Transcriptome analysis of entomopathogenic fungus Beauveria bassiana-infected bean bug (Riptortus pedestris)

Se Jin Lee,Yi-Ting Yang,Yu-Shin Nai,Jae Su Kim 한국응용곤충학회 2016 한국응용곤충학회 학술대회논문집 Vol.2016 No.04

Beauveria bassiana (Bb) is an entomopathogenic fungus with a wide host range, and is commonly used as an environment-friendly biopesticide. However, the molecular mechanisms of Bb-host interactions are not well understood. Here, RNA isolated from a highly virulent strain of B. bassiana (Bb JEF-007) and Riptortus pedestris (Hemiptera: Alydidae) (bean bug) infected with this strain were subjected to high throughput next generation sequencing (NGS) to analyze and compare transcriptomes. Differentially expressed gene (DEG) analysis showed that 2,381 genes were up-regulated and 2,303 genes were down-regulated upon infection. Most DEGs were classified into the categories of single-organism, cellular and metabolism processes by gene ontology (GO) analysis. Carbon metabolism-related enzymes in the glyoxylate cycle were significantly up-regulated, suggesting a possible role for them in Bb growth in the host. This work provides insight into how entomopathogenic B. bassiana occupies agriculturally harmful bean bug at the late stage, which might be essential during fungal infection.

Transcriptome analysis of entomopathogenic fungus Beauveria bassiana-infected bean bug (Riptortus pedestris) to identify possible fungal virulence factors

Se Jin Lee,Yi-Ting Yang,Yu-Shin Nai,Jae Su Kim 한국응용곤충학회 2015 한국응용곤충학회 학술대회논문집 Vol.2015 No.10

The ascomycete fungus Beauveria bassiana is a wide host range entomo- pathogenic fungus, which is commonly used as an environmental friendly biopesticide. However, the molecular mechanisms of host-pathogen interaction of B. bassiana are not well understood. Here, the high throughput next generation sequencing was performed to analyze the transcriptome of B. bassiana JEF-007 infected bean bug (Riptorus pedestris). Differentially expressed gene (DEG) analysis results showed that total 4,684 genes including 2,381 up and 2,303 down regulated genes were identified. Most of the DEGs were classified into single- organism, cellular and metabolism processes by gene ontology (GO) analysis. Metabolism pathway was the most abound category of DEGs via KEGG pathway mapping. Several possible candidates of virulence factors were dramatically expressed after infection, such as cytotoxic lectin, bacterial-like toxin, and proteins related to cell wall, hyphal growth, nutrient uptake and halogenated compounds synthesis. Furthermore, we also found the highest expression of a novel small RNA virus in the infected bean bug, but the relationship between fungal virulence and the RNA virus was under determination. The functional roles of these possible virulence factors are remained unclear, but this work provides a new insight for further fungal studies. Our results reflect systemic impacts of fungal pathogenesis and these findings represent a significant advance in the fungal functional genomics.

Enhanced replication of novel picorna-like virus (RiPV-1) in Beauveria bassiana JEF-007-infected bean bug (Riptortus pedestris)

Yi-Ting Yang,Yu-Shin Nai,Se Jin Lee,Sihyeon Kim,Jae Su Kim 한국응용곤충학회 2016 한국응용곤충학회 학술대회논문집 Vol.2016 No.04

A novel insect-infecting positive sense single-stranded RNA virus, Riptortus pedestris virus-1 (RiPV-1), was found in the Riptortus pedestris transcriptome data by de novo assembly and further confirmed by RACE method. The genome of RiPV-1 consists of 10,554 nucleotides (nt) excluding the poly(A) tail and contains a single large open reading frame (ORF) of 10,371 nt encoding a 3,456 aa polyprotein and flanked by 71 and 112 nt 5' and 3' noncoding regions, respectively. RiPV-1 genome contains the consensus genome organization of picorna-like RNA helicase, cysteine protease, and RNA-dependent RNA polymerase (RdRp) array in that order from the 5' to the 3' end. From the phylogenetic analysis, RiPV-1 was clustered with unassigned insect RNA viruses, APV and KFV, which suggests that these three insect picorna-like viruses might constitute a novel group of insect-infecting RNA viruses. Tissue tropism analysis revealed that RiPV-1 was relatively abundant in the thorax, abdomen, midgut and fat body. Interestingly, RiPV-1 replication was enhanced by Beauveria bassiana JEF-007 infection that was quantified using qRT-PCR. This study identified a novel insect-infecting virus and provided further insight into the relationship between virus, fungus and host.

