scholarly journals DNA Barcoding in Selected Species and Subspecies of Rye (Secale) Using Three Chloroplast Loci (matK, rbcL, trnH-psbA)

2018 ◽  
Vol 47 (1) ◽  
pp. 54-62 ◽  
Author(s):  
Lidia SKUZA ◽  
Izabela SZUĆKO ◽  
Ewa FILIP ◽  
Anastazja ADAMCZYK
Keyword(s):  
Chloroplast Dna ◽  
Dna Barcoding ◽  
Plant Species ◽  
Plant Material ◽  
High Variability ◽  
Universal Primers ◽  
Neighbor Joining ◽  
Cultivated Species ◽  
Chloroplast Dna Regions

DNA barcoding is a relatively new method of identifying plant species using short sequences of chloroplast DNA. Although there is a large number of studies using barcoding on various plant species, there are no such studies in the genus Secale. In this study the plant material consisted of 10 cultivated and non-cultivated species and subspecies of rye genus. Three chloroplast DNA regions (rbcL, matK, trnH-psbA) were tested for their suitability as DNA barcoding regions. Universal primers were used, and sequenced products were analyzed using Neighbor Joining and the Maximum Likelihood in the MEGA 7.1 program. We did not observe high variability in nucleotide sequences within the matK and rbcL regions. Only 2.2% of the sequences showed polymorphism in the rbcL region, while 6.5% in the matK region. The most variable trnH-psbA (15.6%) intergenic region was the most useful for rye barcoding. Individual application of the studied regions did not provide the expected results. None of the regions used in the study allowed the division of rye species and subspecies according to the adopted classification of the genus Secale. The results confirm that the use of matK and rbcL is insufficient for DNA barcoding in rye species, and better discrimination within the genus Secale can be obtained only in combination with the non-coding trnH-psbA sequence. Our results also indicate the necessity of using a different region. All of the new sequences have been deposited in Genbank.

scholarly journals Evaluation of Genetic Diversity by DNA Barcoding of Local Lotus Populations from Thua Thien Hue Province

2020 ◽  
Author(s):  
Dang Thanh Long ◽  
Hoang Thi Kim Hong ◽  
Le Ly Thuy Tram ◽  
Nguyen Thi Quynh Trang
Keyword(s):  
Genetic Diversity ◽  
Chloroplast Dna ◽  
Dna Barcoding ◽  
Plant Species ◽  
High Variability ◽  
Universal Primers ◽  
Minimum Evolution ◽  
Intergenic Regions ◽  
Chloroplast Dna Regions

Background: DNA barcoding is a relatively new method of identifying plant species using short sequences of chloroplast DNA. Although there is a large number of studies using barcoding on various plant species, there are no such studies in the genus Nelumbo. Methods: Three chloroplast DNA regions (rbcL, matK, trnH-psbA) were tested for their suitability as DNA barcoding regions of thirty three lotus samples which were collected in Thua Thien Hue province, Vietnam. Universal primers were used and sequenced products were analyzed using Minimum Evolution method in the MEGA 7.0 program.Result: We did not observe high variability in nucleotide sequences within the rbcL region (0.135%). White Nelumbo, while, the most encoding matK (8.013%) and variable trnH-psbA (with different number of repeating regions TAAAA) intergenic regions was the most useful for Nelumbo barcoding. Individual application of the studied regions did not provide the expected results. None of the regions used in the study allowed the division of white and pink lotus varieties of N. nucifera specie according to the adopted classification of the genus Nelumbo. The results confirm that the use of matK, rbcL and trnH-psbA or combine all three regions together is insufficient for DNA barcoding in white and pink lotus varieties of N. nucifera specie and better discrimination within the genus Nelumbo. Our results also indicate the necessity of using a different region. All of the new sequences have been deposited in GeneBank under the following accession numbers: rbcL (MN011708 to MN068956); matK (MN011719 to MN068978) and trnH-psbA (MN011730 to MN086252). 

scholarly journals A phylogenetic analysis and new delimitation of Crepidiastrum (Asteraceae, tribe Cichorieae)

Phytotaxa ◽  
2014 ◽  
Vol 159 (4) ◽  
pp. 241 ◽  
Author(s):  
Yu-lan Peng ◽  
Yu Zhang ◽  
Xin-fen Gao ◽  
Lin-jing Tong ◽  
Liang Li ◽  
...  
Keyword(s):  
Phylogenetic Analysis ◽  
Maximum Likelihood ◽  
Chloroplast Dna ◽  
Maximum Parsimony ◽  
Phylogenetic Analyses ◽  
Systematic Position ◽  
Likelihood Inference ◽  
Nuclear Its ◽  
Chloroplast Dna Regions

