Colonization study of gfp-tagged Achromobacter marplatensis strain in sugar beet

YingWu Shi,Chun Li,HongMei Yang,Tao Zhang,Yan Gao,Min Chu,Jun Zeng,Qing Lin,OuTiKuEr,YuGuo Li,Xiangdong Huo,Kai Lou 한국미생물학회 2017 The journal of microbiology Vol.55 No.4

This study details the introduction of a gfp marker into anendophytic bacterial strain (Achromobacter marplatensisstrain 17, isolated from sugar beet) to monitor its colonizationof sugar beet (Beta. vulgaris L.). Stability of the plasmidencoding the gfp was confirmed in vitro for at least 72 h ofbacterial growth and after the colonization of tissues, undernonselective conditions. The colonization was observed usingfluorescence microscopy and enumeration of culturableendophytes in inoculated sugar beet plants that grew for 10or 20 days. gfp-Expressing strains were re-isolated from theinner tissues of surface-sterilized roots and stems of inoculatedplants, and the survival of the Achromobacter marplatensis17:gfp strain in plants 20 days after inoculation, evenin the absence of selective pressure, suggests that it is goodcolonizer. These results also suggest that this strain could bea useful tool for the delivery of enzymes or other proteinsinto plants. In addition, the study highlights that sugar beetplants can be used effectively for detailed in vitro studies onthe interactions between A. marplatensis strain 17 and its host,particularly if a gfp-tagged strain of the pathogen is used.

Disturbance Observer-based Nonsingular Fast Terminal Sliding Mode Fault Tolerant Control of a Quadrotor UAV with External Disturbances and Actuator Faults

Fang Wang,Zhigang Ma,Hongmei Gao,Chao Zhou,Changchun Hua 제어·로봇·시스템학회 2022 International Journal of Control, Automation, and Vol.20 No.4

This paper addresses a nonsingular fast terminal sliding mode fault tolerant control (NFTSMFTC) scheme based on disturbance observer (DO) for a quadrotor UAV in the presence of external disturbances and actuator additive faults. Firstly, by analyzing the dynamic characteristics of a quadrotor UAV, the model is decoupled into outer-loop (position subsystem) and inner-loop (attitude subsystem). Secondly, the DO is utilized to deal with external disturbances and actuator additive faults effectively. Furthermore, nonsingular fast terminal sliding mode control (NFTSMC) can avoid the chattering problems of traditional sliding mode control (SMC) and singularity problems of terminal SMC. Moreover, compared with nonsingular terminal SMC, it behaves the faster convergence rate. So the NFTSMC method is applied to design controller. Finally, in the framework of Lyapunov theory, the stability of the closed-loop system is analyzed, and the tracking error is asymptotically stable. The effectiveness of the proposed control scheme is verified by simulation results.

Transcriptome-wide characterization of the WRKY family genes in Lonicera macranthoides and the role of LmWRKY16 in plant senescence

Cao Zhengyan,Wu Peiyin,Gao Hongmei,Xia Ning,Jiang Ying,Tang Ning,Liu Guohua,Chen Zexiong 한국유전학회 2022 Genes & Genomics Vol.44 No.2

Background: Lonicera macranthoides is an important woody plant with high medicinal values widely cultivated in southern China. WRKY, one of the largest transcription factor families, participates in plant development, senescence, and stress responses. However, a comprehensive study of the WRKY family in L. macranthoides hasn't been reported previously. Objective: To establish an extensive overview of the WRKY family in L. macranthoides and identify senescence-responsive members of LmWRKYs. Methods: RNA-Seq and phylogenetic analysis were employed to identify the LmWRKYs and their evolutionary relationships. Quantitative real-time (qRT-PCR) and transgenic technology was utilized to investigate the roles of LmWRKYs in response to developmental-, cold-, and ethylene-induced senescence. Results: A total of 61 LmWRKY genes with a highly conserved motif WRKYGQK were identified. Phylogenetic analysis of LmWRKYs together with their orthologs from Arabidopsis classified them into three groups, with the number of 15, 39, and 7, respectively. 17 LmWRKYs were identified to be differentially expressed between young and aging leaves by RNA-Seq. Further qRT-PCR analysis showed 15 and 5 LmWRKY genes were significantly induced responding to tissue senescence in leaves and stems, respectively. What's more, five LmWRKYs, including LmWRKY4, LmWRKY5, LmWRKY6, LmWRKY11, and LmWRKY16 were dramatically upregulated under cold and ethylene treatment in both leaves and stems, indicating their involvements commonly in developmental- and stress-induced senescence. In addition, function analysis revealed LmWRKY16, a homolog of AtWRKY75, can accelerate plant senescence, as evidenced by leaf yellowing during reproductive growth in LmWRKY16-overexpressing tobaccos. Conclusion: The results lay the foundation for molecular characterization of LmWRKYs in plant senescence.

