DNA barcoding of the Thai species of terrestrial earthworms in the genera Amynthas and Metaphire (Haplotaxida: Megascolecidae)

Jeratthitikul, Ekgachai; Bantaowong, Ueangfa; Panha, Somsak

doi:10.1016/j.ejsobi.2017.06.004

Cited by 19 publications

(8 citation statements)

References 52 publications

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“…The cryptic speciation events have also been reported in many earthworm species using either mitochondrial data (King et al 2008;James et al 2010;Novo et al 2010) or nuclear data (Rougerie et al 2009). The COI barcode was also applied to discriminate earthworm species and to suggest cryptic forms (Jeratthitikul et al 2017).…”

Section: Remarksmentioning

confidence: 99%

“…The K2P species divergences were reported, but were different for earthworm groups, for example, 13-15% for Allolobophora Eisen, 1874 (King et al 2008), 13.2% for Eisenia fetida/andrei complex (Römbke et al 2016), more than 14% in Octolasion lacteum Örley, 1885, Lumbricus rubellus Hoffmeister, 1843 (Klarica et al 2012), more than 18.7% in pheretimoid species (Chang et al 2008), 19.8% in the genus Lumbricus Linnaeus, 1758 (James et al 2010), 8.9-22.9% in the genus Pheretima Kinberg, 1867, 14.2-21.9% in the genus Amynthas Kinberg, 1867, 7.5-18.0% in the genus Pithemera Sim & Easton, 1972 and 11.7-21.0% in the genus Polypheretima Michaelsen, 1934(Aspe & James 2018. Recently, Jeratthitikul et al (2017) also calculated the average species distance of 20% for Amynthas and Metaphire species in Thailand. Our calculation of 16% only applies to the Metaphire species.…”

Section: Molecular Analysismentioning

confidence: 99%

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Notes on the earthworm species, Metaphire anomala (Michaelsen, 1907) (Clitellata, Megascolecidae) in Southern Vietnam, with descriptions of two new species

Nguyen

Lam

Nguyen³

2021

EJT

782

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Integrative taxonomy was employed to exploit the differences between the known Metaphire anomala (Michaelsen, 1907) and other specimens collected in Vietnam. The results brought to light two new species, namely Metaphire iranomala sp. nov. and Metaphire decemtheca sp. nov. The former is easily recognised by having male pores on xix and four pairs of spermathecal pores on 5/6/7/8/9 while the latter is distinguished by having five pairs of spermathecal pores on 4/5/6/7/8/9. The K2P distances of the fragment of the cytochrome c oxidase subunit I (COI) gene are 13.1% between M. iranomala sp. nov. and M. anomala (Michaelsen, 1907) and 18% between M. decemtheca sp. nov. and Metaphire grandiverticulata Nguyen & Lam, 2017. The intraspecific divergences are 1.5%–10.6% for M. iranomala sp. nov. and 2.1%–11.4% for M. decemtheca sp. nov.

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Section: Remarksmentioning

confidence: 99%

Section: Molecular Analysismentioning

confidence: 99%

Notes on the earthworm species, Metaphire anomala (Michaelsen, 1907) (Clitellata, Megascolecidae) in Southern Vietnam, with descriptions of two new species

Nguyen

Lam

Nguyen³

2021

EJT

782

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“…Moreover, many earthworm taxonomists emphasized that integrating morpho-anatomical features with barcoding data provide more contrasting conclusions. These integrative approaches were utilized to discriminate among species and taxa that are new to science (Shekhovtsov et al 2014;Jeratthitikul et al 2017;Lone et al 2020). Furthermore, compared to morpho-anatomical features that require exhaustive work, species discrimination using DNA barcoding is relatively rapid and identification measure is progressed (Gregory 2005).…”

Section: Dna Barcoding and Clitellate Species Identificationmentioning

confidence: 99%

Disentangling earthworm taxonomic stumbling blocks using molecular markers

et al. 2021

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Taxonomic classification of earthworms based on anatomical features has created several challenges for systematics and population genetics. This study examines the application of molecular markers, in particular mitochondrial cytochrome oxidase (COI), to facilitate discrimination of closely related earthworm species. Molecular markers have also provided insights into population genetics by aiding assessment of genetic diversity, lineage sorting, and genealogical distributions of populations for several species. Phylogeography—a study that evaluates the geographical distribution of these genealogical lineages and the role of historical processes in shaping their distribution—has also provided insights into ecology and biodiversity. Such studies are also essential to understand the distribution patterns of invasive earthworm species that have been introduced in non-native ecosystems globally. The negative consequences of these invasions on native species include competition for food resources and altered ecosystems. We anticipate that molecular markers such as COI and DNA barcoding offer potential solutions to disentangling taxonomic impediments in earthworms and advancing their systematics and population genetics.

