Risk Factors of Perioperative Brain Injury in Children Under Two Years Undergoing Coarctation Repair.

BACKGROUND: To investigate clinical manifestations and factors of perioperative brain injury (PBI) after surgical repair of coarctation of the aorta (CoA) combined with other heart malformations under cardiopulmonary bypass (CPB) in children under two years. METHODS: The clinical data of 100 children undergoing CoA repair were retrospectively reviewed between January 2010 and September 2021. Univariate and multivariate analyses were performed to identify factors of PBI development. Hierarchical and K-means cluster analyses were conducted to evaluate the association between hemodynamic instability and PBI. RESULTS: Eight children developed postoperative complications, and all of them had a favorable neurological outcome one year after surgery. Univariate analysis revealed eight risk factors associated with PBI. Multivariate analysis indicated operation duration (P = 0.04, odds ratio [OR], 2.93; 95% confidence interval [CI], 1.04 to 8.28) and pulse pressure (PP) minimum (P = 0.01; OR, 0.22; 95% CI, 0.06 to 0.76) were independently associated with PBI. The following three parameters emerged for cluster analysis: PP minimum, mean arterial pressure (MAP) dispersion, and systemic vascular resistance (SVR) average. Using cluster analysis, PBI mainly occurred in subgroups 1 (12%, three of 26) and 2 (10%, five of 48). The mean value of PP and MAP in subgroup 1 was significantly higher than in subgroup 2. The mean SVR in subgroup 1 was the highest. The lowest PP minimum, MAP, and SVR were observed in subgroup 2. CONCLUSION: Lower PP minimum and longer operation duration were independent risk factors for developing PBI in children under two years during CoA repair. Unstable hemodynamics should be avoided during CPB.

Meditation Using a Mobile App Improves Surgery Trainee Performance: A Simulation-Based Randomized Controlled Trial.

PURPOSE: To primarily investigate: (1) whether a 10-minute instant meditation practice using a mobile app could enhance arthroscopy performance and (2) whether a 10-day app-based meditation could reduce short-term arthroscopic skills deterioration. METHODS: Orthopaedic residents with no previous experience in arthroscopy and meditation were randomly assigned to groups A, B, and C. After initial standard competency-based arthroscopy training on the simulator on day 1, a pretest was performed via the simulator by all participants to assess their initial level of performance, then groups A and B were required to practice app-based mindfulness meditation 10 min/day for 10 consecutive days while group C did nothing. On day 11, all participants returned to perform a posttest. Before the posttest, the participants in group A practiced app-based meditation (10 minutes), whereas groups B and C had no intervention. RESULTS: In total, 43 participants were included and reached similar level of performance after initial training phase in day 1. On day 11, participants in group A had statistically a better instant arthroscopy performance than group B, with greater total score (mean difference [MD] 3.57; P < .001), less completion time (MD -42.89 seconds; P = .001), shorter camera (MD -23.38 cm; P < .001) and grasper (MD -15.23 cm; P = .002) path length, and less cartilage injury (MD -1.07%; P = .012). Participants in group B had less skills deterioration than group C, with better total score (MD -5.42; P < .001), less completion time (MD 51.96s; P = .002), camera path length (MD 28.41 cm; P = .007), and cartilage injury (MD 1.19%; P = .038). CONCLUSIONS: Meditation training using a mobile app enhanced instant simulation-based arthroscopy performance and reduced short-term skills deterioration of orthopaedic residents with no arthroscopy hands-on experience. CLINICAL RELEVANCE: A meditation using mobile app for clinicians and educators should be incorporated into simulation-based arthroscopy curriculums and perhaps clinical settings to improve arthroscopy performance and mental health of orthopaedic residents without any previous arthroscopy experience.

Learning and Short-Term Retention of Simulation-Based Arthroscopic Skills.

