The complete chloroplast genome sequence of the Chinese endemic species Sorbus setschwanensis (Rosaceae) and its phylogenetic analysis

Sorbus setschwanensis Koehne is a pinnate-leaved Sorbus s.str. species endemic to China with narrow distribution and intriguing phylogeny that needs wider attention. In this paper, the complete chloroplast (cp) genome of S. setschwanensis is reported, and its phylogenetic position is analyzed. The complete cp genome of S. setschwanensis is 160 064 bp in size with 36.50% GC content. It has a typical quadripartite structure including a pair of inverted repeat regions (IRs) of 26 378 bp that separates a large single copy (LSC) region of 86 013 bp and a small single copy (SSC) region of 19 295 bp. The cp genome encodes 108 genes, comprising 76 protein-coding genes, 28 tRNA genes and 4 rRNA genes. Additionally, 52 simple sequence repeats (SSRs) and 43 dispersed repeats were identified. Comparison of the whole cp genome with those of other Sorbus species showed an overall high degree of sequence similarity, but there are six highly variable regions (trnR-atpA, petN-psbM, ndhC-trnV, trnE-trnT, trnT-trnL and rpl32-trnL) located in intergenic spacers that may be useful as molecular markers in future population genetic and phylogenetic studies in the genus. Phylogenetic analyses based on 108 coding genes from 25 species in Rosaceae revealed that S. setschwanensis is nested within Sorbus sect. Sorbus together with other pinnately leaved species, but does not form a sister lineage to S. rufopilosa belonging to the same series Multijugae. Thus, the systematic position of S. setschwanensis and relationships with other species in the genus needs to be further studied.     

The complete chloroplast genomes of three Hamamelidaceae species: Comparative and phylogenetic analyses

Hamamelidaceae is an important group that represents the origin and early evolution of angiosperms. Its plants have many uses, such as timber, medical, spice, and ornamental uses. In this study, the complete chloroplast genomes of Loropetalum chinense (R. Br.) Oliver, Corylopsis glandulifera Hemsl., and Corylopsis velutina Hand.‐Mazz. were sequenced using the Illumina NovaSeq 6000 platform. The sizes of the three chloroplast genomes were 159,402 bp (C. glandulifera), 159,414 bp (C. velutina), and 159,444 bp (L. chinense), respectively. These chloroplast genomes contained typical quadripartite structures with a pair of inverted repeat (IR) regions (26,283, 26,283, and 26,257 bp), a large single‐copy (LSC) region (88,134, 88,146, and 88,160 bp), and a small single‐copy (SSC) region (18,702, 18,702, and 18,770 bp). The chloroplast genomes encoded 132–133 genes, including 85–87 protein‐coding genes, 37–38 tRNA genes, and 8 rRNA genes. The coding regions were composed of 26,797, 26,574, and 26,415 codons, respectively, most of which ended in A/U. A total of 37–43 long repeats and 175–178 simple sequence repeats (SSRs) were identified, and the SSRs contained a higher number of A + T than G + C bases. The genome comparison showed that the IR regions were more conserved than the LSC or SSC regions, while the noncoding regions contained higher variability than the gene coding regions. Phylogenetic analyses revealed that species in the same genus tended to cluster together. Chunia Hung T. Chang, Mytilaria Lecomte, and Disanthus Maxim. may have diverged early and Corylopsis Siebold & Zucc. was closely related to Loropetalum R. Br. This study provides valuable information for further species identification, evolution, and phylogenetic studies of Hamamelidaceae plants.     

Molecular phylogeny ofForsythia (Oleaceae) based on chloroplast DNA variation

Phylogenetic relationships of ten wild species and several cultivars ofForsythia were reconstructed based on the chloroplast (cp) DNA variation. A total of 216 cpDNA variants, 44 of which were potentially phylogenetically informative, was detected using 24 restriction endonucleases. Phylogenetic analysis usingFontanesia andAbeliophyllum as outgroups revealed four well defined species groups in the genus: 1)F. suspensa, 2)F. europaea — F. giraldiana, 3)F. ovata — F. japonica — F. viridissima, and 4)F. koreana — F. manshurica — F. saxatilis. The amount of support for each monophyletic group was evaluated by various methods including character number, decay analysis, parsimony bootstrapping, Neighbour-Joining (NJ) — bootstrapping, NJ-jackknifing, and the topology-dependent permutation tail probability (T-PTP) test. The data do not support the hybrid origin ofF. intermedia fromF. suspensa andF. viridissima. The disjunctly distributed European species,F. europaea, was identified as a sister species of the ChineseF. giraldiana and it was probably derived through recent long distance dispersal.     

