Structure Of cpDNA Research Articles

Comparative Analysis of Chloroplast Genome Variation and Evolutionary Analysis of the Endangered Species Dendrobium flexicaule and its Relative Species

Globally, the genus Dendrobium (Family: Orchidaceae) exhibits widespread distribution, predominantly occupying the tropical and subtropical areas of Asia. Although the chloroplast genomes (cpDNAs) of certain Dendrobium species have been documented, the evolutionary relationships among these species remain largely unexplored. In this pioneering investigation, a comprehensive analysis of the complete cpDNAs from Dendrobium species is presented, including Dendrobium flexicaule along with three closely related taxa (D. nobile, D. officinale, and D. huoshanense). The size of the novel complete cpDNAs ranged from 150,529 to 152,588 bp, and their cpDNA structure exhibits a unique quadrifoliate structure reminiscent of female quadrivalents, including large single copy, small single copy (SSC), and inverted repeat regions. A total of 175 simple sequence repeats were identified in the D. flexicaule cpDNA. A few pseudogenes of the NA(D)H dehydrogenase-like (NDH) genes were found in the annotated cpDNAs of the four Dendrobium species. Within the shared pseudogenes ndhF and ndhG, ndhF frequently undergoes mutations. Additionally, the phylogenetic analysis showed that D. flexicaule and D. scoriarum were close sister species. The research findings provide crucial genetic information for the molecular phylogenetic system of D. flexicaule.

The size diversity of the Pteridaceae family chloroplast genome is caused by overlong intergenic spacers

BackgroundWhile the size of chloroplast genomes (cpDNAs) is often influenced by the expansion and contraction of inverted repeat regions and the enrichment of repeats, it is the intergenic spacers (IGSs) that appear to play a pivotal role in determining the size of Pteridaceae cpDNAs. This provides an opportunity to delve into the evolution of chloroplast genomic structures of the Pteridaceae family. This study added five Pteridaceae species, comparing them with 36 published counterparts.ResultsPoor alignment in the non-coding regions of the Pteridaceae family was observed, and this was attributed to the widespread presence of overlong IGSs in Pteridaceae cpDNAs. These overlong IGSs were identified as a major factor influencing variations in cpDNA size. In comparison to non-expanded IGSs, overlong IGSs exhibited significantly higher GC content and were rich in repetitive sequences. Species divergence time estimations suggest that these overlong IGSs may have already existed during the early radiation of the Pteridaceae family.ConclusionsThis study reveals new insights into the genetic variation, evolutionary history, and dynamic changes in the cpDNA structure of the Pteridaceae family, providing a fundamental resource for further exploring its evolutionary research.

Geographic patterns of genetic variation in nuclear and chloroplast genomes of two related oaks (Quercus aliena and Q. serrata) in Japan: implications for seed and seedling transfer

In this study, we assessed geographic patterns of genetic variations in nuclear and chloroplast genomes of two related native oaks in Japan, Quercus aliena and Q. serrata, in order to facilitate development of genetic guidelines for transfer of planting stocks for each species. A total of 12 populations of Q. aliena and 44 populations of Q. serrata were analyzed in this study. Genotyping of nuclear microsatellites in Q. aliena was done with only nine populations (n = 212) due to limited numbers of individuals in two populations, while all 12 populations (n = 89) were used in sequencing chloroplast DNA (cpDNA). In Q. serrata, 43 populations (n = 1032) were genotyped by nuclear microsatellite markers, while cpDNA of 44 populations (n = 350) was sequenced. As anticipated, geographic patterns detected in the variations of Q. aliena’s nuclear genome and its chloroplast haplotype distribution clearly distinguished northern and southern groups of populations. However, those of Q. serrata were inconsistent. The geographic distribution of its chloroplast haplotypes tends to show the predicted differentiation between northern and southern lineages, but geographic signals in the genetic structure of its nuclear microsatellites are weak. Therefore, treating northern and southern regions of Japan as genetically distinct transferrable zones for planting stocks is highly warranted for Q. aliena. For Q. serrata, the strong NE-SW geographic structure of cpDNA should be considered.

Relative importance of pollen and seed dispersal across a Neotropical mountain landscape for an epiphytic orchid

Populations of many species are isolated within narrow elevation bands of Neotropical mountain habitat, and how well dispersal maintains genetic connectivity is unknown. We asked whether genetic structure of an epiphytic orchid, Epidendrum firmum, corresponds to gaps between Costa Rican mountain ranges, and how these gaps influence pollen and seed flow. We predicted that significant genetic structure exists among mountain ranges due to different colonization histories and limited gene flow. Furthermore, we predicted that pollen movement contributes more to gene flow than seeds because seeds are released into strong winds perpendicular to the narrow northwest-southeast species distribution, while the likely pollinators are strong fliers. Individuals from 12 populations and three mountain ranges were genotyped with nuclear microsatellites (nDNA) and chloroplast sequences (cpDNA). Genetic diversity was high for both markers, while nDNA genetic structure was low (FSTn =0.020) and cpDNA structure was moderate (FSTc =0.443). Significant cpDNA barriers occurred within and among mountain ranges, but nDNA barriers were not significant after accounting for geographic distance. Consistent with these contrasting patterns of genetic structure, pollen contributes substantially more to gene flow among populations than seed (mp /ms =46). Pollinators mediated extensive gene flow, eroding nDNA colonization footprints, while seed flow was comparatively limited, possibly due to directional prevailing winds across linearly distributed populations. Dispersal traits alone may not accurately inform predictions about gene flow or genetic structure, supporting the need for research into the potentially crucial role of pollinators and landscape context in gene flow among isolated populations.

