Identification and tissue distribution of carboxylesterase (CXE) genes in Athetis lepigone (Lepidoptera: Noctuidae) by RNA-seq

Ya-Nan Zhang,Zhao-Qun Li,Xiu-Yun Zhu,Jia-Li Qian,Zhi-Ping Dong,Lu Xue,Peng He 한국응용곤충학회 2017 Journal of Asia-Pacific Entomology Vol.20 No.4

Some of the metabolic enzyme carboxylesterases (CXEs) belong to the odorant-degrading enzymes (ODEs) family in insect species, and these play a key role in the degradation of acetate sex pheromones and host plant volatiles. Athetis lepigone is one of the most important agricultural insect pests in the world, can damage> 30 species of host plants, and has caused serious declines in the yield of summer corn in North China since 2011. According to our previous studies, the sex pheromone component of the pest is a binary blend of Z7–12:OAc and Z9–14:OAc at a ratio of 3:7. However, there are no reports regarding the degradation mechanism for these two sex pheromones. Herein, we firstly identified 20 candidate CXE genes in A. lepigone using our previous adult antennal RNA-seq data. Then, we constructed a phylogenetic tree and further conducted tissue distribution analyses to determine the possible functions of these genes. Our results showed that some AlepCXEs displayed adult antennae- predominant, male antennae-biased, or leg/wing-biased expression, indicating these AlepCXEs may have distinct physiological functions and play distinct roles in the degradation of sex pheromones, host plant volatiles, and/or other xenobiotics. These findings will help us to elucidate the exact functions of these genes in the future, and also provide possible target genes for the prevention and control of A. lepigone.

Molecular characterization of the sans fille gene in Antheraea pernyi: A highly conserved gene during the evolution of animals

Ya-Jie Li,Rui Mi,Nan Meng,Zhi-Xin Wen,Xue-Jun Li,Mo Chen,Yan-Qun Liu,Shu-Ying Li 한국응용곤충학회 2013 Journal of Asia-Pacific Entomology Vol.16 No.3

Sans-fille (SNF) is the Drosophila homologue ofmammalian general splicing factors U1A and U2B″, and plays an important role in sex determination in Drosophilamelanogaster. In this study, the snf gene fromAntheraea pernyi (Lepidoptera: Saturniidae), an economically important insect, was isolated and characterized. The obtained 925 bp cDNA sequence contains an open reading frame of 669 bp encoding a polypeptide of 222 amino acids,showing 78% sequence identity to that from D. melanogaster. A database search revealed that SNF protein homologs are present in many animals, including invertebrates and vertebrates, with more than 70% amino acid sequence identities, suggesting that they were highly conserved during the evolution of animals. Phylogenetic analysis revealed that A. pernyi SNF was closely related to Bombyx mori SNF. Quantitative real-time PCR (qRT-PCR) analysis showed that the A. pernyi snf gene was transcribed during five larval developmental stages,and in six tested tissues (ovaries, testes, silk glands, fat body, integument, and hemolymph),with the most abundance determined in the gonads (ovaries or testes). Investigation of expression changes throughout embryonic development indicated that A. pernyi snfmRNAwas expressed at a lowlevel fromdays 0 to 4, and reached amaximum level at day 10, but decreased to a low level before hatching. These results suggest that the product of the snf gene may play important roles in the development of A. pernyi.

Characterization and expression of calcium-activated potassium channels (Slo) in ovary of the mud crab, Scylla paramamosain

Xue-Liang Liu,Hai-Hui Ye,Hui-Yang Huang,Jie Gong,Ya-Nan Yang 한국유전학회 2014 Genes & Genomics Vol.36 No.1

Large conductance calcium-activated potassiumchannels (Slo) play important roles in controllingneuronal excitability. At present, very little is known aboutthe function of Slo channels on ovarian development. Wecloned the SPSlo gene from the mud crab, Scylla paramamosain. This gene shows 91 and 93 % sequence identityto PISlo from the spiny lobster, Panulirus interruptus andCBSlo from the jonah crab, Cancer borealis, respectively. We isolated six variants of the SPSlo cDNA within S. paramamosain ovary tissue. Sequence analysis indicatedthat there were at least seven alternative sites in SPSlo,each with multiple alternative segments. Real-time PCRshowed that the SPSlo gene was expressed in various tissues,and highly expressed in brain and ovary. In addition,the expression of SPSlo changed throughout ovariandevelopment, highest at the early-developing stage (StageII) followed by a slow decrease in subsequent stages. Theseresults suggested that SPSlo channels may be implicated inthe ovarian development of the mud crab.

