猕猴桃属植物叶绿体基因PCR-RFLP分析

分别用2个不同限制性内切酶对猕猴桃属27个种和15个栽培品种的叶绿体基因(rbcL基因和psbA基因)的PCR扩增产物进行酶切分析,共得到25个限制性位点,其中24个具有多态性。确立了该属植物的单元型分布,对该属植物的系统发育方式和部分重要种类的亲缘关系进行了探讨,拓展了可用于该属植物分子系统学研究的遗传信息。     

基于线粒体和核基因序列的蜜蜂属系统发育分析

文章测定了中国分布的蜜蜂属(Apis)5种蜜蜂22个样本的线粒体基因ND2、CO2、16S rRNA以及核基因ITPR的序列,对序列的碱基组成和蜜蜂种间的遗传距离进行了分析。结合下载的蜜蜂属其他4个种的相关序列,采用最大简约法、邻接法和最大似然法重建了蜜蜂属系统发育关系。系统发育分析结果支持蜜蜂属划分为3个类群,即小蜜蜂类群(包括小蜜蜂和黑小蜜蜂)、大蜜蜂类群(包括大蜜蜂和黑大蜜蜂)和穴居蜜蜂类群(西方蜜蜂、东方蜜蜂、沙巴蜂、苏拉威西蜂、绿努蜂),且小蜜蜂类群较早分化。结果还显示,我国海南岛的大蜜蜂和大陆的大蜜蜂之间可能存在较大的遗传分歧。     

木兰族和含笑族的系统学关系——基于叶绿体matK基因序列分析

采用PCR直接测序法,对木兰属国产组(各1种)和含笑属各组(各1种)的叶绿体DNA,matK基因序列进行测定分析,结果表明：木兰属和含笑属不能明显区分开,故不支持以顶生花和腋生花来划分木兰族和含笑族。     

编码叶绿体蛋白质的某些核基因

高等植物叶绿体的一些重要蛋白质基因位于细胞核的基因组内。这些基因的细微结构已研究得较为清楚,并对不同植物的编码叶绿体蛋白质的核基因进行了比较研究。     

楤木属及其近缘类群叶绿体基因组比较与系统发育分析

分别对楤木属（Aralia）和羽叶参属（Pentapanax）各两个物种进行叶绿体基因组测序、组装和注释,并进行结构分析;结合NCBI下载的近缘类群叶绿体组序列,进行系统发育分析。结果显示,4个物种的叶绿体基因组均为环状四分体结构,长度为155 744 ~ 156 201 bp,GC含量为38.1%,均包含132个基因,其中蛋白质编码基因87个,rRNA基因8个,tRNA基因37个。边界分析发现IR（Inverted repeat）区均未发生收缩和扩张。SSR（Simple sequence repeat）序列数量在39 ~ 43个,多为单核苷酸和二核苷酸重复,位置多在非编码区。序列差异多出现在LSC（Large single copy）和SSC（Small single copy）区的非编码区。最大似然树揭示出两个高度支持的单系分支：第1分支包括羽叶参属、浓紫龙眼独活（Aralia atropurpurea Franch.）、食用土当归（Aralia cordata Thunb.）和东北土当归（Aralia continentalis Kitagawa）;第2分支则均由楤木属物种组成。总体来说,楤木属和羽叶参属植物的叶绿体基因组序列比较保守;系统发育分析结果支持羽叶参属置入楤木属内部以及楤木属的各个种成单系。     

基于叶绿体psaA和psbA基因的中国熊野藻属植物系统发育分析

以中国产熊野藻属Kumanoa的两个种, 绞扭熊野藻K. intorta (=绞扭串珠藻Batrachospermum intortum), 弯形熊野藻K. curvata (=弯形串珠藻B. curvatum)和其他6种淡水红藻为实验材料, 对其psaA和psbA基因进行扩增和测序, 并与GenBank中相近序列进行比对分析, 以贝叶斯法、最大似然法和邻接法分别构建了单基因和联合基因系统发育树. 结果表明, 3种方法构建的系统树具有相似的拓扑结构, 反映的系统发育关系基本一致, 熊野藻属中的两个种聚为一支, 与串珠藻属相分离, 支持该属的建立; 中国产的熊野藻属分子学研究结果与来自南美洲及澳洲的该属植物结果一致, 说明该属的建立具有广泛的地理适用性. 系统发育树聚类结果也明确反映了熊野藻属与串珠藻属较近的亲缘关系, 根据果胞枝形态特点, 推测熊野藻属进化地位晚于串珠藻属植物, 而早于顶丝藻目和红索藻目. 此外, 胶串珠藻与其他串珠藻组植物分离, 支持将其单独分组, 红索藻目植物与串珠藻目植物分离, 支持红索藻目的建立. 同时也表明psaA和psbA基因用于淡水红藻分析, 能够较好地反映其系统发育关系.       

