mtDNA Regions Research Articles

Morphological, ecological and genetic characteristics of the sandy sculpin (<i>Leocottus kesslerii</i>, Cottidae) from the lakes Baikal, Arakhlei and Gusinoe, Transbaikalia

Studies on local forms of the Sandy sculpin, Leocottus kesslerii, in the lakes Baikal, Arakhlei and Gusinoye show that the differences between lake forms in terms of external morphology lie in the head length to width ratio. In the Baikal form, the head is wider than long, vs the opposite in both Apakhlei and Gusinoe forms. In the Baikal and Gusinoe forms, there is a gap between the dorsal fins, a gap being absent from the Arakhlei form and the dorsal fins are in touch with each other. In the Arakhlei form, the anal fin is long (37.4–43%, on average 39.3% SL). In the Baikal form, the length of the A base reaches 32.9% SL (29.0–34.5%). In the Baikal and Arakhlei forms, small and sparse bone prickles are present on the body, these being large and dense in the Gusinoe form. The differences identified show a low level of morphological isolation of the forms. More significant differences are established in ecological features: growth rate, food spectrum, reproductive characteristics, and fecundity values. The differences in ecological features are due to the adaptation of the Sandy sculpin forms to specific conditions of the lakes they inhabit. Studies on genetic differentiation were carried out using the control region (CR) of mtDNA. The study forms of the Sandy sculpin revealed weak genetic isolation. The p-distances between the L. kesslerii forms averaged from 0.33 ± 0.12 to 0.51 ± 0.24%. Such low values of genetic distances clearly lie within the limits of intraspecific variability. The data obtained on the morphological, ecological and genetic differentiation of lake forms of the Sandy sculpin show that fish from the lakes Baikal, Arakhlei and Gusinoe belong to the same nominative subspecies, Leocottus kesslerii kesslerii.

Identification of sequence polymorphism in the D-loop region of mitochondrial DNA as a risk factor for breast cancer.

Mitochondrial DNA (mtDNA) variations affect the efficiency of the electron transport chain and production of reactive oxygen species, contributing to carcinogenesis. The D-loop region of mtDNA has emerged as a variation hotspot region in human neoplasia; however, the potential contribution of these variations in breast cancer risk prediction remains unknown. We investigated the relationship between germline single nucleotide polymorphisms (SNPs) in the entire D-loop region and breast cancer risk in Chinese women. Peripheral blood-isolated mtDNA from 2329 patients with breast cancer and 2328 cancer-free controls was examined for SNPs. In the combined cohort, we used traditional risk factors, susceptibility germline polymorphisms, and logistic regression analysis to evaluate the predictive value of susceptibility variants for breast cancer risk. We calculated the area under the receiver operating characteristic curve (AUC) as a measure. We also measured the content of 8-hydroxy-2'-deoxyguanosine (8-OHdG). Individual polymorphisms SNP573 were significantly associated with breast cancer risk in both the discovery and validation cohorts. In the combined cohort, the AUC of the traditional risk factors was 64.3%; after adding susceptibility variants, the AUC was 64.9% (DeLong test, p = 0.007). 8-OHdG levels were significantly higher in patients with breast cancer than in controls and higher in individuals with SNP573 than in those negative for this variation. Overall, oxidative stress might be associated with the risk of breast cancer, and SNP573 might be associated with oxidative stress. Our results indicate the risk potential of polymorphisms in the D-loop region in breast cancer in Southern China.

A preliminary study on detecting human DNA in aquatic environments: Potential of eDNA in forensics