Insecticidal activities and biochemical properties of Pinellia ternata extracts against the beet armyworm Spodoptera exigua

Ya-Nan Zhang,Peng He,Jian-Ping Xue,Qing Guo,Xiu-Yun Zhu,Li-Ping Fang,Jin-Bu Li 한국응용곤충학회 2017 Journal of Asia-Pacific Entomology Vol.20 No.2

The beet armyworm, Spodoptera exigua Hübner (Lepidoptera: Noctuidae), is a key pest of various agricultural crops in many countries throughout the world. The pest requires extensive use of pesticides and field-evolved resistances to conventional insecticides in China and other countries. Pinellia ternata Breit is native to the eastern part of Asia and foundmainly in China,which has been used in traditional Chinese medicines for N1000 years, but few studies have focused on the insecticidal activity of the P. ternate. In order to find natural products that could be used to control the pest in an safe, efficient and ecofriendlymanner,we first time analyzed the components of the anhydrous ethanolic extracts fromthe tubers of Pinellia ternate by using GC–MS method, and then investigated the insecticidal activities and biochemical mechanisms of the extracts against S. exigua. The result of GC–MS showed that 2-Methoxy-4-vinylphenolmay be an insecticidal active component, and we also found the extracts had notable insecticidal activity and disturbed the regular metabolismof S. exiguamainly through altering the activities of detoxification enzymes, digestive enzymes and protective enzymes. These properties suggest that the anhydrous ethanolic extracts from P. ternate can serve as a potential, efficient and ecofriendly S. exigua-control biopesticide.

Multiple down-regulated cytochrome P450 monooxygenase genes contributed to synergistic interaction between chlorpyrifos and imidacloprid against Nilaparvata lugens

Lu Xue,Guanghua Luo,Yang Sun,Shuijin Huang,De-Jin Xu,Guang-Chun Xu,Zhao-Jun Han,Zhong-Yan Gu,Ya-Nan Zhang 한국응용곤충학회 2020 Journal of Asia-Pacific Entomology Vol.23 No.1

Insecticide mixtures are an effective strategy in pest resistance management. The synergistic chlorpyrifos and imidacloprid mixture could significantly increase toxicity against rice pest, Nilaparvata lugens, despite their high levels of resistance. However, synergism mechanisms to explain this phenomenon remain unknown. Chlorpyrifos and imidacloprid at a 1:0.5 ratio showed significant synergism on N. lugens with a combination index value of 0.18 after topical exposure. We constructed a genetic database of the genes expressed in individual and synergistic chlorpyrifos and imidacloprid treatments of N. lugens using Illumina Hiseq™ X Ten, and 17 co-downregulated genes putatively involved in synergism were detected by comparative transcriptome analyses. Expression patterns of the 17 candidate synergistic genes matched with transcriptome sequencing data by quantitative real-time PCR analyses. Feeding of dsRNAs further reduced the expression levels of 10 of these candidate synergistic genes (from 1.68 to 4.13-fold). Nymphs fed with only dsRNAs of CYP4DE1, CYP6AY1v2, CYP353D1, and CYP439A1 experienced more high mortality rates (81.45–90.34%) to improve synergism between chlorpyrifos and imidacloprid. Multiple reductive expressed P450 genes were potentially associated with synergism of a mixture of chlorpyrifos and imidacloprid, as confirmed by comparative transcriptome analyses and RNAi assays. Our findings suggested that synergistic interactions between chlorpyrifos and imidacloprid might be controlled by P450s.

XIAP Associated Factor 1 (XAF1) Represses Expression of X-linked Inhibitor of Apoptosis Protein (XIAP) and Regulates Invasion, Cell Cycle, Apoptosis, and Cisplatin Sensitivity of Ovarian Carcinoma Cells

Zhao, Wen-Jing,Deng, Bo-Ya,Wang, Xue-Mei,Miao, Yuan,Wang, Jian-Nan Asian Pacific Journal of Cancer Prevention 2015 Asian Pacific journal of cancer prevention Vol.16 No.6