Comparative phylogeography of red maple (Acer rubrum L.) and silver maple (Acer saccharinum L.): impacts of habitat specialization, hybridization and glacial history

Aim We analysed variation in chloroplast DNA (cpDNA) in red maple (Acer rubrum L.) and silver maple (Acer saccharinum L.) across a large part of their geographic ranges. Acer rubrum is one of the most common and morphologically variable deciduous trees of eastern North America, while its sister species A. saccharinum has a more restricted habitat distribution and displays markedly less morphological variation. Our objective was to infer the impact of biogeographic history on cpDNA diversity and phylogeographic structure in both species. Location Deciduous forests of eastern North America. Methods We sequenced 1289 to 1645 bp of non-coding cpDNA from A. rubrum (n = 258) and A. saccharinum (n = 83). Maximum parsimony networks and spatial analysis of molecular variance (SAMOVA) were used to analyse phylogeographic structure. Rarefaction analyses were used to compare genetic diversity. Results A total of 40 cpDNA haplotypes were recovered from A. rubrum (38 haplotypes) and A. saccharinum (7 haplotypes). Five of the seven A. saccharinum haplotypes were shared with nearby samples of A. rubrum. SAMOVA recovered four phylogeographic groups for A. rubrum in: (1) south-eastern USA, (2) the Gulf and south-eastern Coastal Plain, (3) the lower Mississippi River Valley, and (4) the central and northern regions of eastern North America. Acer saccharinum had significantly lower haplotype diversity than A. rubrum, and novel haplotypes in post-glaciated northern limits of its range were shared with A. rubrum. Main conclusions This is the first study of A. rubrum to report a distinct phylogeographic group centred on the lower Mississippi River, and the first to examine data comparatively with A. saccharinum. We hypothesized that A. rubrum would display stronger phylogeographic structure and greater haplotype diversity than A. saccharinum because of its greater geographic range, and ecological and morphological variation. This hypothesis was supported by the cpDNA analysis. The sharing of cpDNA and chloroplast simple sequence repeat (cpSSR) haplotypes in areas of geographic overlap provides evidence of introgression, which led to an increase in haplotype diversity in both species, and to novel phylogeographic structure in A. rubrum. We recommend that introgression be considered, along with other potential causes, as an explanation for the phylogeographic structure of cpDNA in plants.

Multiple Pleistocene refugia in the widespread Patagonian tree Embothrium coccineum (Proteaceae)

Embothrium coccineum J.R.Forst. & G.Forst is an endemic tree of the Patagonian temperate forest. The objective of this study is to evaluate the impact of last glaciation events on the genetic structure of this widespread and ecologically tolerant species, to postulate possible refugial areas. Phylogeographic analyses were performed using chloroplast DNA sequences (trnL-trnF spacer and ndhC-trnV spacer) from individuals collected in 34 populations along the total range of the species, and these results were compared with a similar study in Nothofagus. A total of 22 haplotypes were found, three of which were widely distributed while 13 were found at only one location. Historical demography suggests a long period of stable effective population size, decreasing gradually towards the Last Glacial Maximum (LGM), followed by an increase in population size that stabilised 2500 years ago. The phylogeographic analyses reflect recent events of colonisation after the LGM from multiple refugia. In the northern area of its distribution probably the species survived in several pockets within the Andes mountain range and in Cordillera de la Costa in Chile. In the south, it is suggested that Embothrium survived the glacial period at the edge of the glaciers. These findings are in agreement with the fossil pollen record that shows 10 000-year-old grains in the south, suggesting colonisation from nearby areas when ice retreated. Embothrium is a coloniser that naturally occurs as scattered individuals within mixed forests. Hence, the shallow phylogeographic structure reported here reflects a Pleistocene signature highly impacted by drift resulting in the randomly fixation of new variants reducing the cpDNA structure.

RecA maintains the integrity of chloroplast DNA molecules in Arabidopsis.

Although our understanding of mechanisms of DNA repair in bacteria and eukaryotic nuclei continues to improve, almost nothing is known about the DNA repair process in plant organelles, especially chloroplasts. Since the RecA protein functions in DNA repair for bacteria, an analogous function may exist for chloroplasts. The effects on chloroplast DNA (cpDNA) structure of two nuclear-encoded, chloroplast-targeted homologues of RecA in Arabidopsis were examined. A homozygous T-DNA insertion mutation in one of these genes (cpRecA) resulted in altered structural forms of cpDNA molecules and a reduced amount of cpDNA, while a similar mutation in the other gene (DRT100) had no effect. Double mutants exhibited a similar phenotype to cprecA single mutants. The cprecA mutants also exhibited an increased amount of single-stranded cpDNA, consistent with impaired RecA function. After four generations, the cprecA mutant plants showed signs of reduced chloroplast function: variegation and necrosis. Double-stranded breaks in cpDNA of wild-type plants caused by ciprofloxacin (an inhibitor of Escherichia coli gyrase, a type II topoisomerase) led to an alteration of cpDNA structure that was similar to that seen in cprecA mutants. It is concluded that the process by which damaged DNA is repaired in bacteria has been retained in their endosymbiotic descendent, the chloroplast.

