喜马红景天叶绿体基因组特征及其系统发育分析

以青藏高原特有药用植物——喜马红景天（Rhodiola himalensis）为试验材料,利用高通量测序技术对喜马红景天进行叶绿体基因组测序、组装和注释,获得完整的叶绿体基因组。结果显示：喜马红景天叶绿体基因组全长为151 074 bp,GC含量为37.8%,具有1个长单拷贝区、1个短单拷贝区和1对反向重复区的典型四分体结构,其序列长度分别为82 309、17 017、25 874 bp;叶绿体基因组共编码130个基因,其中编码蛋白的基因86个、编码tRNA的基因37个、编码rRNA的基因7个;叶绿体基因组共检测出25 513个密码子,其中编码亮氨酸（Leu）的密码子占比最大;喜马红景天IRa和IRb区的rps19和ycf1基因缺失,长单拷贝区的trnH基因收缩;喜马红景天与圣地红景天（R. sacra）亲缘关系最近;短单拷贝区域的单核苷酸多态性（SNP）变异频率最高。本研究报道了喜马红景天的叶绿体基因组,并对其进行了组装、注释和序列分析,为今后开展喜马红景天的遗传多样性研究和合理开发利用提供理论依据。     

鼠尾草叶绿体基因组序列分析

鼠尾草（Salvia japonica）是唇形科（Labiatae）鼠尾草属（Salvia）的一种多年生草本植物,具有十分重要的药用和经济价值。本文采用第二代测序技术Illumina Hiseq平台对鼠尾草的叶绿体基因组进行测序,同时以鼠尾草近缘物种丹参叶绿体基因组作为参考,组装得到完整叶绿体基因组序列。结果表明,鼠尾草叶绿体基因组序列全长153 995 bp,呈典型的四段式结构,其中LSC区长84 573 bp,SSC区长19 874 bp,两个IR区分别长24 774 bp;鼠尾草叶绿体基因组成功注释13组叶绿体基因,基因的种类、数目及GC含量等与唇形科中其它物种较为类似。这些研究结果丰富了鼠尾草属的叶绿体基因组数据,为今后鼠尾草属植物系统发育关系重建积累了基础性数据。     

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。     

藏波罗花叶绿体基因组分析

藏波罗花（Incarvillea younghusbandii Sprague）是一种传统的补益类中药。其根作草药使用,用于滋补强壮,治产后少乳、久病虚弱、头晕、贫血等症。但目前关于藏波罗花分子遗传信息的研究很少。本研究基于高通量测序技术对藏波罗花叶绿体基因组进行测序、组装和注释,并对其序列特征、密码子偏好性、重复序列、系统发育和分化时间进行分析。结果表明,藏波罗花叶绿体基因组全长为159 323 bp,包含1个大单拷贝区（80 197 bp）、1个小单拷贝区（9 030 bp）和2个反向重复区（35 048 bp）;共注释出120个基因,包括77个蛋白编码基因、8个rRNA基因和35个tRNA基因;密码子偏好性分析显示,AAA是藏波罗花叶绿体基因组中使用最频繁的密码子;从藏波罗花叶绿体基因组中共检测到42个简单重复序列（simple sequence repeats,SSR）;系统发育分析表明,藏波罗花与密生波罗花（Incarvillea compacta）的亲缘关系最近,且在大概466万年前产生分化。本研究对藏波罗花相关资源的科学保护和开发具有重要的现实意义,也可以为后续角蒿属（Incarvillea）的物种鉴定、紫葳科（Bignoniaceae）的种群遗传多样性研究提供基本的遗传资源。     

直刺变豆菜叶绿体全基因组及其特征

直刺变豆菜(Sanicula orthacantha)是中国广泛分布的多年生草本植物, 也是一味著名的民族药。本文通过二代高通量测序平台Illumina HiSeq PE150对直刺变豆菜叶绿体全基因组进行测序, 并通过生物信息学方法对其结构特征进行分析。结果表明: 直刺变豆菜叶绿体全基因组大小为157,163 bp, 包括大单拷贝区(large single copy, LSC)、小单拷贝区(small single copy, SSC)和2个反向重复序列(inverted repeat sequence, IRa和IRb), 长度分别为87,547 bp、17,122 bp和26,247 bp, 具有典型被子植物叶绿体基因组环状四分体结构; 共注释得到129个基因, 包括8个核糖体RNA (rRNA)基因、37个转运RNA (tRNA)基因和84个蛋白质编码基因。直刺变豆菜在叶绿体基因组结构、基因种类、排列顺序上与其他伞形科植物基本一致。直刺变豆菜叶绿体全基因组测序的成功为变豆菜属植物完整叶绿体基因组组装及其特征分析提供了新的方法。     