Background: X-linked inhibitor of apoptosis protein (XIAP) associated factor 1 (XAF1) exhibits aberrantly low or absent expression in various human malignancies, closely associated with anti-apoptosis and overgrowth of cancer cells. However, limited attention has been directed towards the contribution of XAF1 to invasion, apoptosis, and cisplatin (DDP)-resistance of epithelial ovarian cancer (EOC) cells. This study aimed to evaluate the potential effects of XAF1 on invasion, cell cycle, apoptosis, and cisplatin-resistance by overexpressing XAF1 in SKOV-3 and SKOV-3/DDP cells. Methods and Results: The pEGFP-C1-XAF1 plasmid was transfected into SKOV-3 and SKOV-3/DDP cells, and the expression of XAF1 at both mRNA and protein levels was analyzed by reverse transcription-PCR and Western blotting. Overexpression of XAF1 suppressed XIAP expression in both SKOV-3 and SKOV-3/DDP cells. Transwell invasion assays demonstrated that XAF1 exerted a strong anti-invasive effect in XAF1-overexpressing cells. Moreover, flow cytometry analysis revealed that XAF1 overexpression arrested the cell cycle at G0/G1 phase, and cell apoptosis analysis showed that overexpression of XAF1 enhanced apoptosis of SKOV-3 and SKOV-3/DDP cells apparently by activating caspase-9 and caspase-3. Furthermore, MTT assay confirmed a dose-dependent inhibitory effect of cisplatin in the tested tumor cells, and overexpression of XAF1 increased the sensitivity of SKOV-3 and SKOV-3/DDP cells to cisplatin-mediated antiproliferative effects. Conclusions: In summary, our data indicated that overexpression of XAF1 could suppress XIAP expression, inhibit invasion, arrest cell cycle, promote apoptosis, and confer cisplatin-sensitivity in SKOV-3 and SKOV-3/DDP cells. Therefore, XAF1 may be further assessed as a potential target for the treatment of both cisplatin-resistant and non-resistant EOCs.

Remarkable impact of steam temperature on ginsenosides transformation from fresh ginseng to red ginseng

Xu, Xin-Fang,Gao, Yan,Xu, Shu-Ya,Liu, Huan,Xue, Xue,Zhang, Ying,Zhang, Hui,Liu, Meng-Nan,Xiong, Hui,Lin, Rui-Chao,Li, Xiang-Ri The Korean Society of Ginseng 2018 Journal of Ginseng Research Vol.42 No.3

Background: Temperature is an essential condition in red ginseng processing. The pharmacological activities of red ginseng under different steam temperatures are significantly different. Methods: In this study, an ultrahigh-performance liquid chromatography quadrupole time-of-flight tandem mass spectrometry was developed to distinguish the red ginseng products that were steamed at high and low temperatures. Multivariate statistical analyses such as principal component analysis and supervised orthogonal partial least squared discrimination analysis were used to determine the influential components of the different samples. Results: The results showed that different steamed red ginseng samples can be identified, and the characteristic components were 20-gluco-ginsenoside Rf, ginsenoside Re, ginsenoside Rg1, and malonyl-ginsenoside Rb1 in red ginseng steamed at low temperature. Meanwhile, the characteristic components in red ginseng steamed at high temperature were 20R-ginsenoside Rs3 and ginsenoside Rs4. Polar ginsenosides were abundant in red ginseng steamed at low temperature, whereas higher levels of less polar ginsenosides were detected in red ginseng steamed at high temperature. Conclusion: This study makes the first time that differences between red ginseng steamed under different temperatures and their ginsenosides transformation have been observed systematically at the chemistry level. The results suggested that the identified chemical markers can be used to illustrate the transformation of ginsenosides in red ginseng processing.

Flavobacterium zhairuonensis sp. nov., a gliding bacterium isolated from marine sediment of the East China Sea

Sanjit Chandra Debnath,Ahmed Mohammed Abdo Miyah,Can Chen,Huan Sheng,Xue-Wei Xu,Yue-Hong Wu,Dao-Qiong Zheng,Jin-Zhong Xu,Ya-Nan Di,Pin-Mei Wang,Li Shen 한국미생물학회 2019 The journal of microbiology Vol.57 No.12

