用聚乙二醇诱导选定的成对原生质体间的融合.pdf

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植物学报 1994, 36 (7): 489—493
Acta B otanica S inica
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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
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
POLYETHYL ENE GLYCOL - INDUCED FUSION OF
SELECTED PA IRS OF SINGLE PROTOPLASTS
Sun M eng xiang Yang Hong yuan and Zhou Chang
L aboratory of P lant R ep rod uctive B iology College of L if e S cience W uhan U niversity W uhan 430072
Abstract
A new m ethod for polyethylene glycol PEG induced fusion between single pairs of
selected p rotop lastsw as developed. T he p rotop lastsw ere p repared from o leaves. U nder
an inverted m icroscope two defined p rotop lasts w ere selected w ith a hand m ade m icrop ipette
and transferred into a drop let of fusion solution containing 25% PEG M. W. 6000 0 1
mol L m annitol and 0 01 mol L CaC l2· 2H 2O pH 5 6 . 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% PEG 0 35 mol L sucrose and 0 01 mol L CaC l2 · 2H 2O pH
5 6 for app roximately 10 m in follow ed by subsequent w ashing w ith a solution contain