{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2026,10,4]],"date-time":"2026-10-04T18:43:04Z","timestamp":1791139384795,"version":"4.1.0"},"reference-count":33,"publisher":"MDPI AG","issue":"11","license":[{"start":{"date-parts":[[2013,11,19]],"date-time":"2013-11-19T00:00:00Z","timestamp":1384819200000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/3.0\/"}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Hot pepper is one of the economically important crops in Asia. A large number of gene sequences, including expressed sequence tag (EST) and genomic sequences are publicly available. However, it is still a daunting task to determine gene function due to difficulties in genetic modification of a pepper plants. Here, we show the application of the virus-induced gene silencing (VIGS) repression for the study of 459 pepper ESTs selected as non-host pathogen-induced cell death responsive genes from pepper microarray experiments in Nicotiana benthamiana. Developmental abnormalities in N. benthamiana plants are observed in the 32 (7%) pepper ESTs-silenced plants. Aberrant morphological phenotypes largely comprised of three groups: stunted, abnormal leaf, and dead.  In addition, by employing the combination of VIGS and Agrobacterium-mediated transient assays, we identified novel pepper ESTs that involved in Bax or INF1-mediated cell death responses. Silencing of seven pepper ESTs homologs suppressed Bax or INF1-induced  cell death, five of which suppressed both cell death responses in N. benthamiana. The  genes represented by these five ESTs encode putative proteins with functions in  endoplasmic reticulum (ER) stress and lipid signaling. The genes represented by the other two pepper ESTs showing only Bax-mediated cell death inhibition encode a CCCH-type zinc finger protein containing an ankyrin-repeat domain and a probable calcium-binding protein, CML30-like. Taken together, we effectively isolated novel pepper clones that are involved in hypersensitive response (HR)-like cell death using VIGS, and identified silenced clones that have different responses to Bax and INF1 exposure, indicating separate signaling pathways for Bax- and INF1-mediated cell death.<\/jats:p>","DOI":"10.3390\/ijms141122782","type":"journal-article","created":{"date-parts":[[2013,11,19]],"date-time":"2013-11-19T12:33:03Z","timestamp":1384864383000},"page":"22782-22795","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":11,"title":["Identification of Novel Pepper Genes Involved in Bax- or  INF1-Mediated Cell Death Responses by High-Throughput Virus-Induced Gene Silencing"],"prefix":"10.3390","volume":"14","author":[{"given":"Jeong","family":"Lee","sequence":"first","affiliation":[{"name":"Infection and Immunity Research Center, Korea Research Institute of Bioscience & Biotechnology (KRIBB), 125 Gwahak-ro, Yusung-gu, Daejeon 305-600, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Young","family":"Kim","sequence":"additional","affiliation":[{"name":"Infection and Immunity Research Center, Korea Research Institute of Bioscience & Biotechnology (KRIBB), 125 Gwahak-ro, Yusung-gu, Daejeon 305-600, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Doil","family":"Choi","sequence":"additional","affiliation":[{"name":"Infection and Immunity Research Center, Korea Research Institute of Bioscience & Biotechnology (KRIBB), 125 Gwahak-ro, Yusung-gu, Daejeon 305-600, Korea"},{"name":"Department of Plant Science, Seoul National University, Seoul 151-921, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]},{"given":"Jeong","family":"Park","sequence":"additional","affiliation":[{"name":"Infection and Immunity Research Center, Korea Research Institute of Bioscience & Biotechnology (KRIBB), 125 Gwahak-ro, Yusung-gu, Daejeon 305-600, Korea"}],"role":[{"vocabulary":"crossref","role":"author"}]}],"member":"1968","published-online":{"date-parts":[[2013,11,19]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"109","DOI":"10.1016\/S1369-5266(99)80022-3","article-title":"Fast forward genetics based on virus-induced gene silencing","volume":"2","author":"Baulcombe","year":"1999","journal-title":"Curr. Opin. Plant Biol"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"937","DOI":"10.1105\/tpc.10.6.937","article-title":"Initiation and maintenance of virus-induced gene