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SSRN 2024年

作者： Seo, Hyeon Ung Jang, Cheol Seong Plant Genomics Laboratory Interdisciplinary Program in Smart Agriculture Graduate School Kangwon National University Chuncheon24341 Korea Republic of

Salinity is a major problem due to the continuous increase in the salinization of agricultural lands, particularly, paddy fields. Using a forward genetics approach, salt-insensitive TILLING (targeting-induced local lesions in genomes) line 3, sitl3, was selected from a core population induced by gamma-ray irradiation. Under salt stress, sitl3 had greater fresh weight and chlorophyll content, and lower H2O2 and Na+ contents than the wild-type. In the gene (LOC_Os07g46180) with two PWWP domains (OsPWWP4) of sitl3, a premature stop was caused by an SNP, and was named ***462* (a stop gain occurred from the 462th amino acid residue). The OsPWWP4 and substrate proteins (OsEULS2, OsEULS3, and OsEULD2) were identified using yeast two-hybrid, bimolecular fluorescence complementation, in vitro pull-down, and in vitro methyltransferase assays. Subcellular localization of OsPWWP4 and ***462*GFP-tagged proteins revealed they were both localized in the nucleus, while OsEULS2, OsEULS3, and OsEULD2 GFP-tagged proteins were found in the nucleus and cytosol of rice protoplasts. The expression levels of OsEULS2, OsEULS3, OsEULD2 under salt stress were higher in sitl3 than in wild-type plants. In contrast, OsPWWP4 expression was higher in the latter. Genes involved in the salt overly sensitive (SOS) pathway showed higher expression in the aerial tissues of silt3 than in the wild-type. Our data suggest that TILLING line sitl3 is a valuable genetic resource for understanding protein post-translational regulation-related salinity tolerance mechanisms such as methyltransferase activities, and for improving salt tolerance in rice through breeding. © 2024, The Authors. All rights reserved.

关键词： Genes

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Molecular Dissection of a Rice Plant with Mutation of the Leucine Carboxyl Methyl Transferase Gene

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SSRN 2023年

作者： Seo, Hyeon Ung Jang, Cheol Seong Plant Genomics Laboratory Interdisciplinary Program in Smart Agriculture Graduate School Kangwon National University Chuncheon24341 Korea Republic of

Drought is a major environmental factor that negatively affects crop growth and productivity. Rice is a drought-sensitive crop, and its development and yield are greatly affected by stress. Using a forward genetics approach, a drought-insensitive TILLING line, ditl2, induced using gamma-ray irradiation, was selected from 150 rice mutant lines. Under drought treatment conditions, the ditl2 mutant showed a high survival rate, fresh weight, relative water content, soluble sugar content, and low hydrogen peroxide (H2O2) content. Whole-genome re-sequencing of ditl2 showed a single-base adenine deletion of the OsLCMT (LOC_Os03g56370) gene, annotated as leucine carboxyl methyl transferase, resulting in a frameshift (***163fs) with a short peptide of 163 amino acids. The substrate gene OsSnRK1A of OsLCMT was identified through an experiment using yeast two-hybrid and BiFC assays. The subcellular localization of the two genes was confirmed to be in the cytosol. The methyltransferase activities of OsLCMT and ***163fs were examined using the target protein OsSnRK1A in an in vitro assay. Consequently, the methyltransferase activity of ***163fs was lower than that of the wild-type gene. High expression of starch hydrolysis pathway genes, such as amylases, amylomaltases, and plastidic sugar transporters, was found in dilt2 compared to wild-type (WT) plants. The ditl2 mutant is useful as a genetic resource for understanding the relationship between protein methylation and drought insensitivity and improving drought tolerance in rice mutant breeding. © 2023, The Authors. All rights reserved.