OBJECTIVE: The study aimed to examine the learning curve and short-term retention of arthroscopic skills acquired on a simulator. DESIGN: Cohort study. SETTING: Clinical Skills Training Center of Zhujiang Hospital of Southern Medical University PARTICIPANT AND METHODS: Orthopaedic residents (n = 14) without previous arthroscopy experience were included. After basic information was collected and an initial arthroscopy knowledge level test was administered, the subjects received standardised training on the simulator (day 1); then, they completed tasks on the simulator, including guided diagnostics (4 times), triangulation (5 times) and loose body removal (7 times). A learning curve for each skill was generated based on the total scores. The score of the last repetition of each task was the training level. RESULTS: A total of 14 orthopedic residents were enrolled. All participants completed the training and testing. There was a learning curve over the course of training for all 3 arthroscopic skills (p < 0.001). On day 8 after the training, the mean score for guided diagnostics decreased from 49.9 to 48.9 (p = 0.001), and the retention rate was 97.8%. For triangulation, the mean total score decreased from 58.9 to 53.6 (p < 0.001), and the retention rate was 90.8%. For loose body removal, the mean total score decreased from 87.1 to 80.7 (p < 0.001), and the retention rate was 92.7%. CONCLUSIONS: Orthopaedic residents' arthroscopic skills learned through simulator training declined significantly in 1 week after the training, especially more difficult skills.

The frequency of assessment tools in arthroscopic training: a systematic review.

BACKGROUND: Multiple assessment tools are used in arthroscopic training and play an important role in feedback. However, it is not fully recognized as to the standard way to apply these tools. Our study aimed to investigate the use of assessment tools in arthroscopic training and determine whether there is an optimal way to apply various assessment tools in arthroscopic training. METHODS: A search was performed using PubMed, Embase and Cochrane Library electronic databases for articles published in English from January 2000 to July 2021. Eligible for inclusion were primary research articles related to using assessment tools for the evaluation of arthroscopic skills and training environments. Studies that focussed only on therapeutic cases, did not report outcome measures of technical skills, or did not mention arthroscopic skills training were excluded. RESULTS: A total of 28 studies were included for review. Multiple assessment tools were used in arthroscopic training. The most common objective metric was completion time, reported in 21 studies. Technical parameters based on simulator or external equipment, such as instrument path length, hand movement, visual parameters and injury, were also widely used. Subjective assessment tools included checklists and global rating scales (GRS). Among these, the most commonly used GRS was the Arthroscopic Surgical Skill Evaluation Tool (ASSET). Most of the studies combined objective metrics and subjective assessment scales in the evaluation of arthroscopic skill training. CONCLUSIONS: Overall, both subjective and objective assessment tools can be used as feedback for basic arthroscopic skill training, but there are still differences in the frequency of application in different contexts. Despite this, combined use of subjective and objective assessment tools can be applied to more situations and skills and can be the optimal way for assessment. LEVEL OF EVIDENCE: Level III, systematic review of level I to III studies. Key messagesBoth subjective and objective assessment tools can be used as feedback for basic arthroscopic skill training.Combined use of subjective and objective assessment tools can be applied to more situations and skills and can be the optimal way for assessment.

Comparative Mitochondrial Analysis of Cnaphalocrocis exigua (Lepidoptera: Crambidae) and Its Close Relative C. medinalis.

