trnL-trnF Intergenic Spacer Research Articles

Drepananthus khaosoi sp. nov. (Annonaceae) from southern Thailand, with molecular phylogenetic reconstructions

Drepananthus khaosoi sp. nov., a new species from southern Thailand is described and illustrated. It is morphologically most similar to D. ridleyi, especially in the very narrow and elongated petals, but differs in leaf blade size, leaf base, monocarp shape and appearance, monocarp width, length of monocarp stipe, pericarp thickness and seed arrangement. Molecular phylogenetic analyses using chloroplast DNA regions (matK and rbcL exons; trnL intron; atpB-rbcL, psbA-trnH and trnL-trnF intergenic spacers) and including 18 Drepananthus species confirm the monophyly of Drepananthus, but the relationships within the genus are largely unresolved, including a failure to elucidate a sister group of D. khaosoi. The conservation status of the new species is provisionally assessed as Critically Endangered.

Evaluating the effectiveness of DNA-based methods to detect mislabelling and adulteration of chestnut honey

Honey is a naturally sweet and viscous substance that is consumed worldwide for its specific flavour, taste and biological properties, which in turn depend on the type of plant from which bees collect nectar. The quality of honey and, in particular, the information on the product label determines consumer choice and market price. In this study, the true botanical composition of chestnut monofloral honeys produced in Calabria (South Italy) was determined by DNA barcoding, PCR-RFLP and LAMP approaches, all based on the chloroplast trnL-trnF intergenic spacer region. All three DNA-based techniques were reliable in unambiguously identifying the floral composition of the different honeys tested at species level. However, the LAMP assay has the advantage over DNA barcoding and PCR-RFLP of greatly simplifying and speeding up the identification of Castanea sativa as the dominant plant species of the Calabrian chestnut honeys. This may encourage the adoption of this method by the food industry for the rapid detection of intentionally or accidentally mislabelled ingredients in this valuable natural product, thus ensuring consumer safety and protection.

Development of a two-layer machine learning model for the forensic application of legal and illegal poppy classification based on sequence data

Poppies are beneficial plants with a variety of applications, including medicinal, edible, ornamental, and industrial purposes. Some Papaver species are forensically significant plants because they contain opium, a narcotic substance. Internationally trafficked species of illegal poppies are being identified by DNA barcoding employing multiple markers in response to their forensic value. However, effective markers for precise species identification of legal and illegal poppies are still under discussion, with research on illegal poppies focusing on Papaver somniferum L., and species identification studies of Papaver bracteatum and Papaver setigerum DC. still lacking. As a result, in order to evaluate the performance of genetic markers and classify their DNA sequences in the genus Papaver, this study developed the first machine learning-based two-layer model, in which the first layer classifies legal and illegal poppies from the given sequence and the second layer identifies species of illegal poppies using their sequences. We constructed the dataset and investigated biological features from four markers, internal transcribed spacer 1 (ITS1), internal transcribed spacer 2 (ITS2), transfer RNA Leucine (trnL), transfer RNA Leucine - transfer RNA Phenylalanine intergenic spacer (trnL–trnF intergenic spacer) and their combination, using four machine learning algorithms, K-nearest neighbor (KNN), Naïve Bayes (NB), extreme gradient boost (XGBoost) and Random Forest (RF). According to our findings, for Layer 1 to classify legal and illegal poppies, KNN-based models using combined ITS region achieved the greatest performance of accuracy 0.846 and 0.889 using training and test sets, respectively. Additionally, for Layer 2 to identify illegal poppy species, KNN-based models using combined ITS region achieved the best performance of 0.833 and 1.000 for using training and test sets, respectively. To validate the model, the combined ITS region, which includes ITS 1 and 2 sequences, from blind poppy samples were used as a case study, with the Layer 1 correctly classifying legal and illegal poppies with over 0.830 accuracy. Layer 2 correctly identified P. setigerum DC., however, only one of the three P. somniferum L. species was accurately identified. Nevertheless, our research shows that machine learning can be used to classify and identify legal and illegal poppy species using DNA barcodes which can then be used as an efficient and effective forensic tool for improved law enforcement and a safer society.

