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The development of fungal fruiting bodies from a hyphal thallus is inducible under low temperature (cold stress). The molecular mechanism has been subject to surprisingly few studies. Analysis of gene expression level has become an important means to study gene function and its regulation mechanism. But identification of reference genes (RGs) stability under cold stress have not been reported in famous medicinal mushroom-forming fungi Cordyceps militaris. Herein, 12 candidate RGs had been systematically validated under cold stress in C. militaris. Three different algorithms, geNorm, NormFinder and BestKeeper were applied to evaluate the expression stability of the RGs. Our results showed that UBC and UBQ were the most stable RGs for cold treatments in short and long periods, respectively. 2 RGs (UBC and PP2A) and 3 RGs (UBQ, TUB and CYP) were the suitable RGs for cold treatments in short and long periods, respectively. Moreover, target genes, two-component-system histidine kinase genes, were selected to validate the most and least stable RGs under cold treatment, which indicated that use of unstable expressed genes as RGs leads to biased results. Our results provide a good starting point for accurate reverse transcriptase quantitative polymerase chain reaction normalization by using UBC and UBQ in C. militaris under cold stress and better support for understanding the mechanism of response to cold stress and fruiting body formation in C. militaris and other mushroom-forming fungi in future research.

Cordyceps militaris is a famous traditional edible and medicinal fungus in Asia, and its fruiting body has rich medicinal value. The molecular mechanism of fruiting body development is still not well understood in C. militaris. In this study, phylogenetically analysis and protein domains prediction of the 14 putative chitinases were performed. The transcription level and enzyme activity of chitinase were significant increased during fruiting body development of C. militaris. Then, two chitinase genes (Chi1 and Chi4) were selected to construct gene silencing strain by RNA interference. When Chi1 and Chi4 genes were knockdown, the differentiation of the primordium was blocked, and the number of fruiting body was significantly decreased approximately by 50% compared to wild-type (WT) strain. The length of the single mature fruiting body was shortened by 27% and 38% in Chi1- and Chi4-silenced strains, respectively. In addition, the chitin content and cell wall thickness were significantly increased in Chi1- and Chi4-silenced strains. These results provide new insights into the biological functions of chitinase in fruiting body development of C. militaris.

Ganoderic acids (GAs) are well recognized as important pharmacological components of the medicinal species belonging to the basidiomycete genus Ganoderma . However, transcription factors directly regulating the expression of GA biosynthesis genes remain poorly understood. Here, the genome of Ganoderma lingzhi is de novo sequenced. Using DNA affinity purification sequencing, we identify putative targets of the transcription factor sterol regulatory element-binding protein (SREBP), including the genes of triterpenoid synthesis and lipid metabolism. Interactions between SREBP and the targets are verified by electrophoretic mobility gel shift assay. RNA-seq shows that SREBP targets, mevalonate kinase and 3-hydroxy-3-methylglutaryl coenzyme A synthetase in mevalonate pathway, sterol isomerase and lanosterol 14-demethylase in ergosterol biosynthesis, are significantly upregulated in the SREBP overexpression (OE::SREBP) strain. In addition, 3 targets involved in glycerophospholipid/glycerolipid metabolism are upregulated. Then, the contents of mevalonic acid, lanosterol, ergosterol and 13 different GAs as well as a variety of lipids are significantly increased in this strain. Furthermore, the effects of SREBP overexpression on triterpenoid and lipid metabolisms are recovered when OE::SREBP strain are treated with exogenous fatostatin, a specific inhibitor of SREBP. Taken together, our genome-wide study clarify the role of SREBP in triterpenoid and lipid metabolisms of G. lingzhi . An integration of genomic, DAP-seq, RNA-seq, metabolomic and genetic approaches identifies SREBP as a key transcription factor regulating triterpenoid and lipid metabolisms in the medicinal fungus Ganoderma lingzhi .

Triterpenoids are well-known pharmacological components of Sanghuangporus fungi, such as Sanghuangporus lonicericola . This study investigates the inductive effects of paclobutrazol (PBZ) on triterpenoid biosynthesis in the submerged fermentation of S. lonicericola and explores the induction mechanisms via multi-omics and genetic methods. The addition of 100 mg/L PBZ significantly increases the triterpenoid yield by 151.39%. A total of 29 triterpenoids are tentatively identified, of which 18 are newly presented only under PBZ induction. Moreover, 30 genes involved in the MVA pathway and 31 genes encoding cytochrome P450 monooxygenases assumed to be responsible for decoration are identified. Finally, a MYB transcription factor (SlMYB) is identified and found to be downregulated under paclobutrazol induction. Genetic manipulation of SlMYB demonstrates its negative regulatory effect on four putative target genes, including ACAT , MVD , IDI , and FDPS . Electrophoretic mobility gel shift assays verify the direct interactions with the promoters of MVD , IDI , and FDPS . Taken together, PBZ acts as an effective inducer of triterpenoid biosynthesis in S. lonicericola , and the transcription factor SlMYB is negatively regulated. Mechanistic exploration of paclobutrazol induction reveals the role of the MYB transcription factor SlMYB and its direct regulatory interactions with key genes in the MVA pathway, shedding light on the transcriptional regulation underlying enhanced triterpenoid biosynthesis in Sanghuangporus lonicericola .