Rice leaffolders are important pests on rice in Asia, Oceania, and Africa, causing serious loss to rice production. There are two main rice leaffolders in China, namely Cnaphalocrocis medinalis (Guenée) and C. exigua (Butler) with the former having the ability of long-distance migration. To reveal the differences in the mitochondrial genomes (mitogenome) between them, we compared the completed mitogenome of C. exigua with three C. medinalis individuals. Although phylogenetic analysis based on the mitogenomic data strongly supported the close relationship between these two species, many differences were still being revealed. The results showed that the mitogenome of C. exigua was shorter in length (15,262 bp) and slight lower in AT content than that of C. medinalis. Except for the different start codons of nad3 and nad6 gene, we also found the cox1 gene had a typical start codon 'ATG' which suggested that the starting position of this gene must be reconsidered in the entire superfamily Pyraloidea. All tRNAs have a typical clover-leaf structure, except for the dihydrouridine (DHU) stem losing of trnS1, which has the atypical anticondon 'TCT' instead of 'GCT' in C. medinalis and most Pyraloidea species. Two intergenic regions (between trnY and cox1, nad3 and trnA) featured by AT repeats were only found in C. medinalis and even rarely appeared in reported Pyraloidea species. Furthermore, regardless of interspecific comparison or intraspecific comparison of these two species, protein coding genes, especially the atp8 genes, had quite different evolutionary rates.

Simulation-Based Arthroscopic Skills Using a Spaced Retraining Schedule Reduces Short-Term Task Completion Time and Camera Path Length.

PURPOSE: To investigate whether acquiring basic knee arthroscopic skills via a spaced retraining schedule could prevent skills deterioration and achieve further skills improvement. METHODS: In the learning phase, 16 residents with no previous hands-on experience in practicing arthroscopic skills were asked to perform basic arthroscopic tasks on a simulator until they attained perfect scores in each task. Immediately after completing the learning phase, a pretest was performed to assess their performance. Next, they were randomly assigned into 2 groups. The spaced retraining group, which undertook a spaced repetitive training phase with a fixed-time interval, returned on days 2, 4 and 6 to repeat the same tasks for 20 minutes per day, whereas the control group did nothing. On day 7, all participants performed a posttest. A 2 × 2 mixed analysis of variance model was used for statistical analysis. RESULTS: Significant differences between the 2 groups were found in task completion time (P = .003) and camera path length (P = .043) but not cartilage injury (P = .186). Residents in the spaced retraining group decreased their task completion time (163.2 ± 23.9 seconds) whereas the task time in the control group increased (351.3 ± 25.5 seconds). The same pattern was found with the camera path length. CONCLUSIONS: Implementing a spaced retraining schedule in 1 week resulted in a reduced task completion time and camera path length but no significant reduction in cartilage injury. It appears that introducing a spaced retraining schedule to retain arthroscopic skills acquired through massed learning may be advantageous. CLINICAL RELEVANCE: In consideration of the training time available to residents and the trend toward massed learning, this spaced retraining schedule may offer a cost-effective and convenient way for residents to maintain and improve their basic arthroscopic skills with no significant increase in time invested.

Genomic Analysis of Wolbachia from Laodelphax striatellus (Delphacidae, Hemiptera) Reveals Insights into Its "Jekyll and Hyde" Mode of Infection Pattern.

Wolbachia is a widely distributed intracellular bacterial endosymbiont among invertebrates. The wStriCN, the Wolbachia strain that naturally infects an agricultural pest Laodelphax striatellus, has a "Jekyll and Hyde" mode of infection pattern with positive and negative effects: It not only kills many offspring by inducing cytoplasmic incompatibility (CI) but also significantly increases host fecundity. In this study, we assembled the draft genome of wStriCN and compared it with other Wolbachia genomes to look for clues to its Jekyll and Hyde characteristics. The assembled wStriCN draft genome is 1.79 Mb in size, which is the largest Wolbachia genome in supergroup B. Phylogenomic analysis showed that wStriCN is closest to Wolbachia from Asian citrus psyllid Diaphorina citri. These strains formed a monophylogentic clade within supergroup B. Compared with other Wolbachia genomes, wStriCN contains the most diverse insertion sequence families, the largest amount of prophage sequences, and the most ankyrin domain protein coding genes. The wStriCN genome encodes components of multiple secretion systems, including Types I, II, IV, VI, Sec, and Tac. We detected three pairs of homologs for CI factors CifA and CifB. These proteins harbor the catalytic domains responsible for CI phenotypes but are phylogenetically and structurally distinct from all known Cif proteins. The genome retains pathways for synthesizing biotin and riboflavin, which may explain the beneficial roles of wStriCN in its host planthoppers, which feed on nutrient-poor plant sap. Altogether, the genomic sequencing of wStriCN provides insight into understanding the phylogeny and biology of Wolbachia.

