1.Omics for deciphering oral microecology.
Yongwang LIN ; Xiaoyue LIANG ; Zhengyi LI ; Tao GONG ; Biao REN ; Yuqing LI ; Xian PENG
International Journal of Oral Science 2024;16(1):2-2
		                        		
		                        			
		                        			The human oral microbiome harbors one of the most diverse microbial communities in the human body, playing critical roles in oral and systemic health. Recent technological innovations are propelling the characterization and manipulation of oral microbiota. High-throughput sequencing enables comprehensive taxonomic and functional profiling of oral microbiomes. New long-read platforms improve genome assembly from complex samples. Single-cell genomics provides insights into uncultured taxa. Advanced imaging modalities including fluorescence, mass spectrometry, and Raman spectroscopy have enabled the visualization of the spatial organization and interactions of oral microbes with increasing resolution. Fluorescence techniques link phylogenetic identity with localization. Mass spectrometry imaging reveals metabolic niches and activities while Raman spectroscopy generates rapid biomolecular fingerprints for classification. Culturomics facilitates the isolation and cultivation of novel fastidious oral taxa using high-throughput approaches. Ongoing integration of these technologies holds the promise of transforming our understanding of oral microbiome assembly, gene expression, metabolites, microenvironments, virulence mechanisms, and microbe-host interfaces in the context of health and disease. However, significant knowledge gaps persist regarding community origins, developmental trajectories, homeostasis versus dysbiosis triggers, functional biomarkers, and strategies to deliberately reshape the oral microbiome for therapeutic benefit. The convergence of sequencing, imaging, cultureomics, synthetic systems, and biomimetic models will provide unprecedented insights into the oral microbiome and offer opportunities to predict, prevent, diagnose, and treat associated oral diseases.
		                        		
		                        		
		                        		
		                        			Humans
		                        			;
		                        		
		                        			Phylogeny
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		                        			Biomimetics
		                        			;
		                        		
		                        			Dysbiosis
		                        			;
		                        		
		                        			Homeostasis
		                        			;
		                        		
		                        			Mass Spectrometry
		                        			
		                        		
		                        	
2.Identification and expression analysis of citrate synthase 3 gene family members in apple.
Xinrui LI ; Wenfang LI ; Jiaxing HUO ; Long LI ; Baihong CHEN ; Zhigang GUO ; Zonghuan MA
Chinese Journal of Biotechnology 2024;40(1):137-149
		                        		
		                        			
		                        			As one of the key enzymes in cell metabolism, the activity of citrate synthase 3 (CS3) regulates the substance and energy metabolism of organisms. The protein members of CS3 family were identified from the whole genome of apple, and bioinformatics analysis was performed and expression patterns were analyzed to provide a theoretical basis for studying the potential function of CS3 gene in apple. BLASTp was used to identify members of the apple CS3 family based on the GDR database, and the basic information of CS3 protein sequence, subcellular localization, domain composition, phylogenetic relationship and chromosome localization were analyzed by Pfam, SMART, MEGA5.0, clustalx.exe, ExPASy Proteomics Server, MEGAX, SOPMA, MEME, WoLF PSORT and other software. The tissue expression and inducible expression characteristics of 6 CS3 genes in apple were determined by acid content and real-time fluorescence quantitative polymerase chain reaction (qRT-PCR). Apple CS3 gene family contains 6 members, and these CS3 proteins contain 473-608 amino acid residues, with isoelectric point distribution between 7.21 and 8.82. Subcellular localization results showed that CS3 protein was located in mitochondria and chloroplasts, respectively. Phylogenetic analysis divided them into 3 categories, and the number of genes in each subfamily was 2. Chromosome localization analysis showed that CS3 gene was distributed on different chromosomes of apple. The secondary structure of protein is mainly α-helix, followed by random curling, and the proportion of β-angle is the smallest. The 6 members were all expressed in different apple tissues. The overall expression trend from high to low was the highest relative expression content of MdCS3.4, followed by MdCS3.6, and the relative expression level of other members was in the order of MdCS3.3 > MdCS3.2 > MdCS3.1 > MdCS3.5. qRT-PCR results showed that MdCS3.1 and MdCS3.3 genes had the highest relative expression in the pulp of 'Chengji No. 1' with low acid content, and MdCS3.2 and MdCS3.3 genes in the pulp of 'Asda' with higher acid content had the highest relative expression. Therefore, in this study, the relative expression of CS3 gene in apple cultivars with different acid content in different apple varieties was detected, and its role in apple fruit acid synthesis was analyzed. The experimental results showed that the relative expression of CS3 gene in different apple varieties was different, which provided a reference for the subsequent study of the quality formation mechanism of apple.
		                        		
