{"status":"ok","message-type":"work","message-version":"1.0.0","message":{"indexed":{"date-parts":[[2025,10,11]],"date-time":"2025-10-11T02:39:38Z","timestamp":1760150378346,"version":"build-2065373602"},"reference-count":46,"publisher":"MDPI AG","issue":"22","license":[{"start":{"date-parts":[[2023,11,20]],"date-time":"2023-11-20T00:00:00Z","timestamp":1700438400000},"content-version":"vor","delay-in-days":0,"URL":"https:\/\/creativecommons.org\/licenses\/by\/4.0\/"}],"funder":[{"name":"JSPS KAKENHI (Grant-in-Aid for Scientific Research(B)","award":["22H02315","22101490-0","23-231038606","04B2020"],"award-info":[{"award-number":["22H02315","22101490-0","23-231038606","04B2020"]}]},{"name":"NEDO Moonshot Program","award":["22H02315","22101490-0","23-231038606","04B2020"],"award-info":[{"award-number":["22H02315","22101490-0","23-231038606","04B2020"]}]},{"name":"JST-Mirai Program","award":["22H02315","22101490-0","23-231038606","04B2020"],"award-info":[{"award-number":["22H02315","22101490-0","23-231038606","04B2020"]}]},{"name":"Joint Research Program of Arid Land Research Center, Tottori University","award":["22H02315","22101490-0","23-231038606","04B2020"],"award-info":[{"award-number":["22H02315","22101490-0","23-231038606","04B2020"]}]}],"content-domain":{"domain":[],"crossmark-restriction":false},"short-container-title":["IJMS"],"abstract":"<jats:p>Feeder cells and the synthetic auxin 2,4-dichlorophenoxyacetic acid (2,4-D) in a culture medium promote mitosis and cell division in cultured cells. These are also added to nutrient medium for the cultivation of highly active in mitosis and dividing zygotes, produced in vitro or isolated from pollinated ovaries. In the study, an in vitro fertilization (IVF) system was used to study the precise effects of feeder cells and 2,4-D on the growth and development of rice (Oryza sativa L.) zygote. The elimination of 2,4-D from the culture medium did not affect the early developmental profiles of the zygotes, but decreased the division rates of multicellular embryos. The omission of feeder cells resulted in defective karyogamy, fusion between male and female nuclei, and the subsequent first division of the cultured zygotes. The culture of zygotes in a conditioned medium corrected developmental disorders. Proteome analyses of the conditioned medium revealed the presence of abundant hydrolases possibly released from the feeder cells. Exogenously applied \u03b1-amylase ameliorated karyogamy and promoted zygote development. It is suggested that hydrolytic enzymes, including \u03b1-amylase, released from feeder cells may be involved in the progression of zygotic development.<\/jats:p>","DOI":"10.3390\/ijms242216541","type":"journal-article","created":{"date-parts":[[2023,11,20]],"date-time":"2023-11-20T10:32:32Z","timestamp":1700476352000},"page":"16541","update-policy":"https:\/\/doi.org\/10.3390\/mdpi_crossmark_policy","source":"Crossref","is-referenced-by-count":1,"title":["The Feeder Effects of Cultured Rice Cells on the Early Development of Rice Zygotes"],"prefix":"10.3390","volume":"24","author":[{"given":"Yoriko","family":"Watanabe","sequence":"first","affiliation":[{"name":"Department of Biological Sciences, Tokyo Metropolitan University, Minami-Osawa 1-1, Hachioji 192-0397, Tokyo, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Yuko","family":"Nobe","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Tokyo Metropolitan University, Minami-Osawa 1-1, Hachioji 192-0397, Tokyo, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"ORCID":"https:\/\/orcid.org\/0000-0001-5554-4951","authenticated-orcid":false,"given":"Masato","family":"Taoka","sequence":"additional","affiliation":[{"name":"Department