silencing","volume":"10","author":"Ruiz","year":"1998","journal-title":"Plant Cell"},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"455","DOI":"10.5511\/plantbiotechnology.22.455","article-title":"High-throughput functional screening of plant and pathogen genes in planta","volume":"22","author":"Terauchi","year":"2005","journal-title":"Plant Biotechnol"},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"41","DOI":"10.1094\/MPMI-20-0041","article-title":"Identification and characterization of plant genes involved in Agrobacterium-mediated plant transformation by virus-induced silencing","volume":"20","author":"Anand","year":"2007","journal-title":"Mol. Plant Microbe Interact"},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"2291","DOI":"10.1093\/jxb\/erj200","article-title":"High-throughput virus-induced gene-silencing approach to assess the functional relevance of a moisture stress-induced cDNA homologous to lea4","volume":"57","author":"Udayakumar","year":"2006","journal-title":"J. Exp. Bot"},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"5690","DOI":"10.1093\/emboj\/cdg546","article-title":"High throughput virus-induced gene silencing implicates heat shock protein 90 in plant disease resistance","volume":"22","author":"Lu","year":"2003","journal-title":"EMBO J"},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"e63533","DOI":"10.1371\/journal.pone.0063533","article-title":"Multiple classes of immune-related proteases associated with the cell death response in pepper plants","volume":"8","author":"Bae","year":"2013","journal-title":"PLoS One"},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"782","DOI":"10.1111\/j.1469-8137.2007.02225.x","article-title":"A systematic study to determine the extent of gene silencing in Nicotiana benthamiana and other Solanaceae species when heterologous gene sequences are used for virus-induced gene silencing","volume":"176","author":"Hema","year":"2007","journal-title":"New Phytol"},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"201","DOI":"10.1111\/j.1462-5822.2004.00361.x","article-title":"The role and regulation of programmed cell death in plant-pathogen interactions","volume":"6","author":"Greenberg","year":"2004","journal-title":"Cell. Microbiol"},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"305","DOI":"10.1038\/nrm1358","article-title":"Controlled cell death, plant survival and development","volume":"5","author":"Lam","year":"2004","journal-title":"Nat. Rev. Mol. Cell Biol"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"7956","DOI":"10.1073\/pnas.96.14.7956","article-title":"Bax-induced cell death in tobacco is similar to the hypersensitive response","volume":"96","author":"Lacomme","year":"1999","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"12295","DOI":"10.1073\/pnas.211423998","article-title":"Mammalian Bax-induced plant cell death can be down-regulated by overexpression of Arabidopsis Bax Inhibitor-1 (AtBI-1)","volume":"98","author":"Jin","year":"2001","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"425","DOI":"10.1046\/j.1365-313X.2003.01639.x","article-title":"Overexpression of Bax inhibitor suppresses the fungal elicitor-induced cell death in rice (Oryza sativa L.) cells","volume":"33","author":"Matsumura","year":"2003","journal-title":"Plant J"},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"1413","DOI":"10.1105\/tpc.10.9.1413","article-title":"Resistance of nicotiana benthamiana to phytophthora infestans is mediated by the recognition of the elicitor protein INF1","volume":"10","author":"Kamoun","year":"1998","journal-title":"Plant Cell"},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"13","DOI":"10.1094\/MPMI.1997.10.1.13","article-title":"Extracellular protein elicitors from Phytophthora: Host-specificity and induction of resistance to fungal and bacterial phytopathogens","volume":"10","author":"Kamoun","year":"1993","journal-title":"Mol. Plant Microbe Interact"},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"977","DOI":"10.1007\/s00425-008-0797-y","article-title":"NbLRK1, a lectin-like receptor kinase protein of Nicotiana benthamiana, interacts with Phytophthora infestans INF1 elicitin and mediates INF1-induced cell death","volume":"228","author":"Kanzaki","year":"2008","journal-title":"Planta"},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"706","DOI":"10.1105\/tpc.008680","article-title":"Nicotiana