		                        		
		                        		
		                        			Citric Acid
		                        			;
		                        		
		                        			Malus/genetics*
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		                        			Citrate (si)-Synthase
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		                        			Phylogeny
		                        			;
		                        		
		                        			Citrates
		                        			
		                        		
		                        	
3.Identification of banana ADA1 gene family members and their expression profiles under biotic and abiotic stresses.
Qiqi ZHAO ; Wenhui REN ; Huifei ZHU ; Qiuzhen WU ; Chunyu ZHANG ; Xiaoqiong XU ; Binbin LUO ; Yuji HUANG ; Yukun CHEN ; Yuling LIN ; Zhongxiong LAI
Chinese Journal of Biotechnology 2024;40(1):190-210
		                        		
		                        			
		                        			The Spt-Ada-Gcn5-acetyltransferase (SAGA) is an ancillary transcription initiation complex which is highly conserved. The ADA1 (alteration/deficiency in activation 1, also called histone H2A functional interactor 1, HFI1) is a subunit in the core module of the SAGA protein complex. ADA1 plays an important role in plant growth and development as well as stress resistance. In this paper, we performed genome-wide identification of banana ADA1 gene family members based on banana genomic data, and analyzed the basic physicochemical properties, evolutionary relationships, selection pressure, promoter cis-acting elements, and its expression profiles under biotic and abiotic stresses. The results showed that there were 10, 6, and 7 family members in Musa acuminata, Musa balbisiana and Musa itinerans. The members were all unstable and hydrophilic proteins, and only contained the conservative SAGA-Tad1 domain. Both MaADA1 and MbADA1 have interactive relationship with Sgf11 (SAGA-associated factor 11) of core module in SAGA. Phylogenetic analysis revealed that banana ADA1 gene family members could be divided into 3 classes. The evolution of ADA1 gene family members was mostly influenced by purifying selection. There were large differences among the gene structure of banana ADA1 gene family members. ADA1 gene family members contained plenty of hormonal elements. MaADA1-1 may play a prominent role in the resistance of banana to cold stress, while MaADA1 may respond to the Panama disease of banana. In conclusion, this study suggested ADA1 gene family members are highly conserved in banana, and may respond to biotic and abiotic stress.
		                        		
		                        		
		                        		
		                        			Musa/genetics*
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		                        			Phylogeny
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		                        			Fungal Proteins
		                        			;
		                        		
		                        			Cell Nucleus
		                        			;
		                        		
		                        			Histones
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		                        			Stress, Physiological/genetics*
		                        			
		                        		
		                        	
4.Screening and evaluation of the biocontrol efficacy of a Trichoderma brevicompactum strain and its metabolite trichodermin against banana Fusarium wilt.
Xiajun YAO ; Jin XIE ; Yanhua QI ; Bin WANG ; Wenxia FANG ; Gang TAO ; Xiliang JIANG
Chinese Journal of Biotechnology 2024;40(1):211-225
		                        		
		                        			
		                        			The banana Fusarium wilt (BFW) caused by Fusarium oxysporum f. sp. cubense tropical race4 (FocTR4) is difficult to control worldwide, which causes a huge economic losse to banana industry. The purpose of this study was to screen Trichoderma strains with antagonistic activity against FocTR4, to isolate and purify the active compound from the fermentation broth, so as to provide important biocontrol strains and active compound resources. In this work, Trichoderma strains were isolated and screened from the rhizosphere soil of crops, and the strains capable of efficiently inhibiting FocTR4 were screened by plate confrontation, and further confirmed by testing inhibition for the conidial germination and mycelial growth of FocTR4. The phylogenetic tree clarified the taxonomic status of the biocontrol strains. Moreover, the active components in the fermentation broth of the strains were separated and purified by column chromatography, the structure of the most active component was analyzed by nuclear magnetic resonance spectroscopy (NMR), the BFW control effect was tested by pot experiments. We obtained a strain JSHA-CD-1003 with antagonistic activity against FocTR4, and the inhibition rate from plate confrontation was 60.6%. The fermentation broth of JSHA-CD-1003 completely inhibited the germination of FocTR4 conidia within 24 hours. The inhibition rate of FocTR4 hyphae growth was 52.6% within 7 d. A phylogenetic tree was constructed based on the ITS and tef1-α gene tandem sequences, and JSHA-CD-1003 was identified as Trichoderma brevicompactum. Purification and NMR identification showed that the single active compound was trichodermin, and the minimum inhibitory concentration (MIC) was 25 μg/mL. Pot experiments showed that the fermentation broth of strain JSHA-CD-1003 was effective against BFW. The control rate of leaf yellowing was 47.4%, and the rate of bulb browning was 52.0%. Therefore, JSHA-CD-1003 effectively inhibited FocTR4 conidial germination and mycelium growth through producing trichodermin, and showed biocontrol effect on banana wilt caused by FocTR4, thus is a potential biocontrol strain.
		                        		
