ISOLATION AND CHARACTERIZATION OF VACUOLES FROM MELILOTUS-ALBA MESOPHYLL
ISOLATION AND CHARACTERIZATION OF VACUOLES FROM MELILOTUS-ALBA MESOPHYLL
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DOI：
10.1104/pp.68.6.1354
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发表时间：
1981-01-01
期刊：
影响因子：
7.4
通讯作者：
ALIBERT, G
中科院分区：
文献类型：
--
作者：
BOUDET, AM;CANUT, H;ALIBERT, G
甜三叶(M.alba Desr.)叶肉组织原生质体和液泡的制备方法都被描述了。液泡是用一种新的方法获得的，该方法包括在通过二乙氨基乙基葡聚糖层对原生质体进行短暂的离心过程中裂解质膜。该方法将液泡的释放和纯化一步到位。通过酶标记和显微镜检查，发现液泡制剂的污染程度很低。所描述的方法是快速的，并且在不引起渗透压变化的情况下提供了很好的空泡产量。在三叶草的液泡中发现了几种水解酶，并检测了它们的氨基酸含量。
Methods for the preparation of protoplasts and vacuoles from mesophyll tissues of sweet clover (M. alba Desr.) are described. Vacuoles are obtained using a new procedure which involves lysis of the plasmalemma during a brief centrifugation of protoplasts through a diethylaminoethyl dextran layer. This method combines the release of vacuoles and their purification in 1 step. The contamination of vacuole preparations was found to be low, as judged by enzymic markers and microscopic inspection. The method described is rapid and gives a good yield of vacuoles without causing changes in osmotic pressure. Several hydrolases were located in vacuoles from sweet clover, which were also examined for their amino content.