Development of DNA Barcode and Species-Specific Markers forHelopeltis antoniiSignoret andPachypeltis maesarum(Kirkaldy) (Heteroptera: Miridae), Pests of Cashew in India

2012 ◽  
Vol 122 (2) ◽  
pp. 173-182
Author(s):  
R. Asokan ◽  
K. B. Rebijith ◽  
N. K. Krishna Kumar ◽  
K. K. Srikumar ◽  
P. Shivarama Bhat
Keyword(s):  
2020 ◽  
Vol 23 ◽  
pp. 100376
Author(s):  
Amit Kumar ◽  
Vereena Rodrigues ◽  
Kuppusamy Baskaran ◽  
Ashutosh K. Shukla ◽  
Velusamy Sundaresan

2021 ◽  
Vol 43 (3) ◽  
pp. 2177-2188
Author(s):  
Hakjoon Choi ◽  
Wan Seok Kang ◽  
Jin Seok Kim ◽  
Chang-Su Na ◽  
Sunoh Kim

Scutellaria L. (family Lamiaceae) includes approximately 470 species found in most parts of the world and is commonly known as skullcaps. Scutellaria L. is a medicinal herb used as a folk remedy in Korea and East Asia, but it is difficult to identify and classify various subspecies by morphological methods. Since Scutellaria L. has not been studied genetically, to expand the knowledge of species in the genus Scutellaria L., de novo whole-genome assembly was performed in Scutellaria indica var. tsusimensis (H. Hara) Ohwi using the Illumina sequencing platform. We aimed to develop a molecular method that could be used to classify S.indica var. tsusimensis (H. Hara) Ohwi, S. indica L. and three other Scutellaria L. species. The assembly results for S.indica var. tsusimensis (H. Hara) Ohwi revealed a genome size of 318,741,328 bp and a scaffold N50 of 78,430. The assembly contained 92.08% of the conserved BUSCO core gene set and was estimated to cover 94.65% of the genome. The obtained genes were compared with previously registered Scutellaria nucleotide sequences and similar regions using the NCBI BLAST service, and a total of 279 similar nucleotide sequences were detected. By selecting the 279 similar nucleotide sequences and nine chloroplast DNA barcode genes, primers were prepared so that the size of the PCR product was 100 to 1000 bp. As a result, a species-specific primer set capable of distinguishing five species of Scutellaria L. was developed.


2017 ◽  
Vol 7 (1) ◽  
Author(s):  
Priyanka Mishra ◽  
Amit Kumar ◽  
Gokul Sivaraman ◽  
Ashutosh K. Shukla ◽  
Ravikumar Kaliamoorthy ◽  
...  

Abstract The steno-endemic species of genus Decalepis are highly threatened by destructive wild harvesting. The medicinally important fleshy tuberous roots of Decalepis hamiltonii are traded as substitute, to meet the international market demand of Hemidesmus indicus. In addition, the tuberous roots of all three species of Decalepis possess similar exudates and texture, which challenges the ability of conventional techniques alone to perform accurate species authentication. This study was undertaken to generate DNA barcodes that could be utilized in monitoring and curtailing the illegal trade of these endangered species. The DNA barcode reference library was developed in BOLD database platform for candidate barcodes rbcL, matK, psbA-trnH, ITS and ITS2. The average intra-specific variations (0–0.27%) were less than the distance to nearest neighbour (0.4–11.67%) with matK and ITS. Anchoring the coding region rbcL in multigene tiered approach, the combination rbcL + matK + ITS yielded 100% species resolution, using the least number of loci combinations either with PAUP or BLOG methods to support a character-based approach. Species-specific SNP position (230 bp) in the matK region that is characteristic of D. hamiltonii could be used to design specific assays, enhancing its applicability for direct use in CITES enforcement for distinguishing it from H. indicus.


2021 ◽  
Vol 0 (0) ◽  
Author(s):  
Hanan Al-Adilah ◽  
Dhia Al-Bader ◽  
Mohammed Elkotb ◽  
Ioanna Kosma ◽  
Puja Kumari ◽  
...  

Abstract Even though seaweeds have been considered a nutrient-rich dietary source of minerals in other parts of the world, there is little knowledge about trace element accumulation in seaweeds of the Arabian Gulf. The Arabian Gulf is of particular interest due to being an extreme environment, as it features some of the highest temperatures and salinities observed in any marine waters in the world. This study determined the minerals contents using inductively-coupled plasma-mass spectrometry (ICP-MS) in 10 of the most common seaweeds of this region (Iyengaria stellata, Padina boergesenii, Chondria sp., Feldmannia indica, Codium papillatum, Sargassum aquifolium, Ulva chaugulii, Ulva tepida and Ulva sp.) supported by morphological and molecular (DNA barcode)-based identification. The finding of U. chaugulii reported here is a new record both for Kuwait and the Arabian Gulf. Most of the seaweeds were rich in essential minerals including Ca, Mg, Na, K, Fe and Zn and their contents were higher than those of other mineral-rich foods. Principal component analysis revealed species-specific distributions of minerals in seaweeds. U. tepida and I. stellata were found to be exceptionally rich in most of the macro- and trace elements along with low As and Se, and thus can be utilized for food and feed applications.