Wolbachia supplement biotin and riboflavin to enhance reproduction in planthoppers.

Symbiont-mediated nutritional mutualisms can contribute to the host fitness of insects, especially for those that feed exclusively on nutritionally unbalanced diets. Here, we elucidate the importance of B group vitamins in the association of endosymbiotic bacteria Wolbachia with two plant-sap feeding insects, the small brown planthopper, Laodelphax striatellus (Fallén), and the brown planthopper, Nilaparvata lugens (Stål). Infected planthoppers of both species laid more eggs than uninfected planthoppers, while the experimental transfer of Wolbachia into uninfected lines of one planthopper species rescued this fecundity deficit. The genomic analysis showed that Wolbachia strains from the two planthopper species encoded complete biosynthesis operons for biotin and riboflavin, while a metabolic analysis revealed that Wolbachia-infected planthoppers of both species had higher titers of biotin and riboflavin. Furthermore, experimental supplementation of food with a mixture of biotin and riboflavin recovered the fecundity deficit of Wolbachia-uninfected planthoppers. In addition, comparative genomic analysis suggested that the riboflavin synthesis genes are conserved among Wolbachia supergroups. Biotin operons are rare in Wolbachia, and those described share a recent ancestor that may have been horizontally transferred from Cardinium bacteria. Our research demonstrates a type of mutualism that involves a facultative interaction between Wolbachia and plant-sap feeding insects involving vitamin Bs.

Identification of Wolbachia-Responsive miRNAs in the Small Brown Planthopper, Laodelphax striatellus.

Laodelphax striatellus is naturally infected with the Wolbachia strain wStri, which induces strong cytoplasmic incompatibility of its host. MicroRNAs (miRNAs) are a class of endogenous non-coding small RNAs that play a critical role in the regulation of gene expression at post-transcriptional level in various biological processes. Despite various studies reporting that Wolbachia affects the miRNA expression of their hosts, the molecular mechanism underlying interactions between Wolbachia and their host miRNAs has not been well understood. In order to better understand the impact of Wolbachia infection on its host, we investigated the differentially expressed miRNAs between Wolbachia-infected and Wolbachia-uninfected strains of L. striatellus. Compared with uninfected strains, Wolbachia infection resulted in up-regulation of 18 miRNAs and down-regulation of 6 miRNAs in male, while 25 miRNAs were up-regulated and 15 miRNAs were down-regulated in female. The target genes of these differentially expressed miRNAs involved in immune response regulation, reproduction, redox homeostasis and ecdysteroidogenesis were also annotated in both sexes. We further verified the expression of several significantly differentially expressed miRNAs and their predicted target genes by qRT-PCR method. The results suggested that Wolbachia appears to reduce the expression of genes related to fertility in males and increase the expression of genes related to fecundity in females. At the same time, Wolbachia may enhance the expression of immune-related genes in both sexes. All of the results in this study may be helpful in further exploration of the molecular mechanisms by which Wolbachia affects on its hosts.

Multi-terrane structure controls the contrasting lithospheric evolution beneath the western and central-eastern Tibetan plateau.

The Tibetan plateau is manifested by contrasting along-strike lithospheric structures, but its formation mechanism and the relationship with the heterogeneous multi-terrane configuration is a challenging problem. Here we conduct systematic numerical modeling to explore the roles of width, density, and rheological properties of the multiple terranes in the lithospheric evolution of the Tibetan plateau, which reveals two distinct collision modes. In Mode-I, the lithospheric mantles of both the strong and weak terranes in the Tibetan plate are completely detached, followed by the underthrusting of Indian lithosphere beneath the whole plateau. Alternatively, Mode-II is characterized by full detachment of the weak terranes, but (partial) residue of the strong terranes during collision. These two contrasting modes, broadly consistent with the lithospheric structures of western and central-eastern Tibetan plateau, respectively, are strongly dependent on the along-strike variation of the width of the strong Lhasa-Qiangtang terranes.