Human environmental DNA (eDNA) application have not been fully applied or adequately considered in the fields of eDNA and forensics. Nonetheless, this technique holds great potential as a complementary tool for detecting human DNA in aquatic environments, particularly in cases involving crimes connect to such environments. However, the detectability or stability of eDNA can vary depending on several factors. Therefore, this preliminary study investigates the detection and degradation rates of human eDNA, as well as the recovery of nuclear short tandem repeat (STR) profiles and mitochondrial DNA (mtDNA) sequencing, using water samples from both saltwater and freshwater sources. To conduct the experiment, whole human blood was spiked into the water samples. Water samples were then filtered using a 5 µm pore size filter, and samples were collected at various time intervals up to 23 days. A human specific qPCR assay targeting HV1 region of human mtDNA was used to detect human eDNA. Results demonstrated that human eDNA remains detectable for up to 36 hours in freshwater samples and up 84 hours in saltwater samples. The limit of detection (LOD) of human eDNA, (205 copies/µl), was achieved after 60 hours in freshwater and 180 hours in saltwater samples. Partial STR profiles could be recovered up to 24 hours for freshwater and saltwater. Results from mtDNA sequencing indicate that full mtDNA profiles could be recovered from freshwater samples up to 48 hours and remained detectable up to 72 hours in saltwater. Overall, the findings of this study underscore the importance of considering and incorporating human eDNA analysis as a valuable tool in forensic practice. By harnessing the power of eDNA, law enforcement agencies can enhance their investigation capabilities, improve the accuracy of forensic reconstructions, and ultimately contribute to the resolution of cases involving aquatic environments. Further research and validation are needed to optimize and expand the utilization of eDNA techniques in forensic investigations.

Pseudoperonospora cubensis Causing Downy Mildew on Luffa aegyptiaca in Korea

In November 2021, sponge gourd (<i>Luffa aegyptiaca</i>) plants with typical downy mildew symptom were observed in Korea. Based on morphological characteristics of sporangiophores and sporangia, combined with molecular phylogenetic analysis of the internal transcribed spacer rDNA and <i>cox</i>2 mtDNA regions, the causal pathogen was identified as <i>Pseudoperonospora cubensis</i> (Oomycota). This is the first report of downy mildew caused by <i>P. cubensis</i> on <i>L. aegyptiaca</i> in Korea. Given that <i>P. cubensis</i> exhibits high disease incidence and severity on many Cucurbitaceae crops, it could pose a significant threat to sponge gourd cultivation.

Mitochondrial tRNAAsn, tRNACys and tRNATyr encoding genes mutations landscape in colorectal cancer patient

The colorectal cancer (CRC) is one of the significant diseases implicated in high mortality rates in the world. Multi factors contributed in the cancer development and progression like tRNA encoding genes which have been linked with colorectal cancer. The present study aimed to report Mitochondrial tRNAAsn, tRNACys and tRNATyr encoding genes mutations landscape in colorectal cancer patients. A case control design depended on using PCR-sequencing of mtDNA region consisting of three tRNA genes. Data analysis was implemented using multiple alignment seq, tRNA secondary structure construction and phylogenic tree construction. The results showed that one case (female) was 25 years old and cancer stage T3N1MX has shown 6 mutations in tRNAAsn and 3 mutations in both tRNACys and tRNATyr while other samples did not show variations in the patients and control groups. The secondary structure of varied tRNAAsn, tRNACys and tRNATyr in this case might have been affected. The phylogenetic tree showed the conservative sequence of all CRC patients except the case that has a single cluster. The present finding concluded that there was non-association between tRNAAsn, tRNACys and tRNATyr encoding genes in mtDNA with colorectal cancer. One patient has mutations in tRNAs that may be affected in tRNA secondary structure and may be contributed for cancer progression.

Selection promotes age-dependent degeneration of the mitochondrial genome.

Somatic mutations in mitochondrial genomes (mtDNA) accumulate exponentially during aging. Using single cell sequencing, we characterize the spectrum of age-accumulated mtDNA mutations in mouse and human liver and identify directional forces that accelerate the accumulation of mutations beyond the rate predicted by a neutral model. "Driver" mutations that give genomes a replicative advantage rose to high cellular abundance and carried along "passenger" mutations, some of which are deleterious. In addition, alleles that alter mtDNA-encoded proteins selectively increased in abundance overtime, strongly supporting the idea of a "destructive" selection that favors genomes lacking function. Overall, this combination of selective forces acting in hepatocytes promotes somatic accumulation of mutations in coding regions of mtDNA that are otherwise conserved in evolution. We propose that these selective processes could contribute to the population prevalence of mtDNA mutations, accelerate the course of heteroplasmic mitochondrial diseases and promote age-associated erosion of the mitochondrial genome.

