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Optimized Methods for the Isolation of Arabidopsis Female Central Cells and Their Nuclei

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dc.contributor.authorPark, Kyunghyuken
dc.contributor.authorFrost, Jennifer M.-
dc.contributor.authorAdair, Adam James-
dc.contributor.authorKim, Dong Min-
dc.contributor.authorYun, Hyein-
dc.contributor.authorBrooks, Janie S.-
dc.contributor.authorFischer, Robert L.-
dc.contributor.authorChoi, Yeonhee-
dc.date.accessioned2017-04-19T00:19:51Z-
dc.date.available2017-07-30T19:24:59Z-
dc.date.issued2016-10-28-
dc.identifier.citationMOLECULES AND CELLS Vol.39 No.10, pp. 768-775-
dc.identifier.issn1016-8478-
dc.identifier.urihttps://hdl.handle.net/10371/116892-
dc.description.abstractThe Arabidopsis female gametophyte contains seven cells with eight haploid nuclei buried within layers of sporophytic tissue. Following double fertilization, the egg and central cells of the gametophyte develop into the embryo and endosperm of the seed, respectively. The epigenetic status of the central cell has long presented an enigma due both to its inaccessibility, and the fascinating epigenome of the endosperm, thought to have been inherited from the central cell following activity of the DEMETER demethylase enzyme, prior to fertilization. Here, we present for the first time, a method to isolate pure populations of Arabidopsis central cell nuclei. Utilizing a protocol designed to isolate leaf mesophyll protoplasts, we systematically optimized each step in order to efficiently separate central cells from the female gametophyte. We use initial manual pistil dissection followed by the derivation of central cell protoplasts, during which process the central cell emerges from the micropylar pole of the embryo sac. Then, we use a modified version of the Isolation of Nuclei TAgged in specific Cell Types (INTACT) protocol to purify central cell nuclei, resulting in a purity of 75-90% and a yield sufficient to undertake downstream molecular analyses. We find that the process is highly dependent on the health of the original plant tissue used, and the efficiency of protoplasting solution infiltration into the gametophyte. By isolating pure central cell populations, we have enabled elucidation of the physiology of this rare cell type, which in the future will provide novel insights into Arabidopsis reproduction.en
dc.language.isoenen
dc.publisherKOREAN SOCIETY FOR MOLECULAR AND CELLULAR BIOLOGYen
dc.subjectArabidopsis central cellen
dc.subjectembryo sacen
dc.subjectnuclei isolation-
dc.subjectprotoplast-
dc.titleOptimized Methods for the Isolation of Arabidopsis Female Central Cells and Their Nucleien
dc.typeArticleen
dc.contributor.AlternativeAuthor박경혁-
dc.contributor.AlternativeAuthor김동민-
dc.contributor.AlternativeAuthor윤혜인-
dc.contributor.AlternativeAuthor최연희-
dc.identifier.doi10.14348/molcells.2016.0209-
dc.description.srndOAIID:RECH_ACHV_DSTSH_NO:T201620049-
dc.description.srndRECH_ACHV_FG:RR00200001-
dc.description.srndADJUST_YN:-
dc.description.srndEMP_ID:A076495-
dc.description.srndCITE_RATE:2.67-
dc.description.srndFILENAME:molce-39-10-768.pdf-
dc.description.srndDEPT_NM:생명과학부-
dc.description.srndEMAIL:yhc@snu.ac.kr-
dc.description.srndSCOPUS_YN:Y-
dc.description.srndFILEURL:https://srnd.snu.ac.kr/eXrepEIR/fws/file/4cccba36-07e6-4a85-9eda-27f8dd164627/link-
dc.description.srndCONFIRM:Y-
dc.identifier.rimsid51566-
dc.identifier.srndT201620049-
dc.type.rimsART-
Appears in Collections:
College of Natural Sciences (자연과학대학)Dept. of Biological Sciences (생명과학부)Journal Papers (저널논문_생명과학부)
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