Too Warm and Too Dry—Decline and Threat of the Subarctic-Subalpine Liverwort Hygrobiella laxifolia in a Low Mountain Range in Central Europe under the Conditions of Climate Change

Using the liverwort Hygrobiella laxiflora as an example, this study aims to investigate the impact of climate change on montane species and their ecosystems. The study area is the Elbe Sandstone Mountains in Germany and the Czech Republic, where Hygrobiella laxifolia has an isolated occurrence in deeply carved gorges characterized by a specific microclimate (cellar climate). The focus is on determining the rate at which populations are affected. After little change in the population over a long period, a sudden sharp decline has been observed in recent years. The decline correlates with the mass proliferation of the European spruce bark beetle (Ips typographus) observed since 2017, which has led to a strong thinning of the forests in the area. In many areas of occurrence, the forests now have a much sparser tree layer or none at all. Competition from other, more competitive species also plays an important role as a cause of decline, as these have spread more widely on the sites due to the now higher light levels. The severe bark beetle infestation and the large-scale forest fire of 2022 both represent events that led to the almost complete deforestation of large parts of the study area, which was previously unknown on this scale. Since both the bark beetle infestation and the forest fire are strongly intensified by climate change, we assume that climate change is the main reason for the decline of Hygrobiella laxifolia in the area. The populations of Hygrobiella laxifolia of the Elbe Sandstone Mountains show some morphological peculiarities. As two new species have recently been described within Hygrobiella, we have integrated a sample from the area into a molecular phylogeny of Hygrobiella based on the plastid sequences of trnL-trnF intergenic spacer and matK to clarify the relationships. The sample from Elbe Sandstone Mountains fits well into the clade of Hygrobiella laxifolia.

Development of a Genus Identification Method for Poisonous Plants Using Real-Time PCR

We have developed a rapid genus identification method for poisonous plants. The real-time PCR using the TaqMan® probe method was employed for detection, with the amplified targets being the "trnL (UAA)-intron" or "trnL-trnF intergenic spacer" regions of chloroplast DNA. The targeted plants were selected six genera (Aconitum, Colchicum, Veratrum, Brugmansia, Scopolia and Narcissus), which have been implicated in many instances of food poisoning in Japan. A tissue lysis solution was used for DNA extraction, which can be completed within approximate 30 min. A master mix corresponding to the tissue lysis solution was used for real-time PCR reagents. As a result, we were able to complete the entire process from DNA extraction to genus identification in 4 to 5 hr. The detection sensitivity was estimated at approximately 1 pg of DNA for all six plant genera. Remarkably, an amplification plot was discerned even with the crude cell lysates of all samples. It was also possible to obtain amplification curves for three plant samples that had been subjected to simulated cooking (boiling). This study suggests that the developed method can rapidly identify six genera of poisonous plants.

Genetic and ecological differentiation of Caragana jubata (Pall.) Poir.

Caragana jubata (Pall.) Poir. is a cold-tolerant mountain shrub and important medicinal species of Fabaceae family with a disjunctive pan-Asian range. Sequences of the atpH-atpI and trnL-trnF intergenic spacers of chloroplast DNA and of the internal transcribed spacer region, ITS-1, of the nuclear ribosomal DNA were used to investigate the intraspecific evolution and phylogeography of this species. Eleven cpDNA haplotypes were identified. The population subdivision is very high (GST = 0.743, NST = 0.943), suggesting a distinct phylogeographic structure (NST > GST, P < 0.001). Phylogenetic analyses of the eleven haplotypes were clustered into four clades, consistent with their respective distributions in four separate regions: Tibet, Tien Shan, Buryatia and Magadan Region. This pronounced phylogeographic gap is also confirmed by nrITS data, however, there is some mixture of genotypes in the Magadan and Buryat populations. Genetic evidence suggests that the center of origin of C. jubata was Tibet and adjacent mountain systems, from where the species spread westwards along continuous mountain chains towards the Tien Shan and northeastwards to the coast of the Sea of Okhotsk and further northwards to Arctic. Divergence of Tien Shan populations of C. jubata may have been caused by climate change in the Pleistocene associated with increased aridity and expansion of deserts, while subsequent climate-induced cycles of range contraction/expansion increased geographic isolation and fragmentation of more northern populations. Marginal northern populations now occupy sites with the most favourable environmental conditions. Special attention is paid to the Okhotsk populations of this species preserved in Magadan Region.