荷花玉兰叶绿体全基因组高通量测序及结构解析

荷花玉兰是重要的药用、观赏及园林绿化植物.应用454高通量测序技术对荷花玉兰叶绿体全基因组进行测序,解析了其基因组结构,并与近缘物种基因组进行了比较分析.荷花玉兰叶绿体基因组全长为159623bp,两个反向互补重复区(IRs)长26563bp,被分隔的大单拷贝区(LSC)和小单拷贝区(SSC)长度分别为87757和18740bp.成功注释129个叶绿体基因,其中18个基因含有内含子.基因的种类、数目以及GC含量等与其他木兰科物种相类似.生物信息学分析获得218个SSR位点,大多位点富含A-T,具有碱基偏好性.木兰科物种的重复基序类型和丰度相对保守,有利于开发叶绿体基因组载体.木兰亚纲植物叶绿体基因组的大小及IR区边界的变化与ycf1的长度密切相关.采用30个物种叶绿体基因组的66个共有蛋白编码基因构建系统发育树,对木兰属在被子植物中的进化位置进行了探讨.荷花玉兰叶绿体全基因组序列的获得和结构解析对优良品种培育、叶绿体基因组工程、木兰科物种分子标记开发及系统发育关系的研究具有重要价值.     

多花海棠叶绿体基因组分析

多花海棠（Malus floribunda Siebold.）是世界范围内广泛栽培的苹果属物种,具有较高的观赏价值和育种意义。对其进行叶绿体基因组比较分析,有利于完善苹果属系统进化以及种质利用的研究内容。基于全基因组测序数据,组装获得一个完整的具有四分体结构的多花海棠叶绿体基因组。该基因组包括大单拷贝区（88 142 bp）、反向重复区B （26 353 bp）、小单拷贝区（19 189 bp）与反向重复区A （26 353 bp）,共计160 037 bp。多花海棠叶绿体全基因组共注释到111个基因,包括78个蛋白编码基因、29个tRNA基因和4个rRNA基因。此外,在其基因组中识别到大量的重复序列,与三叶海棠和变叶海棠略有差异。通过计算相对同义密码子使用度,发现其高频密码子共30种,并且密码子具有偏向A/T结尾的使用模式。种间序列比对、边界分析的结果表明,大单拷贝区序列变异较大,8种苹果属植物SC区与IR区扩张收缩情况整体上较为相似。基于叶绿体基因组序列的系统进化分析,将多花海棠、湖北海棠和变叶海棠聚为一类。多花海棠叶绿体基因组的研究可为今后遗传标记开发与种质资源利用等提供数据支持。     

红花变豆菜叶绿体基因组结构及同属种间关系研究

红花变豆菜（Sanicula rubriflora F. Schmidt）是有药用价值的植物,全株干燥后与其他药用同属植物易混淆,种间关系存在争议,通过高通量测序技术对红花变豆菜叶绿体基因组测序,利用生物信息学方法对测序数据进行拼接、注释,首次报道红花变豆菜叶绿体基因组结构及特点,利用叶绿体基因组数据,提供种间分类新证据,并且分析相关类群的进化关系。S. rubriflora叶绿体基因组序列的长度为155 721 bp,其中包括一个85 981 bp的大单拷贝区（large single copy,LSC）和一个17 060 bp的小单拷贝区（small single-copy region,SSC）,它们被两个26 340 bp的反向重复区（inverted repeat sequence,IRs）隔开。红花变豆菜叶绿体基因组GC含量为38.20%,包含129个基因,其中84个蛋白质编码基因,37个tRNA基因和8个rRNA基因。红花变豆菜叶绿体基因组结构具有高度保守性,其中编码基因共有51 907个密码子,最多编码5 095个亮氨酸,最少编码689个色氨酸,简单重复序列分析共发现32个位点,大多数是单碱基重复的A/T类型。叶绿体基因组聚类结果支持天胡荽亚科（Hydrocotyloideae）是伞形科（Umbelliferae）内比较原始的类群;变豆菜亚科（Saniculoideae）和芹亚科（Apioideae）为姊妹类群,是伞形科较进化的类群;变豆菜属植物是一个相对自然的类群;红花变豆菜与黄花变豆菜（S. flavovirens）为近缘姊妹种,但是两者形态和地理分布差异较大。该研究结果为变豆菜属属下种间鉴定及其种间演化奠定基础。     