The first complete chloroplast genome sequence of a lycophyte, Huperzia lucidula (Lycopodiaceae)

We used a unique combination of techniques to sequence the first complete chloroplast genome of a lycophyte, Huperzia lucidula. This plant belongs to a significant clade hypothesized to represent the sister group to all other vascular plants. We used fluorescence-activated cell sorting (FACS) to isolate the organelles, rolling circle amplification (RCA) to amplify the genome, and shotgun sequencing to 8× depth coverage to obtain the complete chloroplast genome sequence. The genome is 154,373 bp, containing inverted repeats of 15,314 bp each, a large single-copy region of 104,088 bp, and a small single-copy region of 19,657 bp. Gene order is more similar to those of mosses, liverworts, and hornworts than to gene order for other vascular plants. For example, the Huperzia chloroplast genome possesses the bryophyte gene order for a previously characterized 30 kb inversion, thus supporting the hypothesis that lycophytes are sister to all other extant vascular plants. The lycophyte chloroplast genome data also enable a better reconstruction of the basal tracheophyte genome, which is useful for inferring relationships among bryophyte lineages. Several unique characters are observed in Huperzia, such as movement of the gene ndhF from the small single copy region into the inverted repeat. We present several analyses of evolutionary relationships among land plants by using nucleotide data, inferred amino acid sequences, and by comparing gene arrangements from chloroplast genomes. The results, while still tentative pending the large number of chloroplast genomes from other key lineages that are soon to be sequenced, are intriguing in themselves, and contribute to a growing comparative database of genomic and morphological data across the green plants.     

The complete mitochondrial genome of Rhynchocypris oxycephalus (Teleostei: Cyprinidae) and its phylogenetic implications

Rhynchocypris oxycephalus (Teleostei: Cyprinidae) is a typical small cold water fish, which is distributed widely and mainly inhabits in East Asia. Here, we sequenced and determined the complete mitochondrial genome of R. oxycephalus and studied its phylogenetic implication. R. oxycephalus mitogenome is 16,609 bp in length (GenBank accession no.: MH885043), and it contains 13 protein‐coding genes (PCGs), two rRNA genes, 22 tRNA genes, and two noncoding regions (the control region and the putative origin of light‐strand replication). 12 PCGs started with ATG, while COI used GTG as the start codon. The secondary structure of tRNA‐Ser (AGN) lacks the dihydrouracil (DHU) arm. The control region is 943bp in length, with a termination‐associated sequence, six conserved sequence blocks (CSB‐1, CSB‐2, CSB‐3, CSB‐D, CSB‐E, CSB‐F), and a repetitive sequence. Phylogenetic analysis was performed with maximum likelihood and Bayesian methods based on the concatenated nucleotide sequence of 13 PCGs and the complete sequence without control region, and the result revealed that the relationship between R. oxycephalus and R. percnurus is closest, while the relationship with R. kumgangensis is farthest. The genus Rhynchocypris is revealed as a polyphyletic group, and R. kumgangensis had distant relationship with other Rhynchocypris species. In addition, COI and ND2 genes are considered as the fittest DNA barcoding gene in genus Rhynchocypris. This work provides additional molecular information for studying R. oxycephalus conservation genetics and evolutionary relationships.     

猴欢喜叶绿体全基因组及杜英科系统地位分析

最新的分子系统发育(APG IV)研究中以猴欢喜属(Sloanea L.)为代表的杜英科(Elaeocarpaceae)所在的酢浆草目(Oxalidales)被置于豆类分支(Fabids),且与卫矛目(Celastrales)、金虎尾目(Malpighiales)组成一支(COM分支),但支持率较低.为提高COM分支支...     

