基于流式细胞仪分析的欧洲云杉原胚团细胞核提取液筛选

打开文本图片集

中图分类号：Q942 文献标识码：A 文章编号：1000-3142（2025）08-1475-09

Abstract：Nuclearsuspension isan important foundation foravarietyof molecularbiology researches.Toefectively preparethe nuclear suspensions of Norway spruce （Picea abies） proembryogenic masses （PEM），PEMs of Norway spruce embryoniccelines PaVII，PI3，andI5 wereusedas the materialsfornucleus extraction with sixlysis bufrs： （LB01，Otto's，Galbraith's，GPB，WPB and Tris·MgCl2 ）.Based on detecting DNA content by flow cytometry，the quantityand qualityof the nucleus suspensions were evaluated to screen fortheoptimal extraction buffer.Further，the stability of optimal bufer and the integrity of nucleus were measured.Theresults wereas folows：（1） Compared with the otherlysates，the numberof nuclei obtainedofPaVI linePEMs from LBOlwas the highest，with loweramountof cell debris.（2）In addition，coefficients of variation（ CVs ）of GO/G1 and G2/M nuclei，obtained from LBO1，were not more than 5% . The CVs were relatively low.（3）The nuclei of different lines PEMs were extracted by LBO1.There was no significant diffrenceinthe numberofnuclei extractedandtheproportionofcelldebris among thedifferent lines.（4） The nuclear membrane of most cells maintained in a high integrity when the nuclei of Norway spruce PEMs were extracted via LBOl through morphology investigation.To sumup，compared with otherbuffers，LBOlis theoptimal one for Norway spruce PEMs nucleus extraction，which may provide reference for nucleus extraction from gymnosperms.

Key words： conifer， Picea abies，proembryogenic masses，plant nucleus extraction， flow cytometry

细胞核是真核生物最重要的细胞器之一，是亲本遗传信息的主要载体和执行者（Gumberetal.，2019）。（剩余15842字）