用细菌褐藻胶酶分离的裙带菜（Laminariales，Phaeophyceae）原生质体的再生。

Regeneration from Laminaria japonica Areschoug (Laminariales, Phaeophyceae) protoplasts isolated with bacterial alginase.

机构信息

Laboratory of Microbiology, Department of Food Science and Technology, Faculty of Fisheries, Hokkaido University, 3-1-1 Minato-cho, Hakodate, 041, Hokkaido, Japan.

出版信息

Plant Cell Rep. 1996 Sep;15(12):892-5. doi: 10.1007/BF00231582.

DOI:10.1007/BF00231582

PMID:24178269

Abstract

The conditions for effective isolation of viable protoplasts from Laminaria japonica with an alginase produced by marine bacterium Alteromonas sp. and a commercially available cellulase were investigated. The highest yields of viable protoplasts (7.9∼10.4x10(6) cells g(-1) FW) were obtained with a hypertonic solution containing 50 % seawater, 25 mM MgCl2, 5 mM HEPES buffer system, and 0.5 M mannitol. Protoplasts were not obtained from thalli of L. japonica when an abalone alginase (abalone acetone powder; AAP: Sigma) was used instead of the bacterial alginase. The isolated protoplasts were cultured in an PESI medium at 5 °C. Complete cell wall formation was observed within 7 days, and dividing cells were first observed in a 9-day-old culture. Some protoplasts regenerated into sheet-shaped thalli and rhizoid structures were also observed on some thalli after 30 to 40 days in culture. This is the first report of protoplast regeneration into plantlets of L. japonica Areschoug (Laminariales, Phaeophyceae).

摘要

从海洋细菌交替单胞菌产生的褐藻胶酶和市售纤维素酶中有效分离海带可行原生质体的条件进行了研究。在含有 50%海水、25 mM MgCl2、5 mM HEPES 缓冲系统和 0.5 M 甘露醇的高渗溶液中获得了最高产量的可行原生质体（7.9∼10.4x10(6) 个细胞/g FW）。当使用鲍鱼褐藻胶酶（鲍鱼丙酮粉；AAP：Sigma）代替细菌褐藻胶酶时，从海带藻体中未获得原生质体。分离的原生质体在 PESI 培养基中于 5°C 下培养。在 7 天内观察到完整细胞壁的形成，并且在 9 天龄培养物中首次观察到分裂细胞。在培养 30 至 40 天后，一些原生质体再生为片状藻体，并且在一些藻体上也观察到根状结构。这是第一个关于海带原生质体再生为植物的报告（Laminariales，褐藻）。