七里香蔷薇叶绿体基因组测序及结构分析

七里香蔷薇是蔷薇属木香花的一个变种,在秦岭山区广泛分布,具有重要的观赏价值,同时也是蔷薇属花卉育种的重要种质资源。本研究首次以秦巴山区七里香蔷薇为实验材料,采用Illumina HiSeq4000测序系统对其叶绿体基因组DNA建库测序,建库类型为350 bp DNA小片段文库,测序深度为59×。分析其基因组结构,并且与已报道蔷薇科植物叶绿体基因组进行比较。七里香蔷薇叶绿体基因组序列全长156 544 bp,具有典型被子植物叶绿体基因组环状四分体结构。叶绿体基因组包括大单拷贝区85 678 bp、小单拷贝区18 782 bp和2个反向重复序列26 042 bp。GC含量为37.22%;检测到116个串联重复序列43个散在重复序列;此外,还检测到72个简单重复序列位点。对其功能基因比对分析,该植物的叶绿体基因组共注释得到129个基因,包括86蛋白编码基因、35个tRNA基因和8个rRNA基因。用MEGA6.0软件通过邻近法基于叶绿体全基因组序列对所有已知叶绿体基因组的蔷薇科22个物种进行聚类分析,其与七里香蔷薇和大花香水月季、缫丝花聚在一起。研究结果对于七里香蔷薇野生植物资源的育种及引种驯化具有参考依据。     

两种珍稀白蝶兰属(兰科)叶绿体基因组比较分析

为理解珍稀濒危兰科植物龙头兰(Pecteilis susannae)和景洪白蝶兰(P.hawkesiana)的叶绿体基因组的基本特征,开发用于物种鉴定、保护遗传学和系统发育分析的分子标记,该研究利用二代测序技术对龙头兰和景洪白蝶兰进行浅层基因组测序,采用生物信息学分析方法进行叶绿体基因组的拼接、组装和注释,并与其他近缘物种进行比较基因组分析和系统发育分析。结果表明：(1)龙头兰和景洪白蝶兰的叶绿体基因组大小分别为154 407 bp和153 891 bp,由一对26 550 bp和26 523 bp的反向重复序列(IR)、84 204 bp和83 756 bp的大单拷贝区(LSC)、17 103 bp和17 089 bp的小单拷贝区(SSC)组成;均注释了111个唯一基因,包括77个蛋白质编码基因、30个tRNA基因和4个rRNA基因。(2)在叶绿体基因组中分别鉴定出94个和92个简单重复序列(SSRs)。(3)二者之间存在706个单核苷酸多态性(SNPs)位点和152个插入缺失(InDels)位点,其中cpInDel 067等可以区分2个物种。(4)观察到1个差异较大的基因(accD...     

The complete chloroplast genome of the spring ephemeral plant Alyssum desertorum and its implications for the phylogenetic position of the tribe Alysseae within the Brassicaceae

Alyssum desertorum (Alysseae, Brassicaceae) is an annual spring ephemeral plant whose life cycle is only 2–3 months. It typically has high photosynthetic capacity and a high growth rate. However, little was known about the chloroplast (cp) genome structure of this species. Furthermore, the phylogenetic position of the tribe Alysseae relative to other tribes in the Brassicaceae has not been established and there appear to be inconsistences between different DNA markers. This study is the first report on a cp genome of the genus Alyssum and discusses the phylogenetic relationships of the tribe Alysseae relative to other tribes in the family. The complete cp genome of A. desertorum was 151 677 bp in size and is thus the smallest cp genome of Brassicaceae sequenced to date. The genome includes a large single‐copy region of 81 551 bp, a small single‐copy region of 17 804 bp, and two inverted repeats of 26 161 bp each. The genome contains 132 genes, including 86 protein‐coding genes (PCGs), 38 tRNA genes and 8 rRNA genes. A total of 16 genes contained introns, including 10 PCGs and 6 tRNA genes; the ycf3 and clpP genes contained two introns, and the remaining genes each contained one. Compared to the cp genomes of 21 other Brassicaceae species, the cp genome of Alyssum desertorum was the smallest, as due to variation in gene content and gene length, such as a lack of the rps16 gene and the deletion of some coding genes. Additionally, deletions of introns and intergenic spacers were observed, but their total length was not significantly shorter than those of other taxa. Phylogenetic analysis at the tribal level based on a cp genome dataset revealed that the tribe Alysseae is an early‐diverging lineage that is sister to other species within subclade B of clade II.     