木犀科松林丁香复合体的分类修订

对松林丁香S. pinetorum W. W. Sm.复合体内各学者发表的5个新种进行了分类修订。根据居群取样、性状分析和主坐标分析结果, 圆叶丁香S. wardii W. W. Sm.、S. mairei (H. Lév.) Rehder、S. rugulosa McKelvey和川西丁香S. chuanxiensis S. Z. Qu &; X. L. Chen被处理为松林丁香的异名, 其中S. mairei为新异名。此复合体只有一种, 即松林丁香。     

The chloroplast genome of mulberry: complete nucleotide sequence, gene organization and comparative analysis

The complete nucleotide sequence of mulberry (Morus indica cv. K2) chloroplast genome (158,484 bp) has been determined using a combination of long PCR and shotgun-based approaches. This is the third angiosperm tree species whose plastome sequence has been completely deciphered. The circular double-stranded molecule comprises of two identical inverted repeats (25,678 bp each) separating a large and a small single-copy region of 87,386 bp and 19,742 bp, respectively. A total of 83 protein-coding genes including five genes duplicated in the inverted repeat regions, eight ribosomal RNA genes and 37 tRNA genes (30 gene species) representing 20 amino acids, were assigned on the basis of homology to predicted genes from other chloroplast genomes. The mulberry plastome lacks the genes infA, sprA, and rpl21 and contains two pseudogenes ycf15 and ycf68. Comparative analysis, based on sequence similarity, both at the gene and genome level, indicates Morus to be closer to Cucumis and Lotus, phylogenetically. However, at genome level, inclusion of non-coding regions brings it closer to Eucalyptus, followed by Cucumis. This may reflect differential selection pressure operating on the genic and intergenic regions of the chloroplast genome.Electronic supplementary material Supplementary material is available in the online version of this article at and is accessible for authorized users.Communicated by Y. Tsumura     

The complete nucleotide sequence of the hornwort (Anthoceros formosae) chloroplast genome: insight into the earliest land plants

It is generally believed that bryophytes are the earliest land plants. However, the phylogenetic relationships among bryophytes, including mosses, liverworts and hornworts, are not clearly resolved. To obtain more information on the earliest land plants, we determined the complete nucleotide sequence of the chloroplast genome from the hornwort Anthoceros formosae. The circular double-stranded DNA of 161 162 bp is the largest genome ever reported among land plant chloroplasts. It contains 76 protein, 32 tRNA and 4 rRNA genes and 10 open reading frames (ORFs), which are identical with the chloroplast genome of the other green plants analyzed. The major difference is a larger inverted repeat than that of the liverwort Marchantia, Anthoceros contains an excess of ndhB and rps7 genes and the 3′ exon of rps12. The genes matK and rps15, commonly found in the chloroplast genomes of land plants, are pseudogenes. The intron of rrn23 is the first finding in the known chloroplast genomes of land plants. A striking feature of the hornwort chloroplast is that more than half of the protein-coding genes have nonsense codons, which are converted into sense codons by RNA editing. Maximum-likelihood (ML) analysis, based on 11 518 amino acid sites of 52 proteins encoded in the chloroplast genomes of the green plants, placed liverworts as the sister to all other land plants.     

The complete nucleotide sequence of wild rice (Oryza nivara) chloroplast genome: first genome wide comparative sequence analysis of wild and cultivated rice

We determined the complete nucleotide sequence of the chloroplast genome of wild rice, Oryza nivara and compared it with the corresponding published sequence of relative cultivated rice, Oryza sativa. The genome was 134,494 bp long with a large single-copy region of 80,544 bp, a small single-copy region of 12,346 bp and two inverted repeats of 20,802 bp each. The overall A+T content was 61.0%. The O. nivara chloroplast genome encoded identical functional genes to O. sativa in the same order along the genome. On the other hand, detailed analysis revealed 57 insertion, 61 deletion and 159 base substitution events in the entire chloroplast genome of O. nivara. Among substitutions, transversions were much higher than transitions with the former even more frequent than the latter in the coding region. Most of the insertions/deletions were single-base but a few large length mutations were also detected. The frequency of insertion/deletion events was more in the coding region within inverted repeats. In contrast, a very few substitution events were identified in the coding region. Polymorphism was observed among rice cultivars at loci of large insertion/deletion events. This is the first report describing comparative and genome wide chloroplast analysis between a wild and cultivated crop.     

