桤木属7种植物的核型分析

利用改良去壁低渗法对分布于欧美地区的桦木科(Betulaceae)桤木属(Alnus Mill.)7种植物进行染色体数目与核型分析。结果显示:所有材料染色体形态比较一致,多为由中部(m)及近中部(sm)着丝点染色体组成。意大利桤木(A.cordata)为六倍体,体细胞染色体数为2n=6x=42,核型公式为2n=6x=42=36m+6sm;绿桤木(A.viridis)为八倍体,体细胞染色体数为2n=8x=56,核型公式为2n=8x=56=46m+10sm(SAT);薄叶桤木(A.tenuifolia)、灰桤木(A.incana)、欧洲桤木(A.glutinosa)、裂叶桤木(A.sinuata)和红桤木(A.rubra)均为四倍体,体细胞染色体数均为2n=4x=28,其核型公式分别为2n=4x=28=16m(1SAT)+12sm、2n=4x=28=22m+6sm、2n=4x=28=24m+4sm、2n=4x=28=24m+4sm、2n=4x=28=26m+2sm。其中红桤木(A.rubra)的核型属于1B型,其余均为2B型。     

5种桤木属植物的核型分析

利用改良去壁低渗法对桦木科（Betulaceae）桤木属（Ainus Mill.）东亚地区5种植物进行染色体数目与核型研究。结果显示：所试验物种中染色体形态比较一致,多是由中部（m）及近中部（sm）着丝点染色体组成。其中A.nitida染色体数为2n=28,核型公式为K（2n）=28=22m 4sm(2SAT) 2st(2SAT);A.hirsuta染色体数为2n=42,核型公式为K(2n)=42=36m 6sm; A.cremastogyne,A.formosana 染色体数均为2n=56,核型公式分别为K(2n)=56=2M 40m(1SAT) 14sm(1SAT)和K(2n)=56=46m 8sm 2st;A.firma染色体数为2n=112,核型公式为K(2n)=112=80m 28sm 4st。     

基于ITS序列的栓菌属部分种的分子分类初步研究

栓菌属 Trametes 的一些近缘种宏观和微观形态学非常相近,传统分类学方法难于对其进行准确分类定位。测定了 34 个分类单元的 ITS(包括 5.8SrDNA)序列,并对得到的 43 个分类单元的 ITS 序列进行系统发生分析,构建了聚类分析树状图。该树状图显示,栓菌属类群与其他属类群明显分开,Trametes versicolor 聚类到一个高支持率的独立分支。形态学上定名为 T. hirsuta 和 T. pubescens 物种聚类到同一高支持率的独立分支,试验分析表明这两个种应视为同一物种。     

中国薄叶卷柏复合群的物种划分

卷柏属(Selaginella)是石松类植物中最大的属, 也是分类难度较大的类群之一。该属的物种划分主要基于形态特征, 但许多近缘种在形态上很难区分。近年来, 已有大量分子证据被用于各植物类群的分类学研究, 但目前未发现一套适合卷柏属物种鉴定的分子标记。薄叶卷柏复合群(S. delicatula group)是卷柏属下鉴定较为困难的类群, 包括了薄叶卷柏(S. delicatula)、黑顶卷柏(S. picta)和瓦氏卷柏(S. wallichii) 3个物种, 主要分布于亚洲的热带和亚热带地区。为了探讨薄叶卷柏复合群内物种的亲缘关系和评估不同分子标记在卷柏属分类学研究中的应用价值, 本研究对该复合群物种进行广泛取样, 共收集到73个个体, 并选取3个叶绿体基因(rbcL, psbA和atpI)和2个核基因(26S nrDNA和pgiC)片段进行系统树的构建及叶绿体单倍型分析。研究发现, 基于叶绿体和核基因构建的系统发生关系存在冲突: 叶绿体基因树上薄叶卷柏个体分为两个分支(A和B), 薄叶卷柏B分支与薄叶卷柏A-S. picta分支呈姐妹关系, 并且rbcL单倍型分析结果也表明薄叶卷柏A和B两个分支存在明显分化; 而核基因结果则支持该复合群3个物种各自的单系性, 其中, S. delicatula分支与S. picta分支为姐妹群, S. wallichii与S. delicatula-S. picta分支为姐妹关系。在对复合群分布区大量标本的观察以及野外群体调查的基础上, 评估了植株茎和枝的分枝方式、孢子叶、营养叶(侧叶、中叶和腋叶)和孢子表面纹饰等形态性状的分类学价值。结果表明, 薄叶卷柏A和B分支的样本仅在植株分枝方式和大孢子表面纹饰上存在差异, 但无法依靠小孢子表面纹饰、孢子叶穗和营养叶形态等特征进行区分。基于现有证据, 薄叶卷柏复合群至少可划分为薄叶卷柏、黑顶卷柏和瓦氏卷柏3种, 但彻底澄清该复合群的物种划分还需要获取模式标本产地的材料和细胞学证据。最后, 建议在未来卷柏属的分类学研究集中于该属分类复杂的复合群, 结合使用形态学、细胞学、分子生物学(同时使用核基因和叶绿体分子标记)及地理分布等整合证据来进行物种划分。     