The systematic position of Paraixeris humifusa (Asteraceae) is hard to define, because the circumscription of Paraixeris, Youngia and Crepidiastrum, three closely related genera in subtribe Crepidinae (Cichorieae), is not clear. This paper reports on the relationships between 30 species in subtribe Crepidinae, based on an analysis of nucleotides from one nuclear (ITS) and three chloroplast DNA regions ( trnL-F, rps16 and atpB-rbcL). The phylogenetic analyses used maximum parsimony with maximum likelihood inference. The monophyly of Crepidiastrum in the most recent generic classification of Shih & Kilian (2011) is explored. The results show that 12 species in Crepidiastrum constitute a monophyletic group, and that Paraixeris humifusa should be treated as Youngia humifusa.

scholarly journals Forensic botany and forensic chemistry working together: application of plant DNA barcoding as a complement to forensic chemistry—a case study in Brazil

Genome ◽  
2019 ◽  
Vol 62 (1) ◽  
pp. 11-18 ◽  
Author(s):  
Renato T.F. Paranaiba ◽  
Carlos B.V. Carvalho ◽  
Jorge M. Freitas ◽  
Levy H. Fassio ◽  
Élvio D. Botelho ◽  
...  
Keyword(s):  
Dna Barcoding ◽  
Plant Species ◽  
Plant Material ◽  
Forensic Analysis ◽  
Forensic Chemistry ◽  
Artemisia Absinthium ◽  
Genetic Sequencing ◽  
Plant Dna ◽  
Plant Matter ◽  
Working Together

Recently, Brazilian Federal Police used forensic chemistry and forensic botany techniques on a case. Two packets containing fragmented plant matter were seized and sent for forensic analysis. Forensic chemistry, the gold standard for evaluating plant material suspected to contain illicit substances, did not find illicit materials. Gas chromatography coupled mass spectrometry (GC-MS) identified thujone in the botanical material. Thujone is a chemical compound naturally found in many plant species, notably Artemisia absinthium. Because doubt remained, we next used plant DNA barcoding methods. Total DNA from plant tissue fragments was extracted and five different DNA regions were amplified, sequenced, and analyzed using plant DNA barcoding methods. Genetic analysis yielded 30 good quality sequences representing five taxa. Most specimens were identified as A. absinthium. Few studies focus on practical forensic applications of plant DNA barcoding methods using a case solved in a forensic laboratory with its difficulties and limitations. To the best of our knowledge, this is the first study to report an effective joint effort of forensic chemistry and botany techniques to assess plant material in Brazil. The availability of a new technical approach for the genetic sequencing of plant species will enhance many forensic investigations and inspire similar initiatives.

Real-Time PCR Quantification of Chloroplast DNA Supports DNA Barcoding of Plant Species

2016 ◽  
Vol 58 (3) ◽  
pp. 212-219 ◽  
Author(s):  
Hitomi S. Kikkawa ◽  
Kouichiro Tsuge ◽  
Ritsuko Sugita
Keyword(s):  
Chloroplast Dna ◽  
Real Time ◽  
Dna Barcoding ◽  
Plant Species ◽  
Real Time Pcr

scholarly journals IDENTIFIKASI SPESIES MENGGUNAKAN DNA BARCODING DALAM MENUNJANG BUDIDAYA DAN KONSERVASI TERIPANG DI PERAIRAN LAMPUNG

2021 ◽  
Vol 16 (1) ◽  
pp. 31
Author(s):  
Anna Rejeki Simbolon ◽  
Masteria Yunovilsa Putra ◽  
Ismiliana Wirawati
Keyword(s):  
Dna Barcoding ◽  
Sea Cucumber ◽  
Phylogenetic Reconstruction ◽  
Scuba Diving ◽  
Coi Gene ◽  
Universal Primers ◽  
Neighbor Joining ◽  
Accurate Identification ◽  
Sea Cucumbers ◽  
The Right