大粒香水稻叶绿体基因组特征分析

大粒香作为贵州重要的优质稻资源,推广种植面积大,并且在乡村振兴过程中为社会带来了较高的经济效益。然而,目前对大粒香基因组学的理论研究报道较少。为揭示大粒香水稻叶绿体基因组(cpDNA)特征及系统发育关系,该研究对大粒香叶绿体进行测序,并分析其基因组特征。结果表明:(1)大粒香水稻cpDNA全长134 563 bp,包括1个大单拷贝区(LSC,80 864 bp)、1个小单拷贝区(SSC,12 347 bp)和2个反向重复序列区(IR_s,20 676 bp)。(2)注释到129个基因,可分为蛋白编码、tRNA和rRNA三类基因,数量分别为85个、36个和8个。(3)密码子偏好分析显示,大粒香cpDNA密码子偏好A碱基或U(T)碱基,亮氨酸密码子使用了1 944次,使用次数最多; 半胱氨酸的密码子仅使用198次,使用次数最少。(4)检测到129个SSR,其中有95个单核苷酸重复且大部分SSR序列由A/T碱基组成。(5)系统发育分析结果显示,大粒香与热带粳稻亲缘关系最近,聚为一类。该研究结果揭示了大粒香叶绿体基因组信息,并确定了大粒香系统发育所属分支。     

Mitochondrial genome of Amblyomma javanense: a hard tick parasite of the endangered Malayan pangolin (Manis javanica)

Amblyomma javanense is an important ectoparasite of Manis javanica, although the population genetics, molecular biology and systematics of A. javanense remain poorly understood. In the present study, the mitochondrial genome of A. javanense was sequenced using the Illumina HiSeq sequencing platform (Illumina, San Diego, CA, U.S.A.) and compared with the genomes of two closely related species: Amblyomma fimbriatum and Amblyomma americanum. The intraspecies and interspecies relationships of A. javanense and another 21 selected species were investigated by constructing a maximum-likelihood tree and a neighbour-joining tree. The mitochondrial genome of A. javanense was 14 780 bp in length and contained 13 protein-coding genes, 22 transfer RNA genes, two ribosomal RNA genes and two control regions. The results of the comparisons indicate that there is great similarity among these three species, and both trees indicate that A. javanense is a member of the Amblyomminae. The study of A. javanense of pangolins also indicates the premise and foundation of the relationship between the parasite and other species.     

Characterization of the complete mitochondrial genome sequence of Spirometra erinaceieuropaei (Cestoda: Diphyllobothriidae) from China

Sparganosis, caused by the plerocercoid larvae of members of the genus Spirometra, can cause significant public health problem and considerable economic losses. In the present study, the complete mitochondrial DNA (mtDNA) sequence of Spirometra erinaceieuropaei from China was determined, characterized and compared with that of S. erinaceieuropaei from Japan. The gene arrangement in the mt genome sequences of S. erinaceieuropaei from China and Japan is identical. The identity of the mt genomes was 99.1% between S. erinaceieuropaei from China and Japan, and the complete mtDNA sequence of S. erinaceieuropaei from China is slightly shorter (2 bp) than that from Japan. Phylogenetic analysis of S. erinaceieuropaei with other representative cestodes using two different computational algorithms [Bayesian inference (BI) and maximum likelihood (ML)] based on concatenated amino acid sequences of 12 protein-coding genes, revealed that S. erinaceieuropaei is closely related to Diphyllobothrium spp., supporting classification based on morphological features. The present study determined the complete mtDNA sequences of S. erinaceieuropaei from China that provides novel genetic markers for studying the population genetics and molecular epidemiology of S. erinaceieuropaei in humans and animals.     

极小种群濒危植物广西火桐、丹霞梧桐的叶绿体基因组特征

广西火桐(Firmiana kwangsiensis)和丹霞梧桐(F. danxiaensis)是我国南方特有物种, 其分布范围狭窄, 种群数量少。为了解其叶绿体基因组结构及系统发生关系, 本文通过高通量测序方法获得广西火桐和丹霞梧桐的浅层基因组数据, 通过生物信息学方法对叶绿体全基因组进行组装, 并对其结构特征进行分析。结果表明: 广西火桐和丹霞梧桐的叶绿体基因组大小分别为160,836 bp和161,253 bp, 具有典型被子植物叶绿体基因组环状四分体结构, 包含长度分别为89,700 bp、90,142 bp的大单拷贝区(large single copy, LSC), 长度分别为19,970 bp、20,067 bp的小单拷贝区(small single copy, SSC)及长度分别为25,583 bp、25,522 bp的2个反向重复序列区(inverted repeat sequence, IR)。两个物种的叶绿体基因组共注释得到131个基因, 包括86个蛋白编码基因、37个tRNA基因和8个rRNA基因。广西火桐的叶绿体基因组中共检测出26个正向重复序列、2个反向重复序列、21个回文重复序列、21个串联重复序列和98个简单重复序列; 丹霞梧桐叶绿体基因组中共检测出23个正向重复序列、5个反向重复序列、21个回文重复序列、30个串联重复序列和107个简单重复序列。系统发生分析结果表明5种梧桐属(Firmiana)植物构成两个强烈支持的分支(支持率100%), 一个分支为广西火桐、美丽火桐(F. pulcherrima)和火桐(F. colorata), 其中广西火桐与美丽火桐构成姐妹群; 另一分支是互为姐妹群的丹霞梧桐和云南梧桐(F. major)。综上所述, 广西火桐和丹霞梧桐的叶绿体基因组结构、基因排列及重复序列具有较高的相似性, 系统进化树将5种梧桐属物种分为两个分支, 其中广西火桐和美丽火桐最近; 而丹霞梧桐与云南梧桐关系最近。本研究鉴定的SSR位点可为梧桐属物种系统发生、进化关系的研究提供遗传信息。     