of Chemistry, Tokyo Metropolitan University, Minami-Osawa 1-1, Hachioji 192-0397, Tokyo, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]},{"given":"Takashi","family":"Okamoto","sequence":"additional","affiliation":[{"name":"Department of Biological Sciences, Tokyo Metropolitan University, Minami-Osawa 1-1, Hachioji 192-0397, Tokyo, Japan"}],"role":[{"role":"author","vocabulary":"crossref"}]}],"member":"1968","published-online":{"date-parts":[[2023,11,20]]},"reference":[{"key":"ref_1","doi-asserted-by":"crossref","first-page":"472","DOI":"10.1002\/j.1537-2197.1964.tb06658.x","article-title":"A cytochemical study of embryo development in Stellaria medis","volume":"51","author":"Pritchard","year":"1964","journal-title":"Botany"},{"key":"ref_2","doi-asserted-by":"crossref","first-page":"807","DOI":"10.1002\/j.1537-2197.1968.tb07438.x","article-title":"Capsella embryogenesis: The egg, zygote, and young embryo","volume":"55","author":"Schulz","year":"1968","journal-title":"Am. J. Bot."},{"key":"ref_3","doi-asserted-by":"crossref","first-page":"3","DOI":"10.5586\/4955","article-title":"Rape embryogenesis: I. The proembryo development","volume":"45","author":"Tykarska","year":"1976","journal-title":"Acta Soc. Bot. Pol."},{"key":"ref_4","doi-asserted-by":"crossref","first-page":"2842","DOI":"10.1139\/b84-379","article-title":"Interactions between embryo and endosperm during early developmental stage of maize caryopses (Zea mays)","volume":"62","author":"Schel","year":"1984","journal-title":"Can. J. Bot."},{"key":"ref_5","doi-asserted-by":"crossref","first-page":"359","DOI":"10.1093\/jxb\/44.2.359","article-title":"Embryogenesis: A question of pattern","volume":"259","author":"Lindsey","year":"1993","journal-title":"J. Exp. Bot."},{"key":"ref_6","doi-asserted-by":"crossref","first-page":"23","DOI":"10.1093\/pcp\/pci501","article-title":"Rice Plant development: From zygote to spikelet","volume":"46","author":"Itoh","year":"2005","journal-title":"Plant Cell Physiol."},{"key":"ref_7","doi-asserted-by":"crossref","first-page":"211","DOI":"10.1007\/s00497-009-0129-9","article-title":"Asymmetric cell division of rice zygotes located in embryo sac and produced by in vitro fertilization","volume":"23","author":"Sato","year":"2010","journal-title":"Sex Plant Reprod."},{"key":"ref_8","doi-asserted-by":"crossref","first-page":"12","DOI":"10.1007\/BF00194565","article-title":"In vitro fertilization of single, isolated gametes of maize mediated by electrofusion","volume":"4","author":"Kranz","year":"1991","journal-title":"Sex. Plant Reprod."},{"key":"ref_9","doi-asserted-by":"crossref","first-page":"1464","DOI":"10.4161\/psb.5.11.13468","article-title":"Gamete fusion site on the egg cell and autonomous establishment of cell polarity in the zygote","volume":"5","author":"Okamoto","year":"2010","journal-title":"Plant Signal. Behav."},{"key":"ref_10","doi-asserted-by":"crossref","first-page":"581","DOI":"10.1007\/s00425-007-0506-2","article-title":"Establishment of an in vitro fertilization system in rice (Oryza sativa L.)","volume":"226","author":"Uchiumi","year":"2007","journal-title":"Planta"},{"key":"ref_11","doi-asserted-by":"crossref","first-page":"739","DOI":"10.2307\/3869611","article-title":"In vitro fertilization with isolated, single gametes results in zygotic embryogenesis and fertile maize plants","volume":"5","author":"Kranz","year":"1993","journal-title":"Plant Cell"},{"key":"ref_12","doi-asserted-by":"crossref","first-page":"327","DOI":"10.1007\/s004250050327","article-title":"Differentiation of isolated wheat zygotes into embryos and normal plants","volume":"205","author":"Kumlehn","year":"1998","journal-title":"Planta"},{"key":"ref_13","doi-asserted-by":"crossref","first-page":"835","DOI":"10.1093\/pcp\/pcy250","article-title":"Establishment of an in vitro fertilization system in wheat (Triticum aestivum L.)","volume":"60","author":"Maryenti","year":"2019","journal-title":"Plant Cell Physiol."