Newly discovered Late Triassic Baqing eclogite in central Tibet indicates an anticlockwise West-East Qiangtang collision.

The Triassic eclogite-bearing central Qiangtang metamorphic belt (CQMB) in the northern Tibetan Plateau has been debated whether it is a metamorphic core complex underthrust from the Jinsha Paleo-Tethys or an in-situ Shuanghu suture. The CQMB is thus a key issue to elucidate the crustal architecture of the northern Tibetan Plateau, the tectonics of the eastern Tethys, and the petrogenesis of Cenozoic high-K magmatism. We here report the newly discovered Baqing eclogite along the eastern extension of the CQMB near the Baqing town, central Tibet. These eclogites are characterized by the garnet + omphacite + rutile + phengite + quartz assemblages. Primary eclogite-facies metamorphic pressure-temperature estimates yield consistent minimum pressure of 25 ± 1 kbar at 730 ± 60 °C. U-Pb dating on zircons that contain inclusions (garnet + omphacite + rutile + phengite) gave eclogite-facies metamorphic ages of 223 Ma. The geochemical continental crustal signature and the presence of Paleozoic cores in the zircons indicate that the Baqing eclogite formed by continental subduction and marks an eastward-younging anticlockwise West-East Qiangtang collision along the Shuanghu suture from the Middle to Late Triassic.

The complete mitochondrial genome of Bactrocera diaphora (Diptera: Tephtitidae).

We sequenced and annotated the complete mitochondrial genome (mitogenome) of Bactrocera diaphora (Diptera: Tephtitidae), which is an economically important pest in the southwest area of China, India, Sri Lanka, Vietnam and Malaysia. This mitogenome is 15 890 bp in length with an A + T content of 74.103%, and contains 37 typical animal mitochondrial genes that are arranged in the same order as that of the inferred ancestral insects. All protein-coding genes (PCGs) start with a typical ATN codon, except cox1 that begins with TCG. Ten PCGs stop with termination codon TAA or TAG, whereas cox1, nad1 and nad5 have single T-- as the incomplete stop codon. All of the transfer RNA genes present the typical clover leaf secondary structure except trnS1 (AGN) with a looping D-arm. The A + T-rich region is located between rrnS and trnI with a length of 946 bp, and contains a 20 bp poly-T stretch and 22 bp poly-A stretch. Except the control region, the longest intergenic spacer is located between trnR and trnN that is 94 bp long with an excessive high A + T content (95.74%) and a microsatellite-like region (TA)13.

Identification of Wolbachia-responsive microRNAs in the two-spotted spider mite, Tetranychus urticae.

BACKGROUND: The two-spotted spider mite, Tetranychus urticae, is infected with Wolbachia, which have the ability to manipulate host reproduction and fitness. MicroRNAs (miRNAs) are small non-coding RNAs that are involved in many biological processes such as development, reproduction and host-pathogen interactions. Although miRNA was observed to involve in Wolbachia-host interactions in the other insect systems, its roles have not been fully deciphered in the two-spotted spider mite. RESULTS: Small RNA libraries of infected and uninfected T. urticae for both sexes (in total four libraries) were constructed. By integrating the mRNA data originated from the same samples, the target genes of the differentially expressed miRNAs were predicted. Then, GO and pathway analyses were performed for the target genes. Comparison of libraries showed that Wolbachia infection significantly regulated 91 miRNAs in females and 20 miRNAs in males, with an overall suppression of miRNAs in Wolbachia-infected libraries. A comparison of the miRNA and mRNA data predicted that the differentially expressed miRNAs negatively regulated 90 mRNAs in females and 9 mRNAs in males. An analysis of target genes showed that Wolbachia-responsive miRNAs regulated genes with function in sphingolipid metabolism, lysosome function, apoptosis and lipid transporting in both sexes, as well as reproduction in females. CONCLUSION: Comparisons of the miRNA and mRNA data can help to identify miRNAs and miRNA target genes involving in Wolbachia-host interactions. The molecular targets identified in this study should be useful in further functional studies.