关键词： Drought

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Transcriptome profiling reveals novel insights into the regulation of calcium ion and detoxification genes driving chlorantraniliprole resistance in Spodoptera exigua

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Heliyon 2024年第23期10卷 e40556页

作者： Han, Changhee Kim, Juil Interdisciplinary Graduate Program in Smart Agriculture Kangwon National University Chuncheon South Korea Agriculture and Life Sciences Research Institute Kangwon National University Chuncheon South Korea

Since the commercialization of diamide insecticides, including chlorantraniliprole, in 2007, the overuse of diamide insecticides for over a decade has resulted in excessive chlorantraniliprole resistance in Spodoptera exigua, causing continuous economic losses. While RyR target-site mutations and detoxification enzymes such as cytochrome P450 have been studied as the leading causes of resistance, previous studies, including functional research and synergistic tests, have not confirmed a clear correlation between these factors and the development of resistance. Thus, transcriptome analysis was employed to investigate alternative strategies beyond mutation(s) in RyR or metabolic factors involving detoxification pathways that allow diamide-resistance S. exigua to counteract the calcium ion imbalances induced by chlorantraniliprole effectively. Diamide-resistant, susceptible strains and its F1-hybrid of S. exigua were used for the RNAseq-based differentially expressed gene (DEG) analysis. In total 4669 genes were differentially expressed, with 2809 upregulated and 1860 downregulated in the resistant strain compared to the susceptible strain. GO, KEGG enrichment and orthologous analyses demonstrated that genes involved in metabolic factors were overrepresented in the resistant strain. In particular, overexpressed endoplasmic reticulum (ER)-related calcium ion homeostasis and cell stability-associated genes were newly identified in resistant strain. The selected differentially expressed genes were validated then with qPCR. These genes were inferred to induce cell stability to overcome ER stress derived from calcium ion imbalance caused by chlorantraniliprole. These results provide advanced insights into the critical roles of calcium ion homeostasis- and cell stability-related genes in conferring diamide insecticide resistance. © 2024 The Authors

关键词： Calcium ion imbalance Chlorantraniliprole Endoplasmic reticulum stress RNA sequencing Spodoptera exigua

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Genome-wide analysis of detoxification genes conferring diamide insecticide resistance in Spodoptera exigua identifies CYP9A40

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Chemosphere 2024年 367卷 143623页

作者： Han, Changhee Rahman, Md-Mafizur Kim, Juil Lueke, Bettina Nauen, Ralf Interdisciplinary Graduate Program in Smart Agriculture Kangwon National University Chuncheon24341 Korea Republic of Department Biotechnology and Genetic Engineering Faculty of Biological Science Islamic University Kushtia7003 Bangladesh Agriculture and Life Science Research Institute Kangwon National University Chuncheon24341 Korea Republic of Bayer AG Crop Science Division R&D Monheim40789 Germany

For over a decade, diamide insecticides have been effective against lepidopteran pests like beet armyworm, Spodoptera exigua (Hübner, 1808). However, the evolution of resistance poses a challenge to their sustainable use. We identified an I4790 M mutation in the S. exigua ryanodine receptor (RyR) gene, but its correlation with resistance varied across the field-collected Korean populations of S. exigua. RNA sequencing and differential gene expression analysis were performed to investigate other resistance mechanisms. Diamide-resistant and susceptible strains and F1 hybrids were compared by mapping RNA-seq reads to the S. exigua reference genome. CYP9A40 was identified as a critical gene in diamide resistance due to its high expression in the resistant strains. Synergist bioassays with piperonyl butoxide supported the role of P450s in diamide metabolic resistance in S. exigua. A strong positive correlation between CYP9A40 over-expression levels (up to 80-fold) and diamide LC50 values was obtained for field-collected populations uniformly showing a 100% frequency of the RyR I4790 M target-site resistance allele. To validate the function of CYP9A40 in diamide detoxification, we recombinantly expressed the gene and tested its ability to bind and degrade chlorantraniliprole as a substrate. The results confirmed its catalytic role in diamide metabolism. CYP9A40 has been identified and validated to confer metabolic resistance in Korean S. exigua populations. It works alongside the RyR target-site I4790 M mutation to enhance diamide resistance. These mechanisms offer insights for resistance monitoring and support insecticide resistance management programs to improve control strategies for S. exigua. © 2024 The Author(s)