benthamiana gp91phox homologs NbrbohA and NbrbohB participate in H2O2 accumulation and resistance to Phytophthora infestans","volume":"15","author":"Yoshioka","year":"2003","journal-title":"Plant Cell"},{"key":"ref_18","doi-asserted-by":"crossref","first-page":"383","DOI":"10.1046\/j.1364-3703.2003.00186.x","article-title":"Cytosolic HSP90 and HSP70 are essential components of INF1-mediated hypersensitive response and non-host resistance to pseudomonas cichorii in Nicotiana benthamiana","volume":"4","author":"Kanzaki","year":"2003","journal-title":"Mol. Plant Pathol"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"196","DOI":"10.1007\/s10142-003-0099-1","article-title":"EST and microarray analyses of pathogen-responsive genes in hot pepper (Capsicum annuum L.) non-host resistance against soybean pustule pathogen (Xanthomonas axonopodis pv. glycines)","volume":"4","author":"Lee","year":"2004","journal-title":"Funct. Integr. Genomics"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"777","DOI":"10.1046\/j.1365-313X.2002.01394.x","article-title":"Virus-induced gene silencing in tomato","volume":"31","author":"Liu","year":"2002","journal-title":"Plant J"},{"key":"ref_21","doi-asserted-by":"crossref","first-page":"377","DOI":"10.1016\/S1016-8478(23)13054-8","article-title":"A method of high frequency virus-induced gene silencing in chili pepper (Capsicum annuum L. cv Bukang)","volume":"17","author":"Chung","year":"2004","journal-title":"Mol. Cells"},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"375","DOI":"10.1046\/j.1365-313X.2002.01454.x","article-title":"NPP1 a Phytophthora-associated trigger of plant defense in parsley and Arabidopsis","volume":"32","author":"Fellbrich","year":"2002","journal-title":"Plant J"},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"3916","DOI":"10.1046\/j.1432-1327.2001.02301.x","article-title":"Transcriptionally and post-transcriptionally regulated response of 13 calmodulin genes to tobacco mosaic virus-induced cell death and wounding in tobacco plant","volume":"268","author":"Yamakawa","year":"2001","journal-title":"Eur. J. Biochem"},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"414","DOI":"10.1093\/pcp\/pcm011","article-title":"Pathogen-induced calmodulin isoforms in basal resistance against bacterial and fungal pathogens in tobacco","volume":"48","author":"Takabatake","year":"2006","journal-title":"Plant Cell Physiol"},{"key":"ref_25","doi-asserted-by":"crossref","first-page":"1148","DOI":"10.1093\/pcp\/pcm088","article-title":"The CCCH-type zinc finger proteins AtSZF1 and AtSZF2 regulate salt stress responses in Arabidopsis","volume":"48","author":"Sun","year":"2007","journal-title":"Plant Cell Physiol"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"1012","DOI":"10.1007\/BF01923455","article-title":"BiP (GRP78) an essential hsp70 resident protein in the endoplasmic reticulum","volume":"50","author":"Haas","year":"1994","journal-title":"Experientia"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"20","DOI":"10.1016\/j.tcb.2003.11.001","article-title":"A trip to the ER: Coping with stress","volume":"14","author":"Rutkowski","year":"2004","journal-title":"Trends Cell Biol"},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"353","DOI":"10.1104\/pp.121.2.353","article-title":"Overexpression of a gene that encodes the first enzyme in the biosynthesis of asparagine-linked glycans makes plants resistant to tunicamycin and obviates the tunicamycin-induced unfolded protein response","volume":"121","author":"Koizumi","year":"1999","journal-title":"Plant Physiol"},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"223","DOI":"10.1104\/pp.010397","article-title":"Differential regulation of mRNA levels of acyl carrier protein isoforms in Arabidopsis","volume":"128","author":"Bonaventure","year":"2002","journal-title":"Plant Physiol"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"11436","DOI":"10.1074\/jbc.M806991200","article-title":"Down-regulation of mitochondrial acyl carrier protein in mammalian cells compromises protein lipoylation and respiratory complex I and results in cell death","volume":"284","author":"Feng","year":"2009","journal-title":"J. Biol. 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