Insects ◽  
2019 ◽  
Vol 10 (12) ◽  
pp. 447 ◽  
Author(s):  
Linyu Zheng ◽  
Yue Zhang ◽  
Wenzhao Yang ◽  
Yiying Zeng ◽  
Fan Jiang ◽  
...  

Tephritidae fruit flies (Diptera: Tephritidae) are regarded as important damage-causing species due to their ability to cause great economic losses in fruit and vegetable crops. Bactrocera minax and Bactrocera tsuneonis are two sibling species of the subgenus Tetradacus of Bactrocera that are distributed across a limited area of China, but have caused serious impacts. They share similar morphological characteristics. These characteristics can only be observed in the female adult individuals. The differences between them cannot be observed in preimaginal stages. Thus, it is difficult to distinguish them in preimaginal stages morphologically. In this study, we used molecular diagnostic methods based on cytochrome c oxidase subunit I and species-specific markers to identify these two species and improve upon the false-positive results of previous species-detection primers. DNA barcode sequences were obtained from 900 individuals of B. minax and 63 individuals of B. tsuneonis. Based on these 658 bp DNA barcode sequences of the cytochrome c oxidase subunit I gene, we successfully designed the species-specific primers for B. minax and B. tsuneonis. The size of the B. minax specific fragment was 422 bp and the size of the B. tsuneonis specific fragment was 456 bp. A series of PCR trials ensured the specificity of these two pairs of primers. Sensitivity assay results demonstrated that the detection limit for the DNA template concentration was 0.1~1 ng/μL for these two species. In this study, we established a more reliable, rapid, and low-cost molecular identification method for all life stages of B. minax and B. tsuneonis. Species-specific PCR can be applied in plant quarantine, monitoring and control of B. minax and B. tsuneonis.


2021 ◽  
Vol 4 ◽  
Author(s):  
Judit Fekete ◽  
Dominik Buchner ◽  
Florian Leese ◽  
Judit Padisák ◽  
Gábor Várbíró

The aim of this pilot study was to investigate the potential of eDNA techniques to detect the presence of the two dragonfly species Cordulegaster heros and Cordulegaster bidentata. Both species are classified as “near threatened” according to the IUCN Red List and are strictly protected in several countries. Monitoring these species with traditional sampling methods is often difficult, time-consuming and invasive. In this pilot study, we first collected tissue samples from C. heros and C. bidentata to sequence the traditional DNA-barcode gene fragment COI. We then collected further dragonfly COI sequences from BOLD to design species-specific primers. This, however, was impossible given the enormous variability of COI. Therefore, we refrained from species-specific eDNA assays and followed eDNA metabarcoding protocol using universal (BF2/BF2) and a newly designed dragonfly specific primer. For the evaluation of the method, we took water samples from places where Cordulegaster specimens are known to occur. After the extraction, we used two sequential PCR steps for obtaining the desired amplicon (two-step PCR) using universal primers in the first step, and group (dragonfly) specific primers or universal primers. Amplicons were sequenced on an Illumina MiSeq platform and then analysed the data with the JAMP pipeline. With the newly designed primers and we could effectively detect the targeted dragonfly species from tissue samples, and also from filtered environmental samples. The detection of the species with the traditional method is time consuming and involves the destruction of the specimens. In comparison, with the eDNA method we could easily detect these near threatherned odonates and other dragonfly species in a non-invasive way.


Author(s):  
Lu Yang ◽  
Hua Zhou ◽  
Huili Lai ◽  
Fei Fu ◽  
Wenru Wu

Background: Dendrobium officinale is not only an ornamental plant, but also a valuable medicinal herb that is both effective and widely used in traditional Chinese medicine. However, distinguishing D. officinale from other Dendrobium species is usually a difficult task that need much time and complex technologies due to their very similar external morphologies. The aim of this study is to develop a fast, even on-spot approach to identify D. officinale. Methods: We used DNA barcode-based loop-mediated isothermal amplification (LAMP) method with species-specific LAMP primers targeting the internal transcribed spacer (ITS) region of the rDNA of D. officinale. LAMP reaction time and temperature were optimized and the specificity and sensitivity of LAMP species-specific primers were assessed. Results: This technique showed a high specificity and sensitivity to amplify the genomic DNA of D. officinale and allowed for rapid amplification (within 40 min) of the ITS region under a constant and mild temperature range of 65 °C without using thermocyclers. Besides, by using SYBR® Green I dye as the color developing agent, the color change was easily observed with naked eye. Reaction mixture containing DNA of D. officinale changed from orange to green, while the other Dendrobium species and the negative control retained original orange color. The specificity of this LAMP-based method was confirmed by testing 17 samples of D. officinale and 32 adulterant samples from other Dendrobium species. Conclusions: This LAMP-based rapid identification method does not require expensive equipment or specialized techniques and can be used in field surveys for accurate and fast on site identification.