关键词： Spodoptera exigua Diamide P450 CYP9A40 Heterologous expression RNA-Seq

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Genome-Wide Analysis of Detoxification Genes Conferring Diamide Insecticide Resistance in Spodoptera Exigua Identifies Cyp9a40

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SSRN 2024年

作者： Kim, Juil Han, Changhee Rahman, Md-Mafizur Lueke, Bettina Nauen, Ralf Interdisciplinary Graduate Program in Smart Agriculture Kangwon National University Chuncheon24341 Korea Republic of Department Biotechnology and Genetic Engineering Faculty of Biological Science Islamic University Kushtia7003 Bangladesh Agriculture and Life Science Research Institute Kangwon National University Chuncheon24341 Korea Republic of Bayer AG Crop Science Division Monheim40789 Germany

For over a decade, diamide insecticides have been effective against lepidopteran pests like beet armyworm, Spodoptera exigua (Hübner, 1808). However, the evolution of resistance poses a challenge to their sustainable use. We identified an I4790M mutation in the S. exigua ryanodine receptor (RyR) gene, but its correlation with resistance varied across the field-collected Korean populations of S. exigua. RNA sequencing and differential gene expression analysis were performed to investigate other resistance mechanisms. Diamide-resistant and susceptible strains and F1 hybrids were compared by mapping RNA-seq reads to the S. exigua reference genome. CYP9A40 was identified as a critical gene in diamide resistance due to its high expression in the resistant strains. Synergist bioassays with piperonyl butoxide supported the role of P450s in diamide metabolic resistance in S. exigua. A strong positive correlation between CYP9A40 over-expression levels (up to 80-fold) and diamide LC50 values was obtained for field-collected populations uniformly showing a 100% frequency of the RyR I4790M target-site resistance allele. To validate the function of CYP9A40 in diamide detoxification, we recombinantly expressed the gene and tested its ability to bind and degrade chlorantraniliprole as a substrate. The results confirmed its catalytic role in diamide metabolism. CYP9A40 has been identified and validated to confer metabolic resistance in Korean S. exigua populations. It works alongside the RyR target-site I4790M mutation to enhance diamide resistance. These mechanisms offer insights for resistance monitoring and support insecticide resistance management programs to improve control strategies for S. exigua. © 2024, The Authors. All rights reserved.

关键词： RNA

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Development of novel gene carrier using modified nano hydroxyapatite derived from equine bone for osteogenic differentiation of dental pulp stem cells

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Bioactive Materials 2021年第9期6卷 2742-2751页

作者： Myung Chul Lee Hoon Seonwoo Kyoung Je Jang Shambhavi Pandey Jaewoon Lim Sangbae Park Jae Eun Kim Yun-Hoon Choung Pankaj Garg Jong Hoon Chung Department of Biosystems&Biomaterials Science and Engineering Seoul National UniversitySeoul08826Republic of Korea Department of Biosystems Engineering Seoul National UniversitySeoul08826Republic of Korea Department of Industrial Machinery Engineering Sunchon National University315 Maegok-dongSunchon57922Republic of Korea Interdisciplinary Program in IT-Bio Convergence System Sunchon National UniversitySuncheon57922Republic of Korea Division of Agro-system Engineering Gyeongsang National University501 Jinju-daeroJinju52828Republic of Korea Institute of Agriculture&Life Science Gyeongsang National UniversityJinju52828Republic of Korea Research Institute for Agriculture and Life Sciences Seoul National UniversitySeoul08826Republic of Korea Department of Otolaryngology Ajou University School of MedicineSuwon16499Republic of Korea Ajou University Graduate School of Medicine Bk21 Plus Research Center for Biomedical SciencesSuwon16499Republic of Korea BK21 Global Smart Farm Educational Research Center Seoul National UniversitySeoul08826Republic of Korea