De novo assembly and characterization of the complete chloroplast genome of radish (Raphanus sativus L.)

Radish (Raphanus sativus L.) is an edible root vegetable crop that is cultivated worldwide and whose genome has been sequenced. Here we report the complete nucleotide sequence of the radish cultivar WK10039 chloroplast (cp) genome, along with a de novo assembly strategy using whole genome shotgun sequence reads obtained by next generation sequencing. The radish cp genome is 153,368 bp in length and has a typical quadripartite structure, composed of a pair of inverted repeat regions (26,217 bp each), a large single copy region (83,170 bp), and a small single copy region (17,764 bp). The radish cp genome contains 87 predicted protein-coding genes, 37 tRNA genes, and 8 rRNA genes. Sequence analysis revealed the presence of 91 simple sequence repeats (SSRs) in the radish cp genome.     

Characterization of the Complete Chloroplast Genome of Apple (Malus × domestica,Rosaceae)*

Apple (Malus × domestica) is one of the most important temperate fruits. To better understand the molecular basis of this species, we characterized the complete chloroplast (cp) genome sequence downloaded from Genome Database for Rosaceae. The cp genome of apple is a circular molecule of 160068bp in length with a typical quadripartite structure of two inverted repeats (IRs) of 26352bp, separated by a small single copy region of 19180bp (SSC) and a large single copy region (LSC) of 88184bp. A total of 135 predicted genes (115 unique genes, and another 20 genes were duplicated in the IR) were identified, including 81 protein coding genes, four rRNA genes and 30 tRNA genes. Three genes of ycf15, ycf68 and infA contain several internal stop codons, which were interpreted as pseudogenes. The genome structure, gene order, GC content and codon usage of apple are similar to the typical angiosperm cp genomes. Thirty repeat regions (≥30bp) were detected, twenty one of which are tandem, six are forward and three are inverted repeats. Two hundred thirty seven simple sequence repeat (SSR) loci were revealed and most of them are composed of A or T, contributing to a distinct bias in base composition. Additionally, average 10000bp non coding region contains 24 SSR sites, while protein coding region contains five SSR sites, indicating an uneven distribution of SSRs. The complete cp genome sequence of apple reported in this paper will facilitate the future studies of its population genetics, phylogenetics and chloroplast genetic engineering.     

First reported chloroplast genome sequence of <Emphasis Type="Italic">Punica granatum</Emphasis> (cultivar Helow) from Jabal Al-Akhdar,Oman: phylogenetic comparative assortment with <Emphasis Type="Italic">Lagerstroemia</Emphasis>

Pomegranate (Punica granatum L.) is one of the oldest known edible fruits. It has grown in popularity and is a profitable fruit crop due to its attractive features including a bright red appearance and its biological activities. Scientific exploration of the genetics and evolution of these beneficial traits has been hampered by limited genomic information. In this study, we sequenced the complete chloroplast (cp) genome of the native P. granatum (cultivar Helow) cultivated in the mountains of Jabal Al-Akhdar, Oman. The results revealed a P. granatum cp genome length of 158,630 bp, characterized by a relatively conserved structure containing 2 inverted repeat regions of 25,466 bp, an 18,686 bp small single copy regions, and an 89,015 bp large single copy region. The 86 protein-coding genes included 37 transfer RNA genes and 8 ribosomal RNA genes. Comparison of the P. granatum whole cp genome with seven Lagerstroemia species revealed an overall high degree of sequence similarity with divergence among intergenic spacers. The location, distribution, and divergence of repeat sequences and shared genes of the Punica and Lagerstroemia species were highly similar. Analyses of nucleotide substitution, insertion/deletions, and highly variable regions in these cp genomes identified potential plastid markers for taxonomic and phylogenetic studies in Myrtales. A phylogenetic study of the cp genomes and 76 shared coding regions generated similar cladograms. The complete cp genome of P. granatum will aid in taxonomical studies of the family Lythraceae.     

