Aluminum-tolerant Protoplasts from Roots Can be Collected with Positively Charged Silica Microbeads : a Method Based on Differences in Surface Negativity : ENVIRONMENTAL AND STRESS RESPONSES : MEMBRANES AND BIOENERGETICS

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A new technique was developed for the collection of Al-tolerant plant cells. The procedure takes advantage of the difference in surface charge between Al-tolerant and Al-sensitive protoplasts. Equal volumes of freshly prepared, positively charged silica microbeads (0.05% w/v) and purified protoplasts derived from root tips (2×10^5 ml^<-1>) of rice, maize or pea were mixed and then centrifuged on a discontinuous Ficoll gradient. Intact protoplasts from the Al-tolerant plant were recovered mostly in the bottom fraction, those from the Al-sensitive plant were recoverd at the uppermost interface, and those from the plant with intermediate tolerance to Al were collected at the middle interface. Moreover, the pattern of fractionation was unrelated to the diameter of the protoplasts. The technique should be useful in future investigations of the physiological and biochemical properties of Al-tolerant cells or their plasma membranes.
日本植物生理学会の論文