Hydroxyapatite(HA)is a representative substance that induces bone *** research team extracted nanohydroxyapatite(EH)from natural resources,especially equine bones,and developed it as a molecular biological ***(PEI)was used to coat the EH to develop a gene *** verify that PEI is well coated in the EH,we first observed the morphology and dispersity of PEI-coated EH(pEH)by electron *** pEH particles were well distributed,while only the EH particles were not distributed and ***,the existence of nitrogen elements of PEI on the surface of the pEH was confirmed by EDS,calcium concentration measurement and fourier transform infrared spectroscopy(FT-IR).Additionally,the pEH was confirmed to have a more positive charge than the 25 kD PEI by comparing the zeta *** a result of pGL3 transfection,pEH was better able to transport genes to cells than 25 kD *** verification as a gene carrier for pEH,we induced osteogenic differentiation of DPSCs by loading the BMP-2 gene in pEH(BMP-2/pEH)and delivering it to the *** a result,it was confirmed that osteogenic differentiation was promoted by showing that the expression of osteopontin(OPN),osteocalcin(OCN),and runt-related transcription factor 2(RUNX2)was significantly increased in the group treated with BMP-2/*** conclusion,we have not only developed a novel nonviral gene carrier that is better performing and less toxic than 25 kD PEI by modifying natural HA(the agricultural byproduct)but also proved that bone differentiation can be effectively promoted by delivering BMP-2 with pEH to stem cells.

关键词： Nanocomposites Biomedical applications Hydroxyapatite Polyethyleneimine Gene carrier Osteogenic differentiation

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JOURNAL OF SPINAL CORD MEDICINE 2021年第SUPPL 1期44卷 S278-S324页

作者： [Anonymous] Department of Medicine Division of Physical Medicine and Rehabilitation University of Toronto Toronto ON CA KITE - Toronto Rehabilitation Institute University Health Network Toronto ON CA Institute for Health Policy Management and Evaluation University of Toronto Toronto ON CA Rehabilitation Sciences Institute University of Toronto Toronto ON CA Institute of Medical Sciences University of Toronto Toronto ON CA. Rehabilitation Sciences Institute University of Toronto ON CA KITE Research Institute-Toronto Rehab-University Health Network Toronto ON CA Physical Therapy Department University of Toronto ON CA Praxis Spinal Cord Institute Vancouver BC Université de Montréal (École de réadaptation) Montreal QC CA Institut de réadaptation Gingras-Linsay-de-Montréal Montreal QC CA Division of Physical Medicine & Rehabilitation Department of Medicine Faculty of Medicine & Dentistry University of Alberta Edmonton AB CA University of Calgary Calgary AB CA Alberta Health Services Centre for Family Medicine Kitchener ON CA McMaster University Department of Family Medicine Hamilton ON CA Dept. of Physical Therapy University of Toronto Toronto ON CA KITE-University Health Network Toronto ON CA Praxis Spinal Cord Institute Vancouver BC CA Center for Interdisciplinary Research in Rehabilitation and Social Integration (CIRRIS) Quebec City QC CA Dept. of Rehabilitation Laval University Quebec City QC CA Institute of Health Policy Management and Evaluation University of Toronto Toronto ON CA School of Rehabilitation Science University of Saskatchewan Saskatoon SK CA Ontario Neurotrauma Foundation Toronto ON CA Canadian Spinal Research Organization Toronto ON CA CIUSSS du Centre-Sud-de-l ‘Île-de-Montréal (Installation Gingras-Lindsay) Montréal QC CA École de réadaptation Université de Montréal Montréal QC CA First Steps Wellness Centre Regina SK CA Stan Cassidy Centre for Rehabilitation Horizon Health Network Fredericton NB CA Departmen

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