},{"key":"ref_14","doi-asserted-by":"crossref","first-page":"927","DOI":"10.1104\/pp.71.4.927","article-title":"Auxin Requirements of Sycamore Cells in Suspension Culture","volume":"71","author":"Moloney","year":"1983","journal-title":"Plant Physiol."},{"key":"ref_15","doi-asserted-by":"crossref","first-page":"594","DOI":"10.1007\/BF00270061","article-title":"Effects of culture density, conditioned medium and feeder cultures on microspore embryogenesis in Brassica napus L. cv. Topas","volume":"8","author":"Huang","year":"1990","journal-title":"Plant Cell Rep."},{"key":"ref_16","doi-asserted-by":"crossref","first-page":"229","DOI":"10.1016\/0168-9452(95)04146-L","article-title":"Effect of nurse culture and conditioned medium on colony formation and plant regeneration from Lolium perenne protoplasts","volume":"108","author":"Folling","year":"1995","journal-title":"Plant Sci."},{"key":"ref_17","doi-asserted-by":"crossref","first-page":"939","DOI":"10.1104\/pp.104.053843","article-title":"Auxin-dependent cell division and cell elongation. 1-naphthaleneacetic acid and 2,4-dichlorophenoxyacetic acid activate different pathways","volume":"137","author":"Campanoni","year":"2005","journal-title":"Plant Physiol."},{"key":"ref_18","doi-asserted-by":"crossref","unstructured":"Wo\u00f3jcik, A.M., Wo\u00f3jcikowska, B., and Gaj, M.D. (2020). Current Perspectives on the Auxin-Mediated Genetic Network that Controls the Induction of Somatic Embryogenesis in Plants. Int. J. Mol. Sci., 21.","DOI":"10.3390\/ijms21041333"},{"key":"ref_19","doi-asserted-by":"crossref","first-page":"395","DOI":"10.1007\/BF00392367","article-title":"Microcallus formation from maize protoplasts prepared from embryogenic callus","volume":"168","author":"Hodges","year":"1986","journal-title":"Planta"},{"key":"ref_20","doi-asserted-by":"crossref","first-page":"515","DOI":"10.1007\/BF00232786","article-title":"An improved procedure for plant regeneration from indica and japonica rice protoplasts","volume":"14","author":"Jain","year":"1995","journal-title":"Plant Cell Rep."},{"key":"ref_21","first-page":"23","article-title":"Secreted molecules and their role in embryo formation in plants: A mini-review","volume":"47","year":"2005","journal-title":"Acta Biol. Crac. Ser. Bot."},{"key":"ref_22","doi-asserted-by":"crossref","first-page":"1880","DOI":"10.1104\/pp.125.4.1880","article-title":"N-acetylglucosamine and glucosamine-containing arabinogalactan proteins control somatic embryogenesis","volume":"125","author":"Tadesse","year":"2001","journal-title":"Plant Physiol."},{"key":"ref_23","doi-asserted-by":"crossref","first-page":"1533","DOI":"10.1104\/pp.114.236059","article-title":"Dynamics of male and female chromatin during karyogamy in rice zygotes","volume":"165","author":"Ohnishi","year":"2014","journal-title":"Plant Physiol."},{"key":"ref_24","doi-asserted-by":"crossref","first-page":"206","DOI":"10.1104\/pp.15.01953","article-title":"Development of polyspermic rice zygotes","volume":"171","author":"Toda","year":"2016","journal-title":"Plant Physiol."