Phylogenetic analysis of six different species of Saraca L. (Fabaceae, Caesalpinioideae) based on chloroplast matK gene

Sil S, De KK, Ghosh A. 2021. Phylogenetic analysis of six different species of Saraca L. (Fabaceae, Caesalpinioideae) based on chloroplast matK gene. Biodiversitas 22: 3880-3889. Saraca L. is one of the most important genera, with several horticultural and therapeutic values. Specific taxonomic and phylogenetic knowledge of Saraca through molecular data is essential for accessing its true medicinal benefits. Nineteen different Partial matK gene sequences of the chloroplast genome of six different species of Saraca, including four amplified and 15 retrieved from the NCBI gene bank, were place in a sequence alignment. The resulting data were examined to determine their phylogenetic and evolutionary interrelationships. The comparative analysis of different sequences of each of the species revealed intra-specific molecular diversity, and the comparison of the matK sequences of six different species defined their inter-specific molecular diversity. The analysis of partial matK sequences revealed the presence of 87 variable sites, 14 parsimony informative sites, 54 singleton sites, and 237 quadri-fold degenerate sites. The approximate nucleotide composition was A-31.02%, T-37.46%, C-16.06%, and G-15.46%. The value of transition/transversion bias was 0.90. About 522 codons were analyzed and the presence of 34 variable sites, 8 parsimony informative sites, and 25 singleton sites was observed within their respective amino acid sequences. The average pair-wise distance was 0.0444, and 189 segregating sites and 0.018809 nucleotide diversity were observed. The evolution of different species of Saraca and their phylogenetic interrelationships were observed by analyzing their matK sequences. The relative homogeneity of S. indica is quite low. S. dives had the earliest evolutionary trends while S. declinata had the most recent. S. asoca and S. indica are quite similar on the molecular level but can be treated as different species while the difference between S. declinata and one of its synonyms, S. palembanica, indicates the possibility of separating them into different species.

Endorsement and Phylogenetic Analysis of some Fabaceae Plants based on DNA Barcoding gene MatK

DNA barcodes have been considered as a tool to facilitate species identification based on their simplicity and high-level accuracy compression to the complexity and subjective biases linked to morphological identification of taxa. MaturaseK gene “ MatK” of the chloroplast is very crucial in the plant system which is involved in the group II intron splicing. The main objective of this current study is determining the relative utility of the “ MatK” chloroplast gene for barcoding in fifteen legume trees by both single region and multiregional approaches. The chloroplast “ MatK” gene sequences were submitted to GenBank and accession numbers (GenBank: LC602060, LC602154, LC602263, LC603347, LC603655, LC603845, LC603846, LC603847, LC604717, LC604718, LC605994, LC604799, LC605995, LC606468, LC606469) were obtained with sequence length ranging from 730 to 1545 nucleotides. These DNA sequences were aligned with database sequence using PROMALS server , Clustal Omega server and Bioedit program. Also, the maximum likelihood and neighbor-joining algorithms for phylogenetic reconstruction using the MEGA-X program were employed. Overall, these results indicated that the phylogenetic tree analysis and the evolutionary distances of an individual dataset of each species were agreed with a phylogenetic tree of all each other consisting of two clades, the first clade comprising (Enterolobium contortisiliquum, Albizia lebbek), Acacia saligna , Leucaena leucocephala, Dichrostachys Cinerea, (Delonix regia, Parkinsonia aculeata), (Senna surattensis, Cassia fistula, Cassia javanica) and Schotia brachypetala were more closely to each other, respectively. The remaining four species of Erythrina humeana, (Sophora secundiflora, Dalbergia Sissoo, Tipuana Tipu) constituted the second clade. Therefore, MatK gene is considered promising a candidate for DNA barcoding in plant family Fabaceae and providing a clear relationship between the families. Moreover, their sequences could be successfully utilized in single nucleotide polymorphism (SNP) or part of the sequence as DNA fragment analysis utilizing polymerase chain reaction (PCR) in plant systematic.

