs, and effective migration ranged from low (N(e)m=0.3) to high (N(e)m=4.5). Deviation from Hardy-Weinberg equilibrium was regularly observed indicating significant departures from random mating. Instantaneous measures of inbreeding within these populations ranged from near zero to moderate (median F=0.16) and overall inbreeding levels averaged F-IS=0.18. Estimates of genetic diversity, Phi(ST), and genetic distance were highest between Michigan and all other broodstock groups and lowest between New York and Ohio. Genetic differentiation among groups did not correlate with geographic distance. Overall, the patterns of variation exhibited by the captive (Michigan and Ohio) populations were similar to patterns exhibited by the other wild populations, indicating that spawning and management practices to date have not significantly reduced levels of genetic variation. (C) 2007 Elsevier B.V. All rights reserved.