Discovery of Root-Lesion Nematode (Pratylenchus scribneri) on Corn in Hainan Province of China

Corn (Zea mays L.) is a very important cereal crop and serves as food, feed, and industrial material (Liu et al. 2016). The root-lesion nematode (RLN) is considered one of the most important plant-parasitic nematodes and can cause economic losses in agriculture worldwide (Jones et al. 2013). In January 2020, five samples were collected from a corn field in Lingshui Lizu Autonomous County, Hainan Province, China. The collected corn plants (cv. Denghai 685) were growing poorly and roots showed distinct lesions and rot. Corn fields with symptoms of stunted plants, and brown lesions on roots were widespread. This corn disease was severe in Lingshui Lizu Autonomous County. RLN were extracted from soil samples by the modified Baermann funnel (Hooper et al. 2005). All the samples contained RLN ranging from 9 to 82 (average 39) RLN per 100 cm3 of soil and 113 to 257 (average 194) RLN per 5 g roots. The extracted RLN were sterilized and cultured on carrot disks at 25°C for 90 days. Afterwards, seeds of corn (cv. Denghai 685) were sown in pots containing 1.8 liters of sterilized soil. Eight plants, one per pot, were infested with 1,000 RLN, eight pots of noninfested corn plants were used as controls, and plants were kept in a greenhouse at 25°C. At 75 days after inoculation, symptoms were like those initially observed in corn fields, whereas no symptoms were observed in the control plants. Nematodes in the soil and roots were extracted using the same method as previously described (Hooper et al. 2005). The average number of RLN per pot was approximately 4,250 in soil and 820 in roots, the reproduction factor (final number of nematodes/initial number of nematodes) was 5.07, no RLN were found in the control. The experiment was conducted twice. The morphological and molecular studies of RLN were examined to confirm species identification. The main morphological measurements of adult females (n = 15) included body length = 526.0 μm ± 17.1 (standard error) (range = 498.0 to 560.5 μm), stylet = 16.0 μm ± 0.3 (15.5 to 16.5 μm), tail length = 29.0 μm ± 1.5 (26.5 to 31.0 μm), a = 23.6 ± 0.6 (22.6 to 24.4), b = 5.6 ± 0.3 (5.2 to 6.0), c = 18.3 ± 0.9 (16.4 to 19.7), V = 78.2% ± 0.6 (77.4 to 79.2%), lip region with two annules. No males were found in the samples. This population was identified as Pratylenchus scribneri, based on the morphological characters (Castillo and Vovlas, 2007). DNA was isolated from individual nematodes followed by proteinase K-based lysis (Wang et al. 2011). The D2/D3 expansion region of the 28S rRNA gene, rDNA-internal transcribed spacer (ITS) region, and mitochondrial cytochrome oxidase I (mtDNA-COI) gene were amplified with primers D2A/-D3B (De Ley et al. 1999), TW81/ AB28 (Vovlas et al. 2011) and JB3/ JB5 (Liu et al. 2018), respectively. The PCR products were purified and ligated into pJET 1.2/blunt cloning vectors and transformed to Escherichia coli strain DH5α, and then sequenced. The obtained 28S rRNA gene D2/D3 region sequences (785bp), ITS sequences (886 bp) and mtDNA-COI (447bp) in this study were submitted to GenBank. The D2/D3 region of the 28S rRNA sequences of the RLN collected in Lingshui (GenBank accession no. MZ701843) showed 99.75% identity with P. scribneri sequences available in the GenBank (KX842628 and KX842625). The ITS sequences of the RLN collected in this study (MZ701842) showed the highest identity of 97.06% with P. scribneri sequences available in the GenBank (KX842626). The mtDNA-COI sequences of the RLN collected in this study (OK040228) showed 100% identity with P. scribneri (MN366409). Both morphological and molecular data confirmed the identity of P. scribneri. P. scribneri has been reported on corn in Inner Mongolia, Hebei, Shanxi, Shandong, Henan, Jiangsu, and Liaoning provinces of China (Li et al. 2019). As far as we know, this is the first report of P. scribneri on corn in Hainan Province, China. Since the RLN can cause considerable damage to corn, strategic measures should be taken to prevent the spread of P. scribneri to other regions in China.