		                        		
		                        		
		                        			Fusarium
		                        			;
		                        		
		                        			Musa
		                        			;
		                        		
		                        			Phylogeny
		                        			;
		                        		
		                        			Trichodermin
		                        			;
		                        		
		                        			Hypocreales
		                        			
		                        		
		                        	
5.Identification and expression analysis of TCP family members in tobacco (Nicotiana tabacum L.).
Shize WANG ; Yun LI ; Yucui HAN ; Shizhou YU ; Shuang WANG ; Yong LIU ; Xiaohu LIN
Chinese Journal of Biotechnology 2024;40(1):226-238
		                        		
		                        			
		                        			TCP family as plant specific transcription factor, plays an important role in different aspects of plant development. In order to screen TCP family members in tobacco, the homologous sequences of tobacco and Arabidopsis TCP family were identified by genome-wide homologous alignment. The physicochemical properties, phylogenetic relationships and cis-acting elements were analyzed by bioinformatics. The homologous genes of AtTCP3/AtTCP4 were screened, and RT-qPCR was used to detect the changes of gene expression upon 20% PEG6000 treatment. The results show that tobacco contains 63 TCP family members. Their amino acid sequence length ranged from 89 aa to 596 aa, and their protein hydropathicity grand average of hydropathicity (GRAVY) ranged from -1.147 to 0.125. The isoelectric point (pI) ranges from 4.42 to 9.94, the number of introns is 0 to 3, and the subcellular location is all located in the nucleus. The results of conserved domain and phylogenetic relationship analysis showed that the tobacco TCP family can be divided into PCF, CIN and CYC/TB1 subfamilies, and each subfamily has a stable sequence. The results of cis-acting elements in gene promoter region showed that TCP family genes contain low docile acting elements (LTR) and a variety of stress and metabolic regulation related elements (MYB, MYC). Analysis of gene expression patterns showed that AtTCP3/AtTCP4 homologous genes (NtTCP6, NtTCP28, NtTCP30, NtTCP33, NtTCP42, NtTCP57, NtTCP63) accounted for 20% PEG6000 treatment significantly up-regulated/down-regulated expression, and NtTCP30 and NtTCP57 genes were selected as candidate genes in response to drought. The results of this study analyzed the TCP family in the tobacco genome and provided candidate genes for the study of drought-resistance gene function and variety breeding in tobacco.
		                        		
		                        		
		                        		
		                        			Nicotiana/genetics*
		                        			;
		                        		
		                        			Phylogeny
		                        			;
		                        		
		                        			Plant Breeding
		                        			;
		                        		
		                        			Amino Acid Sequence
		                        			;
		                        		
		                        			Arabidopsis
		                        			;
		                        		
		                        			Polyethylene Glycols
		                        			
		                        		
		                        	
6.Characteristics of the chloroplast genome of Camellia insularis.
Jin ZHANG ; Yongbiao DENG ; Bo ZHAO
Chinese Journal of Biotechnology 2024;40(1):280-291
		                        		
		                        			
		                        			In this study, the chloroplast genome of Camellia insularis Orel & Curry was sequenced using high-throughput sequencing technology. The results showed that the chloroplast genome of C. insularis was 156 882 bp in length with a typical tetrad structure, encoding 132 genes, including 88 protein-coding genes, 36 tRNA genes, and 8 rRNA genes. Codon preference analysis revealed that the highest number of codons coded for leucine, with a high A/U preference in the third codon position. Additionally, 67 simple sequence repeats (SSR) loci were identified, with a preference for A and T bases. The inverted repeat (IR) boundary regions of the chloroplast genome of C. insularis were relatively conserved, except for a few variable regions. Phylogenetic analysis indicated that C. insularis was most closely related to C. fascicularis. Yellow camellia is a valuable material for genetic engineering breeding. This study provides fundamental genetic information on chloroplast engineering and offers valuable resources for conducting in-depth research on the evolution, species identification, and genomic breeding of yellow Camellia.
		                        		