Cordyceps militaris is a famous traditional edible and medicinal fungus in Asia, and its fruiting body has rich medicinal value. The molecular mechanism of fruiting body development is still not well understood in C. militaris. In this study, phylogenetically analysis and protein domains prediction of the 14 putative chitinases were performed. The transcription level and enzyme activity of chitinase were significant increased during fruiting body development of C. militaris. Then, two chitinase genes (Chi1 and Chi4) were selected to construct gene silencing strain by RNA interference. When Chi1 and Chi4 genes were knockdown, the differentiation of the primordium was blocked, and the number of fruiting body was significantly decreased approximately by 50% compared to wild-type (WT) strain. The length of the single mature fruiting body was shortened by 27% and 38% in Chi1- and Chi4-silenced strains, respectively. In addition, the chitin content and cell wall thickness were significantly increased in Chi1- and Chi4-silenced strains. These results provide new insights into the biological functions of chitinase in fruiting body development of C. militaris.

To the Editor： Endoscopic submucosal dissection （ESD） has been widely applied in clinical practice for resecting gastrointestinal mucosal lesions. However, the risk of post-ESD complications is relatively high, and the bleeding is one of the most common complications after ESD, which needs timely detection and treatment. At recent, several new endoscopic imaging techniques have been developed to improve the diagnostic efficiency. Linked color imaging （LCI）, which is a newly developed endoscopic technique, could enhance the color contrast and thus have advantage of identifying bleeding points during ESD. So far, whether the application of LCI in ESD procedure is feasible and sate has not been ever explored. In this report, we described a patient with rectal neuroendocrine tumor who was successfully treated by ESD using LCI.

Aim： The aim of the present study was to further improve the therapeutic effects for human hepatocellular carcinoma （HCC） and reduce the damage in normal cells using a novel chemo-gene-virotherapeutic strategy. Methods： An oncolytic adenoviral vector （ZD55） similar to the typical oncolytic adenovirus ONYX-015, with a deletion of E1B-55K gene, was employed to express the second mitochon- dria-derived activator of caspases （Smac） protein by constructing a recombinant virus ZD55-Smac. The enhanced cytotoxicity of the combined treatment of ZD55- Smac with cisplatin or 5-fluorouracil （5-FU） was evaluated in several HCC cell lines. Moreover, the negative effects on normal cells have been tested in human normal liver cell lines L-02 and QSG-7701 cell lines by 3-（4,5-dimethylthiazol-2-yl）- 2,5-diphenyltetrazolium bromide assay and apoptotic cell staining. Results： According to our observation, ZD55-Smac is superior to ONYX-015 in sensitizing chemotherapy, ZD55-Smac used in conjunction with chemotherapy was found to exhibit obviously enhanced cytotoxicity in HCC cells, yet significantly abolished the negative toxicity in normal cells by utilizing the tumor selective replication vector and reducing the dosage. Conclusion： This chemo-gene-virotherapeutic （cisplatin or 5-FU＋ZD55-Smac） strategy is superior to the conventional chemo- gene or chemo-viro approach.

This study aims to optimize the node deployment of underwater wireless sensor networks (UWSNs) using intelligent optimization algorithms and robot collaboration technology to enhance network performance and coverage. The study employs the chemical reaction optimization (CRO) algorithm, which combines the advantages of genetic algorithms, simulated annealing algorithms, and ant colony algorithms. The CRO algorithm is enhanced through a structure correction function to determine the optimal node deployment scheme to achieve effective and optimal coverage control of the UWSN. Additionally, the flexibility and autonomy of robots are leveraged to improve the efficiency of node deployment and address the unique challenges posed by the underwater environment. Furthermore, the study conducts a comparative analysis of different intelligent optimization algorithms and demonstrates the effectiveness and advantages of the enhanced CRO algorithm in optimizing node deployment for UWSNs. The study findings reveal that the improved algorithm achieves an average coverage rate of 95.66%, significantly outperforming traditional intelligent optimization algorithms. The coverage of UWSNs can be significantly improved by utilizing the enhanced CRO algorithm and robot collaboration technology for node deployment optimization, which offers an effective approach for achieving optimal node deployment. Moreover, the rational deployment of nodes enhances the monitoring capability, resource utilization efficiency, and accuracy of environmental monitoring in underwater networks. The results of this study hold great practical significance for underwater environment monitoring, marine resource exploration, and marine scientific research.

The MYB gene family plays a vital regulatory role in plant metabolism, stress response, and floral color. The R2R3-MYB gene family of C. goeringii was identified, and its expression was analyzed using bioinformatics in this article. The R2R3-MYB genes of Arabidopsis thaliana were used as a reference to determine 104 CgMYB genes and categorize them into 22 subfamilies. Exon/intron organizations and conserved motif analysis revealed that the majority of CgMYB genes were highly conserved, and chromosome localization and collinearity analysis provided evidence of tandem duplication and segmental duplication events, indicating the phenomenon of gene family expansion and contraction. The function of CgMYB genes was analyzed by cis -acting element and gene ontology (GO) enrichment. In addition, we selected CgMYB91 and CgMYB32 for RT–qPCR, suggesting that CgMYB91 and CgMYB32 are associated with anthocyanin formation. In short, this study provides a comprehensive and specific function of the R2R3-MYB transcription factors (TFs) in orchids.

To tackle the challenges of extensive data transmission and high redundancy in wireless sensor networks (WSNs), this study proposes a novel data collection scheme based on expected network coverage and clustered compressive sensing (CS). First, the K-medoids clustering algorithm organizes nodes within the WSN coverage area into clusters. Combined with an optimized network coverage algorithm, a node scheduling strategy is introduced to focus on critical observation areas while minimizing overall energy consumption. Next, by analyzing the relationship between network clustering and node deployment, a weakly correlated observation matrix is designed to mitigate the impact of packet loss on data collection. Finally, the sparrow search algorithm is employed to enhance the accuracy of CS data reconstruction at the cluster head. Simulation results demonstrate that, compared to existing data collection schemes, the proposed approach significantly reduces WSN transmission overhead, ensures accurate recovery of raw data, decreases data reconstruction error, and extends network lifetime.