天门冬科黄精族细胞学研究进展

在全面收集和整理黄精族染色体数据的基础上,对国内外有关黄精族各类群间的染色体数目和倍性的变化规律进行了总结,并从染色体的多倍化和非整倍化与系统发育关系和地理分布方面探讨了黄精族内各属的起源和演化关系问题。黄精族包括黄精属、舞鹤草属、异黄精属和竹根七属,共约100余种,其中舞鹤草属(x=18)、异黄精属(x=16)和竹根七属(x=20)的染色体基数稳定,而黄精属染色体基数波动较大,主要为x=8~16,既有多倍化也有非整倍化现象。染色体数据表明黄精族4个属的染色体进化模式各不相同,揭示了黄精族内染色体从高基数向低基数演化的规律;各属内染色体的演化主要是体现在二倍体水平上的核型变异,多倍化在本族中不占主导地位;仅黄精属内伴有非常强烈的非整倍化现象;细胞学证据与分子系统发育的结果比较吻合,为黄精族内属间以及属下的系统发育与进化提供了重要的参考资料。     

叶表皮及种皮特征在黄精族系统学研究中的应用

通过光学显微镜和扫描电子显微镜对黄精族trib．Polygonateae 7属79种以及相关类群12属15种 的叶下表皮形态及种皮微形态进行了观察。结果表明广义黄精族植物的叶表皮形态和种皮形态可分别 分为4种类型和6种类型。在黄精族中,鹿药属Smilacina和黄精属Polygonatum的叶表皮和种皮特征在 属内表现出一定的多样性,据此可将黄精属植物分为两类：第一类多表现为叶表皮细胞形状不规则,其垂周壁为波曲形或无皱褶但弯曲,种皮表面浅穴状;另一类叶表皮细胞形状为长方形或菱形,其垂周壁 直或无皱褶但弯曲,种皮表面具脊状突起或网状结构。其中,叶表皮细胞垂周壁无皱褶但弯曲为过渡 类型,在两类植物中均有表现。本研究结果还显示出竹根七属Disporopsis和黄精属的互叶类以及鹿药属 同具有波状垂周壁的叶表皮细胞和穴型种皮。舞鹤草属Maianthemum和鹿药属的S．stellata,S．trifolia 等的种皮特征相似。万寿竹属Disporum的叶表皮特征在属内表现得相当一致,但种皮特征在东亚分布 的种和北美分布的种之间区别明显。扭柄花属Streptopus叶表皮和种皮特征在属内没有分化。卵叶扭 柄花S．ovalis的叶表皮和种皮特征与属内其它种之间没有区别,确证了它在本属中的位置。扭柄花属、 万寿竹属、七筋菇属Clintonia与黄精族其它类群差别较大,但前两者与Uvulariaceae科的油点草属Tricyr- tis和细钟花属Uvudaria较为接近,从而支持了Dahlgren将其移至Uvulariaceae的观点。而在与外类群的关系中,铃兰族的铃兰属Convallaria与黄精族具相近的叶表皮和种皮特征。     

中国舞鹤草属新记录种

报道中国舞鹤草属一新记录种——两色鹿药[Maianthemum bicolor(Nakai)Cubey],经解剖、切片和扫描电镜观察,采自吉林省通化县白鸡峰上的两色鹿药与目前国内的舞鹤草属近缘种存在显著形态差异。     

安徽黄精属(Polygonatum)植物的分支分析

以形态学为依据 ,结合细胞分类学、叶表皮和花粉形态的研究成果 ,用分支分析的方法探讨了安徽产黄精属 11种植物的种间演化关系。在分支分析中 ,选择万寿竹属、舞鹤草属和竹根七属作为复合外类群。根据复合外类群比较原则和一般演化规律 ,确定性状极性。结果表明 :( 1)琅琊黄精与长苞黄精为姊妹群 ,并与其余 9种黄精明显分为 2大支 ;( 2 )另一支 9种黄精中 ,玉竹、长梗黄精和金塞黄精亲缘关系密切并与其余 6种再分为 2支 ;( 3)剩下 6种黄精中 ,距药黄精和多花黄精 2种互生叶黄精极为密切并与 4种轮生叶黄精分为 2组 ;( 4 ) 4种轮生叶黄精中 ,黄精、轮叶黄精和安徽黄精亲缘关系最为密切     

