Identification of odorant-binding proteins and functional analysis of antenna-specific BhorOBP28 in Batocera horsfieldi (Hope).

BACKGROUND: The important wood-boring pest Batocera horsfieldi has evolved a sensitive olfactory system to locate host plants. Odorant-binding proteins (OBPs) are thought to play key roles in olfactory recognition. Therefore, exploring the physiological function of OBPs could facilitate a better understanding of insect chemical communications. RESULTS: In this research, 36 BhorOBPs genes were identified via transcriptome sequencing of adults' antennae from B. horsfieldi, and most BhorOBPs were predominantly expressed in chemosensory body parts. Through fluorescence competitive binding and fluorescence quenching assays, the antenna-specific BhorOBP28 was investigated and displayed strong binding affinities forming stable complexes with five volatiles, including (+)-α-Pinene, (+)-Limonene, ß-Pinene, (-)-Limonene, and (+)-Longifolene, which could also elicit conformation changes when they were interacting with BhorOBP28. Batocera horsfieldi females exhibited a preference for (-)-Limonene, and a repellent response to (+)-Longifolene. Feeding dsOBP19 produced by a bacteria-expressed system with a newly constructed vector could lead to the knockdown of BhorOBP28, and could further impair B. horsfieldi attraction to (-)-Limonene and repellent activity of (+)-Longifolene. The analysis of site-directed mutagenesis revealed that Leu7, Leu72, and Phe121 play a vital role in selectively binding properties of BhorOBP28. CONCLUSION: By modeling the molecular mechanism of olfactory recognition, these results demonstrate that BhorOBP28 is involved in the chemoreception of B. horsfieldi. The bacterial-expressed dsRNA delivery system gains new insights into potential population management strategies. Through the olfactory process concluded that discovering novel behavioral regulation and environmentally friendly control options for B. horsfieldi in the future. © 2024 Society of Chemical Industry.

Biomechanical Comparison of Six Different Root-Analog Implants and the Conventional Morse Taper Implant by Finite Element Analysis.

Taper implants differ greatly from anatomical teeth in shape. In this study, seven three-dimensional finite element models were established, including a conventional taper implant and six root-analog implants with different root numbers and shapes. Vertical, horizontal, and oblique instantaneous loads of 100 N were applied to the models to obtain stress distribution in the implant, mucosa, cortical bone, and cancellous bone. ANSYS was used to perform the analysis under hypothetical experimental conditions. We find the stresses in all the implants and surrounding tissues varied by loading direction, the sequence of stress magnitude is vertical load, oblique load, and then horizontal load. The maximum stress values in root-analog implants were significantly less than in the taper implant. Moreover, stress distribution in the former was equalized contrary to the concentrated stress in the latter. Root-analog implants with different root geometry also revealed a pattern: stresses in multiple-root implant models were lower than those in single-root implants under the same load. The implant with a long and rounded root distributed the stress more uniformly, and it was mainly concentrated on the implant itself and cancellous bone. However, the opposite effect was observed in the short implant on mucosa and cortical bone. The root geometry of anatomical teeth can modify their functions. A uniform-shaped implant can hardly meet their functional requirements. Thus, the root-analog implant could be a possible solution.

Transcriptome analysis of genes associated with autolysis of Coprinus comatus.

Coprinus comatus, widely known as "Jituigu", is an important commodity and food in China. The yield of C. comatus, however, is substantially reduced by the autolysis of the fruiting bodies after harvest. To gain insight into the molecular mechanism underlying this autolysis, we divided the growth of C. comatus fruiting bodies into four stages: infant stage (I), mature stage (M), discolored stage (D), and autolysis stage (A). We then subjected these stages to de novo transcriptomic analysis using high-throughput Illumina sequencing. A total of 12,946 unigenes were annotated and analyzed with the Gene Ontology (GO), Clusters of Orthologous Groups of proteins (COG), and Kyoto Encyclopedia of Genes and Genomes (KEGG). We analyzed the differentially expressed genes (DEGs) between stages I and M, M and D, and D and A. Because the changes from M to D are thought to be related to autolysis, we focused on the DEGs between these two stages. We found that the pathways related to metabolic activity began to vary in the transition from M to D, including pathways named as autophagy-yeast, peroxisome, and starch and sucrose metabolism. This study also speculates the possible process of the autolysis of Coprinus comatus. In addition, 20 genes of interest were analyzed by quantitative real-time PCR to verify their expression profiles at the four developmental stages. This study, which is the first to describe the transcriptome of C. comatus, provides a foundation for future studies concerning the molecular basis of the autolysis of its fruiting bodies.