},{"key":"ref_25","doi-asserted-by":"crossref","unstructured":"Deushi, R., Toda, E., Koshimizu, S., Yano, K., and Okamoto, T. (2021). Effect of paternal genome excess on the developmental and gene expression profiles of polyspermic zygotes in rice. Plants, 10.","DOI":"10.3390\/plants10020255"},{"key":"ref_26","doi-asserted-by":"crossref","first-page":"3167","DOI":"10.1073\/pnas.0630309100","article-title":"Close identity of a pressure-stabilized intermediate with a kinetic intermediate in protein folding","volume":"100","author":"Kitahara","year":"2003","journal-title":"Proc. Natl. Acad. Sci. USA"},{"key":"ref_27","doi-asserted-by":"crossref","first-page":"277","DOI":"10.1016\/j.jmb.2005.01.052","article-title":"NMR snapshots of a fluctuating protein structure: Ubiquitin at 30 bar-3 kbar","volume":"347","author":"Kitahara","year":"2005","journal-title":"J. Mol. Biol."},{"key":"ref_28","doi-asserted-by":"crossref","first-page":"3192","DOI":"10.5504\/BBEQ.2012.0056","article-title":"Identification, molecular cloning, and recombinant gene expression of an extracellular a-amylase from Dactylis Glomerata L. embryogenic suspension cultures","volume":"26","author":"Rakleova","year":"2012","journal-title":"Biotech. Biotechnol. Eq."},{"key":"ref_29","doi-asserted-by":"crossref","first-page":"811","DOI":"10.1007\/s00425-005-0027-9","article-title":"Monoclonal antibody against a cell wall marker protein for embryogenic potential of Dactylis glomerata L. suspension cultures","volume":"222","author":"Tchorbadjieva","year":"2005","journal-title":"Planta"},{"key":"ref_30","doi-asserted-by":"crossref","first-page":"1225424","DOI":"10.3389\/fpls.2023.1225424","article-title":"Secretome analysis revealed that cell wall remodeling and starch catabolism underlie the early stages of somatic embryogenesis in Pinus nigra","volume":"14","author":"Pernis","year":"2023","journal-title":"Front. Plant Sci."},{"key":"ref_31","first-page":"425","article-title":"A carrot somatic embryo mutant is rescued by chitinase","volume":"4","author":"Cordewener","year":"1992","journal-title":"Plant Cell"},{"key":"ref_32","doi-asserted-by":"crossref","first-page":"56","DOI":"10.1007\/s004250050296","article-title":"Extracellular b-1,3-glucanases are induced during early somatic embryogenesis in Cichorium","volume":"205","author":"Helleboid","year":"1998","journal-title":"Planta"},{"key":"ref_33","doi-asserted-by":"crossref","first-page":"69","DOI":"10.1016\/S1016-8478(23)15005-9","article-title":"Purification, characterization, and cDNA cloning of rice class III chitinase","volume":"3","author":"Park","year":"2002","journal-title":"Mol. Cells"},{"key":"ref_34","doi-asserted-by":"crossref","first-page":"615","DOI":"10.2307\/3869804","article-title":"Rhizobium lipooligosaccharides rescue a carrot somatic embryo mutant","volume":"5","author":"Heidstra","year":"1993","journal-title":"Plant Cell"},{"key":"ref_35","doi-asserted-by":"crossref","first-page":"285","DOI":"10.1007\/s11240-006-9139-6","article-title":"Embryo production through somatic embryogenesis can be used to study cell differentiation in plants","volume":"86","year":"2006","journal-title":"Plant Cell Tissue Organ Cult."},{"key":"ref_36","doi-asserted-by":"crossref","first-page":"909","DOI":"10.1007\/s00709-019-01349-3","article-title":"Gene expression and localization of arabinogalactan proteins during the development of anther, ovule, and embryo in rice","volume":"256","author":"Ma","year":"2019","journal-title":"Protoplasma"},{"key":"ref_37","doi-asserted-by":"crossref","first-page":"2061","DOI":"10.1093\/jxb\/erj159","article-title":"Localization of arabinogalactan proteins in egg cells, zygotes, and two-celled proembryos and effects of b-D-glucosyl Yariv reagent on egg cell fertilization and zygote division in Nicotiana tabacum L.","volume":"57","author":"Qin","year":"2006","journal-title":"J. Exp. Bot."