Rediscovery and redescription of Niphargus enslini Karaman, 1932 (Amphipoda, Niphargidae) in southern Germany

Niphargus enslini Karaman, 1932 was collected only once in 1905 from the Falkensteiner Höhle (Baden-Württemberg, Germany). Two years after its description, the species was synonymized with Niphargus virei and not studied any more. During recent surveys on German niphargids, further samples collected in this cave did not yield N. enslini specimens, but this species was collected in the Blätterteighöhle and in the Schwarzer Brunnen, two caves located in Baden-Württemberg and intercepting the same karstic aquifer feeding Falkensteiner Höhle. In an integrative taxonomic approach, we carefully studied the morphology of the newly collected specimens and sequenced two molecular markers (fragments of the cytochrome c oxidase subunit I (COI) and of the nuclear 28S rRNA gene) to test for possible conspecificity of N. enslini with N. virei. Morphological analysis confirmed that N. enslini is distinct from the N. virei species complex. We provide a redescription of newly collected material, together with new drawings of a more than 100 years old topotypic female. We briefly discuss the putative origin of N. enslini and the age of its split from the N. virei species complex.

Descriptions of Trichodorus lownsberyi sp. n., Nanidorus minor (Colbran, 1956) Siddiqi, 1974 (Nematoda: Trichodoridae) and Longidorus quercus sp. n. (Nematoda: Longidoridae) from Mexico

Summary A new species of stubby root nematode, Trichodorus lownsberyi sp. n., collected from soil around Buxus sempervirens at Montecillo Campus, Colegio de Postgraduados, Mexico, is described and illustrated. Trichodorus lownsberyi sp. n. is characterised by the abundant males with a characteristic spicule shape, the narrower part about mid-blade having visible bristles in most specimens, and the presence and position of the three ventromedian cervical papillae, which are all anterior to the secretory-excretory pore. Females possess a rhomboid-shaped vagina and oval to rounded triangular vaginal sclerotised pieces with a pore-like vulva. The phylogenetic relationships of T. lownsberyi sp. n. with related species were constructed using the ITS2 rRNA and the D2-D3 expansion segments of 28S rRNA gene sequences. Trichodorus lownsberyi sp. n. was a sister species to T. viruliferus in the phylogenetic trees. A Mexican population of Nanidorus minor from a peach orchard is also described and illustrated. The new needle nematode, Longidorus quercus sp. n., was recovered from soil around roots of oak, Quercus crassipes, from Cerro Jusda ‘El Diablo’ in Mexico State. Longidorus quercus sp. n. females are characterised by the C-shaped posterior end of the body after fixation, L = 4.9 (3.9-5.6) mm, a = 71 (60.8-93.2), lip region rounded, sometimes slightly flattened, marked by depression, odontostylet 170 (144-206) μm long, hemispherical to bluntly conoid tail and no males. The new species was characterised using the D2-D3 of 28S rRNA and COI gene sequences. The phylogenetic relationships of L. quercus sp. n. with other Longidorus species were reconstructed using the D2-D3 of 28S rRNA gene sequences.