2021 ◽  
Vol 15 (1) ◽  
pp. 1-22
Author(s):  
Vladimir A. Lukhtanov ◽  
Alexander V. Dantchenko

The Polyommatus (Agrodiaetus) damone (Eversmann, 1841) species complex comprises from 5 to 8 species distributed in southeastern Europe and southern Siberia. Here we used chromosomal and DNA-barcode markers in order to test the taxonomic hypotheses previously suggested for this complex. We revealed that all taxa within this group demonstrate chromosomal stasis and share the same or very similar haploid chromosome number (n = 66 or n = 67). This finding is unexpected since the karyotypes are known to be very diverse and species-specific within the other taxa of the subgenus Agrodiaetus Hübner, 1822. Analysis of the mitochondrial gene COI revealed six diverged clusters of individuals within the complex. Each cluster has a specific geographic distribution and is characterized by distinct morphological features in the wing pattern. The clusters mostly (but not always) correlate with traditionally recognized species. As a result of our study, we describe a new subspecies P. (A.) iphigenides zarmitanussubsp. nov. from Uzbekistan and Tajikistan and show that the taxon originally described as Lycaena kindermanni var. melania Staudinger, 1886 represents a subspecies P. (A.) iphigenides melanius (Staudinger, 1886). Polyommatus (A.) samusi Korb, 2017 (syn. nov.) and P. (A.) melanius komarovi Korb, 2017 (syn. nov.) are considered here as junior subjective synonyms of P. (A.) iphigenides iphigenides (Staudinger, 1886).


Bothalia ◽  
2018 ◽  
Vol 48 (1) ◽  
Author(s):  
Johan J. Spies ◽  
Paula Spies

Background: Clivia is a genus of the family Amaryllidaceae endemic to South Africa and Swaziland. Six species and one natural hybrid have been described. Some morphological traits overlap between some species, thus causing taxonomic confusion.Objectives: The discriminatory power of the core DNA barcodes (matK and rbcLa) was evaluated, and the current taxonomy of Clivia was assessed.Method: Seventy-four two-locus DNA barcodes from 4 to 18 specimens per species were generated.Results: The matK region had a higher mean intraspecific variation of 0.21 compared with the 0.02 of rbcLa. The two-locus barcodes have an aligned length of 1335 base pairs. Three species, Clivia mirabilis, Clivia nobilis and Clivia caulescens, are monophyletic in the Bayesian Inference (BI) cladogram. The remaining Clivia species (Clivia miniata, Clivia gardenii, Clivia robusta and their affinities) are paraphyletic. Clivia is divided into 17 haplogroups with those of C. mirabilis and C. nobilis being unique. Clivia caulescens has three haplotypes. The Clivia species from the north-eastern distribution range of the Eastern Cape and KwaZulu-Natal provinces have 11 haplogroups and no species-specific DNA barcodes. These groups have no correlation with the current taxonomy or geographical distribution.Conclusions: Only 37.33% of the species can be correctly identified with the ‘best match’ option in SpeciesIdentifier. Clivia mirabilis, C. nobilis and C. caulescens have unique DNA barcodes to identify them. Specimens from the Ngome area in KwaZulu-Natal have a unique DNA barcode, separating them from the rest of C. gardenii. A taxonomic revision is suggested.


2021 ◽  
Vol 34 (1) ◽  
Author(s):  
IRIS SEGURA-GARCIA ◽  
SABAI SOE ◽  
NYO-NYO TUN ◽  
STEPHEN BOX

Flatfishes in the family Cynoglossidae are an important coastal fishery in Myanmar. Due to the overlapping morphologies of multiple tonguesole species, caught both as bycatch from trawl fisheries and targeted specifically by small scale fishers, they are all marketed under a single local name, “khwayshar”. This presents a management challenge given the potential differences in the species-specific life-histories, population dynamics, fishing vulnerability and harvest rates. This study investigated the species diversity of tonguesole landings from coastal communities of the Tanintharyi Region of southern Myanmar. DNA barcoding was used to distinguish potentially 10 different species, of which five were identified to species level and five at the genus level. Unconfirmed genetic identifications were based on external morphology. The poor efficacy of DNA barcoding for tonguesole species identification resulted from the limited DNA barcode reference sequences available for the family Cynoglossidae in public databases. An asymmetric occurrence and relative abundance of the identified species in landing sites where samples were collected suggested that the most common species was Cynoglossus oligolepis (Bleeker, 1855), a new species record for Myanmar, followed by Cynoglossus lingua Hamilton, 1822. The results of the present study provide new information to characterise the tonguesole fishery as a first step in the development of management plans for the coastal fishery in Myanmar.


Sign in / Sign up

Export Citation Format

Share Document