基于叶绿体DNA序列分析东亚七筋姑的遗传进化

通过对东亚七筋姑(Clintoniaudensis Trautv.etMey.)19个居群的cpDNA基因间隔区(cpDNA intergenic regions)rpl20-rps12和trnL-F进行序列分析,结果表明,当空位(Gap)作缺失处理时,trnL-F全序列排序后的长度为754个位点,G C含量为35.2%～35.8%,rpl20-rps12序列排序后长度为814bp,G C含量为34.1%～34.3%.聚类结果显示四倍体居群没有单独聚为一支,其多倍体可能是同源多倍体,起源方式属于异地多起源.     

铃兰族(广义)花粉形态与叶表皮特征的研究

对铃兰族(广义)7个属分别作了花粉(17种)扫描电镜观察和叶表皮(12种)的光学显微镜和扫描电镜观察。花粉可分为8个类型。在狭义的铃兰族的4个属内,花粉全为远极单槽,舟状。它们的外壁除夏须草属外,都具细孔。夏须草属的花粉外壁则为细皱。 Hutchinson(1934)的蜘蛛抱蛋族花粉形态变异很大,其中开口箭属和万年青属的花粉为远极单槽,舟形,外壁具穿孔或网纹,而蜘蛛抱蛋属的花粉则为球状,无萌发孔。其间的显著差异支持Nakai为前两个属建立万年青族(Rohdeae)。表1归纳了7个属的花粉形态;图1是我们对铃兰族(广义)花粉形态演化的见解。叶表皮观察表明,气孔器为无规则型,上表皮角质层主要为条纹加厚,或均匀加厚,而铃兰属的角质层秕糠状加厚。7个属的叶表皮特征归纳于表2。     

基于线粒体COⅡ和Cytb基因序列的8种漠甲系统发育关系

对8种漠甲昆虫线粒体Cytb基因579bp和COⅡ基因585bp的序列片段进行联合分析。结果表明,8种甲虫在长度为1164bp的序列中碱基T,C,A,G的平均含量分别为33.7%,21.2%,33.5和11.6%,A+T平均含量明显高于G+C含量。密码子第三位点A+T含量高达77.2%,而该位点G的平均含量仅为4.0%。序列中碱基替换多发生于第三位点,转换略多于颠换,转换主要以C←T为主,颠换以A←T为主。以土甲族的Eumylada potanini为外群构建的系统发育树表明:漠王和漠甲以很高的置信值聚为一支,二者的关系与Bouchard最新的分类观点相吻合,即漠王族并入漠甲族成为一族;鳖甲族在支序图上另成一支,可视为一个单系群。     

后丽盲蝽属作为属级分类单元的DNA分子证据

用蛋白酶K消化法提取并正反链双向测得半翅目盲蝽科盲蝽族(Mirini)中后丽盲蝽属(Apolygus)和草盲蝽属(Lygus)9个种的线粒体DNA细胞色素b基因片段的序列,应用Mega计算该两个属7个种11个Cyt b 432 bp片段序列的碱基组成、核苷酸的种间变异率、所编码的144个氨基酸的变异率。其核苷酸变异率：后丽盲蝽属内小于0．02,草盲蝽属内小于0．08(个别种例外);两个属种类之间平均变异率都大于0．10,平均为0．15。氨基酸变异率：后丽盲蝽属内种间无变异,而草盲蝽属内只有L．gemellatus与其他种间有13个氨基酸变异,仅占9．03％;两属各种类间却存在8～11个氨基酸的变异。表明在分子水平上两属之间较之于属内种间具有明显的差异,支持将后丽盲蝽从亚属提升为属级分类单元的分类观点。     

