Polymorphisms of Mitochondrial COII and16S rRNA Gene in Meloidogyne enterolobii on Mulberry

This study explores the genetic diversity and polymorphisms of Meloidogdyne enterolobii (M enterolobii) on mulberry in China. The sequence of cytochrome oxidase subunit II (COII) and 16S rRNA gene in M enterolobii populations in Guangdong, Guangxi, and Hunan Provinces were PCR-amplified, sequenced, and analyzed for genetic diversity. The haplotypes (Hap) numbers, the total number of mutations, the average number of nucleotide differences (K), haplotype diversity (Hd), and nucleotide diversity (π) of mtCOII gene were 14, 25, 3.563, 0.942 and 0.00429, respectively. The significant differences in Fst value (0.125) and a high level of gene flow (2.83) were detected among the 19 M enterolobii populations. High genetic variation within each population and a small genetic distance among populations was observed. Both phylogenetic analyses and network mapping of the 14 haplotypes revealed a dispersed distribution pattern of the 19 M enterolobii populations. There was an absence of branches strictly corresponding to the 19 range sampling sites. The analysis of molecular variance (AMOVA) revealed that the genetic differentiation of M enterolobii populations was mainly contributed by the variation within each of the defined geographical groups. No significant correlation was found between the genetic distance and geographical distance of 19 M enterolobii populations. This study provides theoretical basis for the future control of M enterolobii and also provides a guarantee for the production of other hosts of M enterolobii.

Genetic diversity of indigenous catfish from Indonesia based on mitochondrial Cytochrome Oxidase Subunit II gene

Budiariati V, Susmiati T, Waroh S, Putri RCA, Widayanti R. 2021. Genetic diversity of indigenous catfish from Indonesia based on mitochondrial Cytochrome Oxidase Subunit II gene. Biodiversitas 22: 593-600. Catfish is one of the most demanding fish in Indonesia and served in a variety of traditional culinary. Due to their identical morphology and close relation between species in the order of Siluriformes, it is quite tricky to distinguish the species. This can be a threat to develop catfish production in Indonesia since there is a wide variety of catfish species in this mega biodiversity country. The study aimed to analyze the genetic diversity of Indonesian indigenous catfish especially those known as Baung fish by local people based on COII gene. The study also aimed to determine the phylogenetic relationship between the samples and compare them with the GenBank data. A total of 24 samples used in this study from 8 different rivers from 3 different islands and two samples were collected from coastal areas. The study results showed that there is genetic diversity of the Indonesian indigenous catfish based on COII gene. The sequences among 24 samples showed that from 691 nucleotides of COII gene, there were very subtle nucleotides differences of samples that originated from Bojonegoro, Magelang, and samples collected from Baru Beach, Yogyakarta. Based on COII amino acid sequences, 6 polymorphic amino acid sites were on-site number 64, 115, 123, 129, 144, and 165. The samples encoded LLB1 and LPB1 from Baru Beach, Yogyakarta, showed highest different amino acids in the six sites. Samples from the river of Central Java, Sumatra, and Kalimantan belonged to Bagridae family and consist of two different species Hemibagrus sp. and Mystus sp while samples from East Java belonged to Pangasiidae family. The Samples from coastal belonged to Ariidae family.

First Detection of Meloidogyne luci (Nematoda: Meloidogynidae) Parasitizing Potato in the Azores, Portugal

Potato is the third most important crop in the world after rice and wheat, with a great social and economic importance in Portugal as it is grown throughout the country, including the archipelagos of Madeira and the Azores. The tropical root-knot nematode (RKN) Meloidogyne luci is a polyphagous species with many of its host plants having economic importance and the ability to survive in temperate regions, which pose a risk to agricultural production. In 2019, M. luci was detected from soil samples collected from the council of Santo António in Pico Island (Azores). Bioassays were carried out to obtain females, egg masses, and second-stage juveniles to characterize this isolate morphologically, biochemically, and molecularly. The observed morphological features and morphometrics showed high similarity and consistency with previous descriptions. Concerning the biochemical characterization, the esterase (EST) phenotype displayed a pattern with three bands similar to the one previously described for M. luci and distinct from M. ethiopica. Regarding the molecular analysis, an 1800 bp region of the mitochondrial DNA between cytochrome oxidase subunit II (COII) and 16S rRNA genes was analyzed and the phylogenetic tree revealed that the isolate grouped with M. luci isolates (99.17%). This is the first report of M. luci parasitizing potato in the Azores islands, contributing additional information on the distribution of this plant-parasitic nematode.