菊科紫菀属3种植物的核型分析

菊科紫菀属密毛系(Aster Series Vestiti Ling)含3个物种:密毛紫菀(A.vestitus Franch.)、灰枝紫菀(A.poliothamnus Diels)和西固紫菀(A.sikuensis W.W.Smith et Farr.),新近的分子系统学研究对密毛系是否单系提出质疑。该文采用常规根尖压片法,对紫菀属密毛系3种植物4个居群的核型进行观察分析。结果显示:3种植物的核型公式均为2n=2x=18=14m+4sm(2SAT);密毛紫菀居群2个的核型属于1A型,灰枝紫菀和西固紫菀为2A型。密毛紫菀和西固紫菀的核型资料为首次报道。灰枝紫菀的染色体数目、基数和不对称性等核型资料与先前唯一的相关报道结果(体细胞染色体数目为32,核型公式2n=2x=32=26sm+6st,属4A型,染色体中未发现随体)迥然不同。3种植物的核型结果不支持保留密毛系,而核型不对称性类型支持该实验室先前提出的亚属划分。3种植物随体的大小属于紫菀亚族型,这支持欧亚紫菀属与北美紫菀类没有密切亲缘关系的观点。     

广州市景观湖微囊藻的分类学研究

于2008年3月至2009年4月,对广州市区若干景观湖水体的微囊藻属(Microcystis)进行分类学研究。共观察到11种微囊藻,分别是铜绿微囊藻(M. aeruginosa Kützing)、放射微囊藻(M. botrys Teiling)、坚实微囊藻(M. firma Kützing)、水华微囊藻(M. flosaquae Wittrock)、鱼害微囊藻(M. ichthyoblabe Kützing)、挪氏微囊藻(M. novacekii Komárek)、苍白微囊藻(M. pallida (Farlow) Lemm.)、假丝微囊藻(M. pseudofilamentosa Crow)、史密斯微囊藻(M. smithii Komárek & Anagnostidis)、绿色微囊藻(M. viridis A. Braun)和惠氏微囊藻(M. wesenbergii Komárek),对它们的形态学特征进行描述,并比较这些种类间的形态区别。     

紫色甘薯品系6种酶的同工酶分析

采用聚丙烯酰胺凝胶电泳技术,选用过氧化物酶(POD)、超氧化物歧化酶(SOD)、细胞色素氧化酶(CYT)、淀粉酶(AMY)、酯酶(EST)、苹果酸脱氢酶(MDH)等六种酶的同工酶系统对14个紫色甘薯品种(系)进行了同工酶分析.结果表明:过氧化物酶(POD)、细胞色素氧化酶(CYT) 、超氧化物歧化酶(SOD)等三种酶的同工酶酶谱很丰富,分别有13、11、10条酶带;淀粉酶(AMY)、酯酶(EST)、苹果酸脱氢酶(MDH)等三种酶的同工酶酶带相对较少,分别为4、7、7条.聚类分析结果表明,所有供试紫色甘薯品系或品种分成7类:I类包括品系A1、A2、A7;II类包括品系B1～B3;III类包括品系B7和B8;IV类只有品系B9;V类包括品系A3、A4、A6;VI类只有品系A5;VII类只有品种"山川紫".     