Lattice-based Threshold Ring Signature with Message Block Sharing

( Jiangshan Chen ),( Yupu Hu ),( Wen Gao ),( Hongmei Liang ) 한국인터넷정보학회 2019 KSII Transactions on Internet and Information Syst Vol.13 No.2

Threshold ring signature scheme enables any t entities from N ring members to spontaneously generate a publicly verifiable t-out-of-N signature anonymously. The verifier is convinced that the signature is indeed generated by at least t users from the claimed group, but he cannot tell them apart. Threshold ring signatures are significant for ad-hoc groups such as mobile ad-hoc networks. Based on the lattice-based ring signature proposed by Melchor et al. at AFRICRYPT’13, this work presents a lattice-based threshold ring signature scheme, employing the technique of message block sharing proposed by Choi and Kim. Besides, in order to avoid the system parameter setup problems, we proposed a message processing technique called “pad-then-permute”, to pre-process the message before blocking the message, thus making the threshold ring signature scheme more flexible. Our threshold ring signature scheme has several advantages: inherits the quantum immunity from the lattice structure; has considerably short signature and almost no signature size increase with the threshold value; provable to be correct, efficient, indistinguishable source hiding, and unforgeable.

Interaction of the Lysophospholipase PNPLA7 with Lipid Droplets through the Catalytic Region

Chang, Pingan,Sun, Tengteng,Heier, Christoph,Gao, Hao,Xu, Hongmei,Huang, Feifei Korean Society for Molecular and Cellular Biology 2020 Molecules and cells Vol.43 No.3

Mammalian patatin-like phospholipase domain containing proteins (PNPLAs) play critical roles in triglyceride hydrolysis, phospholipids metabolism, and lipid droplet (LD) homeostasis. PNPLA7 is a lysophosphatidylcholine hydrolase anchored on the endoplasmic reticulum which associates with LDs through its catalytic region (PNPLA7-C) in response to increased cyclic nucleotide levels. However, the interaction of PNPLA7 with LDs through its catalytic region is unknown. Herein, we demonstrate that PNPLA7-C localizes to the mature LDs ex vivo and also colocalizes with pre-existing LDs. Localization of PNPLA7-C with LDs induces LDs clustering via non-enzymatic intermolecular associations, while PNPLA7 alone does not induce LD clustering. Residues 742-1016 contains four putative transmembrane domains which act as a LD targeting motif and are required for the localization of PNPLA7-C to LDs. Furthermore, the N-terminal flanking region of the LD targeting motif, residues 681-741, contributes to the LD targeting, whereas the C-terminal flanking region (1169-1326) has an anti-LD targeting effect. Interestingly, the LD targeting motif does not exhibit lysophosphatidylcholine hydrolase activity even though it associates with LDs phospholipid membranes. These findings characterize the specific functional domains of PNPLA7 mediating subcellular positioning and interactions with LDs, as wells as providing critical insights into the structure of this evolutionarily conserved phospholipid-metabolizing enzyme family.

Identifying long non-coding RNAs and characterizing their functional roles in swine mammary gland from colostrogenesis to lactogenesis

Shi Lijun,Zhang Longchao,Wang Ligang,Liu Xin,Gao Hongmei,Hou Xinhua,Zhao Fuping,Yan Hua,Cai Wentao,Wang Lixian 아세아·태평양축산학회 2022 Animal Bioscience Vol.35 No.6

Objective: This study was conducted to identify the functional long non-coding RNAs (lncRNAs) for swine lactation by RNA-seq data of mammary gland. Methods: According to the RNA-seq data of swine mammary gland, we screened lncRNAs, performed differential expression analysis, and confirmed the functional lncRNAs for swine lactation by validation of genome wide association study (GWAS) signals, functional annotation and weighted gene co-expression network analysis (WGCNA). Results: We totally identified 286 differentially expressed (DE) lncRNAs in mammary gland at different stages from 14 days prior to (-) parturition to day 1 after (+) parturition, and the expressions of most of lncRNAs were strongly changed from day –2 to day +1. Further, the GWAS signals of sow milk ability trait were significantly enriched in DE lncRNAs. Functional annotation revealed that these DE lncRNAs were mainly involved in mammary gland and lactation developing, milk composition metabolism and colostrum function. By performing weighted WGCNA, we identified 7 out of 12 lncRNA-mRNA modules that were highly associated with the mammary gland at day –14, day –2, and day +1, in which, 35 lncRNAs and 319 mRNAs were involved. Conclusion: This study suggested that 18 lncRNAs and their 20 target genes were promising candidates for swine parturition and colostrum occurrence processes. Our research provided new insights into lncRNA profiles and their regulating mechanisms from colostrogenesis to lactogenesis in swine. Objective: This study was conducted to identify the functional long non-coding RNAs (lncRNAs) for swine lactation by RNA-seq data of mammary gland.Methods: According to the RNA-seq data of swine mammary gland, we screened lncRNAs, performed differential expression analysis, and confirmed the functional lncRNAs for swine lactation by validation of genome wide association study (GWAS) signals, functional annotation and weighted gene co-expression network analysis (WGCNA).Results: We totally identified 286 differentially expressed (DE) lncRNAs in mammary gland at different stages from 14 days prior to (-) parturition to day 1 after (+) parturition, and the expressions of most of lncRNAs were strongly changed from day –2 to day +1. Further, the GWAS signals of sow milk ability trait were significantly enriched in DE lncRNAs. Functional annotation revealed that these DE lncRNAs were mainly involved in mammary gland and lactation developing, milk composition metabolism and colostrum function. By performing weighted WGCNA, we identified 7 out of 12 lncRNA-mRNA modules that were highly associated with the mammary gland at day –14, day –2, and day +1, in which, 35 lncRNAs and 319 mRNAs were involved.Conclusion: This study suggested that 18 lncRNAs and their 20 target genes were promising candidates for swine parturition and colostrum occurrence processes. Our research provided new insights into lncRNA profiles and their regulating mechanisms from colostrogenesis to lactogenesis in swine.