},{"key":"ref_38","doi-asserted-by":"crossref","first-page":"9","DOI":"10.1046\/j.1365-313X.1995.08010009.x","article-title":"Early cytological events after induction of cell division in egg cells and zygote development following in vitro fertilization with angiosperm gametes","volume":"8","author":"Kranz","year":"1995","journal-title":"Plant J."},{"key":"ref_39","doi-asserted-by":"crossref","first-page":"1656","DOI":"10.1093\/pcp\/pcz077","article-title":"Sperm Entry into the Egg Cell Induces the Progression of Karyogamy in Rice Zygotes","volume":"60","author":"Ohnishi","year":"2019","journal-title":"Plant Cell Physiol."},{"key":"ref_40","doi-asserted-by":"crossref","first-page":"515","DOI":"10.1111\/j.1365-313X.2006.02970.x","article-title":"Tobacco zygotic embryogenesis in vitro: The original cell wall of the zygote is essential for maintenance of cell polarity, the apical\u2013basal axis and typical suspensor formation","volume":"49","author":"He","year":"2007","journal-title":"Plant J."},{"key":"ref_41","doi-asserted-by":"crossref","unstructured":"Abiko, M., Furuta, K., Yamauchi, Y., Fujita, C., Taoka, M., Isobe, T., and Okamoto, T. (2013). Identification of proteins enriched in rice egg or sperm cells by single-cell proteomics. PLoS ONE, 8.","DOI":"10.1371\/journal.pone.0069578"},{"key":"ref_42","doi-asserted-by":"crossref","first-page":"37","DOI":"10.1007\/s00497-006-0020-x","article-title":"Isolation of gametes and central cells from Oryza sativa L.","volume":"19","author":"Uchiumi","year":"2006","journal-title":"Sex. Plant Reprod."},{"key":"ref_43","doi-asserted-by":"crossref","first-page":"680","DOI":"10.1038\/227680a0","article-title":"Cleavage of structural proteins during the assembly of the head of bacteriophage T4","volume":"227","author":"Laemmli","year":"1970","journal-title":"Nature"},{"key":"ref_44","doi-asserted-by":"crossref","first-page":"361","DOI":"10.1016\/0003-2697(80)90470-4","article-title":"A simplified ultrasensitive silver stain for detecting proteins in polyacrylamide gels","volume":"105","author":"Oakley","year":"1980","journal-title":"Anal. Biochem."},{"key":"ref_45","doi-asserted-by":"crossref","first-page":"5864","DOI":"10.1074\/jbc.M211509200","article-title":"V-1, a protein expressed transiently during murine cerebellar development, regulates actin polymerization via interaction with capping protein","volume":"278","author":"Taoka","year":"2003","journal-title":"J. Biol. Chem."},{"key":"ref_46","doi-asserted-by":"crossref","first-page":"4847","DOI":"10.1021\/pr500262p","article-title":"Global PROTOMAP profiling to search for biomarkers of early-recurrent hepatocellular carcinoma","volume":"13","author":"Taoka","year":"2014","journal-title":"J. Proteome Res."}],"container-title":["International Journal of Molecular Sciences"],"original-title":[],"language":"en","link":[{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/22\/16541\/pdf","content-type":"unspecified","content-version":"vor","intended-application":"similarity-checking"}],"deposited":{"date-parts":[[2025,10,10]],"date-time":"2025-10-10T21:26:26Z","timestamp":1760131586000},"score":1,"resource":{"primary":{"URL":"https:\/\/www.mdpi.com\/1422-0067\/24\/22\/16541"}},"subtitle":[],"short-title":[],"issued":{"date-parts":[[2023,11,20]]},"references-count":46,"journal-issue":{"issue":"22","published-online":{"date-parts":[[2023,11]]}},"alternative-id":["ijms242216541"],"URL":"https:\/\/doi.org\/10.3390\/ijms242216541","relation":{},"ISSN":["1422-0067"],"issn-type":[{"type":"electronic","value":"1422-0067"}],"subject":[],"published":{"date-parts":[[2023,11,20]]}}}