Molecular phylogenetic identification of Artemisia l. Species from Mongolia

The Artemisia L. genus, one of the largest genera in the Asteraceae family, consists of many medicinally important and phylogenetically unresolved species. To define the phylogenetic relationship of Artemisia species, nucleotide sequences of the nuclear ITS (Intergenic spacer DNA) region, chloroplast trnL-trnF intergenic spacer, partial sequences of plastid rbcL gene were identified from medicinally important 12 species included in 3 subgenera. The phylogenetic tree was constructed through the Neighbor-Joining and Maximum Parsimony analysis, respectively. The results of study revealed that the combination of the nucleotide sequences from the ITS and rbcL region was more efficient in determining the phylogenic relationship of species.

The search for hybrids between Myriophyllum sibiricum and M. spicatum (Haloragaceae) in North Eurasia allowed to clarify their morphological differences and distribution

Myriophyllum sibiricum and M. spicatum are sympatric in large part of their vast ranges in Eurasia and North America and have different habitat preferences only in East Europe and Siberia. Such absence of both geographic and ecological isolation should trigger extensive hybridization between these species that is indeed observed in North America. The hybrid could be reliably documented only with genetic markers, and in Eurasia this was done only in China. Using genetic markers (ITS2 region of nuclear ribosomal DNA and trnL-trnF intergenic spacer of cpDNA) we documented hybrids between M. sibiricum and M. spicatum on the North Asian Pacific islands with dynamic geological history and climate (Commander and the southern Kuril islands). The hybrid also originated in East Europe (Ivanovo Region) as a result of ecological isolation break due to rapid artificial habitat change. Using genetically verified material throughout temperate Eurasia we clarified morphological characters discriminating M. sibiricum and M. spicatum and, basing on them, refined the species distribution. For the first time we provide a full morphological characteristic of the hybrid between M. sibiricum and M. spicatum and suggest a name for it, M. × hybridum.

Authentication of Wax Apple (Syzygium samarangense (Blume) Merr. & L.M Perry) Delima and Citra Cultivars by Morphological and Molecular Approach

Wax apple Delima and Citra Cultivars are two superior non-climacteric tropical fruit commodities from Demak Regency which have similar morphological characters but have different fruit characteristics. Identification wax apple cultivars from Demak regency using DNA barcoding approach has not been researched yet. The aims of this research are to analyze the morphological similarities of wax apple between Delima and Citra cultivars and to identify genetic variations of wax apple Delima and Citra using trnL-trnF intergenic spacer for molecularly authentication. The results shown that there were no differences in the environmental parameters of the wax apple two cultivated area in Demak Regency. According to the morphological approach, the Delima and Citra cultivars in Demak Regency were grouped separately into Delima and Citra clusters with a similarity index of 61.5% (Citra cultivars) and 60.5% (Delima cultivars) and separated based on the origin of their cultivation area. Genetic variations between wax apple Delima and Citra in Demak Regency consisted of deletions (9delA 17delA) and conserved P6 loop in all compared cultivars. The possibility of heteroplasmy R (A,G) found at 73 nucleotide number in Delima Betokan cultivar and Citra cultivars (Betokan and Jungpasir). Meanwhile, Delima Jungpasir cultivar is K (G,T). There were P8 stem-loops with different lengths between Delima cultivars (Betokan and Jungpasir) and the same length in Citra cultivars (Betokan and Jungpasir). Therefore, the trnL-trnF intergenic spacer has not been thoroughly used in wax apple Delima and Citra authentication due to the presence of a secondary structure which causes the loss of the electropherogram signal so that the nucleotide sequence cannot be read.

Molecular phylogenetic and morphological support for the recognition of Friesodielsia lalisae (Annonaceae), a new species from S Thailand

Thirteen species of Friesodielsia Steenis (Annonaceae) and 11 representatives of related genera were included in molecular phylogenetic analyses using up to six plastid DNA regions (psbA-trnH, trnL-trnF intergenic spacers; trnL intron; matK, ndhF, rbcL exons). The results support the recognition of a new species, F. lalisae Damth., Baka & Chaowasku from Narathiwat, S Thailand, as belonging to one of the two major clades of Friesodielsia. The members of this clade show outer petals that separate at anthesis and have a concave basal portion, while members in the other major clade exhibit outer petals that separate early during developmental stages and have a flat base. Friesodielsia lalisae is described and illustrated. It is most morphologically similar to F. argentea (J. Sinclair) Steenis and F. glauca (Hook. f. & Thomson) Steenis but differs from the two by having dissimilar sepal shape, higher proportion of inner petal to outer petal length and longer inner petals. The new species additionally differs from the former by having different leaf base and from the latter by having denser indumentum on young twigs and shorter flowering pedicels. Narathiwat, a province to which the new species is endemic, seems to be one of the most underexplored areas in Thailand as evidenced by a number of species described based on recent material. The conservation status of the new species is provisionally assessed as Critically Endangered. Citation: Damthongdee A., Khunarak N., Kaeokula S., Saengpho C., Wiya C., Ue-aree P., Baka A., Aongyong K. & Chaowasku T. 2023: Molecular phylogenetic and morphological support for the recognition of Friesodielsia lalisae (Annonaceae) a new species from S Thailand. – Willdenowia 53: 45–55. Version of record first published online on 10 April 2023 ahead of inclusion in April 2023 issue.