Phylogenetics of native conifer species in Vietnam based on two chloroplast gene regions rbcL and matK

We used two chloroplast gene regions (matK and rbcL) as a tool for the identification of 33 local conifer species. All 136 sequences, 101 newly generated (14 species for gene matK; 16 species for gene rbcL) and 35 retrieved from the GenBank, were used in the analysis. The highest genetic distance (matK region) was recorded between the species in Cupressaceae with an average of 5% (0.1–8.5), Podocarpaceae with an average of 6% (0–8.5), Taxaceae with an average of 5% (0.2–0.5) and Pinaceae with an average of 20.4% (0.8–54.1). The rbcL region showed a low genetic distance between the species in Cupressaceae 2% (0–3.3), Podocarpaceae 3% (0.6–3.4), Taxaceae 1% (0–2.1) and Pinaceae 1.2% (0–5.82). The phylogenetic analyses using the Maximum likelihood (ML) and Bayesian inference (BI) bootstrap values obtained at the branching nodes of each species ranged from 62 to 100% (Maximum likelihood bootstrap – MLBS and Bayesian posterior probabilities – BPP) for the matK gene; from 66 to 100% (MLBS) and 60 to 100% (BPP) for the rbcL region. The rbcL region was not identified between the species of Taxaceae and Cephalotaxaceae. The matK gene region was very clear in the different species among the families (Cupressaceae, Podocarpaceae, and Cephalotaxaceae) and unsuitable for identifying closely related species in Amentotaxus (Taxaceae) and Pinus (Pinaceae). The gene (matK) is a useful tool as a barcode in the identification of conifer species of Cupressaceae, Podocarpaceae, and Cephalotaxaceae in Vietnam.

Fagopyrum longistylum (Polygonaceae), a new species from Sichuan, China

A new species of Fagopyrum (Polygonaceae), Fagopyrum longistylum, is described and illustrated from Huili County, Sichuan Province, China on the basis of morphological, caryological, and molecular data. F. longistylum is morphologically similar to F. gracilipes from which it differs in having green stem and short-styled flowers with long anther but it is self-compatible. In addition, F. longistylum is a diploid species with 2n=2x=16 while F. gracilipes is tetraploid with 4n=4x=32. Molecular data based on nucleotide sequence polymorphisms of internal transcribed spacers (ITSs) and the maturase K (matK) gene confirm the separation of the new species.

DNA BARCODING EDELWEISS (Anaphalis longifolia) ASAL SUMATERA UTARA MENGGUNAKAN SEKUEN GEN maturase K

Anaphalis longifolia merupakan anggota dari family Asteraceae yang tersebar di dataran tinggi Eropa, Amerika, hingga Asia. Penelitian tentang tanaman ini masih terbatas pada studi habitat, sedangkan penelitian terkait identifikasi molekuler masih belum dilakukan. Penelitian ini bertujuan untuk menganalisis DNA barcode dari A. longifolia menggunakan sekuen matK gene. Sampel yang diperoleh dari Sumatera Utara kemudian di Isolasi DNA, di amplifikasi menggunakan primer spesifik, lalu disequencing. Hasil sequencing dianalisis menggunakan program Molecular Evolution Genetics Analysis (MEGA) Version X. Hasil penelitian menunjukkan bahwa sekuen matK gen berhasil diamplifikasi pada panjang 800-850 kb. Hasil analisis pohon filogenetik menunjukkan bahwa sekuen matK gene dapat mengelompokkan A. longifolia. Pada sekuen matK gene A. longifolia, AT content lebih tinggi dibandingkan dengan GC conten. Jarak genetik yang diperoleh berkisar 0-0.0014. Hasil analisis alignment sekuen matK gene menunjukkan terdapat 1521 karakter yang dapat diamati, 1403 karakter conserved site, 118 karakter variable site, 9 karakter parsimony informative site, dan 7 karakter single nucleotide polymorphism (SNP) site. Sekuen matK gene dapat digunakan sebagai DNA barcoding untuk mengidentifikasi A. longifolia. Hasil penelitian ini diharapkan dapat memberikan informasi penting dalam konservasi A. longifolia.