The complete mitochondrial genomes of six species of Tetranychus provide insights into the phylogeny and evolution of spider mites.

Many spider mites belonging to the genus Tetranychus are of agronomical importance. With limited morphological characters, Tetranychus mites are usually identified by a combination of morphological characteristics and molecular diagnostics. To clarify their molecular evolution and phylogeny, the mitochondrial genomes of the green and red forms of Tetranychus urticae as well as T. kanzawai, T. ludeni, T. malaysiensis, T. phaselus, T. pueraricola were sequenced and compared. The seven mitochondrial genomes are typical circular molecules of about 13,000 bp encoding and they are composed of the complete set of 37 genes that are usually found in metazoans. The order of the mitochondrial (mt) genes is the same as that in the mt genomes of Panonychus citri and P. ulmi, but very different from that in other Acari. The J-strands of the mitochondrial genomes have high (∼ 84%) A+T contents, negative GC-skews and positive AT-skews. The nucleotide sequence of the cox1 gene, which is commonly used as a taxon barcode and molecular marker, is more highly conserved than the nucleotide sequences of other mitochondrial genes in these seven species. Most tRNA genes in the seven genomes lose the D-arm and/or the T-arm. The functions of these tRNAs need to be evaluated. The mitochondrial genome of T. malaysiensis differs from the other six genomes in having a slightly smaller genome size, a slight difference in codon usage, and a variable loop in place of the T-arm of some tRNAs by a variable loop. A phylogenic analysis shows that T. malaysiensis first split from other Tetranychus species and that the clade of the family Tetranychoidea occupies a basal position in the Trombidiformes. The mt genomes of the green and red forms of T. urticae have limited divergence and short evolutionary distance.

The complete mitochondrial genome sequence of Sogatella furcifera (Horváth) and a comparative mitogenomic analysis of three predominant rice planthoppers.

The white-backed planthopper (WBPH), Sogatella furcifera (Horváth), is one of the most destructive pests of rice crops in many Asian countries. Using long-PCR and shotgun library methods, we sequenced the entire mitochondrial genomes (mt-genomes) of two WBPH individuals. Total lengths of the mt-genome of the two WBPH individuals were 16,612 bp and 16,654 bp with an identical AT content of 76.19%. Among the 13 protein coding genes (PCGs), only nad5 used an atypical initiation codon GTG. Most of the tRNA genes had the typical cloverleaf secondary structure except that the dihydrouridine (DHU) arms in two trnS genes and the TΨC arm of trnG gene did not form a stable stem-loop structure. Similar to the brown planthopper (BPH), Nilaparvata lugens (Stål), and the small brown planthopper (SBPH), Laodelphax striatellus (Fallén), some extraordinary features were observed in the WBPH mt-genome. These include similar gene rearrangement pattern, unusually short length of the atp8 gene and variable numbers of tandem repeat (VNTR) structure in control region. Interestingly, the same tandem repeat unit with stable secondary structure appeared in two different planthoppers, WBPH and SBPH, which belong to two different genera of the Delphacidae. This peculiar feature provides a direct evidence for the close relationship between the two planthoppers and updates our understanding of the evolutionary characteristics of mitochondrial control region. Comparison with two other predominant rice planthoppers (BPH and SBPH) revealed that different PCGs of mitochondria exhibit different evolutionary patterns.