中国蜘蛛抱蛋属二新记录种

首次报道了黄瓣蜘蛛抱蛋(Aspidistra lutea H.-J.Tillich)和吉婆岛蜘蛛抱蛋(A.arnautovii H.-J.Tillich)在中国的分布。这两种植物分别被发现于广西西南部的靖西县、龙州县。文中提供了形态学描述和图片。凭证标本存放于广西植物标本馆(IBK)。     

七种圈养鹤的部分血液生化指标分析

为探索圈养鹤类的血液生化指标差异,本研究对白鹤(Grus leucogeranus)、丹顶鹤(G.japonensis)、黑冠鹤(Balearica pavonina)、黑颈鹤(G.nigricollis)、灰冠鹤(B.regulorum)、蓝鹤(Anthropoides paradiseus)和白枕鹤(G.vipio)的10项血液生化指标(总胆固醇、甘油三酯、总蛋白、白蛋白、球蛋白等)进行统计分析,并与国际物种信息系统(ISIS)提供的数据进行比较。结果表明,1)不同鹤类之间总胆固醇差异显著,而甘油三酯和血糖没有显著性差异,除了黑冠鹤和灰冠鹤的总胆固醇以及蓝鹤的血糖外,其余鹤类的常值均高于ISIS的平均值。2)不同鹤类之间的总蛋白、球蛋白和白蛋白的浓度差异显著,但总蛋白、球蛋白和白蛋白均在ISIS的正常范围内。3)不同鹤类间肌酐和尿酸不具有显著性差异,其中尿酸低于ISIS平均值,但仅尿素氮差异显著。不同鹤类之间血液生化指标存在差异,与圈养鹤的营养水平和环境有关。因此血液生化指标对于圈养鹤类营养状态和健康情况的判断具有重要参考价值,对圈养动物饲养管理具有重要的指导作用。     

中国马勃类真菌新记录种记述

本文报道了中国马勃类真菌5个属的15个新记录种,分别为灰球菌属Bovista的铜色灰球菌B. aenea、棕灰球菌B. brunnea、坎氏灰球菌B. cunninghamii、泥灰球菌B. limosa、黑灰球菌B. nigrescens、毛灰球菌B. tomentosa;秃马勃属Calvatia的粗皮秃马勃C. turneri;脱盖马勃属Disciseda的异脱盖马勃D. anomala、白脱盖马勃D. candida;马勃属Lycoperdon的兰宾马勃L. lambinonii、裂纹马勃L. rimulatum;隔马勃属Vascellum的精致隔马勃V. delicatum、透明隔马勃V. hyalinum、间隔马勃V. intermedium、南美隔马勃V. pampeanum。部分标本保存在首都师范大学生命科学学院标本馆(BJTC)。     

福建省放线菌结瘤植物共生菌Frankia遗传多样性研究

[目的]了解福建省放线菌结瘤植物共生固氮菌Frankia的遗传多样性.[方法]利用16S-23SrDNA间隔区(rrn)和nifD-K基因间隔区的PCR扩增和RFLP技术,分析了福建省木麻黄、杨梅、桤木、胡颓子等共生Frankia纯培养菌株的遗传差异.[结果]17个菌株获得rrn扩增片段,2个杨梅菌株和1个胡颓子菌株扩增未成功,酶切图谱经聚类分析表明6个地点的细枝木麻黄、短枝木麻黄、粗枝木麻黄12个共生Frankia菌株同源性高,属于一个类群,2个地点的4个杨梅菌株和1个四川桤木菌株亲缘关系近,为另一类群.25个Frankia菌株的,nifD-K基因间隔区PCR-RFLP分析结果显示,7个地点的3种木麻黄14个菌株聚类为一个类群,4个地点的7个杨梅菌株、2个地点的2个四川桤木菌株以及1个台湾桤木菌株聚类为另一个类群,胡颓子菌株则为独立的类群.[结论]研究结果表明福建省共生Frankia遗传多样性丰富.     

黑长臂猿栖息地旱冬瓜和潺槁木姜子种群分布格局和动态

旱冬瓜（Alnus nepalensis）和潺槁木姜子（Litsea glutinosa）是云南省无量山黑长臂猿（Nomascus concolor）栖息地中的两种常见乔木.利用最近邻体法和Heygi单木竞争指数模型,对两个种群的分布格局和竞争情况进行分析.结果表明,旱冬瓜在不同生长阶段均呈现随机分布,而伴生的潺槁木姜子种群则呈现随机分布 聚集分布 均匀分布的变化趋势.种间竞争和物种自身生物学特性对两个种群的分布格局具有显著影响.旱冬瓜种群的龄级结构为衰退型,潺槁木姜子种群则为成长型.     