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“…Additionally, this technique can be used to identify dead earthworm specimens found in the environment and poorly preserved specimens in laboratories. DNA barcoding is a modern technique that acquires the extra benefit of being simple, accurate, and rapid in characterizing various micro and macro-organisms 19,20) . Several mitochondrial gene sequences, including NADH subunit 5, 18S rRNA, 16S rRNA, 12S rRNA, COII, and cytochrome c oxidase subunit I (COI) , are used as genetic markers for species identification in a variety of in-vertebrates and vertebrate taxa.…”

Section: Introductionmentioning

confidence: 99%

“…Several genetic phylogenies of Megascolecidae have been constructed and published over the last few decades 24) to resolve systematic issues among closely related species with no apparent morphological differences 25,26) . Similarly, in earthworms belonging to the pheretimoid complex, the COI gene was used to distinguish morphologically different earthworms such as Metaphire houlleti, Amynthas arenulus, and A. longiculicaulatus 19) as well as to assess the phylogenetic relationships in Metaphire and Amynthas 27) . Phylogenetic analysis indicated that a single gene or multiple genes might be useful for identifying cryptic soil invertebrate species such as earthworms 15,28,29) .…”

Section: Introductionmentioning

confidence: 99%

DNA Barcoding: Molecular Identification and Phylogenetic Analysis of Pheretimoid Earthworm (<i>Metaphire</i> sp. and <i>Amynthas</i> sp.) Based on Mitochondrial Partial COI Gene from Sialkot, Pakistan

Hussain¹,

Liaqat²,

Mubin

et al. 2022

J. Oleo Sci.

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The extremely difficult and challenging process is identifying pheretimoid species, genus Metaphire and Amynthas involving increased homoplasy in various morphological characteristics. The molecular identification, phylogenetic relationships, and evolutionary divergence time of earthworms belonging to the pheretimoid complex were investigated in this study using partial mitochondrial COI (cytochrome C oxidase subunit I) gene sequences ranging from 550-680 bp. Results revealed that 86 pheretimoid earthworms were morphologically different from a total of 342 mature worms. Moreover, 11 pheretimoid species were molecularly identified, including Metaphire posthuma (02), M. anomala (01), M. houlleti (02), M. californica (01), M. birmanica (02), Amynthas minimus (01), A. morrisi (01), and M. bununa (01). A phylogenetic tree was constructed with bootstrap values of 95%, which supported a monophyletic lineage of two well-supported clades formed by 12 partial COI sequences and 48 GenBank sequences using Hirudo medicinalis as an outgroup. The monophyly of these obtained genera indicated overall similarity at species level. Today, species like Amynthas, Metaphire and Pheretima have worm diversity in the form of pheretimoid earthworms, which dates to the Late Miocene (11.2-5.3 Mya) and the Pliocene (5.3-2.4 Mya).Compared to all relevant pheretimoid species, genetic p-distance values ranged from 0.0% to 0.57% (less than 1%). These low range values demonstrated that both genera Metaphire and Amynthas, supported the theory, which states that there are shared similarities among the species, despite different morphology. The current study is the first attempt in Pakistan to identify earthworms through DNA barcoding thus providing a genomic stamp. The work explored the significance of COI gene sequences to construct molecular tools that will be useful to overcome the different obstacles in morphologically similar earthworm identification and their phylogenetic study.

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DNA barcoding of the Thai species of terrestrial earthworms in the genera Amynthas and Metaphire (Haplotaxida: Megascolecidae)

Cited by 19 publications

References 52 publications

Notes on the earthworm species, Metaphire anomala (Michaelsen, 1907) (Clitellata, Megascolecidae) in Southern Vietnam, with descriptions of two new species

Notes on the earthworm species, Metaphire anomala (Michaelsen, 1907) (Clitellata, Megascolecidae) in Southern Vietnam, with descriptions of two new species

Disentangling earthworm taxonomic stumbling blocks using molecular markers

DNA Barcoding: Molecular Identification and Phylogenetic Analysis of Pheretimoid Earthworm (<i>Metaphire</i> sp. and <i>Amynthas</i> sp.) Based on Mitochondrial Partial COI Gene from Sialkot, Pakistan

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