Genetic diversity and intraspecific mitochondrial DNA variations in the Georgian Mountain breed of Bos taurus reveal admixture, introgression and potential parallel vs. convergent evolution patterns

This study elucidates the haplotype diversity and mechanisms of evolutionary divergence for a broad population of the Georgian Mountain breed (GMB) of Bos taurus, using the sequencing and analysis of its mitochondrial DNA (mtDNA). In the evolutionary analyses, sequences of the targeted mtDNA region, involving the D-loop, CYTB, tRNA-Thr, and tRNA-Pro encoding genetic loci were analyzed using MEGA11, DnaSP, and SplitsTree software packages. A total of 25 haplotypes were determined among 82 individuals of GMB, belonging predominantly to the haplogroups T (T3, T1, T2, T4) or Q (Q1). Ten singleton haplotypes could also be determined in the GMB population. In the maximum likelihood evolutionary analysis, the singleton haplotype SNGT-9 appeared to be most closely related to the Bos indicus sub-haplogroup I1a. The haplotype diversity (0.997), nucleotide diversity (0.00636) and the overall mean distance within a population (0.01) calculated for GMB were greater as compared to the respective estimates (0.930, 0.00482 and 0.00) determined for its closest cattle relatives globally, suggesting stronger selection. It is suggested that the GMB diversity has been shaped by both parallel and convergent evolution, as well as by possible introgression, while pinpointing this breed’s ancient origin collectively.

DNA Barcoding of Freshwater Eels Anguilla spp from Kuari River Based on Cytochrome C Oxidase Subunit I (COI) Gene

Graphical Abstract Highlight Research Freshwater eels in Kuari River consisted of two species marmorata and A. bengalensis Anguilla marmorata had the highest similarity of 99.82%-100%, while bengalensis indicated the highest identity of 99.84%-100% to the same species in the GenBank database. Phylogenetic species indicated that Anguilla marmorata and Anguilla bengalensis form two different sub-clusters. The water quality in the Bengkulu Kuari River were still in tolerance for fish growth and survival. Abstract Freshwater eels (Anguilla spp.) are classified in the family Anguillidae, and included in the catadromous group. This study aimed to determine the COI gene sequences of the mitochondrial DNA, analyze the genetic distances, and phylogenetics, and characterize the physical and chemical parameters of between freshwater eels habitat in the Kuari River Bengkulu. This research was conducted from November 2020 – April 2021. The methods used in barcoding eel species were DNA isolation, DNA amplification using PCR (Polymerase Chain Reaction), electrophoresis, and sequencing of COI gene regions in mtDNA. The COI mtDNA gene fragments were obtained from PCR results with an annealing temperature of 50°C for 30 seconds in 35 cycles. BLASTn analysis of eel samples AM3 and AM4 had the highest similarity of 99.82%-100% to Anguilla marmorata, and samples AB2-AB5 indicated the highest identity of 99.84%-100% to Anguilla bengalensis. Phylogenetic species indicated that Anguilla marmorata and Anguilla bengalensis form two different sub-clusters. The water qualities in the Bengkulu Kuari River were temperatures 26.5-27.5oC, pH 7.1-8.7, dissolved oxygen 6.19-9.54 mg L-1, brightness 21-47 cm, ammonia 0.16-0.41 mg L-1, total alkalinity 20–52 mg L-1, TDS 33–49 mg L-1, salinity 0.3-0.4 ppt and water velocity 0.5-0.8 ms-1. The COI gene in DNA barcoding is very appropriate to be used for the identification of Anguilla spp species by comparing the DNA sequence of the COI gene with the existing database in the Genbank.

The Fast Evolution of the Stenobothrini Grasshoppers (Orthoptera, Acrididae, and Gomphocerinae) Revealed by an Analysis of the Control Region of mtDNA, with an Emphasis on the Stenobothrus eurasius Group.