MANGIFERA KEMANGA BLUME (ANACARDIACEAE) TAXONOMIC ASSESSMENT FOR GENETIC DIVERSITY BASED ON MOLECULAR SUBSTANTIATION

Mangifera kemanga Blume is a wild relative of mango (Mangifera indica) and a local fruit with various potential uses. The community uses its fruit and leaves for food and its wood as a building material. However, the genetic diversity and taxonomic status of M. kemanga species still need more exploration and have a dispute with M. caesia Jack. The presented study aimed to determine the taxonomic status of M. kemanga and its relationship with M. caesia through a molecular approach and to recognize the species’ delimitation. The recorded data analysis used the trnL-trnF intergenic spacer sequence. Carrying out DNA isolation employed the Geneaid Genomic DNA Mini Kit (Plant) protocol, with the DNA sequences analyzed for kinship using Maximum Parsimony and Neighbor-Joining methods and genetic diversity analysis performed using DnaSP 6. A total of 27 trnL-trnF intergenic spacer sequence identification and authentication resulted from BLAST on NCBI as sequences derived from the genus Mangifera. The phylogenetic tree revealed that the accessions of M. kemanga and M. caesia are very similar, hence, considered not as independent species. Thus, based on the trnL-trnF intergenic spacer sequences, M. kemanga is a synonym of M. caesia with taxonomic status as a variety of M. caesia. This molecular-based taxonomic evidence is significant for determining the valid species status of M. kemanga so that it can provide basic information for further studies on biodiversity and germplasm conservation.

The Complete Chloroplast Genome of An Ophiorrhiza baviensis Drake Species Reveals Its Molecular Structure, Comparative, and Phylogenetic Relationships.

Ophiorrhiza baviensis Drake, a flowering medical plant in the Rubiaceae, exists uncertainly within the Ophiorrhiza genus' evolutionary relationships. For the first time, the whole chloroplast (cp) genome of an O. baviensis Drake species was sequenced and annotated. Our findings demonstrate that the complete cp genome of O. baviensis is 154,770 bp in size, encoding a total of 128 genes, including 87 protein-coding genes, 8 rRNAs, and 33 tRNAs. A total of 59 SSRs were screened in the studied cp genome, along with six highly variable loci, which can be applied to generate significant molecular markers for the Ophiorrhiza genus. The comparative analysis of the O. baviensis cp genome with two published others of the Ophiorrhiza genus revealed a high similarity; however, there were some notable gene rearrangements in the O. densa plastome. The maximum likelihood phylogenetic trees were constructed based on the concatenation of the rps16 gene and the trnL-trnF intergenic spacer sequence, indicating a close relationship between the studied O. baviensis and other Ophiorrhiza. This study will provide a theoretical molecular basis for identifying O. baviensis Drake, as well as species of the Ophiorrhiza genus, and contribute to shedding light on the chloroplast genome evolution of Rubiaceae.

Distribution of Biodiversity of Wild Beet Species (Genus Beta L.) in Armenia under Ongoing Climate Change Conditions.

The reported annual temperature increase and significant precipitation drop in Armenia impact the country’s ecosystems and biodiversity. The present study surveyed the geographical distribution of the local wild beet species under the ongoing climate change conditions. We showed that B. lomatogona, B. corolliflora and B. macrorhiza are sensitive to climate change and were affected to various degrees, depending on their location. The most affected species was B. lomatogona, which is at the verge of extinction. Migration for ca. 90 and 200–300 m up the mountain belt was recorded for B. lomatogona and B. macrorhiza, respectively. B. corolliflora was found at 100–150 m lower altitudes than in the 1980s. A general reduction in the beet’s population size in the native habitats was observed, with an increased number of plants within the populations, recorded for B. corolliflora and B. macrorhiza. A new natural hybrid Beta x intermedium Aloyan between B. corolliflora and B. macrorhiza was described and confirmed using chloroplast DNA trnL-trnF intergenic spacer (LF) and partially sequenced alcohol dehydrogenase (adh) of nuclear DNA. An overview of the wild beets reported in Armenia with the taxonomic background, morphological features, and distribution is provided. Conservation measures for preservation of these genetic resources are presented.

