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Multigene phylogeny of root-knot nematodes and molecular characterization of Meloidogyne nataliei Golden, Rose & Bird, 1981 (Nematoda: Tylenchida).

ABSTRACT: The root-knot nematodes of the genus Meloidogyne are highly adapted, obligate plant parasites, consisting of nearly one hundred valid species, and are considered the most economically important group of plant-parasitic nematodes. Six Meloidogyne species: M. arenaria, M. hapla, M. incognita, M. microtyla, M. naasi and M. nataliei were previously reported in Michigan, USA. For this study, Meloidogyne nataliei was isolated from the grapevine Vitis labrusca from the type locality in Michigan, USA, and was characterized using isozyme analysis and ribosomal and mitochondrial gene sequences. No malate dehydrogenase activity was detected using macerate of one, five, six, seven or ten females of M. nataliei per well. However, one strong band (EST?=?S1; Rm: 27.4) of esterase activity was detected when using homogenates of ten egg-laying females per well. Phylogenetic analyses of sequences of the partial 18S ribosomal RNA, D2-D3 of 28S rRNA, internal transcribed spacer of rRNA, mitochondrial cytochrome oxidase subunit I genes and the cytochrome oxidase subunit II-16S rRNA intergeneric fragment from fifty-five valid Meloidogyne species and M. nataliei were conducted using Bayesian inference and maximum likelihood methods. From these results, we infer 11 distinct clades among studied species, with M. nataliei and M. indica composing a basal lineage. Seventy five percent of these species belong to seven clades within the Meloidogyne superclade. Characterization of these clades is provided and evolutionary trends within the root-knot nematodes are discussed.

SUBMITTER: Alvarez-Ortega S

PROVIDER: S-EPMC6692364 | biostudies-literature |

REPOSITORIES: biostudies-literature

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Project description:Meloidogyne polycephannulata n. sp. is described from specimens collected from an area cultivated with carrot cv. Brasilia, in the city of Rio Paranaíba, in the region of Alto Paranaíba, Minas Gerais State, Brazil. The perineal pattern of the female is circular to ovoid with a high dorsal arch that has widely spaced, coarse annulations. The lateral field may have a deep furrow separating the dorsal and ventral arches. The medial lips are short and wide, whereas the lateral lips are large and triangular. The female stylet is 15-16 ?m long with wide knobs, distinctly divided by an indentation in the center. Its tip is slightly curved dorsally. The excretory pore opens 34-65 ?m from the anterior end. Females retain eggs and second-stage juveniles in their body cavity, similar to that of the cyst-forming nematodes. Males are 1.3-1.7 mm long and have a high head cap that is rounded and slopes posteriorly. The labial disc is fused to the medial lips. The head region has several irregular annulations that are similar in appearance to the first or second body annules that are likewise irregular, making the head region appear to be extremely large. The stylet of the male is 21-24 ?m long; it is slender, and has small, rounded knobs, that are distinctly indented medially and appear heart-shaped. The shaft has several tiny projections throughout its length. Mean second-stage juvenile length is 411.7 ?m. The juvenile head cap is elevated, the medial lips are small, and the lateral lips are elongate to triangular-shaped. The head region has several short, incomplete and irregular transverse annulations. The juvenile stylet is 14-23 ?m long with small, rounded, and sloping knobs. The thin tail ends with a short hyaline portion that is variable in size (16-26 ?m) and with a small, rounded tip. Isozyme profiles of esterases from Meloidogyne javanica show 3 strong bands (SB) at Rm 46, 59, and 66; profiles of M. polycephannulata n. sp. show a SB at Rm 47 and a weak band (WB) at Rm 52; M. petuniae has two SB at Rm 44 and 53; M. phaseoli has a SB at 53, 58, and 64 Rm; M. brasilensis has three SB at Rm 40, 58, and 66 and a WB at Rm 71; M. pisi has a SB at Rm 40, 60, and 64 and two WB at 46 and 50 Rm. Data from sequencing the 18S rDNA region of M. polycephannulata n. sp. confirms that it is different from M. arabicida, M. arenaria, M. ethiopica, M. incognita, M. javanica, M. paranaensis, and M. thailandica. Sequence identity among these eight species ranged between 85 to 93.4%. Meloidogyne polycephannulata n. sp. reproduces very well on carrot and tomato; poorly on pepper; and not at all on cotton, peanut, tobacco, watermelon, and sweet corn.

Project description:An unknown root-knot nematode was found at high density on grape roots collected from Yunnan Province. Morphometric traits and measurements, isozyme phenotypes, and molecular analysis clearly differentiated this nematode from previously described root-knot nematodes. This new species is described, illustrated and named Meloidogyne vitis sp. nov. The new species can be distinguished from other Meloidogyne spp. by a unique combination of characters. Females display a prominent neck, an excretory pore is located on the ventral region between 23rd and 25th annule behind lips, an EP/ST ratio of approximately 2.5 (1.98-2.96), a perineal pattern with two large and prominent phasmids, and a labial disc fused with the medial lips to form a dumbbell-shaped structure. Males display an obvious head region, a labial disc fused with the medial lips to form a dumbbell-shaped structure, no lateral lips, a prominent slit-like opening between the labial disc and medial lips, a distinct sunken appearance of the middle of the medial lips, and four incisures in the lateral field. Second-stage juveniles are characterized by a head region with slightly wrinkled mark, a labial disc fused with the medial lips to form a dumbbell-shaped structure, a slightly sunken appearance of the middle of the medial lips, a slit-like amphidial openings between the labial disc and lateral lips, and four incisures in the lateral field. The new species has rare Mdh (N3d) and Est phenotypes (VF1). Phylogenetic analysis based on ITS1-5.8S-ITS2, D2D3 fragments of rDNA, and coxI and coxII fragments of mtDNA sequences clearly separated the new species from other root-knot nematodes, and the closest relative was Meloidogyne mali. Meloidogyne mali was collected for amplifying these sequences as mentioned above, which were compared with the corresponding sequences of new species, the result showed that all of these sequences with highly base divergence (48-210 base divergence). Moreover, sequence characterized amplified region (SCAR) primers for rapid identification of this new species were designed.

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Multigene phylogeny of root-knot nematodes and molecular characterization of Meloidogyne nataliei Golden, Rose & Bird, 1981 (Nematoda: Tylenchida).

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