The two cryptic grasshopper species of the genus Stenobothrus, S. eurasius and S. hyalosuperficies, demonstrate different acoustic behavior despite a strong similarity in morphology. A hybridization between these species is possible in the contact zone; however, there are little molecular data about the relationships of these species. The analysis of the mtDNA control region (CR) reveals that haplotypes of S. hyalosuperficies have more in common with the more distant Stenobothrus species than with the closely related S. eurasius. In the contact zone, S. eurasius has mt-haplotypes shared with S. hyalosuperficies, which might indicate an introgression of mtDNA from S. hyalosuperficies to the S. eurasius gene pool. We also analyze the structure and evolutionary rate of the mtDNA CR for the Stenobothrus genus and estimate the time of divergence of the species within the genus. The phylogenetic tree of the tribe Stenobothrini reconstructed with either the CR or COI gave the same four groups. The phylogenetic tree of the Stenobothrus genus has a star-like topology with each mtDNA haplotype found in any analyzed species, except S. eurasius, which forms a separate branch. The maximum degree of incomplete lineage sorting can demonstrate either ancestral polymorphism or introgression.

Horizontally transferred mitochondrial DNA tracts become circular by microhomology-mediated repair pathways.

The holoparasitic plant Lophophytum mirabile exhibits remarkable levels of mitochondrial horizontal gene transfer (HGT). Gathering comparative data from other individuals and host plants can provide insights into the HGT process. We sequenced the mitochondrial genome (mtDNA) from individuals of two species of Lophophytum and from mimosoid hosts. We applied a stringent phylogenomic approach to elucidate the origin of the whole mtDNAs, estimate the timing of the transfers, and understand the molecular mechanisms involved. Ancestral and recent HGT events replaced and enlarged the multichromosomal mtDNA of Lophophytum spp., with the foreign DNA ascending to 74%. A total of 14 foreign mitochondrial chromosomes originated from continuous regions in the host mtDNA flanked by short direct repeats. These foreign tracts are circularized by microhomology-mediated repair pathways and replicate independently until they are lost or they eventually recombine with other chromosomes. The foreign noncoding chromosomes are variably present in the population and likely evolve by genetic drift. We present the 'circle-mediated HGT' model in which foreign mitochondrial DNA tracts become circular and are maintained as plasmid-like molecules. This model challenges the conventional belief that foreign DNA must be integrated into the recipient genome for successful HGT.

Evaluation of unmodifiedhuman cell-derived extracellular vesicle mitochondrial deoxyribonucleic acid-based biodistribution in rodents.

Recently, extracellular vesicles (EVs) have been developed as therapeutic targets for various diseases. Biodistribution is crucial for EVs intended for therapeutic purposes because it can determine the degree of on- and off-target effects. This study aimed to explore techniques to evaluate the biodistribution of unmodified EVs. We devised a novel quantitative polymerase chain reaction (qPCR)-based assay to detect unmodified EVs by targeting mitochondrial deoxyribonucleic acid (mtDNA), a constituent of EVs. We focused on specific mtDNA regions that exhibited homologous variations distinct from their rodent mtDNA counterparts to establish this analytical approach. Herein, we successfully designed primers and probes targeting human and rodent mtDNA sequences and developed a highly specific and sensitive qPCR method. Furthermore, the quantification range of EVs isolated from various cells differed based on the manufacturer and cell source. IRDye 800CW-labelled Expi293F EV mimetics were administered to the animals via the tail vein to compare the imaging test and mtDNA-qPCR results. The results obtained from imaging tests and mtDNA-qPCR to investigate EV biodistribution patterns revealed differences. The results revealed that our newly developed method effectively determined the biodistribution of unmodified EVs with high sensitivity and reproducibility.

G-quadruplex propensity in H. neanderthalensis, H. sapiens and Denisovans mitochondrial genomes.

Current methods of processing archaeological samples combined with advances in sequencing methods lead to disclosure of a large part of H. neanderthalensis and Denisovans genetic information. It is hardly surprising that the genome variability between modern humans, Denisovans and H. neanderthalensis is relatively limited. Genomic studies may provide insight on the metabolism of extinct human species or lineages. Detailed analysis of G-quadruplex sequences in H. neanderthalensis and Denisovans mitochondrial DNA showed us interesting features. Relatively similar patterns in mitochondrial DNA are found compared to modern humans, with one notable exception for H. neanderthalensis. An interesting difference between H. neanderthalensis and H. sapiens corresponds to a motif found in the D-loop region of mtDNA, which is responsible for mitochondrial DNA replication. This area is directly responsible for the number of mitochondria and consequently for the efficient energy metabolism of cell. H. neanderthalensis harbor a long uninterrupted run of guanines in this region, which may cause problems for replication, in contrast with H. sapiens, for which this run is generally shorter and interrupted. One may propose that the predominant H. sapiens motif provided a selective advantage for modern humans regarding mtDNA replication and function.

