First Detection of Ditylenchus destructor Parasitizing Maize in Northeast China

Maize is one of the most important crops in the world. Heilongjiang province has the largest maize area in China. Plant-parasitic nematodes are important agricultural pests, which cause huge economic losses every year and have attracted global attention. Potato rot nematode Ditylenchus destructor is a plant-parasitic nematode with a wide range of hosts and strong survival ability in different environments, which brings risks to agricultural production. In 2020, D. destructor was detected in seven maize fields in Heilongjiang province. Morphological identification and molecular approach were used to characterize the isolated D. destructor. The observed morphological and morphometric characteristics were highly similar and consistent with the existing description. The DNA sequencing on the D2/D3 region of the ribosomal DNA 28S and the phylogenetic analysis showed that D. destructor population obtained from maize and other isolates infesting carrot, sweet potato, and potato were in subclade I supported by a 96% bootstrap value. Additionally, the phylogenetic analysis of the ITS rRNA gene sequence further indicated that this D. destructor population from maize clustered in a clade I group and belonged to ITS rRNA haplotype C. An inoculation experiment revealed that D. destructor was pathogenic on the maize seedlings in pots and caused the disease symptoms in the stem base of maize seedlings. This is the first report of D. destructor causing stem rot of maize in Heilongjiang province, and contributes additional information on disease control and safe production of maize in the region.

Straw retention efficiently improves fungal communities and functions in the fallow ecosystem

Background Straw retention is a substitute for chemical fertilizers, which effectively maintain organic matter and improve microbial communities on agricultural land. The purpose of this study was to provide sufficient information on soil fungal community networks and their functions in response to straw retention. Hence, we used quantitative real-time PCR (qRT-PCR), Illumina MiSeq (ITS rRNA) and FUNGuild to examine ITS rRNA gene populations, soil fungal succession and their functions under control (CK) and sugarcane straw retention (SR) treatments at different soil layers (0–10, 10–20, 20–30, and 30–40 cm) in fallow fields. Result The result showed that SR significantly enhanced ITS rRNA gene copy number and Shannon index at 0–10 cm soil depth. Fungi abundance, OTUs number and ACE index decreased with the increasing soil depth. The ANOSIM analysis revealed that the fungal community of SR significantly differed from that of CK. Similarly, significant difference was also observed between topsoil (0–20 cm) and subsoil (20–40 cm). Compared with CK, SR decreased the relative abundance of the pathogen, while increased the proportion of saprotroph. Regarding soil depth, pathogen relative abundance in topsoil was lower than that in subsoil. Besides, both sugarcane straw retention and soil depths (topsoil and subsoil) significantly altered the co-occurrence patterns and fungal keystone taxa closely related to straw decomposition. Furthermore, both SR and topsoil had higher average clustering coefficients (aveCC), negative edges and varied modularity. Conclusions Overall, straw retention improved α-diversity, network structure and fungal community, while reduced soil pathogenic microbes across the entire soil profile. Thus, retaining straw to improve fungal composition, community stability and their functions, in addition to reducing soil-borne pathogens, can be an essential agronomic practice in developing a sustainable agricultural system.

Diagnosis of potato rot nematode Ditylenchus destructor using PCR-RFLP

During an investigation of nematodes in the Moscow region of Russia in 2019, a known species Ditylenchus destructor was recovered from tubers of potato plants. The genus Destructor is one of the most problematic genera of plant-parasitic nematodes. The numerous species reported for this genus have been cited from various sources. Due to the morphological similarity of many species and the lack of separation characteristics, the identification of D. destructor is difficult. Molecular taxonomy and phylogeny were used to confirm the identification. In the current study, PCR-RFLP illustrative models for the amplification of the ITS-rRNA gene were provided with two enzymes that could recognize D. destructor in potato tubers. Analysis of the rDNA sequences spanning both ITS1-ITS2 regions was carried out on the collected populations. The digestion of the PCR product of the ITS1-5.8S-ITS2 region with the enzyme TaqI produced three fragments; 100, 190, 550, and with Tru1I, two fragments were produced; 300 and 480 bp. The obtained DNA sequences were compared with those DNA sequences deposited in GenBank of populations isolated in other countries. The results showed no distinction between populations isolated from different host plant species, including populations found in the Russian Federation. New sequences from ITS-rRNA were deposited in the GenBank under accession number MN122076, MN658597, MN658599, MN658637, MN658638.