Differentiation of Basidiobolus spp. Isolates: RFLP of a Diagnostic PCR Amplicon Matches Sequence-Based Classification and Growth Temperature Preferences

The genus Basidiobolus, known since 1886, is primarily associated with reptiles and amphibians. Although globally distributed, rare infections caused by members of this genus mainly occur in tropical and subtropical regions. Morphological and physiological characteristics were used in the past for the description of species. However, some of these characteristics vary depending on culture conditions. Therefore, most species names are regarded as synonyms of B. ranarum as the only pathogenic species. Yet, not all environmental isolates are necessarily pathogenic. This study aimed to analyze if environmental Basidiobolus isolates can be distinguished reliably based on morpho-physiological and molecular characteristics. Eleven isolates originally obtained from feces of south African reptiles and one type strain, Basidiobolus microsporus DSM 3120, were examined morpho-physiologically. Sequence analysis of the 18S and partial 28S rRNA gene and restriction analysis of a diagnostic amplicon (restriction fragment length polymorphism, RFLP) were performed for all 12 strains. Based on the results obtained, morphological features and the 18S rRNA sequence proved insufficient for the reliable differentiation of isolates. However, isolates were distinguishable by growth temperature profiles, which matched isolate clusters established by partial 28S rRNA gene sequence and restriction analysis of a Basidiobolus specific diagnostic PCR amplicon. Our results indicate that RFLP analysis can be used as a fast screening method to identify Basidiobolus isolates with similar physiological characteristics.

An integrated morphological–molecular approach reveals new insights on the systematics of the octocoral Telestula humilis (Thomson, 1927) (Octocorallia:Alcyonacea:Clavulariidae)

Telestula humilis (Thomson, 1927) is a rare deep-sea stoloniferan octocoral distributed in the eastern Atlantic. Here we compared seven putative colonies of this species collected off Spain with the lectotype from the Oceanographic Museum of Monaco and found them to be identical morphologically. Phylogenetic analyses on both full mitogenomes and a concatenated alignment containing two mtDNA genes (mtMutS and Cox1) and nuclear 28S rRNA gene recovered Telestula humilis sister to Incrustatus and Inconstantia rather than to other species of Telestula. This therefore supports its taxonomic reassignment to Pseudotelestula gen. nov. as Pseudotelestula humilis comb. nov. The taxonomic reassignment is also supported by subtle differences observed between the morphology of the colony and the sclerome of Pseudotelestula humilis comb. nov. and the two sister genera. The occurrence of an intrusion tissue with sclerites in the basal part of the gastric cavity of the adult polyps is shared among Telestula and Pseudotelestula gen. nov. However, Pseudotelestula gen. nov. has sclerites arranged in a collaret and points below the tentacles, the sclerites of the calyx wall and the stolon are plump warty spindles, and the intrusion tissue has long sticks and spindles with cone-like spines.

Description of Longidorus barsii Radivojević & De Luca sp. n. (Nematoda: Longidoridae) from Serbia and observations on some taxonomic characters

Summary Longidorus barsii sp. n., from Mt Tara in the Balkan Peninsula, is described and characterised by using a polyphasic approach. The species has numerous males. The female body is 5-7 mm long, rather stout and resembles a large Xiphinema. The lip region is wide, with rounded sides continuous with the neck, frontally flattened and depressed around the oral aperture, amphids are pouch-like and distinctly bi-lobed and the odontostyle is moderately long. The nuclei of the pharyngeal glands are in the normal position, the dorsal nucleus located somewhat posterior to anterior third of bulb. The uteri are long, the distal inner epithelium densely covered with papilla-like outgrowths. The tail is rounded, bluntly conoid and very short. Alpha-numerical identification codes: A4/5, B45, C3, D3, E2, F3, G 1(2), H1, I2, J1, K67. The morphologically most similar species are L. kheirii, L. polyae and L. profundorum. Additional observations are provided on the anterior body region and genital organs in L. barsii sp. n., L. piceicola, L. silvae, and L. uroshis. Selected features are discussed from the taxonomic and functional points of view. The D2-D3 expansion domains of the 28S rRNA gene and the ITS region of L. barsii sp. n. were amplified and sequenced. Phylogenetic analysis using the D2-D3 expansion domains of the 28S rRNA gene revealed close evolutionary relationships with L. polyae, L. athesinus and three unidentified Longidorus spp.