Two New Species of Meiogyne (Annonaceae) from Vietnam, Based on Molecular Phylogeny and Morphology

Meiogyne rubra Jaikhamseub, Damth. & Chaowasku sp. nova and M. vietnamica Jaikhamseub, T.A. Le & Chaowasku sp. nova (Annonaceae) from Vietnam are described and illustrated. Molecular phylogenetic reconstructions based on up to eight plastid DNA regions (matK, ndhF, rbcL and ycf1 exons; trnL intron; ndhF-rpl32, rpl32-trnL and trnL-trnF intergenic spacers) and incorporating 33 accessions (31 species) of Meiogyne were performed. The two new species, as well as M. anomalocarpa, M. caudata, M. chiangraiensis and M. subsessilis are included in molecular phylogenetic analyses for the first time. Two major clades were retrieved within Meiogyne: a small clade consisting of M. anomalocarpa, M. hainanensis and M. kanthanensis, and a much larger clade comprising the remaining accessions. The phylogenetic position of the two new species in the latter clade is obscure. Meiogyne rubra appears to be morphologically most similar to M. monosperma. The former differs from the latter by having a narrower leaf blade, free (vs. basally connate) and longer sepals, longer outer petals and larger inner petals. Meigyne vietnamica is morphologically most similar to M. caudata, but differs from it by having an acute to acuminate (vs. usually distinctly caudate-acuminate) leaf apex, non-prominent (vs. prominent) secondary veins on lower leaf surface and higher number of secondary veins in the leaves. A key to the species of Meiogyne in Vietnam is also provided.

DNA Barcoding of Prunus Species Collection Conserved in the National Gene Bank of Egypt

Two intergenic spacers cpDNA barcoding regions were used to assess the genetic diversity and phylogenetic structure of a collection of 25 Prunus accessions. The trnH-psbA and trnL-trnF intergenic spacers were able to distinguish and identify only four Prunus species. The average aligned length was 316–352 bp and 701–756 bp for trnH-psbA and trnL-trnF, respectively. The overall evolutionary divergence was higher in trnH-psbA than trnL-trnF. The transition/transversion bias (R) recorded as 0.59 in trnL-trnF and 0.89 in trnH-psbA. The number of invariable sites, nucleotide diversity (Pi), and the average number of nucleotide differences (k) was higher in the trnH-psbA region. The trnL-trnF records was above the other region in the number of variable sites, number of singleton variable sites, and the parsimony informative sites. Phylogenetic relationships among the 25 accessions of Prunus species were investigated. Most of the different Prunus species clustered in a homogenized distribution in both regions, except for the plum (P. domestica) accession (African Rose) was assigned with the peach (P. persica) accessions. The two intergenic cpDNA trnH-psbA and trnL-trnF were able to distinguish and identify the four Prunus species accessions.

Clarification of the Position of Linum stelleroides Planch. within the Phylogeny of the Genus Linum L.

The phylogeny of members of the family Linaceae DC. ex Perleb has not been adequately studied. In particular, data on the phylogenetic relationship between Linum stelleroides Planch. and other representatives of the blue-flowered flax are very controversial. In the present work, to clarify this issue, we obtained DNA sequences of three nuclear loci (IGS and ITS1 + 5.8S rDNA + ITS2 of the 35S rRNA gene and the 5S rRNA gene) and eight chloroplast loci (rbcL, the trnL–trnF intergenic spacer, matK, the 3′ trnK intron, ndhF, trnG, the psbA–trnH intergenic spacer, and rpl16) of 10 Linum L. species (L. stelleroides, L. hirsutum, L. perenne, L. leonii, L. lewisii, L. narbonense, L. decumbens, L. grandiflorum, L. bienne (syn. L. angustifolium), and L. usitatissimum) using high-throughput sequencing data. The phylogenetic analysis showed that L. stelleroides forms a basal branch in the blue-flowered flax clade. Previously found inconsistencies in the position of L. stelleroides and some other species in the Linaceae phylogenetic tree resulted from the erroneous species identification of some of the studied plant samples.