基于RAPD技术探讨黄精属部分药用植物系统位置

目的：从分子水平探讨黄精属部分药用植物间的系统位置。方法：采用RAPD分子标记方法,以同科万寿竹属植物宝铎草（Disporum sessile D．Don）为外类群,对百合科黄精属19批（6种）药用植物进行基因组DNA的多态性分析,通过聚类分析探讨黄精种属间亲缘关系。结果：筛选出7条随机引物,聚类结果表明,黄精属植物在属级分类上特征明显,但属内分类有交叉。结论：RAPD技术可以为黄精属的分类和种的鉴别提供一定的理论依据。     

雷州地区马尾藻系统进化分析

通过PCR扩增测序及种间比对,对获得的6种马尾藻18SrRNA、COI和ITS基因部分同源序列进行分析,结果显示:(1)18S rRNA基因同源序列长度为1 673 bp,其中保守位点1 668个,可变位点3个,简约信息位点1个,单变异多态位点2个;T、C、A、G的平均含量分别为26.4％、21.4％、24.4％、27.8％.(2)COI同源序列长度为643 bp,其中保守位点567个,可变位点76个,简约信息位点28个,单变异多态位点47个;T、C、A、G的平均含量分别为40.0％、17.6％、19.1％、23.3％.(3)ITS同源序列长度为1 539 bp,其中保守位点1 272个,可变位点193个,简约信息位点55个,单变异多态位点138个;T、C、A、G的平均含量分别为23.5％、26.5％、17.1％、32.9％.(4)基于Kimnra双参数模型计算遗传距离,选用褐藻纲部分物种为外源,构建18S rRNA,COI和ITS基因序列系统发育树.用COI和ITS基因构建的发育树与利用形态学进行的分类较为一致,真马尾藻亚属(Sargassum)的张氏马尾藻(Sargassum zhangii)、全缘马尾藻(Sargassum integerrimum)、匍枝马尾藻(Sargassum polycystum)、亨氏马尾藻(Sargassum henslowianum)、灰叶马尾藻(Sargassum cinereum)亲缘关系较近,先聚为一支,再与反曲叶亚属的半叶马尾藻(Sargassum hemiphyllum)聚为一支,然后与其他亚属的聚为一大支;而用18S rRNA基因构建的发育树中,亨氏马尾藻先与半叶马尾藻聚为一支,后与真马尾藻亚属的聚为一支,最后与其他亚属的聚为一大支.从序列的保守度分析,18SrRNA基因具有高度保守性,可用于属以上单元鉴别分类;COI和ITS基因多态性较高,可用于种间分类.     

Molecular systematic studies on the tribe Polygonateae (s. l.) in China based on RFLPs data of PCR-amplified chloroplast DNA fragments

Phylogenetic relationships in the tribe Polygonateae s. l. of Liliaceae were investigated with an RFLP analysis of two PCR-amplified chloroplast genome DNA fragments. One fragment is the transfer RNA gene for lysine( trnK gene) including the maturase-encoding gene matK, and the other is the rpl16 gene which codes for a chloroplast ribosomal protein in large subunit. The trnK gene is ca. 2600 bp in length in all the taxa, but the rpl16 gene ranges from ca. 1140 bp to ca. 1320 bp in length among different genera and in Polygonatum. The results suggested that the tribe Polygonateae s. str. consisting of Polygonatum, Disporopsis, Smilacina and Maianthemum is closely related to Convallaria in Convallarieae, and supported the earlier results that Streptopus and Disporum should be removed out from the tribe Polygonateae s. l.. The results also showed that Polygonatum formed a clad with Disporopsis, while Smilacina lumped with Maianthemum in the tribe Polygonateae s. str.. In addition, the present study supports the view that S. ginfoshanicumshould be transferred from Smilacina to Polygonatum.     