Indel variation in the mitochondrial ND5 region supports monophyly of the tribe Hippoglossoidini (sensu Vinnikov et al. 2018) within the family Pleuronectidae

AbstractIndel (insertion–deletion) events observed in the genome represent irreversible mutational processes, making indel regions crucial characteristics for discussing phylogenetic relationships. The tribe Hippoglossoidini is a recently proposed taxonomic group based on the molecular phylogenetic analyses of both mitochondrial (mt) and nuclear DNA sequences. However, no synapomorphic characteristics have been identified within this tribe, either morphologically or molecularly. In the present study, we sequenced the ND5 region of mtDNA in the righteye flounder species and conducted interspecific comparisons. We found a 12 bp indel immediately upstream of the stop codon in the ND5 region. A comparative analysis of this region with outgroup species from the Paralichthyidae revealed that the indel was a unique insertion shared by the common ancestor of the Hippoglossoidini species, providing irreversible evidence to support the monophyly of this taxonomic group (synapomorphic characters).

A mito-y conundrum: is more general or less but specific sequence data optimal? A comparison of the ForenSeq mtDNA whole genome and control region kits

ABSTRACT Mitochondrial DNA (mtDNA) is more prevalent than genomic DNA, making it useful for challenging forensic samples such as degraded bones and hair shafts. A comparison of the ForenSeq whole genome and control region mtDNA kits was undertaken on bones burnt to high temperatures and hair shafts covering a range of hair colours and treatments. Haplogroup predictions were made from the same haplogroup branch per donor for one bone burnt ≥ 700°C and all hair segments. Overall, the whole genome mtDNA kit was both more successful and produced more discriminatory haplogroup predictions than the control region kit.

Mitochondrial DNA as a target for analyzing the biodistribution of cell therapy products.

Biodistribution tests are crucial for evaluating the safety of cell therapy (CT) products in order to prevent unwanted organ homing of these products in patients. Quantitative polymerase chain reaction (qPCR) using intronic Alu is a popular method for biodistribution testing owing to its ability to detect donor cells without modifying CT products and low detection limit. However, Alu-qPCR may generate inaccurate information owing to background signals caused by the mixing of human genomic DNA with that of experimental animals. The aim of this study was to develop a test method that is more specific and sensitive than Alu-qPCR, targeting the mitochondrial DNA (mtDNA) sequence that varies substantially between humans and experimental animals. We designed primers for 12S, 16S, and cytochrome B in mtDNA regions, assessed their specificity and sensitivity, and selected primers and probes for the 12S region. Human adipose-derived stem cells, used as CT products, were injected into the tail vein of athymic NCr-nu/nu mice and detected, 7 d after administration, in their lungs at an average concentration of 2.22 ± 0.69 pg/μg mouse DNA, whereas Alu was not detected. Therefore, mtDNA is more specific and sensitive than Alu and is a useful target for evaluating CT product biodistribution.

Genetic analysis of Octopus cyanea reveals high gene flow in the South-West Indian Ocean.

Octopus cyanea (Gray, 1849), abundant in the South-West Indian Ocean (SWIO), constitutes a vital resource for both subsistence and commercial fisheries. However, despite this socioeconomic importance, and recent indications of overfishing, little is known about the population structure of O. cyanea in the region. To inform sustainable management strategies, this study assessed the spatio-temporal population structure and genetic variability of O. cyanea at 20 sites in the SWIO (Kenya, Tanzania, Mozambique, Madagascar, Mauritius, Rodrigues, and the Seychelle Islands) by complementary analysis of mitochondrial DNA (mtDNA) noncoding region (NCR) sequences and microsatellite markers. MtDNA analysis revealed a shallow phylogeny across the region, with demographic tests suggesting historic population fluctuations that could be linked to glacial cycles. Contrary to expectations, NCR variation was comparable to other mtDNA regions, indicating that the NCR is not a hypervariable region. Both nuclear and mtDNA marker types revealed a lack of genetic structure compatible with high gene flow throughout the region. As adults are sedentary, this gene flow likely reflects connectivity by paralarval dispersal. All samples reported heterozygote deficits, which, given the overall absence of structure, likely reflect ephemeral larval recruitment variability. Levels of mtDNA and nuclear variability were similar at all locations and congruent with those previously reported for harvested Octopodidae, implying resilience to genetic erosion by drift, providing current stock sizes are maintained. However, as O. cyanea stocks in the SWIO represent a single, highly connected population, fisheries may benefit from additional management measures, such as rotational closures aligned with paralarval ecology and spanning geopolitical boundaries.