里白科植物的系统发育和分歧时间估计——基于叶绿体三个基因序列的证据

里白科(Gleicheniaceae)是古老的真蕨类植物,最早的化石记录可追溯到石炭纪。现存类群的属级分类和系统演化关系一直存在很大的分歧,为了进一步探讨该类群的起源演化,文中运用最大简约法和贝叶斯演绎方法对18种代表现存里白科植物全部6属(包括新测的12种)的叶绿体3个编码基因序列(atpB,rbcL和rps4)进行分析,探讨其主要分类群(属级)的系统演化关系。结果显示,里白科植物为一个单系群,由3个分支构成:里白属(Diplopterygium Nakai)和Gleichenia japonica构成一个分支;芒萁属(Dicranopteris Bernh.)和Gleichenella pec-tinata构成另一个分支;假芒萁属(Sticherus C.Presl)与单种属Stromatopteris Mettenius及Gleichenia dicarpa构成第三个分支。用宽松分子钟方法推测里白科主要类群的起源时间为:现代里白科植物起源于早白垩世(111—140Ma),其主要分支类群随后发生多样性分化,里白属和芒萁属的快速辐射演化均发生在古近纪(40—64Ma,36—50Ma)。起源时间的估算结果暗示化石种三叠里白Diplopterygium triassica不应归入现代里白属,其归属需要重新考虑。     

基于叶绿体DNA atpB-rbcL区序列探讨石蒜属种间系统发育关系

摘要：为了探讨石蒜属（Lycoris Herb.）的种间系统发育关系,对石蒜属95个材料包括15种、4变种及2个人工杂种的叶绿体 DNA atpB-rbcL间隔区进行了测序,结合花部形态和核型特征,探讨了石蒜属种间系统关系及其可能的杂交起源,结果表明：在系统发育树上亲缘关系近的材料聚在一起,其中矮小石蒜（L. radiata var. pumila）和换锦花（L. sprengeri）与2个人工杂交种（Hybrid 1、Hybrid 2）、麦秆石蒜（L. straminea）、江苏石蒜（L. houdyshelii）、短蕊石蒜（L. caldwellii）和乳白石蒜（L. albiflora）具有密切的亲缘关系。atpB-rbcL序列揭示的石蒜属种间关系与染色体核型的分类结果部分一致,主要表现在具有近端部着丝粒(A)染色体的种与具有中部(M)和端部(T)着丝粒染色体的种各成一支,与形态和染色体分类结果一致;不同之处在于具有中部、端部和近端部着丝粒染色体的种分散在两个主要分支内,进一步验证了具有中部、端部和近端部3种着丝粒类型染色体组的石蒜如麦秆石蒜、江苏石蒜、短蕊石蒜和乳白石蒜等是杂交起源的假设,结合2个人工杂交种分析,揭示了短蕊石蒜和乳白石蒜的近端部着丝粒染色体来源于换锦花;麦秆石蒜和江苏石蒜近端部着丝粒染色体来源于矮小石蒜。     

基于多基因序列和形态性状的牡丹组种间关系

牡丹被认为是中国的国花,具有很高的医学、观赏和经济价值.野生牡丹被认为是栽培牡丹的野生祖先,因此弄清牡丹组的种间亲缘关系具有重要的理论和实践意义.由于受到信息量的限制,根据单基凼数据或形态数据往往无法对牡丹组的种间关系得到明确的结果.本研究用12份样品代表野生牡丹组(Paeonia section Moutan DC.,Paeoniaceae)8个种,利用包括核基因(Adh1A、Adh2和GPAT)和叶绿体基因(trnS-trnG和rps16-trnQ)的DNA序列以及形态性状的多套数据来探讨野牛牡丹的种间关系.合并分析得到具高支持率的牡丹组物种间的系统发育关系.结果表明,芍药属牡丹组8个野生种分为两个亚组,即肉质花盘亚组subseet.Delavayanae和革质花盘亚组subsect.Vaginatae.肉质花盘亚组包括滇牡丹P delavayi和大花黄牡丹P.ludlowii;革质花盘亚组包括其余6个种.革质花盘亚组中,四川牡丹P.decomposita ssp.decomposita和紫斑牡丹P. rockii ssp. rockii关系密切;卵叶牡丹P.qiui和矮牡丹P. jishanenMs关系密切;银屏牡丹P. suffruticosa ssp.yinpingmudan与风丹P. ostii关系 密切,并且后两个分支为姊妹群.     