20(S)-Ginsenoside Rh2 displays efficacy against T-cell acute lymphoblastic leukemia through the PI3K/Akt/mTOR signal pathway

Ting Xia,Jin Zhang,Chuanxin Zhou,Yu Li,Wenhui Duan,Bo Zhang,Min Wang,Jianpei Fang 고려인삼학회 2020 Journal of Ginseng Research Vol.44 No.5

Background: T-cell acute lymphoblastic leukemia (T-ALL) is a kind of aggressive hematological cancer, and the PI3K/Akt/mTOR signaling pathway is activated in most patients with T-ALL and responsible for poor prognosis. 20(S)-Ginsenoside Rh2 (20(S)-GRh2) is a major active compound extracted from ginseng, which exhibits anti-cancer effects. However, the underlying anticancer mechanisms of 20(S)-GRh2 targeting the PI3K/Akt/mTOR pathway in T-ALL have not been explored. Methods: Cell growth and cell cycle were determined to investigate the effect of 20(S)-GRh2 on ALL cells. PI3K/Akt/mTOR pathway-related proteins were detected in 20(S)-GRh2-treated Jurkat cells by immunoblotting. Antitumor effect of 20(S)-GRh2 against T-ALL was investigated in xenograft mice. The mechanisms of 20(S)-GRh2 against T-ALL were examined by cell proliferation, apoptosis, and autophagy. Results: In the present study, the results showed that 20(S)-GRh2 decreased cell growth and arrested cell cycle at the G1 phase in ALL cells. 20(S)-GRh2 induced apoptosis through enhancing reactive oxygen species generation and upregulating apoptosis-related proteins. 20(S)-GRh2 significantly elevated the levels of pEGFP-LC3 and autophagy-related proteins in Jurkat cells. Furthermore, the PI3K/Akt/mTOR signaling pathway was effectively blocked by 20(S)-GRh2. 20(S)-GRh2 suppressed cell proliferation and promoted apoptosis and autophagy by suppressing the PI3K/Akt/mTOR pathway in Jurkat cells. Finally, 20(S)-GRh2 alleviated symptoms of leukemia and reduced the number of white blood cells and CD3 staining in the spleen of xenograft mice, indicating antitumor effects against T-ALL in vivo. Conclusion: These findings indicate that 20(S)-GRh2 exhibits beneficial effects against T-ALL through the PI3K/Akt/mTOR pathway and could be a natural product of novel target for T-ALL therapy.

Trichel Pulse in Negative DC Corona discharge and Its Electromagnetic Radiations

Yu Zhang,Li-Juan Liu,Jin-Song Miao,Zu-Lin Peng,Ji-Ting Ouyang 대한전기학회 2015 Journal of Electrical Engineering & Technology Vol.10 No.3

We investigate in this paper the radiated electromagnetic waves together with the discharge characteristics of Trichel pulse of negative DC corona discharge in air in pin-to-plate and wire-to-plate configurations. The feature of the current pulse and the frequency spectrum of the electromagnetic radiations were measured under various pressures and gas gaps. The results show that the repetition frequency and the amplitude of Trichel pulse current depend on the discharge conditions, but the rising time of the pulse relates only to the radius of needle or wire and keeps constant even if the other conditions (including the discharge current, the gas gap and the gas pressure) change. There exists the characterized spectrum of electromagnetic waves from negative corona discharge in Trichel pulse regime. These characterized radiations do not change their frequency at a given cathode geometry even if the averaged current, the gas gap or the air pressure changes, but the amplitude of radiations changes accordingly. The characterized electromagnetic radiations from Trichel pulse corona relate to the formation or the rising edge of current pulse. It confirms that the characterized radiations from Trichel pulse supply information of discharge system and provide a potential method for detecting charged targets.

Human lactoferrin efficiently targeted into caprine beta-lactoglobulin locus with transcription activator-like effector nucleases

Yu-Guo Yuan,Shao-Zheng Song,Meng-Ming Zhu,Zheng-Yi He,Rui Lu,Ting Zhang,Fei Mi,Jin-Yu Wang,Yong Cheng 아세아·태평양축산학회 2017 Animal Bioscience Vol.30 No.8

Objective: To create genetically modified goat as a biopharming source of recombinant human lacotoferrin (hLF) with transcription activator-like effector nucleases. Methods: TALENs and targeting vector were transferred into cultured fibroblasts to insert hLF cDNA in the goat beta-lactoglobulin (BLG) locus with homology-directed repair. The gene targeted efficiency was checked using sequencing and TE7I assay. The bi-allelic gene targeted colonies were isolated and confirmed with polymerase chain reaction, and used as donor cells for somatic cell nuclear transfer (SCNT). Results: The targeted efficiency for BLG gene was approximately 10%. Among 12 Bi-allelic gene targeted colonies, five were used in first round SCNT and 4 recipients (23%) were confirmed pregnant at 30 d. In second round SCNT, 7 (53%), 4 (31%), and 3 (23%) recipients were confirmed to be pregnant by ultrasound on 30 d, 60 d, and 90 d. Conclusion: This finding signifies the combined use of TALENs and SCNT can generate bi-allelic knock-in fibroblasts that can be cloned in a fetus. Therefore, it might lay the foundation for transgenic hLF goat generation and possible use of their mammary gland as a bioreactor for large-scale production of recombinant hLF.