Restriction pattern analysis of deoxyribonucleic acid isolated from callus and cell suspension of actinorhizal and non-actinorhizal Betulaceae

Using cell suspensions, a method was elaborated to isolate high-molecular-weight genomic deoxyribonucleic acid (DNA; 65 MDa or more) from members of the Betulaceae: Alnus incana (L.) Moench, Alnus glutinosa (L.) Gaertn. and Betula papyrifera Marsh. The method was also effective for isolation of DNA from callus cells. Based on the chemical lysis of protoplasts, this procedure yielded 130 μg (callus) to 250 μg (cell suspension) of DNA (g fresh cells)⁻¹, with a ratio A₂₀₀/A₂₈ of 1.7–2.0. The purified DNA obtained, formed distinct bands when restricted fragments were electrophoresed. Among the 10 endonucleases used for restriction analysis of Alnus glutinosa, Alnus incana and Betula papyrifera genomes, PvuI1 (EC 3.1.23.33) was unique in giving identical patterns for the two Ainus species. An unusual pattern occurred when Al-2 DNA was restricted with Ava II (EC 3.1.23.4). It formed a ladder with a repeating fragment unit of 181 base pairs long. With the enzymes tested, no differences in restriction patterns were observed among clones of Alnus incana (AI-2 vs AI-2), Betula papyrifera (BP-4 vs BP-8) and subclones of Ainus glutinosa AG-1 (PLFJ709 vs LF1709), suggesting genetic stability of the Betulaceae cultures.     

中国桤木属植物的细胞学研究(Ⅰ)

对桦木科桤木属自然分布于中国西南地区的4个种进行了细胞学研究,结果分别为:川滇桤木K(2n)=56=30m+24sm+2st;桤木K(2n)=56=38m+16sm+2M;蒙自桤木K(2n)=56=28m+26sm+2st;毛桤木K(2n)=56=42m+14sm;其中川滇桤木的核型属于2A型,其余均为2B型;上述种类染色体数目均为2n=56。4种材料的核型均为首次报道。     

Features of the intrageneric Alnus-Frankia specificity

Host specificity between local Frankia strains and native alders [Alnus incana (L.) Moench and A. glutinosa (L.) Gaertn.] was evaluated in inoculation experiments. Pure cultures of Frankia , whether originating from A. incana or A. glutinosa , were infective and effective on both host species. These pure cultures were isolated from spore-negative (Sp⁻) nodules. From spore-positive (Sp⁺) nodules no Frankia isolates were obtained. This strain type resisted all our isolation attempts and therefore crushed nodules had to be used for Sp⁺ type inoculations.
The Sp⁺ type Frankia populations differed in their host specificity. Sp⁺ nodules from A. glutinosa were effective on both alder species, but Sp⁺ nodules from A. incana induced effective nodules only on the original host; on A. glutinosa only small (1-3mm) prenodule-like structures were found. Such A. glutinosa plants died on N-free medium, thus showing that these nodules were ineffective. In the effective nodules the middle cortex was dominated by infected cells filled with vesicle clusters. In the ineffective nodules only a few cortical cells were infected and sporangia predominated in these cells. Surprisingly enough they also contained vesicle-like structures as demonstrated in electron micrographs.     

欧洲赤杨根瘤及其内生菌的形态学研究

对在云南生长的欧洲赤杨自然发生的根瘤及其内生菌──Ｆｒａｎｋｉａ进行了光学显微镜和扫描电镜的观察,结果表明,这种根瘤与原产地生长的欧洲赤杨所结根瘤十分相似;与原产地在云南的蒙自桤木的根瘤结构也很相近。从这种根瘤中分离到的内生菌的纯培养具有典型的Ｆｒａｎｋｉａ的特征。     

De novo discovery and multiplexed amplification of microsatellite markers for black alder (Alnus glutinosa) and related species using SSR-enriched shotgun pyrosequencing

Recent developments in sequencing technologies and bioinformatics analyses provide an unprecedented opportunity for cost and time effective high quality microsatellite marker discovery in nonmodel organisms for which no genomic information is available. Here, we use shotgun pyrosequencing of a microsatellite-enriched library to develop, for the first time, microsatellite markers for Alnus glutinosa, a keystone tree species of European riparian woodland communities. From a total of 17?855 short sequences, we identified 590 perfect microsatellites from which 392 had designed primers. A subset of 48 loci were tested for amplification, 12 of which were polymorphic in A. glutinosa. These 12 loci were successfully coamplified in a single multiplex polymerase chain reaction experiment and validated for population genetics applications. In addition, 10 and 8 of these microsatellites were found to be transferable to the related A. incana and A. cordata species. The developed multiplex of 12 microsatellite markers therefore provides new opportunities for experimental evolutionary and forest genetics research in Alnus.     