猬草及其近缘属植物单拷贝核Pgk1基因序列的系统发育分析

文章对禾本科小麦族猬草属及其近缘属Thinopyrum(Eb)、Lophopyrum(Ee)、拟鹅观草属(St)、新麦草属(Ns)、大麦属(H)、赖草属(NsXm)和披碱草属(StH)植物共23个类群的单拷贝核Pgk1基因序列进行系统发育分析, 探讨猬草属及其近缘属植物的系统发育关系。序列分析发现Pgk1基因序列在L. arenarius和Psa. juncea中有81 bp的Stowaway家族DNA转座元件插入, 而在Hy. duthiei、Hy. duthiei ssp. longearistata和L. akmolinensis中有29 bp Copia家族的反转录转座元件插入。最大似然和贝叶斯推断进行的系统发育分析表明: (1)猬草属模式种Hy. patula与披碱草属、拟鹅观草属和大麦属具有密切的亲缘关系; (2)猬草属的其他物种Hy. duthiei、Hy. duthiei ssp. longearistata、Hy. coreana和Hy. komarovii与新麦草属和赖草属植物亲缘关系密切。研究结果支持将Hy. patula从猬草属组合到披碱草属中, 而Hy. duthiei、Hy. duthiei ssp. longearistata、Hy. coreana和Hy. komarovii应组合到赖草属中。     

基于形态学性状的洪平杏与近缘种种间关系研究

本研究通过野外调查获得洪平杏（Armeniaca hongpingensis T. T. Yu & C. L. Li）、杏（A. vulgaris Lam.）、梅（A. mume Siebold）和杏梅（ A. mume var. bungo Makino）有关的叶、花、果、果核及枝条的21个定量性状和15个定性性状,并对这些数据进行了聚类分析。结果显示,无论是PAM聚类还是UPGMA聚类分析都可清晰地将4种杏属植物分开,说明物种之间具有明显的区分特征。主成分分析的前3个主成分中,一年生枝颜色、被毛情况,叶及果实被毛,叶形指标（叶形、叶尖、叶基）和果核表面特征的绝对权重值都在0.9以上,表明这些性状在4种杏属植物的分类中具有重要的作用。电镜扫描观察结果发现,供试花粉均是单粒花粉,等极,辐射对称,具3孔萌发沟。杏、梅、杏梅的花粉表面纹饰均为条纹状,但洪平杏的表面纹饰为间断条纹,且有不明显穿孔。4种植物叶上表皮细胞均具条纹状角质增厚。与其他种相比,洪平杏细胞表面条纹常弯曲、交叠,无固定走向。根据聚类分析结果和微形态特征,支持将洪平杏作为区别于梅和杏的一个独立种的处理。     

The phylogenetic position of red algae revealed by multiple nuclear genes from mitochondria-containing eukaryotes and an alternative hypothesis on the origin of plastids

