Prairie Regional Laboratory, National Research Council, S7N OW9, Saskatoon, Saskatchewan, Canada.
Planta. 1974 Dec;115(4):355-67. doi: 10.1007/BF00388618.
Protoplasts of Vicia hajastana Grossh. obtained from suspension-culture cells and Pisum sativum L. obtained from leaves adhered tightly to each other in concentrated solutions of high-molecular-weight polyethylene glycol (PEG). The adhesion occurred non-specifically between the free protoplasts from the same species as well as from the different species and genus. It was enhanced by enrichment of the PEG solution with calcium. Very few heteroplasmic fusions occurred during the period when the protoplasts were incubated in the PEG solution. However, many heterokaryons (up to 10%) were formed soon after the PEG solution was diluted out. The same phenomena were also observed in protoplasts from suspension-culture cells of Glycine max L. and from leaves of Hordeum vulgare L. Vicia and soybean protoplasts obtained from cultured cells regenerated cell walls and underwent sustained cell division after such treatment. Some Vicia-pea heterokaryons divided once. Over 10% of the soybean-barley hybrids divided in 7 days. Some divided 4-5 times and formed small clusters of cells in 10 days. The hybrids were recognizable because they contained chloroplasts from the leaf protoplast and exhibited morphological characters typical of the chlorophyll-less cells. None of the protoplasts from pea and barley leaves, either with or without PEG treatment, underwent cell division during the period of observation. The mechanism of adhesion and fusion of the protoplasts has been discussed.
薇菜和豌豆原生质体的获得，是将悬浮培养细胞置于高相对分子质量聚乙烯醇（PEG）溶液中，使细胞彼此紧密相连，然后再将其从细胞中分离出来。这种方法不仅适用于同种细胞，也适用于不同种属的细胞。这种非特异性的黏附作用是通过在 PEG 溶液中添加钙来增强的。在原生质体在 PEG 溶液中孵育的过程中，很少发生异质融合。然而，在 PEG 溶液稀释后不久，就会形成许多异核体（高达 10%）。同样的现象也发生在大豆悬浮培养细胞和大麦叶片的原生质体中。经过这种处理后，从培养细胞中获得的薇菜和大豆原生质体重新形成细胞壁，并进行持续的细胞分裂。一些薇菜-豌豆异核体只分裂了一次。超过 10%的大豆-大麦杂种在 7 天内分裂。有些杂种在 10 天内分裂 4-5 次，并形成小细胞簇。这些杂种可以被识别，因为它们含有来自叶原生质体的叶绿体，并表现出无叶绿素细胞的典型形态特征。在观察期间，无论是用 PEG 处理还是未用 PEG 处理的豌豆和大麦叶片的原生质体都没有进行细胞分裂。已经讨论了原生质体的黏附和融合的机制。