DNA barcoding analysis and phylogenetic relationships of Indian wild coffee species

Wild coffee species are the reservoirs of genetic diversity and could play a critical role in the genetic improvement of coffee. However, the conservation and genetic assessment of wild coffee species has been largely neglected. In the present study, DNA barcoding approaches were employed to assess the phylogenetic relationships between five indigenous wild coffee species from India and compared with two cultivated, and four wild coffee species of African origin. The efficacy of three barcoding loci namely matK,rbcL, and trnL-trnF was investigated using PCR amplification and sequence characterization. The intergenic spacer trnL-trnF is the highly polymorphic loci followed by matK and rbcL chloroplast gene. Among the three barcoding loci, the matK locus has the maximum number of parsimony informative sites, whereas the trnL-trnF locus contains maximum singleton variable sites. Although all the three loci contain a few unique fixed nucleotides (UFNs), no individual barcode locus has the critical nucleotide sequence tags for all the five Indian wild coffee species that help in species discrimination. However, the multilocus combinations are efficient in discriminating the species due to the presence of SNPs and specific sequence tags. The phylogenetic tree constructed using the maximum likelihood analysis of the combined barcoding loci separated all the Indian wild coffee species from African wild coffee species compared to phylogeny inferred using individual barcoding loci. Our study supports the utility of DNA barcoding as a useful tool for coffee species identification, which can be used for conservation purposes.

Pyramidanthe and Mitrella (Annonaceae, Uvarieae) unified: molecular phylogenetic and morphological congruence, with new combinations in Pyramidanthe

The delimitation of the genera Pyramidanthe and Mitrella in the tribe Uvarieae of the family Annonaceae is assessed by molecular phylogenetic analyses and morphological re-evaluation. Using up to six plastid DNA regions (matK, ndhF, rbcL exons; trnL intron; psbA-trnH, trnL-trnF intergenic spacers) and including, among others, two accessions of Pyramidanthe and five accessions of Mitrella, the resulting phylogeny shows that Pyramidanthe and Mitrella are recovered in a clade sister to the Fissistigma clade. The Pyramidanthe-Mitrella clade is composed of a trichotomy: a clade consisting of Pyramidanthe accessions and two clades containing Mitrella accessions. In combination with negligible morphological distinctions between the two genera, they are consequently merged, with 11 new combinations under the chosen name Pyramidanthe: P. beccarii, P. clementis, P. cylindrica, P. dielsii, P. elegans, P. kentii, P. ledermannii, P. mabiformis, P. schlechteri, P. sylvatica and P. tiwiensis. The names M. dielsii (the basionym of P. dielsii) and P. rufa (a heterotypic synonym of P. prismatica) are lectotypified. Pyramidanthe s. lat. possesses the following diagnostic traits: usually indistinct secondary leaf veins with a brochidodromous to brochidodromous-eucamptodromous venation, a reticulate tertiary leaf venation, axillary inflorescences, presence of a basal excavation on an inner side of each outer petal, and inner petals that are much smaller than the outer petals and cohering marginally at anthesis. Citation: Bangkomnate R., Damthongdee A., Baka A., Aongyong K. & Chaowasku T. 2021: Pyramidanthe and Mitrella (Annonaceae, Uvarieae) unified: molecular phylogenetic and morphological congruence, with new combinations in Pyramidanthe. – Willdenowia 51: 383–394. Version of record first published online on xx December 2021 ahead of inclusion in December 2021 issue.

Comparative Genetic Analysis between the Jeju ‘Inchangkyool’ and Chinese ‘Ichangensis’ (Citrus ichangensis) using Internal Chloroplast trnL-trnF Intergenic Spacers and Transcribed Spacer Sequence Regions

Comparative Genetic Analysis between the Jeju ‘Inchangkyool’ and Chinese ‘Ichangensis’ (<i>Citrus ichangensis</i>) using Internal Chloroplast trnL-trnF Intergenic Spacers and Transcribed Spacer Sequence Regions

Sequencing Relationships among Five Species of Hydrocotyle Based on TrnL-TrnF Intergenic Spacer Gene

Sequencing Relationships among Five Species of Hydrocotyle Based on TrnL-TrnF Intergenic Spacer Gene