Route, speed and mode of oak postglacial colonisation across the British Isles: Integrating molecular ecology, palaeoecology and modelling approaches

Summary This paper describes the route, speed and mode of colonisation of oaks by integrating a number of independent analyses using molecular ecology, palaeoecology and simulation modelling approaches. Using a synthetic map of the contemporary distribution of chloroplast DNA (integrating several published and unpublished data sets and describing variation in 1468 trees from 313 autochthonous stands of Q. robur and Q. petraea from Britain and Ireland), and considering the postglacial topographic landscape, the most likely routes of postglacial colonisation across the British Isles are suggested. The overall pattern of these directions agrees with previous interpretations, but several routes, particularly those into Ireland, differ from previous interpretations and benefit here from using a single synthesised data set. Interestingly, the Atlantic oakwoods appear to have been colonised by individuals bearing a single haplotype (type 12). Two palaeoecology data sets, published separately for Britain and Ireland, are synthesised here and used to infer the timing of first arrival of oaks across the British Isles (between 9500 and 6000 years before present). The maximum observed colonisation speed within the British Isles is approximately 500 m year-1 in central and southern England. Outputs from a simulation model, which mimics postglacial colonisation processes, and which has been parameterised for the colonisation rate observed from the pollen core record and contemporary cpDNA structure, predict that the rapid colonisation rate observed, for at least the southern portion of the British Isles, can only be achieved via very rare (an approximate frequency 0.01%), very long distance seed dispersal events (up to 100 km). Potential agents of such dispersal events are birdsor major meteorological disturbances, e.g. hurricanes. Additional simulation modelling and genetic analysis of latitudinally stratified populations indicate that non-synchronous colonisation fronts, topographic barriers and temperature related survival may also have had an effect on the speed of migration and resulting genetic structure. Finally, in an attempt to record predicted long distance seed dispersal events, a novel curve fitting technique is applied to molecular parentage assignment data for field established seedlings from a contemporary population. A notable discrepancy is recorded between contemporary field estimates (just over 1 km) and those predicted by simulation modelling, and is discussed in detail. A concluding section describes future research priorities.

Species‐independent, geographical structuring of chloroplast DNA haplotypes in a montane herb Ipomopsis (Polemoniaceae)

Two hypotheses have been proposed to explain the occurrence of hybrid zones between red‐flowered Ipomopsis aggregata and white‐flowered I. tenuituba. Either local adaptation to hummingbird and hawkmoth pollinators has given rise to sympatric (or parapatric) divergence of flower colour and morphology (primary intergradation at hybrid zones), or alternatively two previously allopatric species are coming into contact at several geographical areas of secondary intergradation. We examined restriction site patterns in nuclear DNA (nrDNA), chloroplast DNA (cpDNA) and mitochondrial DNA (mtDNA) from populations of I. aggregata and I. tenuituba representing seven zones of sympatry. No variation was detected in a 350‐bp fragment of mtDNA and uninformative levels of variation were observed for nrDNA. We detected 22 potentially informative restriction site polymorphisms in cpDNA, all of which united geographical areas containing populations of both species. We detected no informative species‐specific markers. Studies of other species (e.g. oaks) have detected similar species‐independent geographical structure of cpDNA. However, in these cases secondary interegradation could be inferred from species‐specific nuclear alleles. The pattern in Ipomopsis is consistent with both primary intergradation (independent speciation in each area of sympatry) or secondary intergradation involving complete cytoplasmic replacement. Thus, additional data are needed to explain the origin of hybrid zones in Ipomopsis.

Physical map of chloroplast DNA of the onion Allium cepa L., showing the location of photosynthesis-related genes.

To compare the gene order of chloroplast genome among monocotyledonous plants, we constructed a physical map of chloroplast DNA (cpDNA) of onion (Allium cepa L. cv. Shounan red, 2n=16) with four restriction endonucleases, PstI, SalI, XhoI, and SmaI. The cpDNA of Allium cepa was found to be a circular molecule with a total size of ca. 155 kb and containing two inverted repeats of 26 kb each that disrupt the rest of the molecule into small (ca. 16 kb) and large (ca. 86 kb) single copy regions. The endonuclease recognition sites of the physical map were confirmed by Southern hybridizations of total onion cpDNA with homologous and heterologous probes. 16 genes were localized on the physical map of the onion cpDNA. Genome structure of onion cpDNA was similar in terms of genome size and gene order of cpDNA to that of tobacco cpDNA, differing from the cpDNA structure of gramineous plants.