Phylogeny and biogeographic diversification of Maianthemum (Ruscaceae: Polygonatae)

Maianthemum (Ruscaceae) comprises 28-38 species and includes the two traditionally recognized genera: Maianthemum sensu stricto and Smilacina. Thirty-seven samples representing 22 species of Maianthemum and six closely related outgroup taxa were sequenced for eight chloroplast and nuclear markers (trnL-F, rps16, rpl16, psbA-trnH, rbcL, ndhF, trnK, and ITS) with a total length of nearly 10,000 base pairs. Phylogenetic analyses supported the monophyly of Maianthemum with Maianthemum sensu stricto nested within Smilacina. Almost all species from the eastern Himalayan region in SW China except for Maianthemum tatsienense and M. stenolobum form a well supported clade. This clade is characterized morphologically by short filaments and large anthers, relatively large flowers, and pubescent stems and leaves. Maianthemum tatsienense and M. stenolobum from SW to central China form another clade. The other species from eastern Asia (central to NE China and Japan) and the New World fall into several clades. The intercontinental disjunction between eastern Asia and North America in Maianthemum sensu stricto is estimated to be at 1.68 million years ago (mya) with the Bayesian relaxed clock relying on uncorrelated rates. A recent radiation at about 2.04mya is suggested in the high mountains of SW China, corresponding to the geographical heterogeneity in that region after the uplift of the Himalayas. Long distance dispersal by birds may have facilitated the evolution of their intercontinental disjunction and their biogeographic diversifications in SW China.     

Phylogenetic relationships among genera in theLiliaceae-Asparagoideae-Polygonatae s.l. inferred fromrbcL gene sequence data

The chloroplast gene encoding ribulose-1,5-bisphosphate-carboxylase (rbcL) was sequenced for phylogenetic analysis of 13 species (10 genera) in the tribePolygonatae s.l. of theLiliaceae-Asparagoideae. The data were analysed using maximum parsimony and neighbour-joining methods. There were 233 phylogenetically informative sites out of 1368 base pairs compared. The results suggest that there are three monophyletic groups withinPolygonatae s.l. with high bootstrap confidence values. Group A representsPolygonatae s.str., with generaMaianthemum, Smilacina, Convallaria, Disporopsis, andPolygonatum. Group B containsUvularia andDisporum and group C includesStreptopus, Tricyrtis, Clintonia, andProsartes. The study suggests thatPolygonatae s.l. are not a monophyletic group, including at least three groups of different phylogenetic origin. Monophyly of the taxa within groups A, B, and C is supported by the high bootstrap confidence values (85–100%) of the bootstrap replications for both parsimony and neighbour-joining methods. The differences between each group (calculated as 100x base substitutions per site) were 6.99–9.03 for group A and B, 4.92–7.35 for A and C, and 6.66–7.57 for B and C.     

Euglena gracilis chloroplast ribosomal protein operon: a new chloroplast gene for ribosomal protein L5 and description of a novel organelle intron category designated group III

We describe the structure (3840 bp) of a novel Euglena gracilis chloroplast ribosomal protein operon that encodes the five genes rpl16-rpl14-rpl5-rps8-rpl36. The gene organization resembles the spc and the 3'-end of the S10 ribosomal protein operons of E. coli. The rpl5 is a new chloroplast gene not previously reported for any chloroplast genome to date and also not described as a nuclear-encoded, chloroplast protein gene. The operon contains at least 7 introns. We present evidence from primer extension analysis of chloroplast RNA for the correct in vivo splicing of five of the introns. Two of the introns within the rps8 gene flank an 8 bp exon, the smallest exon yet characterized in a chloroplast gene. Three introns resemble the classical group II introns of organelle genomes. The remaining 4 introns appear to be unique to the Euglena chloroplast DNA. They are uniform in size (95-109 nt), share common features with each other and are distinct from both group I and group II introns. We designate this new intron category as 'group III'.     

油菜叶绿体16SrRNA基因的一级结构分析

本文报道了油菜叶绿体16S rRNA基因的全顺序及其5′端上游的156bp和3′端下游的101bp的核苷酸顺序。油菜叶绿体16s rRNA基因长为1491bp,和烟草、玉米相比,同源程度分别为98.5％、96.1％。油菜叶绿体16S rRNA基因5′端上游及3′端下游的顺序能互补而形成一个较大的茎环结构,但与烟草相比,由于3′端下游顺序有79bp的缺失,因此,该结构中的茎部分大小仅为烟草的二分之一。