		                        		
		                        		
		                        			Genome, Chloroplast/genetics*
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		                        			Phylogeny
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		                        			Plant Breeding
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		                        			Camellia/genetics*
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		                        			Chloroplasts/genetics*
		                        			
		                        		
		                        	
7.HIV-1 Transmission among Injecting Drug Users is Principally Derived from Local Circulating Strains in Guangxi, China.
Ping CEN ; Hua Yue LIANG ; Yuan YANG ; Fei ZHANG ; Shi Xiong YANG ; Ju Cong MO ; Yi FENG ; Jie Gang HUANG ; Chuan Yi NING ; Chun Yuan HUANG ; Yao YANG ; Na LIANG ; Bing Yu LIANG ; Li YE ; Hao LIANG
Biomedical and Environmental Sciences 2023;36(5):418-430
		                        		
		                        			OBJECTIVE:
		                        			The mode of human immunodeficiency virus (HIV) transmission via injection drug use (IDU) still exists, and the recent shift in IDU-related transmission of HIV infection is largely unknown. The purpose of this study was to analyze the spatiotemporal sources and dynamics of HIV-1 transmission through IDU in Guangxi.
		                        		
		                        			METHODS:
		                        			We performed a molecular epidemiological investigation of infections across Guangxi from 2009 to 2019. Phylogenetic and Bayesian time-geographic analyses of HIV-1 sequences were performed to confirm the characteristics of transmission between IDUs in combination with epidemiological data.
		                        		
		                        			RESULTS:
		                        			Among the 535 subjects, CRF08_BC (57.4%), CRF01_AE (28.4%), and CRF07_BC (10.7%) were the top 3 HIV strains; 72.6% of infections were linked to other provinces in the transmission network; 93.6% of sequence-transmitted strains were locally endemic, with the rest coming from other provinces, predominantly Guangdong and Yunnan; 92.1% of the HIV transmission among people who inject drugs tended to be transmitted between HIV-positive IDUs.
		                        		
		                        			CONCLUSION
		                        			HIV recombinants were high diversity, and circulating local strains were the transmission sources among IDUs in Guangxi. However, there were still cases of IDUs linked to other provinces. Coverage of traditional prevention strategies should be expanded, and inter-provincial collaboration between Guangxi, Yunnan, and Guangdong provinces should be strengthened.
		                        		
		                        		
		                        		
		                        			Humans
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		                        			HIV-1/genetics*
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		                        			HIV Infections
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		                        			Drug Users
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		                        			Phylogeny
		                        			;
		                        		
		                        			Bayes Theorem
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		                        			China/epidemiology*
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		                        			Genotype
		                        			
		                        		
		                        	
8.Metagenomic Analysis of Environmental Samples from Wildlife Rescue Station at Poyang Lake, China.
Jia LIU ; Xi Yan LI ; Wen Tao SONG ; Xiao Xu ZENG ; Hui LI ; Lei YANG ; Da Yan WANG
Biomedical and Environmental Sciences 2023;36(7):595-603
		                        		
		                        			OBJECTIVE:
		                        			To improve the understanding of the virome and bacterial microbiome in the wildlife rescue station of Poyang Lake, China.
		                        		
		                        			METHODS:
		                        			Ten smear samples were collected in March 2019. Metagenomic sequencing was performed to delineate bacterial and viral diversity. Taxonomic analysis was performed using the Kraken2 and Bracken methods. A maximum-likelihood tree was constructed based on the RNA-dependent RNA polymerase (RdRp) region of picornavirus.
		                        		
		                        			RESULTS:
		                        			We identified 363 bacterial and 6 viral families. A significant difference in microbial and viral abundance was found between samples S01-S09 and S10. In S01-S09, members of Flavobacteriia and Gammaproteobacteria were the most prevalent, while in S10, the most prevalent bacteria class was Actinomycetia. Among S01-S09, members of Myoviridae and Herelleviridae were the most prevalent, while the dominant virus family of S10 was Picornaviridae. The full genome of the pigeon mesivirus-like virus (NC-BM-233) was recovered from S10 and contained an open reading frame of 8,124 nt. It showed the best hit to the pigeon mesivirus 2 polyprotein, with 84.10% amino acid identity. Phylogenetic analysis showed that RdRp clustered into Megrivirus B.
		                        		
		                        			CONCLUSION
		                        			This study provides an initial assessment of the bacteria and viruses in the cage-smeared samples, broadens our knowledge of viral and bacterial diversity, and is a way to discover potential pathogens in wild birds.
		                        		