Role of Hibernation Promoting Factor in Ribosomal Protein Stability during Pseudomonas aeruginosa Dormancy

Pseudomonas aeruginosa is an opportunistic pathogen that causes biofilm-associated infections. P. aeruginosa can survive in a dormant state with reduced metabolic activity in nutrient-limited environments, including the interiors of biofilms. When entering dormancy, the bacteria undergo metabolic remodeling, which includes reduced translation and degradation of cellular proteins. However, a supply of essential macromolecules, such as ribosomes, are protected from degradation during dormancy. The small ribosome-binding proteins, hibernation promoting factor (HPF) and ribosome modulation factor (RMF), inhibit translation by inducing formation of inactive 70S and 100S ribosome monomers and dimers. The inactivated ribosomes are protected from the initial steps in ribosome degradation, including endonuclease cleavage of the ribosomal RNA (rRNA). Here, we characterized the role of HPF in ribosomal protein (rProtein) stability and degradation during P. aeruginosa nutrient limitation. We determined the effect of the physiological status of P. aeruginosa prior to starvation on its ability to recover from starvation, and on its rRNA and rProtein stability during cell starvation. The results show that the wild-type strain and a stringent response mutant (∆relA∆spoT strain) maintain high cellular abundances of the rProteins L5 and S13 over the course of eight days of starvation. In contrast, the abundances of L5 and S13 reduce in the ∆hpf mutant cells. The loss of rProteins in the ∆hpf strain is dependent on the physiology of the cells prior to starvation. The greatest rProtein loss occurs when cells are first cultured to stationary phase prior to starvation, with less rProtein loss in the ∆hpf cells that are first cultured to exponential phase or in balanced minimal medium. Regardless of the pre-growth conditions, P. aeruginosa recovery from starvation and the integrity of its rRNA are impaired in the absence of HPF. The results indicate that protein remodeling during P. aeruginosa starvation includes the degradation of rProteins, and that HPF is essential to prevent rProtein loss in starved P. aeruginosa. The results also indicate that HPF is produced throughout cell growth, and that regardless of the cellular physiological status, HPF is required to protect against ribosome loss when the cells subsequently enter starvation phase.

Molecular phylogeny of several species of Hoplolaimina (Nematoda: Tylenchida) associated with turfgrass in Korea, with comments on their morphology

Summary Seven species belonging to Suborder Hoplolaimina are characterised using integrative taxonomy, considering both morphological and molecular phylogenetic analyses of the 28S-rRNA, ITS-rRNA and COI gene sequences. It is evident that, as more populations of Pratylenchus zeae are continuously characterised, the species continues to display an ever-increasing intraspecific genetic variation within the 28S-rRNA and ITS-rRNA genes. However, the COI gene sequences exhibit minimum intraspecific variation and thus might be the most powerful DNA barcoding marker for the precise identification of P. zeae and should therefore be recommended as a complementary technique in the identification process of the species. Pratylenchus zeae, Meloidogyne graminicola and Heterodera pratensis are characterised herein for the first time in Korea, while the presence in Korea of P. penetrans, P. scribneri, H. avenae, and M. marylandi, is molecularly confirmed.

Molecular identification, phylogeny and phylogeography of the entomopathogenic nematodes of the genus Heterorhabditis Poinar, 1976: a multigene approach