Abstract Red algae are one of the main photosynthetic eukaryotic lineages and are characterized by primitive features, such as a lack of flagella and the presence of phycobiliproteins in the chloroplast. Recent molecular phylogenetic studies using nuclear gene sequences suggest two conflicting hypotheses (monophyly versus non-monophyly) regarding the relationships between red algae and green plants. Although kingdom-level phylogenetic analyses using multiple nuclear genes from a wide-range of eukaryotic lineages were very recently carried out, they used highly divergent gene sequences of the cryptomonad nucleomorph (as the red algal taxon) or incomplete red algal gene sequences. In addition, previous eukaryotic phylogenies based on nuclear genes generally included very distant archaebacterial sequences (designated as the outgroup) and/or amitochondrial organisms, which may carry unusual gene substitutions due to parasitism or the absence of mitochondria. Here, we carried out phylogenetic analyses of various lineages of mitochondria-containing eukaryotic organisms using nuclear multigene sequences, including the complete sequences from the primitive red alga Cyanidioschyzon merolae. Amino acid sequence data for two concatenated paralogous genes (α- and β-tubulin) from mitochondria-containing organisms robustly resolved the basal position of the cellular slime molds, which were designated as the outgroup in our phylogenetic analyses. Phylogenetic analyses of 53 operational taxonomic units (OTUs) based on a 1525-amino-acid sequence of four concatenated nuclear genes (actin, elongation factor-1α, α-tubulin, and β-tubulin) reliably resolved the phylogeny only in the maximum parsimonious (MP) analysis, which indicated the presence of two large robust monophyletic groups (Groups A and B) and the basal eukaryotic lineages (red algae, true slime molds, and amoebae). Group A corresponded to the Opisthokonta (Metazoa and Fungi), whereas Group B included various primary and secondary plastid-containing lineages (green plants, glaucophytes, euglenoids, heterokonts, and apicomplexans), Ciliophora, Kinetoplastida, and Heterolobosea. The red algae represented the sister lineage to Group B. Using 34 OTUs for which essentially the entire amino acid sequences of the four genes are known, MP, distance, quartet puzzling, and two types of maximum likelihood (ML) calculations all robustly resolved the monophyly of Group B, as well as the basal position of red algae within eukaryotic organisms. In addition, phylogenetic analyses of a concatenated 4639-amino-acid sequence for 12 nuclear genes (excluding the EF-2 gene) of 12 mitochondria-containing OTUs (including C. merolae) resolved a robust non-sister relationship between green plants and red algae within a robust monophyletic group composed of red algae and the eukaryotic organisms belonging to Group B. A new scenario for the origin and evolution of plastids is suggested, based on the basal phylogenetic position of the red algae within the large clade (Group B plus red algae). The primary plastid endosymbiosis likely occurred once in the common ancestor of this large clade, and the primary plastids were subsequently lost in the ancestor(s) of the Discicristata (euglenoids, Kinetoplastida, and Heterolobosea), Heterokontophyta, and Alveolata (apicomplexans and Ciliophora). In addition, a new concept of “Plantae” is proposed for phototrophic and nonphototrophic organisms belonging to Group B and red algae, on the basis of the common history of the primary plastid endosymbiosis. The Plantae include primary plastid-containing phototrophs and nonphototrophic eukaryotes that possibly contain genes of cyanobacterial origin acquired in the primary endosymbiosis.     

One species or two? Multilocus analysis of nucleotide variation of Melastoma penicillatum and Melastoma sanguineum (Melastomataceae) in Hainan,China

Melastoma comprises more than 20 shrub species and is distributed in tropical Asia and Oceania. Melastoma penicillatum, the most narrowly distributed taxon of this genus in China, was recently incorporated into another morphologically similar species, M. sanguineum. Based on its distinct morphological traits, unique habitat and flowering time, we propose that M. penicillatum should be a distinct species. In this study, we sequenced three nuclear genes and four chloroplast intergenic spacers of M. sanguineum and M. penicillatum at two locations in Hainan to test this hypothesis. There was no sequence variation at all four chloroplast intergenic spacers within and between M. sanguineum and M. penicillatum, suggesting that they should have diverged recently. However, they showed strong divergence at all three nuclear genes and no shared haplotypes were observed between them. Bayesian clustering-based STRUCTURE analysis revealed that individuals of M. sanguineum and M. penicillatum were clustered into their own clades. Fst calculations showed that both taxa had marked population genetic differentiation, but population differentiation in M. penicillatum was stronger than that in M. sanguineum. Stronger population differentiation in M. penicillatum may be caused by its discontinuous high-elevation and understory habitats, which is disadvantageous for bird-mediated dispersal. Our analysis based on the isolation-with-migration (IM) model revealed bidirectional, asymmetrical gene flow in M. sanguineum and little gene flow in M. penicillatum. Taken together, our results indicate that M. penicillatum is a distinct species and should not be incorporated into M. sanguineum. The two species might have diverged recently, and lineage sorting has been incomplete. Differential adaptation to different elevations might have led to speciation in this case.     