The complete chloroplast genome sequence of the endangered species Syringa pinnatifolia (Oleaceae)

Syringa pinnatifolia is an endangered endemic species in China with important ornamental and medicinal value, and it needs urgent protection. Here, we report the complete chloroplast (cp) genome structure of S. pinnatifolia and its evolution is inferred through comparative studies with related species. The S. pinnatifolia cp genome was 155 326 bp and contained a large single copy region (LSC) of 86 167 bp and a small single copy region (SSC) of 17 775 bp, as well as a pair of inverted repeat regions (IRs) of 25 692 bp. A total of 113 unique genes were annotated, including 79 protein‐coding genes, 30 tRNA genes and four rRNA genes. The GC content of the S. pinnatifolia cp genome was 37.9%, and the corresponding values in the LSC, SSC and IR regions were 36.0, 32.1, 43.2% respectively. Repetitive sequences analysis revealed that the S. pinnatifolia cp genome contained 38 repeats. Microsatellite marker detection analysis identified 253 simple sequence repeats (SSRs), which provides opportunities for future studies of the population genetics and phylogenetic relationships of Syringa. Phylogenetic analysis of 29 selected cp genomes revealed that S. pinnatifolia is closely related to Syringa vulgaris and all 27 Lamiales species formed a clade separate from the two outgroup species. This newly characterized S. pinnatifolia chloroplast genome will provide a useful genomic resource of phylogenetic inference and the development of more genetic markers for species discrimination and population studies in the genus Syringa.     

The complete chloroplast genome sequence of <Emphasis Type="Italic">Dodonaea viscosa</Emphasis>: comparative and phylogenetic analyses

The plant chloroplast (cp) genome is a highly conserved structure which is beneficial for evolution and systematic research. Currently, numerous complete cp genome sequences have been reported due to high throughput sequencing technology. However, there is no complete chloroplast genome of genus Dodonaea that has been reported before. To better understand the molecular basis of Dodonaea viscosa chloroplast, we used Illumina sequencing technology to sequence its complete genome. The whole length of the cp genome is 159,375 base pairs (bp), with a pair of inverted repeats (IRs) of 27,099 bp separated by a large single copy (LSC) 87,204 bp, and small single copy (SSC) 17,972 bp. The annotation analysis revealed a total of 115 unique genes of which 81 were protein coding, 30 tRNA, and four ribosomal RNA genes. Comparative genome analysis with other closely related Sapindaceae members showed conserved gene order in the inverted and single copy regions. Phylogenetic analysis clustered D. viscosa with other species of Sapindaceae with strong bootstrap support. Finally, a total of 249 SSRs were detected. Moreover, a comparison of the synonymous (Ks) and nonsynonymous (Ka) substitution rates in D. viscosa showed very low values. The availability of cp genome reported here provides a valuable genetic resource for comprehensive further studies in genetic variation, taxonomy and phylogenetic evolution of Sapindaceae family. In addition, SSR markers detected will be used in further phylogeographic and population structure studies of the species in this genus.     

Complete chloroplast genome of Myracrodruon urundeuva and its phylogenetics relationships in Anacardiaceae family

Continuous exploratory use of tree species is threatening the existence of several plants in South America. One of these threatened species is Myracroduron urundeuva, highly exploited due to the high quality and durability of its wood. The chloroplast (cp) has been used for several evolutionary studies as well traceability of timber origin, based on its gene sequences and simple sequence repeats (SSR) variability. Cp genome organization is usually consisting of a large single copy and a small single copy region separated by two inverted repeats regions. We sequenced the complete cp genome from M. urundeuva based on Illumina next-generation sequencing. Our results show that the cp genome is 159,883 bp in size. The 36 SSR identified ranging from mono- to hexanucleotides. Positive selection analysis revealed nine genes related to photosystem, protein synthesis, and DNA replication, and protease are under positive selection. Genome comparison a other Anacardiaceae chloroplast genomes showed great variability in the family. The phylogenetic analysis using complete chloroplast genome sequences of other Anacardiaceae family members showed a close relationship with two other economically important genera, Pistacia and Rhus. These results will help future investigations of timber monitoring and population and evolutionary studies. Supplementary InformationThe online version contains supplementary material available at 10.1007/s12298-021-00989-1.