Summary Presently, the genus Heterorhabditis contains 16 valid entomopathogenic nematode species. In this study we used samples from 11 species: H. amazonensis, H. bacteriophora, H. baujardi, H. beicherriana, H. downesi, H. floridensis, H. georgiana, H. indica, H. megidis, H. noenieputensis, and H. zealandica to amplify and sequence five gene fragments: the D2-D3 expansion segments of 28S rRNA, ITS rRNA, COI mtDNA genes and unc-87 and cmd-1 genes encoding thin filament (F-actin)-associated protein and calmodulin, respectively. Fifty new sequences for 11 species were generated. More than 980 sequences of five genes were analysed. Phylogenetic and sequence analysis of these genes using Bayesian inference, maximum likelihood and statistical parsimony confirmed a division of the genus into three clades (groups): ‘Indica’, ‘Bacteriophora’ and ‘Megidis’. The analysis of gene sequences downloaded from GenBank and identified as Heterorhabditis revealed many cases of species misidentifications and presence of reading mistakes in some sequences. Synonymisation of H. somsookae with H. baujardi, H. gerrardi, H. pakistanensis with H. indica, and H. sonorensis with H. taysearae, are confirmed by sequence and phylogenetic analysis. The ITS rRNA and COI genes could be considered as informative markers for species identification, barcoding and phylogeographical studies of Heterorhabditis.

Cactodera solani n. sp. (Nematoda: Heteroderidae), a new species of cyst-forming nematode parasitising tomato in Mexico

Summary A new species of cyst-forming nematode, Cactodera solani n. sp., from Mexico is described. The new species was found in a glasshouse in Mexico City parasitising tomato and common lambsquarter. This cyst nematode has light brown to almost black cysts, presents a small vulval cone with circumfenestra and morphologically it most resembles C. milleri. Cactodera solani n. sp. can be differentiated from C. milleri by having smaller cysts that are 417 (291-581) × 324 (204-505) vs 632 (515-730) × 506 (419-598) μm in size and a longer stylet in the second-stage juveniles of 25 (24-27) vs 22 (21-23) μm. Phylogenetic relationships within populations and species of Cactodera are given based on the analysis of the ITS rRNA and the partial COI gene sequences. The ITS rRNA and COI gene sequences clearly differentiated C. solani n. sp. from other Cactodera species. This new cyst-forming nematode has several generations per year and its life cycle can be completed in 49 days on tomato and in 40 days on common lambsquarter at 20-25°C.

Morphological and molecular characterisation of Distolabrellus veechi (Rhabditida: Mesorhabditidae) from India

Summary A redescription of Distolabrellus veechi (Rhabditida: Mesorhabditidae) from agricultural soils in Uttarakhand and Uttar Pradesh, India, is provided. Specimens were extracted from soil using the Galleria soil baiting technique, with two strains of nematodes named JP1 and JP2 being obtained. Morphological and morphometric studies on the species agree well with previous records. Molecular analyses are provided using 18S, 28S and ITS rRNA sequences, the ITS sequences being obtained for the first time in this species. These analyses show that the material examined agrees well with previously studied populations. Phylogenetic analyses showed Distolabrellus to be the sister group of Crustorhabditis and Teratorhabditis, all having the male spicules fused for more than 50% of their length. Illustrations and phylogenetic trees based on 18S, 28S and ITS rRNA sequences are provided.

Description of Stenomitos kolaenensis and S. hiloensis sp. nov. (Leptolyngbyaceae, Cyanobacteria) with an emendation of the genus

Stenomitos is a recently established cyanobacterial genus, some species of which appear to be cryptic. Here we describe two new species in this genus, Stenomitos kolaensis sp. nov. isolated from the Al-Fe humic podzols of a boreal forest near Nikel town, Murmansk region, Russia and S. hiloensis sp. nov. isolated from a basaltic seep wall on Akeola Road, Hilo, Hawaii, USA. Phylogenetic analyses were conducted on the 16S and 16S-23S ITS rRNA gene regions using Bayesian Inference, and Maximum Likelihood. Phylogenetic analysis of the 16S-23S ITS rRNA region resulted in both S. kolaensis and S. hiloensis forming separate clades from other Stenomitos lineages. Antarctic strains of Stenomitos frigidus (previously reported as “Leptolyngbya frigida”) show that species to be polyphyletic and in need of revision. The structure of the conserved ITS regions (Box-B, D1-D1ʹ, V2 and V3 helices) provided support for separation of the species, and the p-distances among aligned ITS regions further confirmed that a number of species exist within the genus. S. kolaensis and S. hiloensis can be distinguished from other described Stenomitos species (S. rutilans and S. tremulus) by their geographical distribution, habitat preference, 16S rRNA phylogeny, and differences in the secondary structure of the 16S-23S ITS region.