High β-Glucosidase Secretion in Saccharomyces cerevisiae Improves the Efficiency of Cellulase Hydrolysis and Ethanol Production in Simultaneous Saccharification and Fermentation

Hong Ting Tang,Jin Hou,Yu Shen,Lili Xu,Hui Yang,Xu Fang,Xiaoming Bao 한국미생물생명공학회(구 한국산업미생물학회) 2013 Journal of microbiology and biotechnology Vol.23 No.11

Bioethanol production from lignocellulose is considered as a sustainable biofuel supply. However, the low cellulose hydrolysis efficiency limits the cellulosic ethanol production. The cellulase is strongly inhibited by the major end product cellobiose, which can be relieved by the addition of β-glucosidase. In this study, three β-glucosidases from different organisms were respectively expressed in Saccharomyces cerevisiae and the β-glucosidase from Saccharomycopsis fibuligera showed the best activity (5.2 U/ml). The recombinant strain with S. fibuligera β-glucosidase could metabolize cellobiose with a specific growth rate similar to the control strain in glucose. This recombinant strain showed higher hydrolysis efficiency in the cellulose simultaneous saccharification and fermentation, when using the Trichoderma reesei cellulase, which is short of the β-glucosidase activity. The final ethanol concentration was 110% (using Avicel) and 89% (using acid-pretreated corncob) higher than the control strain. These results demonstrated the effect of β-glucosidase secretion in the recombinant S. cerevisiae for enhancing cellulosic ethanol conversion.

Genomic characterization of newly found virus in Riptortus pedestris (Hemiptera: Alydidae)

Se Jin Lee,Yi-Ting Yang,Yu-Shin Nai,Mi Rong Lee,Jae Su Kim 한국농약과학회 2017 한국농약과학회 학술발표대회 논문집 Vol.2017 No.4

A novel picorma-like virus (RiPV-1) found in Beauveria bassiana infected bean bug (Riptortus pedestris)

Se Jin Lee,Yi-Ting Yang,Yu-Shin Nai,Mi Rong Lee,Sihyeon Kim,Jae Su Kim 한국응용곤충학회 2016 한국응용곤충학회 학술대회논문집 Vol.2016 No.10

A viral genome was assembled de novo from next-generation sequencing (NGS) data from bean bugs, Riptortus pedestris, infected with an entomopathogenic fungus, Beauveria bassiana (Bb), and was further confirmed via the RACE method. Based on the phylogenetic analysis of the RdRp sequences, RiPV-1 was clustered in the unassigned insect RNA viruses with two other viruses, APV and KFV. These three viruses were suggested to constitute a new group of insect RNA viruses. Interestingly, RiPV-1 replication was increased dramatically in bean bugs 2 to 6 days after fungal infection. In conclusion, a novel insect RNA virus was found by NGS data assembly. This virus can provide further insight into the interaction between virus, fungus and the host.

Trichel Pulse in Negative DC Corona discharge and Its Electromagnetic Radiations

Zhang, Yu,Liu, Li-Juan,Miao, Jin-Song,Peng, Zu-Lin,Ouyang, Ji-Ting The Korean Institute of Electrical Engineers 2015 Journal of Electrical Engineering & Technology Vol.10 No.3

We investigate in this paper the radiated electromagnetic waves together with the discharge characteristics of Trichel pulse of negative DC corona discharge in air in pin-to-plate and wire-to-plate configurations. The feature of the current pulse and the frequency spectrum of the electromagnetic radiations were measured under various pressures and gas gaps. The results show that the repetition frequency and the amplitude of Trichel pulse current depend on the discharge conditions, but the rising time of the pulse relates only to the radius of needle or wire and keeps constant even if the other conditions (including the discharge current, the gas gap and the gas pressure) change. There exists the characterized spectrum of electromagnetic waves from negative corona discharge in Trichel pulse regime. These characterized radiations do not change their frequency at a given cathode geometry even if the averaged current, the gas gap or the air pressure changes, but the amplitude of radiations changes accordingly. The characterized electromagnetic radiations from Trichel pulse corona relate to the formation or the rising edge of current pulse. It confirms that the characterized radiations from Trichel pulse supply information of discharge system and provide a potential method for detecting charged targets.