DNA barcode of Metroxylon sagu and others palm species using matK gene

Abbas B, Kabes RJ, Mawikere NL, Ruimassa RMR, Maturbong RA. 2020. DNA barcode of Metroxylon sagu and other palm species using matK gene. Biodiversitas 21: 4047-4057. Palm family encompasses numerous species, and they disperse broadly across tropical and subtropical countries. The palm family is generally used as an ornamental plant, food, oil, and industrial raw materials. Species of palm that accumulate large amount carbohydrate in the trunk is sago palm (Metroxylon sagu Rottb). The objectives of this study were to explore the plastid sequence associated with matK genes in the palm family and to reveal DNA barcode of 16 genera and 28 species of the palm. Plant materials used in the studies were derived from Sago Research Center (SRC), and other palm sequences were retrieved from the GenBank, NCBI accessions. The PCR product was sequenced by the 1st Base Asia, Singapore. Sequences of the matK gene that were observed in the plastid genome of M. sagu were registered into the NCBI GenBank as DNA barcode of M. sagu. Percent query cover of Blast analysis range from 98% to 100%, and identity range from 97.70% to 100%. Plastid nucleotides associated with matK genes in the chloroplast genome of palm family were shown substantial differences in inter-genera and small differences in inter-species. Genetic distance among palm family range from 0 to 0.026 with nucleotide variation observed was of 0.008822 value. The result of molecular phylogenetic analysis showed that the palm family separated into three clades and three sub-clades based on the plastid matK gene. Species of M. sagu and M. warburgii were considered closely related as compared to other palm species. The matK gene barcoding method is one powerful tool for identification inter-genera and inter-species of the palm.

Comparative genomics plastomes of the Amaryllidaceae family species

The genus Allium covers more than 800 species, signaling among the largest among monocotyledons. The genus contains many economically important species, including garlic, leeks, onions, chives and Chinese chives. Due to the high conservation of chloroplast genomes compared to nuclear genomes and mitochondrial genome, sequence of chloroplasts in Amaryllidaceae have been consistently used for species identification and various in silico programs and strategies have been used to identify, characterize and compare plastid genome regions. Plastome from 15 species of the Amaryllidaceae family revealed similarity in both sequences and in the organization of their gene regions. The base pairs (bp) number ranged from 145,819 (A. paradoxum) to 159,125 (A. ursinum). In respect the GC content, the species presented a variation between 36.7% (A. schoenoprasum and A. sativum) and 37.5% (A. coddii) and the gene space ranged from 84.760 (A. paradoxum) to 94.766 (A. sativum). The genes that encode proteins showed values between 78 (A. paradoxum) to 89 (A. cepa). Phylogenetic trees acquired through alignment of complete plastomas and the plastidial matK gene revealed similarity to the proposed classification for the family. For the genus Allium, there was the formation of three clades with perfect correspondence of the clusters to the three evolutionary lines of the genus.

Using ITS-rDNA and matK gene nucleotide sequences for identification ginseng species in Panax in Phu Xai Lai Leng, Ky Son, Nghe An

DNA barcoding is a useful tool for species identification using standardized genomic DNA fragments. We used DNA barcodes (ITS-rDNA and matK gene) to explore Panax (32 samples collected from Phu Xai Lai Leng mountain and 19 samples collected from medicinal nursery of TH), and to investigate the phylogenetic taxonomy of Panax. In this study, the PCR success rate for ITS-rDNA and matK region was 100%. The success rate of bidirectional sequencing of PCR product was 100% of ITS-rDNA and matK region with length of 616 bp, 1433 bp, respectively. All 32 samples (Panax TB) of Phu Xai Lai Leng have a close relationship with P. stipuleanatus (MLBS = 99%, BPP = 100%). All 19 samples (Panax TH) of medicinal nursery have a close relationship with P. notoginseng (MLBS = 100%, BPP = 100%). Interspecific genetic distances within and among Panax species was varied from 0.2% to 7.9%, average (4%) (ITS-rDNA gene) and 0.1 to 2.9%, average (1.2%) (matK gene). The genetic relationship of species/gender belonging to the Panax genus showed that they have the same evolutionary origin and discovered that new distributed of P. stipuleanatus in Phu Xai Lai Leng mountain in Vietnam.