A multiplex DNA probe-based method for simultaneous identification of adulteration in meat samples

Meat adulteration and admixing are prevalent malpractices observed in processed and raw meat samples, where the consumption of adulterated meat has been associated with food allergies, financial losses, and consumer distrust. Meat authentication is pivotal to address these concerns. The meat authenticity can be determined through genetic, protein, and immunological markers and advanced detection methods. However, these methods often target a single species and lack the specificity to distinguish closely related species. Here, in the present study, we have developed a multiplex detection method based on the species-specific primers and probes, that can target four meat species in one reaction. The developed method amplifies the mitochondrial genomic regions of chicken, pork, sheep and goat using TaqMan multiplex probe-based RT-qPCR assay. Unique pairs of species-specific primers and probes that target specific mitochondrial DNA (mtDNA) regions of each species were designed and screened for specificity and sensitivity. The detection limit for species identification using the designed primers in real-time qPCR assays was 0.1 picogram per microliter (pg/μL) DNA detected in singleplex reaction and facilitates the simultaneous detection of closely related species, such as goat and sheep. Further, DNA-based probes were utilized in a multiplex real-time qPCR assay to identify chicken, pork, sheep and goat DNA in a single tube reaction. The multiplex assay was validated for raw and processed meat products, demonstrating its applications in ensuring the quality of meat products and safeguarding consumer interests.

Genetic diversity, spatial connectivity, and population structure of Asian silurid catfish Wallago attu (Bloch and Schneider, 1801) in the Ganga River System: insights from mitochondrial DNA analysis.

The Ganga River System (GRS) is a biodiversity hotspot, its ecological richness is shaped by a complex geological history. In this study, we examined the genetic diversity, spatial connectivity, and population structure of the Asian Silurid catfish, Wallago attu, across seven tributaries of the GRS. We employed three mitochondrial DNA (mtDNA) regions: cytochrome c oxidase subunit I (COXI), cytochrome b (Cyt b), and control region (CR). Our comprehensive dataset encompassed 2420bp of mtDNA, derived from 176 W. attu individuals across 19 sampling sites within the seven rivers of GRS. Our findings revealed high gene diversity (Hd:0.99) within W. attu populations. Analysis of Molecular Variance (AMOVA) highlighted that maximum genetic variations were attributed within the populations, and the observed genetic differentiation among the seven populations of W. attu ranged from low to moderate. Network analysis uncovered the presence of three distinct genetic clades, showing no specific association with seven studied rivers. Bayesian skyline plots provided insights into the demographic history of W. attu, suggesting a recent population expansion estimated to have occurred approximately 0.04million years ago (mya) during the Pleistocene epoch. These results significantly enhance our understanding of the genetic diversity and spatial connectivity of W. attu, serving as a vital foundation for developing informed conservation strategies and the sustainable management of this economically valuable resource within the Ganga River System.

First Report of Meloidogyne enterolobii Infecting Yellow pitaya (Selenicereus megalanthus) in China.