Diversity and specificity of Frankia strains in nodules of sympatric Myrica gale, Alnus incana, and Shepherdia canadensis determined by rrs gene polymorphism

The identity of Frankia strains from nodules of Myrica gale, Alnus incana subsp. rugosa, and Shepherdia canadensis was determined for a natural stand on a lake shore sand dune in Wisconsin, where the three actinorhizal plant species were growing in close proximity, and from two additional stands with M. gale as the sole actinorhizal component. Unisolated strains were compared by their 16S ribosomal DNA (rDNA) restriction patterns using a direct PCR amplification protocol on nodules. Phylogenetic relationships among nodular Frankia strains were analyzed by comparing complete 16S rDNA sequences of study and reference strains. Where the three actinorhizal species occurred together, each host species was nodulated by a different phylogenetic group of Frankia strains. M. gale strains from all three sites belonged to an Alnus-Casuarina group, closely related to Frankia alni representative strains, and were low in diversity for a host genus considered promiscuous with respect to Frankia microsymbiont genotype. Frankia strains from A. incana nodules were also within the Alnus-Casuarina cluster, distinct from Frankia strains of M. gale nodules at the mixed actinorhizal site but not from Frankia strains from two M. gale nodules at a second site in Wisconsin. Frankia strains from nodules of S. canadensis belonged to a divergent subset of a cluster of Elaeagnaceae-infective strains and exhibited a high degree of diversity. The three closely related local Frankia populations in Myrica nodules could be distinguished from one another using our approach. In addition to geographic separation and host selectivity for Frankia microsymbionts, edaphic factors such as soil moisture and organic matter content, which varied among locales, may account for differences in Frankia populations found in Myrica nodules.     

不同光环境对桤木幼苗生长和光合特性的影响

通过设置3个光照强度(100%、56.2% 和12.5%),模拟森林幼苗生长的旷地(采伐迹地)、林窗和林下光环境,研究不同光照强度对外来种台湾桤木和乡土种四川桤木幼苗的生长、光合特性以及生物量积累与分配的影响.结果表明： 低光环境限制了两种桤木幼苗形态指标的增长,适当遮荫的林窗环境比旷地更有利于幼苗的生长.台湾桤木幼苗具有较高的比叶面积和相对生长速率,较大的单叶面积、叶长、叶宽、株高和基径,较少的叶片数和较低的叶面积比、叶柄长.低光环境下,台湾桤木幼苗的最大净光合速率、光饱和点和表观光量子效率较高,光补偿点和暗呼吸速率较低.随着光照强度的降低,台湾桤木幼苗具有更高的根生物量比和更低的叶生物量比;四川桤木幼苗则相反,加剧了动物取食和机械损伤的风险.     

Evidence that some Frankia sp. strains are able to cross boundaries between Alnus and Elaeagnus host specificity groups.

Phenotypic and genotypic methods were used to prove the existence of Frankia strains isolated from an Elaeagnus sp. that are able to cross the inoculation barriers and infect Alnus spp. also. Repeated cycles of inoculation, nodulation, and reisolation were performed under axenic conditions. Frankia wild-type strain UFI 13270257 and three of its coisolates did exhibit complete infectivity and effectiveness on Elaeagnus spp. and Hippophaë rhamnoides and variable infectivity on Alnus spp. Microscopical observation of host plant roots showed that these strains are able to infect Alnus spp. by penetrating deformed root hairs. Reisolates obtained from nodules induced on monoxenic Alnus glutinosa, Alnus incana, and Elaeagnus angustifolia resembled the parent strains in host infectivity range, in planta and in vitro morphophysiology, isoenzymes, and nif and rrn restriction fragment length polymorphisms, thus fulfilling Koch's postulates on both host plant genera. Alnus and Elaeagnus group-specific polymerase chain reaction DNA amplifications, DNA-DNA hybridizations, and partial gene sequences coding for 16S rRNA provided evidence for the genetic uniformity of wild-type strains and their inclusion into one and the same genomic species, clearly belonging to the Elaeagnus group of Frankia species.