Identification of Subterranean Termite Genus, Reticulitermes (Blattodea: Rhinotermitidae) in Taiwan

Species identification in the genus Reticulitermes is often difficult because of ambiguous morphological characters. Challenges in Reticulitermes spp. recognition have also been encountered in East Asia, including Taiwan. Because of unknown Reticulitermes taxa in Taiwan and the possible origin of alien Reticulitermes kanmonensis Takematsu in Japan and Korea, reexamining Reticulitermes fauna in Taiwan is imperative. To clarify the Reticulitermes fauna in Taiwan, this study applied two mitochondrial genes (cytochrome oxidase subunit II [COII] and 16S rDNA) and morphological characters for species delimitation. Reticulitermes specimens collected from 63 localities across the main and adjacent islands of Taiwan were analyzed. Phylogenetic analyses, morphological comparisons, and ecological traits suggested the existence of three species in Taiwan: Reticulitermes flaviceps (Oshima), R. kanmonensis, and Reticulitermes leptomandibularis Hsia and Fan. Altitudinal distributions among the three Reticulitermes termites tended to differ: R. flaviceps adapted to low hills, but R. kanmonensis and R. leptomandibularis occurred in medium mountainous areas. The combined data, including haplotype diversities and distribution range, suggest that 1) R. flaviceps is an endemic species and only found in Taiwan; 2) R. kanmonensis and R. leptomandibularis are both native species in Taiwan and China; 3) Japanese R. kanmonensis populations originated from southern China and/or Taiwan and that Korean populations were possibly introduced from Japan.

Two new species of Grylloblatta Walker, 1914 (Grylloblattodea: Grylloblattidae) from western North America, and a neotype designation for G. rothi Gurney 1953

Grylloblatta rothi Gurney, 1953 is redescribed and a neotype is designated from Cultus Mountain in the Oregon Cascades, U.S.A. Two new species of Grylloblatta are described, bringing the total number of Grylloblatta species to 15. Grylloblatta chintimini new species is described from Marys Peak in the Coast Range of Western Oregon, where it occurs on snowpack near the 1250 m summit. Grylloblatta newberryensis new species is described from Newberry Volcano in Central Oregon, where it is associated with snowfields overlying geologically-young lava flows. Morphological characters, primarily derived from male genitalia, are presented to diagnose these species and differentiate them from other Grylloblatta spp. in Oregon, Washington, and California. Molecular sequences from the cytochrome oxidase subunit II gene suggest that significant divergence has occurred among these species and provide a tool to aid identification of juvenile and female specimens.

First Report of Downy Mildew (Hyaloperonospora lunariae) on White Mustard (Sinapis alba) in Brazil

Sinapis alba (Brassicaceae), white mustard, is broadly cultivated for its seed used as component of table mustard (4). In June 2013, a group of diseased S. alba were observed in a vegetable garden on the campus of the Universidade Federal de Viçosa (municipality of Viçosa, state of Minas Gerais, Brazil). Foliage of diseased plants showed numerous chlorotic areas that developed into severe leaf blight with abundant downy mildew growth abaxially. A dried representative specimen has been deposited in the herbarium at the Universidade Federal de Viçosa (accession no. VIC 39743). The fungus had the following morphology: Sporangiophores arborescent, dichotomously branched, 540 to 840 × 8 to 10 μm hyaline, smooth, branches 105 to 210 μm long; esterigmata subacutate and curved, in pairs, 15 to 42 μm long; sporangia globose, 18 to 24 × 15 to 18 μm, hyaline, smooth. DNA was extracted using a Wizard Promega purification kit. The cytochrome oxidase subunit II (COX2) region was amplified with COX2f and COX2r primers (3). The sequence has been deposited in GenBank (Accession No. KJ396953). DNA sequences representing morphologically similar taxa were downloaded from GenBank nucleotide database, aligned in MEGA 5, and analyzed using Bayesian inference and Markov chain Monte Carlo simulation implemented in MrBayes 3.0 with five repetitions. A sequence of Albugo candida was used as outgroup in the analysis. The morphological characteristics places the fungus on S. alba in the complex of species of Pernosporaceae that attack the Brassicaceae. These are notoriously difficult to discriminate by morphology but our COX2-based phylogenetic analysis places it in Hyaloperonospora lunariae (1). This species was previously only known to cause downy mildew on other species of Brassicaceae (Lunaria annua and Erucastrum nasturtiifolium) in Europe (2). To our knowledge, this is the first report of this pathogen-host association in the world. References: (1) O. Constantinescu and J. Fatehi. Nova Hediwigia 74:291, 2002 (2) D. F. Farr and A. Y. Rossman. Fungal Databases, Systematic Mycology and Microbiology Laboratory. Online publication. ARS, USDA, 2013. (3) D. S. S. Hudspeth et al. Mycologia 92:674, 2000. (4) B. B. Simpson and M. C. Ogorzaly. Econonic Botany. McGraw Hill, San Diego, CA, 2001.

Three new species of Grylloblatta Walker (Insecta: Grylloblattodea: Grylloblattidae), from southern Oregon and northern California

Grylloblatta oregonensis new species, Grylloblatta siskiyouensis new species, and Grylloblatta marmoreus new speciesare described from the Klamath Mountains region of Oregon and California, U.S.A. Two species were found at OregonCaves National Monument, with G. oregonensis inhabiting the dark zone of caves and G. siskiyouensis inhabiting surfacehabitat and the twilight zone of caves. The third species, G. marmoreus, is known from Big Foot Cave and Planetary DairyCave, Marble Mountains, California. These three species are recognized on the basis of morphological characters anddistinguished from nearby Grylloblatta species. Analysis of genetic data from the cytochrome oxidase subunit II genesupports the morphological diagnosis of these three species as unique lineages and confirms their genetic divergence from other ice-crawler populations found in Oregon and California.