Yellow pitaya, Selenicereus megalanthus, is a night-blooming, climbing cacti of tropical origin, which has received increasing attention for its potential as a new exotic fruit crop (Lichtenzveig et al. 2000). The crop is grown extensively in Hainan Province, China (3000 ha). In October 2021, a survey was conducted on a farm located in Changjiang (19°21'4″N, 108°47'2″S), Hainan Province, China. Some yellow pitaya plants were found that were stunted and chlorotic, with abnormally thin stems (Fig. 1B), and no symptoms on healthy plants (Fig. 1A). Dead plants were also observed. Many galls and females with egg masses were observed on roots (Figs. 1C & 1D). This is typical of root-knot nematode (RKN) infections, and the incidence of infection was 36.7%. Meloidogyne sp. females and egg masses were dissected from roots of the infected plants. The perineal pattern of females (n= 5) was round to oval-shaped with a high dorsal arch (Figs. 1I & 1J). Second-stage juveniles (J2s) had truncated lips (Figs. 1E & 1F) and long-conical tails with bluntly rounded tips (Figs. 1G & 1H). The J2s body length (n= 24) averaged 416.79 μm (349.21 to 472.76 μm) with a mean width of 15.36 μm (12.47 to 17.52 μm); mean stylet length was 11.16 μm (10.10 to 13.23 μm); tail length averaged 53.73 μm (43.46 to 65.90 μm). The morphological characteristics matched the original description of M. enterolobii (Yang and Eisenback 1983). Males were not found. Genomic DNA was extracted from eight single J2s, and the mitochondrial (mtDNA) region between COII and 16S rRNA gene was amplified with primers C2F3/1108 (Powers and Harris 1993). A 652-bp DNA fragment was obtained, for which the sequence (GenBank accession no. OP122499) was 100% identical to the sequences of M. enterolobii isolates from China（MN269947）and the USA (MN809527). Furthermore, species identification was also confirmed using M. enterolobii specific primers Me-F/Me-R. An amplicon size of ∼230 bp was obtained, which is consistent with those previously reported for M. enterolobii (Fig. 2) (Long et al. 2006). Therefore, this population was identified as M. enterolobii based on morphological and molecular characteristics. Pathogenicity tests were performed in the greenhouse at 26℃ and 80% relative humidity with a 14-h/10-h light/dark photoperiod. Ten RKN-free S. megalanthus seedlings were transplanted into pots containing sterilized soil. After 3 weeks, the roots of 5 plants were inoculated with 3,000 eggs and J2s of M. enterolobii per plant. Five uninoculated plants were used as control plants. After 2 months, no galling or symptoms were observed on the control plants. All inoculated plants had galled roots similar to those observed in the field. Females and egg masses were obtained by dissecting galls. The nematode reproduction factor (RF= final population/initial population) was 1.9. Adult females (n= 5) dissected from inoculated plants were identified as M. enterolobii with sequence-specific primers Me-F/Me-R, thus confirming pathogenicity. The pathogenicity test was carried out twice with similar results. M. enterolobii is one of the most damaging species of RKN, due to its wide host range, high level of pathogenicity, and ability to develop and reproduce on several crops with resistance genes to other RKN (Castagnone-Sereno 2012). To our knowledge, this is the first report of S. megalanthus (yellow pitaya) as a host of M. enterolobii in China. Further studies are needed to develop and evaluate integrated management strategies.

Morphological and molecular characterizations of Pratylenchus coffeae infecting Ming aralia and coffee in Vietnam.

Pratylenchus coffeae, belonging to the root-lesion nematode group, is a highly prevalent and destructive plant-parasitic nematode that is able to infest a wide range of host plants. Although this species' devastating impacts on coffee plantations across the world are widely known on other host plants, its association with Ming aralia has never been reported. Our study characterized two popu lations of P. coffeae (associated with Ming aralia and coffee) and compared them with other popu lations from previous studies in Vietnam and other countries in the world. The identification of P. coffeae in our study was confirmed by the comprehensive analysis encompassing morphological examination, morphometric data, and molecular characterizations of the COI mtDNA and D2D3 of 28S rRNA regions. The cluster and MDS analyses revealed that the two populations of P. coffeae from Vietnam are closely related to those from Japan and Indonesia. The D2-D3 sequences of 28S rRNA and COI mtDNA regions exhibited high similarity among these populations, indicating a stable genetic profile. Our research contributes to a better understanding of the distribution and genetic characterizations of P. coffeae by offering new morphological and molecular insights into the presence of this nematode in Vietnam. Additionally, this nematode species was found to be associated with host plant's symptoms such as chlorotic leaves, stunted growth and root lesion in both hosts. Given the economic significance of both Ming aralia and coffee crops in Vietnam, as well as the damaging potential of P. coffeae, this study emphasizes the need of proactive nematode management measures to control this destructive pest.