Characterization of the complete mitochondrial genome of Antheraea pernyi strain Luhong.

The present study for the first time describes the complete mitochondrial (mt) genome of Antheraea pernyi Guérin-Méneville 1855 strain Luhong, a genetic lethal mutant exhibiting especially red skin color. The mt genome is 15,563 bp in length that is the smallest among the sequenced A. pernyi inbred strains. This genome displays an identical genomic component and gene order to other six known A. pernyi mt genomes. The mt genome-based phylogenetic analysis clustered Luhong with four strains exhibiting yellow skin color, consistent with the traditional view that all of them belonged to the yellow blood lineage.

[Analysis of volatile compounds of Salix babylonica by GC-MS].

OBJECTIVE: To study the volatile compounds of Salix babylonica, and to provide scientific basis for efficacy of attracting natural enemies and medicinal value of Salix babylonica. METHODS: The volatile compounds were collected from Salix babylonica by dy- namic headspace absorption method, and analyzed by gas chromatography-mass spectrum( GC-MS). RESULTS: Forty-one volatile com- pounds were identified from Salix babylonica. Intact plant of Salix babylonica had thirty-three kinds of volatile compounds and aphid-in- fested plant of Salix babylonica had thirty-eight kinds of volatile compounds. The volatile components of intact plant and aphid-infested plant had significant differences in the composition and proportion. Among them,the volatile components of aphid-infested plant added six kinds of vinyl materials,one kind of aldehyde and one kind of ketone. CONCLUSION: Adding volatile components,which included vinyl materials,aldehydes and ketones,might have the efficacy of attracting natural enemies. The volatile components might play an important role in the ecological protection for Salix babylonica. The functional components from the volatile compounds had good medicinal value, which are worthy of further research.

[Analysis of volatile compounds of fresh tea leaves from yaoluoping nature preserve by SDE-GC-MS].

OBJECTIVE: To study the volatile compounds of fresh tea leaves from Yaoluoping Nature Preserve, and to provide scientific basis for the quality and medicinal value of tea from high mountainous area. METHODS: The volatile compounds were extracted from fresh tea leaves by simultaneous distillation and extraction, and analyzed by gas chromatography-mass spectrum (GC-MS). RESULTS: 41 volatile compounds were identified from two tea cultivars, Shifoxiang and Shifocui. Shifoxiang had 32 kinds of volatile compounds and Shifocui had 38 kinds of volatile compounds. The main volatile components of Shifoxiang and Shifocui were green leaf volatiles. The kind and relative content of terpenes of Shifocui were more than that of Shifoxiang. Also the kind and relative content of ketones of Shifocui were more than that of Shifoxiang. There was one kind of heterocyclic compound of Shifocui. CONCLUSION: The main volatile components of Shifoxiang and Shifocui had an important role for the formation of unique aroma of high mountain ous tea. The composition and proportion of volatile components between Shifoxiang and Shifocui had certain differences. Some trace components may be associated with defense in the process of tea plant growth. The functional components from the volatile components had good medicinal value, worthy of further research.

Molecular analysis and its expression of a pou homeobox protein gene during development and in response to salinity stress from brine shrimp, Artemia sinica.