Teripang merupakan komoditas perikanan yang saat ini dibudidayakan dan dieksploitasi di perairan Lampung. Namun terdapat kesulitan dalam mengidentifikasi teripang karena kemiripan morfologis di antara spesies yang ada. Identifikasi yang baik berguna agar proses pembudidayaan dan konservasi dapat tepat sasaran. Penggunaan DNA barcoding dapat digunakan untuk mengidentifikasi jenis-jenis teripang yang ada, jarak genetik, dan keragaman genetik intra/inter spesies. Penelitian ini bertujuan untuk mengidentifikasi teripang di perairan Lampung dengan menggunakan sekuen DNA gen COI. Teripang diambil dengan menggunakan metode jelajah pada saat surut dan dengan scuba diving. Pengamatan DNA menggunakan primer universal ceF, pengeditan dan diurutkan dengan program Geneious ver 9 dan program BLAST. Konstruksi pohon filogenetik dilakukan dengan metode neighbor joining (NJ) pada model Kimura-2. Penelitian ini menunjukkan spesies teripang yang teridentifikasi adalah Holothuria leucospilota, H. atra, Stichopus vastus, dan S. horrens dengan jarak kesamaan 99%-100%. S. vastus dan S. horrens memiliki jarak genetik terendah dengan pengurangan yang tinggi. Rekonstruksi filogenetik memperlihatkan pengelompokkan spesies-spesies ke dalam genus Holothuria dan Stichopus. Stichopus sp. memiliki kesamaan morfologi yang tinggi sehingga kesalahan identifikasi sering terjadi. DNA barcoding dapat mengidentifikasi teripang secara cepat dan akurat sehingga pengelolaan teripang baik secara budidaya maupun pengambilan langsung di alam dapat berkelanjutan. Identifikasi spesies yang tepat menjadi kunci utama dalam upaya pembudidayaan dan konservasi teripang yang tepat sasaran dan berkelanjutan.Sea cucumbers is a highly valued fishery commodity that is currently cultivated and exploited in Lampung waters. However, differentiating a sea cucumber species from another is sometimes difficult due the morphological similarities between the species. Developing an accurate identification method is then critical to ensure successfull farming activities and conservation efforts of sea cucumbers. DNA barcoding could be used to accurately identify sea cucumber species, genetic distance, and genetic diversity between species. This study aimed to identify sea cucumbers existed in Lampung waters using DNA barcoding of the COI gene with ceF and ceR universal primers. Sea cucumbers are taken using the cruising method at low tide and by scuba diving. The DNA sequence was then edited and aligmented using the Geneious ver.9 program and analyzed using the BLAST program. Phylogenetic tree construction was carried out using the neighbor joining (NJ) method on the Kimura-2 model. This study showed that the identified species of sea cucumbers were Holothuria leucospilota, H. atra, Stichopus vastus, and S. horrens with a similarity distance of 99%-100%. S. vastus and S. horrens have the lowest genetic range. Phylogenetic reconstruction shows the classification of species into the genus Holothuria and Stichopus. Stichopus sp. have high morphological similarities within the same genus which often lead to species misidentification. DNA barcoding can identify sea cucumbers quickly and accurately. This method allows the identification of the right sea cucumber species which is the main key in the effort to cultivate and conserve targeted and sustainable sea cucumbers.

Phylogenetic position ofKontumia(Polypodiaceae) inferred from four chloroplast DNA regions

2012 ◽  
pp. n/a-n/a
Author(s):  
Changkyun Kim ◽  
Hong-Guang Zha ◽  
Tao Deng ◽  
Hang Sun ◽  
Su-Gong Wu
Keyword(s):  
Chloroplast Dna ◽  
Phylogenetic Position ◽  
Chloroplast Dna Regions

scholarly journals Molecular variation in chloroplast DNA regions in ancestral species of wheat.

Genetics ◽  
1994 ◽  
Vol 137 (3) ◽  
pp. 883-889 ◽  
Author(s):  
N T Miyashita ◽  
N Mori ◽  
K Tsunewaki
Keyword(s):  
Chloroplast Dna ◽  
Restriction Enzymes ◽  
Triticum Dicoccoides ◽  
The Other ◽  
Major Type ◽  
Aegilops Speltoides ◽  
Triticum Timopheevi ◽  
Restriction Sites ◽  
D Values ◽  
Chloroplast Dna Regions

Abstract Restriction map variation in two 5-6-kb chloroplast DNA regions of five diploid Aegilops species in the section Sitopsis and two wild tetraploid wheats, Triticum dicoccoides and Triticum araraticum, was investigated with a battery of four-cutter restriction enzymes. A single accession each of Triticum durum, Triticum timopheevi and Triticum aestivum was included as a reference. More than 250 restriction sites were scored, of which only seven sites were found polymorphic in Aegilops speltoides. No restriction site polymorphisms were detected in all of the other diploid and tetraploid species. In addition, six insertion/deletion polymorphisms were detected, but they were mostly unique or species-specific. Estimated nucleotide diversity was 0.0006 for A. speltoides, and 0.0000 for all the other investigated species. In A. speltoides, none of Tajima's D values was significant, indicating no clear deviation from the neutrality of molecular polymorphisms. Significant non-random association was detected for three combinations out of 10 possible pairs between polymorphic restriction sites in A. speltoides. Phylogenetic relationship among all the plastotypes (plastid genotype) suggested the diphyletic origin of T. dicoccoides and T. araraticum. A plastotype of one A. speltoides accession was identical to the major type of T. araraticum (T. timopheevi inclusively). Three of the plastotypes found in the Sitopsis species are very similar, but not identical, to that of T. dicoccoides, T. durum and T. aestivum.

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