Wolbachia play an important role in affecting mtDNA variation of Tetranychus truncatus (Trombidiformes: Tetranychidae).

The prevalence of the endosymbiont Wolbachia and its effects on mitochondria variation were analyzed in seven natural populations of Tetranychus truncatus Ehara (Trombidiformes: Tetranychidae) in current study. Five Wolbachia strains (wtru1, wtru5, wtru7, wtru8, and wtru12) were detected based on the surface protein of Wolbachia (wsp) sequence data and the multiple locus sequences typing data, suggesting that multiple separate invasions have occurred. Part of mitochondrial cytochrome oxidase subunit I gene was sequenced from infected individuals revealing 10 different haplotypes. As predicted, the haplotype and nucleotide diversity were lower in infected individuals than that in uninfected individuals. Furthermore, phylogenetic and analysis of molecular variance analyses revealed that the distribution of mtDNA haplotypes is not associated with geography. Rather, it is strongly concordant with infection status. These data support the hypothesis that Wolbachia infection can affect the genetic structure and diversity of the host mites.

Various infection status and molecular evidence for horizontal transmission and recombination of Wolbachia and Cardinium among rice planthoppers and related species.

Wolbachia and Cardinium are widely distributed and are considered important for their ability to disturb reproduction and affect other fitness-related traits of their hosts. By using multilocus sequence typing (MLST), RFLP (restriction fragment length polymorphism) and 16S ribosomal DNA gene sequencing methods, we extensively surveyed Wolbachia and Cardinium infection status of four predominant rice planthoppers and one kind of leafhopper in different rice fields. The results demonstrated that Sogatella furcifera (Horváth) and Laodelphax striatellus (Fallén) were infected with the same Wolbachia strain (wStri), while Nilaparvata lugens (Stål) and its closely related species Nilaparvata muiri China were infected with two phylogeneticlly distant strains, wLug and wMui, respectively. Three new Wolbachia strains (provisionally named wMfas1, wMfas2 and wMfas3) were detected in the leafhopper Macrosteles fascifrons (Stål). Only S. furcifera was co-infected with Cardinium, which indicated that the distribution of Cardinium in these rice planthoppers was narrower than that of Wolbachia. Unambiguous intragenic recombination events among these Wolbachia strains and incongruent phylogenetic relationships show that the connections between different Wolbachia strains and hosts were more complex than we expected. These results suggest that horizontal transmission and host associated specialization are two factors affecting Wolbachia and Cardinium infections among planthoppers and their related species.

The complete mitochondrial genomes of two rice planthoppers, Nilaparvata lugens and Laodelphax striatellus: conserved genome rearrangement in Delphacidae and discovery of new characteristics of atp8 and tRNA genes.