Characterization of the complete mitochondrial genome of Cryptotermes domesticus (Blattodea: Kalotermitidae): Genome description and phylogenetic implications

The complete mitochondrial genome of Cryptotermes domesticus (Haviland) was sequenced and annotated to study its characteristics and the phylogenetic relationship of C. domesticus to other termite species. The mitogenome of C. domesticus is a circular, close, and double-stranded molecule with a length of 15,655 bp. The sequenced mitogenome contains 37 typical genes, which are highly conserved in gene size, organization, and codon usage. Transfer RNA genes (tRNAs) also have typical secondary structures. All of the 13 protein-coding genes (PCGs) start with an ATN codon, except for nad4, which starts with GTG and terminates with the terminal codon TAA and TAG or the incomplete form T-- (cox2 and nad5). Most tRNAs have a typical cloverleaf structure, except for trnS1, in which this form is replaced by a simple loop and lacks the dihydrouridine (DHU) arm. The nucleotide diversity (Pi) and nonsynonymous (Ka)/synonymous (Ks) mutation rate ratios indicate that nad1, cox1, and cox3 are the most conserved genes, and that cox1 has the lowest rate of evolution. In addition, an 89 bp repeated sequence was found in the A + T-rich region. Phylogenetic analysis was performed using Bayesian inference (BI) and maximum likelihood (ML) methods based on 13 PCGs, and the monophyly of Kalotermitidae was supported.     

丝带凤蝶线粒体基因组全序列及其系统学意义(英文)

锯凤蝶类与凤蝶科其他类群的系统发生关系及其分类学地位一直存在争议。本研究采用PCR和long PCR技术测定了属于锯凤蝶类的丝带凤蝶Sericinus montelus线粒体基因组全序列; 结合已有的其他凤蝶科物种的相应序列数据, 基于13个蛋白质编码基因重建了凤蝶科主要类群的系统发生树, 探讨了它们之间的系统发生关系。基因组分析结果表明： 丝带凤蝶线粒体基因组全长15 242 bp, 包括13个编码蛋白基因(ATP6, ATP8, COⅠ-Ⅲ, ND1-6, ND4L和Cytb)、 22个tRNA基因、 16S和12S rRNA基因以及非编码的控制区; 基因组A, T, G和C含量分别为40.1%, 40.8%, 7.4%和11.7%, 表现出明显的AT偏倚。所有的蛋白质编码基因都使用标准的起始密码子(ATN); 除ND4 和 ND4L基因使用单个的T作为终止密码子外, 其余蛋白编码基因都使用了标准的终止密码子(TAA)。除丝氨酸 tRNA的二氢尿苷突环缺失外, 所有tRNA基因都形成典型的三叶草型结构。基因组中共存在12个大小介于2~65 bp之间的基因间隔区以及15个大小介于1~8 bp之间的基因重叠区, 其中, 存在于COⅡ和tRNA^Lys之间的24 bp的间隔区在其他鳞翅目昆虫中未曾见到。以邻接法和最大简约法并基于13个蛋白质编码基因序列对凤蝶科进行了系统发生分析。结果显示, 丝带凤蝶和中华虎凤蝶Luehdorfia chinensis先构成一个支系, 再和冰清绢蝶Parnassius bremeri构成姊妹群; 表明锯凤蝶类应作为族级分类单元归于凤蝶科下的绢蝶亚科。