Abstract
DNA barcoding with a new cytochrome oxidase c subunit 1 primer set generated a 721 to 724 bp fragment used for the identification of 322 Meloidogyne specimens, including 205 new sequences combined with 117 from GenBank. A maximum likelihood analysis grouped the specimens into 19 well-supported clades and four single-specimen lineages. The "major" tropical apomictic species ( Meloidogyne arenaria , Meloidogyne incognita , Meloidogyne javanica ) were not discriminated by this barcode although some closely related species such as Meloidogyne konaensis were characterized by fixed diagnostic nucleotides. Species that were collected from multiple localities and strongly characterized as discrete lineages or species include Meloidogyne enterolobii , Meloidogyne partityla , Meloidogyne hapla , Meloidogyne graminicola , Meloidogyne naasi , Meloidogyne chitwoodi , and Meloidogyne fallax . Seven unnamed groups illustrate the limitations of DNA barcoding without the benefit of a well-populated reference library. The addition of these DNA sequences to GenBank and the Barcode of Life Database (BOLD) should stimulate and facilitate root-knot nematode identification and provide a first step in new species discovery.
Highlights
DNA barcoding with a new cytochrome oxidase c subunit 1 primer set generated a 721 to 724 bp fragment used for the identification of 322 Meloidogyne specimens, including 205 new sequences combined with 117 from GenBank
The objective of this study is to present a primer set used for the amplification of 721 to 724 bp of COI sequence from Meloidogyne
The COI gene region used as a diagnostic marker in this study appears to discriminate many of the described species of Meloidogyne
Summary
DNA barcoding with a new cytochrome oxidase c subunit 1 primer set generated a 721 to 724 bp fragment used for the identification of 322 Meloidogyne specimens, including 205 new sequences combined with 117 from GenBank. Seven unnamed groups illustrate the limitations of DNA barcoding without the benefit of a wellpopulated reference library The addition of these DNA sequences to GenBank and the Barcode of Life Database (BOLD) should stimulate and facilitate root-knot nematode identification and provide a first step in new species discovery. Multiple primer sets for amplification of nematode cytochrome oxidase c subunit 1 (COI) are available (Derycke et al, 2005, 2010; Prosser et al, 2013; Kiewnick et al, 2014; Powers et al, 2014; Janssen et al, 2016). Nucleotide sequences have been submitted to GenBank (accession numbers MH128384–MH128585) and the Barcode of Life Database (BOLD)
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