植物学报1994,36(7):489493 Acta Botan ica s in ica 用聚乙二醇诱导选定的成对 原生质体间的融合 孙蒙祥杨弘远周嫦 (武汉大学生命科学学院植物生殖生物学研究室,武汉430072) 摘要 微吸管选取单对原生质体,在含聚乙二醇(PEG)的微滴中诱导融合。此法克服了常规 的PEG群体融合方法中的盲目性,能排除一方亲本原生质体自相融合和多个原生质体的融 合,以及未融合的原生质体的混杂,保证融合产物来自选定的成对原生质体,从而使PEG融 合技术精确化。此法在植物细胞工程和细胞生物学研究中有广泛的应用前景 关键词原生质体融合;聚乙二醇 POLYETHYL ENE GLY COL-ND UCED FUSON OF SEL ECTED PA IRS OF SNGLE PRO TO PLASTS Sun M eng "xiang, Yang Hong"yuan and Zhou Chang (Laboratory of Plant Reproductie B ology, College of L fe Science, W whan University, W uhan 430072) Abstract A new m ethod for polyethy lene glycol (PEG )-induced fusin betw een single pairs of an inverted m croscope two def med p ro top lasts w ere selected w ith a hand m ade m icrop pette and transferred in to a drop let of fusin so lutin contain ing 25% PEG (M. W. 6000),0. mol mannitol and 0. 01 mol CaCk. 2HO(pH 5. 6). Slightly moving the p pette caused the p top lasts to contact and adhere to each other the fuson pairs w ere then tran sferred to a o lutin contain ing 10% PEG, 0.35 mol sucro se and 0.01 mol CaCk. 2HO(pH 5.6)for app rox m ately 10 m in, fo llow ed by sub sequent w ash ing w ith a solutin con tan ing °收稿日期1993-12-27接受日期1994-01-21 *国家自然科学基金资助课题。 o1994-2008ChinaAcademicJOumalElectronicPublishingHouseAllrightsreservedhttp:/www.cnki.net植 物 学 报 1994, 36 (7): 489—493 Acta B otanica S inica 用聚乙二醇诱导选定的成对 原生质体间的融合3 孙蒙祥 杨弘远 周 嫦 (武汉大学生命科学学院植物生殖生物学研究室, 武汉 430072) 摘 要 用微吸管选取单对原生质体, 在含聚乙二醇(PEG) 的微滴中诱导融合。此法克服了常规 的 PEG 群体融合方法中的盲目性, 能排除一方亲本原生质体自相融合和多个原生质体的融 合, 以及未融合的原生质体的混杂, 保证融合产物来自选定的成对原生质体, 从而使 PEG 融 合技术精确化。此法在植物细胞工程和细胞生物学研究中有广泛的应用前景。 关键词 原生质体; 融合; 聚乙二醇α POLYETHYL ENE GLYCOL- IND UCED FUSION OF SEL ECTED PA IRS OF SINGL E PROTOPLASTS Sun M eng2xiangΚYang Hong2yuan and Zhou Chang ;L aboratory of P lant R ep rod uctive B iologyΨCollege of L if e S cienceΨW uhan U niversityΨW uhan 430072Γ Abstract A new m ethod for polyethylene glycol ; PEGΓ2induced fusion betw een single pairs of selected p rotop lastsw as developed. T he p rotop lastsw ere p repared from tobacco leaves. U nder an inverted m icroscope two defined p rotop lasts w ere selected w ith a hand2m ade m icrop ipette and transferred into a drop let of fusion solution containing 25% PEG ;M. W. 6000ΓΚ 011 molöL m annitol and 0101 molöL CaC l2·2H 2O ;pH 516Γ. Slightly moving the p ipette caused the p rotop lasts to contact and adhere to each other. the fusion pairs w ere then transferred to a solution containing 10% PEGΚ 0135 molöL sucrose and 0101 molöL CaC l2·2H 2O ; pH 516Γfor app roxim ately 10 m inΚfollow ed by subsequent w ashing w ith a solution containing α 收稿日期: 1993212227 接受日期: 1994201221 3 国家自然科学基金资助课题