Brine shrimps of the genus Artemia are aquatic species of economic importance because of their important significance to aquaculture and are used as a model species in physiology and developmental biology. Research on Artemia POU homeobox gene function will enhance our understanding of the physiological and developmental processes of POU homeobox gene in animals. Herein, a full-length cDNA encoding an Artemia POU homeobox protein gene 1 (APH-1) from Artemia sinica (designated as As-APH-1) was cloned and characterized by a reverse-transcription polymerase chain reaction (RT-PCR) and rapid amplification of cDNA end (RACE) method. The As-APH-1 gene encoded a protein of 388 amino acid polypeptide with a calculated molecular mass of 42.85kDa and an isoelectric point of 6.90 and the protein belongs to the POU III family. Multiple sequence alignments revealed that A. sinica As-APH-1 protein sequence shared a conserved POU homeobox domain with other species. The early and persistent expression of As-APH-1 in the naupliar stages by semi-quantitative RT-PCR and whole-mount embryonic immunohistochemistry suggest that As-APH-1 functions very early in the salt gland and may be required continuously in this organ. Later in development, expression of As-APH-1 begins to dramatically decrease and disappear in salt gland of the sub-adult Artemia. In addition, we also discovered that As-APH-1 increased obviously as the salinity increased, indicating that As-APH-1 might be used as a good indicator of salinity stress. In summary, we are the first to identify the As-APH-1 gene and to determine its gene expression patterns in early embryogenesis stages and in different salinity stress in brine shrimp, A. sinica. The result of expression of As-APH-1 affected by salinity changes will provide us further understanding of the underlying mechanisms of osmoregulation in Artemia early embryonic development.

Molecular cloning and its expression of trachealess gene (As-trh) during development in brine shrimp, Artemia sinica.

Basic helix-loop-helix-PAS (bHLH-PAS) family transcription factors are implicated in multiple developmental and physiological regulatory processes. Herein, a full-length cDNA encoding a bHLH-PAS domain transcription factor trachealess gene (designated as As-trh) was cloned and characterized from brine shrimp (Artemia sinica) for the first time. The full-length cDNA of As-trh was 2,698 bp with a 2,319 bp open reading frame encoding a deduced protein of 772 amino acid polypeptide with a calculated molecular mass of 86.02 kDa and an isoelectric point of 5.87. Sequence alignment revealed that As-trh had high homology with other species trh gene, including the D-trh gene in Drosophila melanogaster and Bm-trh in Bombyx mori. The early and persistent expression of As-trh in the naupliar stages by whole-mount embryonic in situ hybridization and immunohistochemistry suggest that As-trh functions very early in the salt gland and may be required continuously in this tissue. Later in development, expression of As-trh begins to decrease and disappear in salt gland of the older nauplius and appears in the thoracic epipods of the sub-adult Artemia. These results indicated that As-trh might play an important role in osmoregulatiory organ development from the larvae stages through adult stages.

Cloning and characterization of p8 homolog cDNA in the Atlantic halibut (Hippoglossus hippoglossus).

The p8 gene encodes a transcription factor known to modulate cell growth, division, and apoptosis and influences gene expression. In this study, an Atlantic halibut (Hippoglossus hippoglossus) homolog of the p8 gene was cloned, sequenced, and characterized. The full-length p8 cDNA consists of 601 bp and encodes 76 amino acids with a molecular mass of 9 kD. The bHLH region is well conserved between Atlantic halibut and other animals. Analysis by RT-PCR showed that the p8 transcript is constitutively expressed in 9 of the 12 tissues tested: pancreas, intestine, stomach, gill, head kidney, heart, liver, ovary, and spleen. A predicted microRNA target site was found in the 3'UTR of Atlantic halibut p8 mRNA. We speculate that the target site may pair to microRNA molecules because the target site resides in a big loop, a space large enough for the binding of microRNA molecules.

In silico cloning and characterization of p8 homolog cDNA from common urchin (Paracentrotus lividus).

The p8 protein is a transcription factor with a basic helix-loop-helix motif and a nuclear localization signal. In the present study, a common sea urchin (Paracentrotus lividus) homolog of p8 cDNA was cloned, sequenced and characterized. The full-length p8 cDNA consists of 896 bp and encodes 71 amino acids with a molecular mass of 8.238 kD. Homology alignments found that several phosphorylation sites and the most negative (Asp and Glu)/positive (Arg) propensities charged residues were well conserved between P. lividus and other species. Analysis by RT-PCR showed that there was no obvious difference in the patterns of expression during embryogenesis from unfertilization egg up to the pluteus stage, but p8 mRNA expression levels varied among tested adult tissues suggesting that p8 has a key function in embryogenesis but non-vital function under physiological conditions. Evolution relationships between P. lividus p8 and other species p8 homologs revealed in the phylogenetic tree were in agreement with the concept of traditional taxonomy.