Probabilistic Analysis Indicates Discordant Gene Trees in Chloroplast Evolution

Abstract Analyses of whole-genome data often reveal that some genes have evolutionary histories that diverge from the majority phylogeny estimated for the entire genome. We present a probabilistic model that deals with heterogeneity among gene trees, implement it via the Gibbs sampler, and apply it to the plastid genome. Plastids and their genomes are transmitted as a single block without recombination, hence homogeneity among gene trees within this genome is expected. Nevertheless, previous work has revealed clear heterogeneity among plastid genes (e.g., Delwiche and Palmer 1996). Other studies, using whole plastid genomes of various algae and land plants, found little additional heterogeneity (Martin et al. 1998; Adachi et al. 2000). We augment the earlier studies by using a data set of 14 taxa: 6 land plants, 2 green algae, a diatom, 2 red algae and a cryptophyte, the cyanelle of the glaucocystophyte Cyanophora, and the blue–green alga Synechocystis as an outgroup. Contrary to the earlier analyses, we cannot find even a single, dominant consensus tree. Therefore, we formulate a probabilistic model that divides the genes into two sets: those that follow the consensus tree and those that have independent gene trees. No particular tree is supported by more than three-fourths of the genes. But the set of genes that follows a certain tree is fairly independent of data processing and the method of analysis. With one possible exception, we find no evidence for collinear or functionally related genes to follow similar trees. The phylogenetic pattern also seems independent of bias in amino acid composition. Among possible explanations for the observed phenomenon, the hypothesis that different genes have different covarion structures is difficult to assess. But gene duplication may be possible through the inverted or direct repeat regions, while horizontal gene transfer seems less likely. In contrast to green algae and land plants, inverted repeat regions in red algae and in Cyanophora show abundant differences among the copies. Thus, genes may get duplicated when they are recruited into the inverted repeat region and one of the two copies may be lost after leaving the inverted repeat region.     

Variability of chloroplast DNA and nuclear ribosomal DNA in cassava (Manihot esculenta Crantz) and its wild relatives

Chloroplast DNA (cp) and nuclear ribosomal DNA (rDNA) variation was investigated in 45 accessions of cultivated and wild Manihot species. Ten independent mutations, 8 point mutations and 2 length mutations were identified, using eight restriction enzymes and 12 heterologous cpDNA probes from mungbean. Restriction fragment length polymorphism analysis defined nine distinct chloroplast types, three of which were found among the cultivated accessions and six among the wild species. Cladistic analysis of the cpDNA data using parsimony yielded a hypothetical phylogeny of lineages among the cpDNAs of cassava and its wild relatives that is congruent with morphological evolutionary differentiation in the genus. The results of our survey of cpDNA, together with rDNA restriction site change at the intergenic spacer region and rDNA repeat unit length variation (using rDNA cloned fragments from taro as probe), suggest that cassava might have arisen from the domestication of wild tuberous accessions of some Manihot species, followed by intensive selection. M. esculenta subspp flabellifolia is probably a wild progenitor. Introgressive hybridization with wild forms and pressures to adapt to the widely varying climates and topography in which cassava is found might have enhanced the crop's present day variability.     

胡颓子科叶绿体基因组系统发育分析与演化趋势推断

【目的】探索胡颓子科叶绿体基因组演化趋势,为胡颓子科植物物种鉴定以及资源开发利用提供理论依据。【方法】该研究从头组装并注释了沙棘属和野牛果属共4个类群的叶绿体基因组,结合已发表的叶绿体基因组序列,比较了胡颓子科各类群叶绿体基因组的基因构成、重复序列和结构特征,建立了系统发育树,并通过高分化区定位了该科叶绿体基因组的潜在DNA条形码区域。【结果】胡颓子科各属叶绿体基因组在四分体结构、基因数量和排列上高度相似;沙棘属和野牛果属的反向重复区(IR)和整个基因组重复序列数目较胡颓子属有扩张和增加的趋势。胡颓子科18个类群的叶绿体全基因组序列的系统发育树中,胡颓子属个体聚为一支,最先分化出来,沙棘属和野牛果属各自聚为一支,具有最近的共同祖先;从长单拷贝区(LSC)和短单拷贝区(SSC)筛选出3个DNA条形码候选区,其中ycf1基因的鉴定效果最佳。【结论】胡颓子科的叶绿体基因组结构保守,但其非编码区序列在各属间存在明显差异,且IR区序列与重复序列在演化过程中分别有扩张和增多的趋势。研究选定的DNA条形码序列能很好区分胡颓子科各属之间以及胡颓子属内物种间关系。     

外源基因在莱茵衣藻叶绿体中的表达

把莱茵衣藻(Chlamydomonas reinhardtii)叶绿体作为生物反应器来表达外源基因具有广阔的应用前景。人们利用莱茵衣藻叶绿体表达体系已成功表达多种重组蛋白,其中包括人类药用蛋白。综述了莱茵衣藻叶绿体转化的方法、影响外源基因表达的主要因素以及外源基因在莱茵衣藻叶绿体表达研究进展。