A yellow pigmented, Gram-stain-negative, aerobic bacterium designated A5.7T was studied to evaluate the taxonomic position following the modern polyphasic approach. The strain was isolated from sediments near Zhairuo Island, which is situated in the East China Sea. Cells were non-spore forming rods without flagella but showed motility by gliding. Growth was observed at 15–35°C (optimum 28°C), pH 6.0–9.0 (optimum pH 6.5) and 0–2% (w/v) NaCl (optimum 0–0.5%) in LB broth. The major respiratory quinone of A5.7T was menaquinone 6. The major polar lipid of A5.7T was phosphatidylethanolamine and the predominant fatty acids (> 5%) were iso-C15:0, iso-C17:0 3-OH, C15:1 ω6c, iso-C15:0 3-OH, iso-C15:1 G, summed feature 3 (C16:1 ω7c and/or C16:1 ω6c) and summed feature 9 (iso-C17:1 ω9c and/or C16:0 10-methyl). Phylogenetic analysis based on 16S rRNA gene sequences showed that the isolate belongs to the genus Flavobacterium and shares the highest sequence similarities with Flavobacterium sharifuzzamanii A7.6T (98.5%), Flavobacterium tistrianum GB 56.1T (98.3%), Flavobacterium nitrogenifigens NXU-44T (97.8%), Flavobacterium anhuiense D3T (97.6%), Flavobacterium ginsenosidimutans THG 01T (97.6%), and Flavobacterium foetidum CJ42T (97.6%). Digital DNA-DNA hybridization and average nucleotide identity values between the strain and its closest phylogenetic neighbors showed the ranges from 19.6 to 34.1% and 73.7 to 87.9%, respectively. Therefore, based on polyphasic characteristics, strain A5.7T represents a novel species of the genus Flavobacterium for which the name Flavobacterium zhairuonensis sp. nov. is proposed. The type strain is A5.7T (= KCTC 62406T = MCCC 1K03494T).

Selection of Reference Genes for Real-time Quantitative PCR Normalization in the Process of Gaeumannomyces graminis var. tritici Infecting Wheat

Li-hua Xie,Xin Quan,Jie Zhang,Yan-yan Yang,Run-hong Sun,Ming-cong Xia,Bao-guo Xue,Chao Wu,Xiao-yun Han,Ya-nan Xue,Li-rong Yang 한국식물병리학회 2019 Plant Pathology Journal Vol.35 No.1

Gaeumannomyces graminis var. tritici is a soil borne pathogenic fungus associated with wheat roots. The accurate quantification of gene expression during the process of infection might be helpful to understand the pathogenic molecular mechanism. However, this method requires suitable reference genes for transcript normalization. In this study, nine candidate reference genes were chosen, and the specificity of the primers were investigated by melting curves of PCR products. The expression stability of these nine candidates was determined with three programs-geNorm, Norm Finder, and Best Keeper. TUBβ was identified as the most stable reference gene. Furthermore, the exopolygalacturonase gene (ExoPG) was selected to verify the reliability of TUBβ expression. The expression profile of ExoPG assessed using TUBβ agreed with the results of digital gene expression analysis by RNA-Seq. This study is the first systematic exploration of the optimal reference genes in the infection process of Gaeumannomyces graminis var. tritici.

Selection of Reference Genes for Real-time Quantitative PCR Normalization in the Process of Gaeumannomyces graminis var. tritici Infecting Wheat

Xie, Li-hua,Quan, Xin,Zhang, Jie,Yang, Yan-yan,Sun, Run-hong,Xia, Ming-cong,Xue, Bao-guo,Wu, Chao,Han, Xiao-yun,Xue, Ya-nan,Yang, Li-rong The Korean Society of Plant Pathology 2019 Plant Pathology Journal Vol.35 No.1

Gaeumannomyces graminis var. tritici is a soil borne pathogenic fungus associated with wheat roots. The accurate quantification of gene expression during the process of infection might be helpful to understand the pathogenic molecular mechanism. However, this method requires suitable reference genes for transcript normalization. In this study, nine candidate reference genes were chosen, and the specificity of the primers were investigated by melting curves of PCR products. The expression stability of these nine candidates was determined with three programs-geNorm, Norm Finder, and Best Keeper. $TUB{\beta}$ was identified as the most stable reference gene. Furthermore, the exopolygalacturonase gene (ExoPG) was selected to verify the reliability of $TUB{\beta}$ expression. The expression profile of ExoPG assessed using $TUB{\beta}$ agreed with the results of digital gene expression analysis by RNA-Seq. This study is the first systematic exploration of the optimal reference genes in the infection process of Gaeumannomyces graminis var. tritici.