		                        		
		                        		
		                        			Animals
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		                        			Animals, Wild/genetics*
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		                        			Lakes
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		                        			Phylogeny
		                        			;
		                        		
		                        			Picornaviridae/genetics*
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		                        			Viruses/genetics*
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		                        			China
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		                        			Metagenomics
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		                        			Genome, Viral
		                        			
		                        		
		                        	
9.Identification of heat shock protein hsp70 family genes from Rana amurensis and its expression profiles upon infection.
Tingting LIU ; Jingjing GUO ; Zhaodong CHEN ; Yufen LIU ; Legang JING ; Peng LIU ; Wenge ZHAO
Chinese Journal of Biotechnology 2023;39(4):1710-1730
		                        		
		                        			
		                        			Heat shock proteins (HSPs) widely exist in all organisms, the structures of which are usually extraordinarily conservative. They are also well-known stress proteins that are involved in response to physical, chemical and biological stresses. HSP70 is an important member of the HSPs family. In order to study the roles of amphibians HSP70 during infection, the cDNA sequence of Rana amurensis hsp70 family genes were cloned by homologous cloning method. The sequence characteristics, three-dimensional structure and genetic relationship of Ra-hsp70s were analyzed by bioinformatics methods. The expression profiles under bacterial infection were also analyzed by real-time quantitative PCR (qRT-PCR). Expression and localization of HSP70 protein were tested by immunohistochemical techniques. The results showed that three conservative tag sequences of HSP70 family, HSPA5, HSPA8 and HSPA13, were found in HSP70. Phylogenetic tree analysis indicated four members are distributed in four different branches, and members with the same subcellular localization motif are distributed in the same branch. The relative expression levels of the mRNA of four members were all significantly upregulated (P < 0.01) upon infection, but the time for up-regulating the expression levels were diverse in different tissues. The immunohistochemical analysis showed that HSP70 was expressed to different degrees in the cytoplasm of liver, kidney, skin and stomach tissue. The four members of Ra-hsp70 family have ability to respond bacterial infection to varying degrees. Therefore, it was proposed that they are involved in biological processes against pathogen and play different biological functions. The study provides a theoretical basis for functional studies of HSP70 gene in amphibians.
		                        		
		                        		
		                        		
		                        			Heat-Shock Proteins/genetics*
		                        			;
		                        		
		                        			Phylogeny
		                        			;
		                        		
		                        			Amino Acid Sequence
		                        			;
		                        		
		                        			HSP70 Heat-Shock Proteins/metabolism*
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		                        			Stress, Physiological
		                        			
		                        		
		                        	
10.Cloning of adipor1 and adipor2 genes in Rana dybowskii and its expression pattern upon infection.
Dongmei XU ; Yiming LIU ; Yugang SUN ; Yufen LIU ; Peng LIU ; Wenge ZHAO
Chinese Journal of Biotechnology 2023;39(4):1731-1746
		                        		
		                        			
		                        			Adiponectin receptor 1 (AdipoR1) and Adiponectin receptor 2 (AdipoR2) can bind to adiponectin (AdipoQ) secreted by adipose tissue to participate in various physiological functions of the body. In order to explore the role of AdipoR1 and AdipoR2 in amphibians infected by Aeromonas hydrophila (Ah), the genes adipor1 and adipor2 of Rana dybowskii were cloned by reverse transcription-polymerase chain reaction (RT-PCR) and analyzed by bioinformatics. The tissue expression difference of adipor1 and adipor2 was analyzed by real-time fluorescence quantitative polymerase chain reaction (qRT-PCR), and an inflammatory model of R. dybowskii infected by Ah was constructed. The histopathological changes were observed by hematoxylin-eosin staining (HE staining); the expression profiles of adipor1 and adipor2 after infection were dynamically detected by qRT-PCR and Western blotting. The results show that AdipoR1 and AdipoR2 are cell membrane proteins with seven transmembrane domains. Phylogenetic tree also shows that AdipoR1 and AdipoR2 cluster with the amphibians in the same branch. qRT-PCR and Western blotting results show that adipor1 and adipor2 were up-regulated at different levels of transcription and translation upon Ah infection, but the response time and level were different. It is speculated that AdipoR1 and AdipoR2 participate in the process of bacterial immune response, providing a basis for further exploring the biological functions of AdipoR1 and AdipoR2 in amphibians.
		                        		
		                        		
		                        		
		                        			Animals
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		                        			Receptors, Adiponectin/metabolism*
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		                        			Phylogeny
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		                        			Adiponectin/metabolism*
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		                        			Cloning, Molecular
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		                        			Ranidae/genetics*
		                        			
		                        		
		                        	
            
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