Features of Recently Transmitted HIV-1 Clade C Viruses that Impact Antibody Recognition: Implications for Active and Passive Immunization

Rademeyer, Cecilia,Korber, Bette,Seaman, Michael S.,Giorgi, Elena E.,Thebus, Ruwayhida,Robles, Alexander,Sheward, Daniel J.,Wagh, Kshitij,Garrity, Jetta,Carey, Brittany R.,Gao, Hongmei,Greene, Kelli M Public Library of Science 2016 PLoS pathogens Vol.12 No.7

<▼1><P>The development of biomedical interventions to reduce acquisition of HIV-1 infection remains a global priority, however their potential effectiveness is challenged by very high HIV-1 envelope diversity. Two large prophylactic trials in high incidence, clade C epidemic regions in southern Africa are imminent; passive administration of the monoclonal antibody VRC01, and active immunization with a clade C modified RV144-like vaccines. We have created a large representative panel of C clade viruses to enable assessment of antibody responses to vaccines and natural infection in Southern Africa, and we investigated the genotypic and neutralization properties of recently transmitted clade C viruses to determine how viral diversity impacted antibody recognition. We further explore the implications of these findings for the potential effectiveness of these trials. A panel of 200 HIV-1 Envelope pseudoviruses was constructed from clade C viruses collected within the first 100 days following infection. Viruses collected pre-seroconversion were significantly more resistant to serum neutralization compared to post-seroconversion viruses (p = 0.001). Over 13 years of the study as the epidemic matured, HIV-1 diversified (p = 0.0009) and became more neutralization resistant to monoclonal antibodies VRC01, PG9 and 4E10. When tested at therapeutic levels (10ug/ml), VRC01 only neutralized 80% of viruses in the panel, although it did exhibit potent neutralization activity against sensitive viruses (IC<SUB>50</SUB> titres of 0.42 μg/ml). The Gp120 amino acid similarity between the clade C panel and candidate C-clade vaccine protein boosts (Ce1086 and TV1) was 77%, which is 8% more distant than between CRF01_AE viruses and the RV144 CRF01_AE immunogen. Furthermore, two vaccine signature sites, K169 in V2 and I307 in V3, associated with reduced infection risk in RV144, occurred less frequently in clade C panel viruses than in CRF01_AE viruses from Thailand. Increased resistance of pre-seroconversion viruses and evidence of antigenic drift highlights the value of using panels of very recently transmitted viruses and suggests that interventions may need to be modified over time to track the changing epidemic. Furthermore, high divergence such as that observed in the older clade C epidemic in southern Africa may impact vaccine efficacy, although the correlates of infection risk are yet to be defined in the clade C setting. Findings from this study of acute/early clade C viruses will aid vaccine development, and enable identification of new broad and potent antibodies to combat the HIV-1 C-clade epidemic in southern Africa.</P></▼1><▼2><P><B>Author Summary</B></P><P>Vaccine and passive immunization prophylactic trials that rely on antibody-mediated protection are planned for HIV-1 clade C epidemic regions of southern Africa, which have amongst the highest HIV-1 incidences globally. This includes a phase 2b trial of passively administered monoclonal antibody, VRC01; as well as a phase 3 trial using the clade C modified version of the partially efficacious RV144 vaccine. The extraordinary diversity of HIV-1 poses a major obstacle to these interventions, and our study aimed to determine the implications of viral diversity on antibody recognition. Investigations using our panel of very early viruses augment current knowledge of vulnerable targets on transmitted viruses for vaccine design and passive immunization studies. Evidence of antigenic drift with viruses becoming more resistant over time suggests that these prevention modalities will need to be updated over time and that combinations of antibodies will be necessary to achieve coverage in passive immunization studies. We further show that it may be more difficult to obtain protection in the genetically diverse clade C epidemic compared to RV144 where the epidemic is less diverse, although it should be noted that the correlates of infection risk are yet to be defined in the clade C setti