BACKGROUND: Nilaparvata lugens (the brown planthopper, BPH) and Laodelphax striatellus (the small brown planthopper, SBPH) are two of the most important pests of rice. Up to now, there was only one mitochondrial genome of rice planthopper has been sequenced and very few dependable information of mitochondria could be used for research on population genetics, phylogeographics and phylogenetic evolution of these pests. To get more valuable information from the mitochondria, we sequenced the complete mitochondrial genomes of BPH and SBPH. These two planthoppers were infected with two different functional Wolbachia (intracellular endosymbiont) strains (wLug and wStri). Since both mitochondria and Wolbachia are transmitted by cytoplasmic inheritance and it was difficult to separate them when purified the Wolbachia particles, concomitantly sequencing the genome of Wolbachia using next generation sequencing method, we also got nearly complete mitochondrial genome sequences of these two rice planthoppers. After gap closing, we present high quality and reliable complete mitochondrial genomes of these two planthoppers. RESULTS: The mitogenomes of N. lugens (BPH) and L. striatellus (SBPH) are 17, 619 bp and 16, 431 bp long with A + T contents of 76.95% and 77.17%, respectively. Both species have typical circular mitochondrial genomes that encode the complete set of 37 genes which are usually found in metazoans. However, the BPH mitogenome also possesses two additional copies of the trnC gene. In both mitochondrial genomes, the lengths of the atp8 gene were conspicuously shorter than that of all other known insect mitochondrial genomes (99 bp for BPH, 102 bp for SBPH). That two rearrangement regions (trnC-trnW and nad6-trnP-trnT) of mitochondrial genomes differing from other known insect were found in these two distantly related planthoppers revealed that the gene order of mitochondria might be conservative in Delphacidae. The large non-coding fragment (the A+T-rich region) putatively corresponding responsible for the control of replication and transcription of mitochondria contained a variable number of tandem repeats (VNTRs) block in different natural individuals of these two planthoppers. Comparison with a previously sequenced individual of SBPH revealed that the mitochondrial genetic variation within a species exists not only in the sequence and secondary structure of genes, but also in the gene order (the different location of trnH gene). CONCLUSION: The mitochondrial genome arrangement pattern found in planthoppers was involved in rearrangements of both tRNA genes and protein-coding genes (PCGs). Different species from different genera of Delphacidae possessing the same mitochondrial gene rearrangement suggests that gene rearrangements of mitochondrial genome probably occurred before the differentiation of this family. After comparatively analyzing the gene order of different species of Hemiptera, we propose that except for some specific taxonomical group (e.g. the whiteflies) the gene order might have diversified in family level of this order. The VNTRs detected in the control region might provide additional genetic markers for studying population genetics, individual difference and phylogeographics of planthoppers.

Diversity of Wolbachia in natural populations of spider mites (genus Tetranychus): evidence for complex infection history and disequilibrium distribution.

Wolbachia are endosymbiotic bacteria that commonly infect arthropods and cause reproductive disorders in host. Within several Tetranychus species, Wolbachia have been detected and shown to affect their reproduction. However, little is known about their transmission and distribution patterns in natural populations of Tetranychus species. Here, we used multilocus sequence typing to confirm Wolbachia infection status and examined the relationship between Wolbachia infection status and host phylogeny, mitochondrial diversity, and geographical range in five Tetranychus species (Tetranychus truncatus, Tetranychus urticae, Tetranychus pueraricola, Tetranychus phaselus, and Tetranychus kanzawai) from 21 populations in China. The prevalence of Wolbachia within the five Tetranychus species ranged from 31.4 to 100 %, and the strains were remarkably diverse. Together, these observations indicate that Wolbachia was introduced to these populations on multiple separate occasions. As in other arthropods, the same Tetranychus species can accommodate very different strains, and identical Wolbachia occasionally infect different species. These observations suggest that Wolbachia are transmitted both vertically and horizontally. Horizontally, transmission is probably mediated by the host plants. The distribution patterns of Wolbachia were quite different among populations of the same species, suggesting that the dynamics of Wolbachia in nature may be affected by ecological and other factors.

Wolbachia strengthens cardinium-induced cytoplasmic incompatibility in the spider mite Tetranychus piercei McGregor.

Wolbachia and Cardinium are maternally inherited intracellular bacteria that can manipulate the reproduction of their arthropod hosts, such as by inducing cytoplasmic incompatibility (CI). Although the reproductive alteration induced by Wolbachia or Cardinium have been well investigated, the effects of these two endosymbionts co-infecting the same host are poorly understood. We found that Tetranychus piercei McGregor is naturally infected with Wolbachia and Cardinium. We performed all possible crossing combinations using naturally infected and cured strains, and the results show that Wolbachia induced a weak level of CI, while Cardinium-infected and doubly infected males caused severe CI. Wolbachia and Cardinium could not rescue CI each other; however, Wolbachia boosted the expression of Cardinium-induced CI. Quantitative PCR results demonstrated that CI was associated with the infection density of Wolbachia and Cardinium.