Isolation of mesophyll and paraveinal mesophyll protoplasts from soybean leaves

Vincent R. Franceschi , Maurice S.B. Ku , Vernon A. Wittenbach
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引用次数: 19

Abstract

Mesophyll (MP) and paraveinal mesophyll protoplasts (PVMP) have been prepared with high yields from primary and trifoliate leaves of soybean, using a Cellulase CEL or CELF and Pectolyase Y-23 enzyme mixture. The PVMP, which are specialized for nitrogen metabolism and storage, are larger than MP and contain very few chloroplasts. This gives rise to considerable difference in the buoyant density of the two protoplast types, which has been exploited to obtain pure preparations of each. The MP and PVMP preparations were free of cellular debri and were stable for several days on ice. The purity of the preparations was further indicated by radial immunodiffusion assay using antibody to a glycoprotein specifically located in the paraveinal mesophyll (PVM) vacuole.

大豆叶肉和叶鞘旁叶肉原生质体的分离
利用纤维素酶CEL或CELF和果胶酶Y-23混合酶,以大豆初生叶和三叶叶为原料,高产制备了叶肉(MP)和叶肉旁原生质体(PVMP)。PVMP是专门用于氮代谢和储存的,比MP大,含有很少的叶绿体。这导致两种原生质体的浮力密度有相当大的差异,这已经被用来获得每一种原生质体的纯制剂。MP和PVMP制剂不含细胞碎屑,并在冰上稳定放置数天。该制剂的纯度进一步通过径向免疫扩散试验表明,抗体针对的糖蛋白特异性位于叶肉旁液泡(PVM)。
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