放线菌共生固氮放线菌原生质体的形成和再生。

Formation and regeneration of protoplasts of the actinorhizal nitrogen-fixing actinomycete frankia.

机构信息

Department of Bacteriology, University of Wisconsin, Madison, Wisconsin 53706.

出版信息

Appl Environ Microbiol. 1987 Jan;53(1):53-6. doi: 10.1128/aem.53.1.53-56.1987.

DOI:10.1128/aem.53.1.53-56.1987

PMID:16347266

原文链接:https://pmc.ncbi.nlm.nih.gov/articles/PMC203601/

Abstract

Procedures for forming and regenerating protoplasts of four Frankia strains are described. Cells obtained from growth medium containing 0.1% glycine were digested with lysozyme (250 mug/ml) in a medium containing 0.5 M sucrose, 5.0 mM CaCl(2), and 5.0 mM MgCl(2). Protoplasts were formed during 15 to 120 min of digestion at 25 degrees C. Optimum conditions for protoplast regeneration involved placing protoplasts on a layer of complex growth medium containing 0.3 M sucrose, 5.0 mM CaCl(2), and 5.0 mM MgCl(2) which was overlaid with a layer of 0.8% low-melting-point agarose containing 0.5 M sucrose, 5.0 mM MgCl(2), and 5.0 mM CaCl(2). The maximum regeneration efficiency was 36.9% for strain CpI1, 1.3% for strain ACN1, 27% for strain EAN1pec, and 20% for strain EuI1c.

摘要

描述了四种弗兰克氏放线菌原生质体制备和再生的程序。用含 0.1%甘氨酸的生长培养基获得的细胞，用 250μg/ml 溶菌酶在含 0.5 M 蔗糖、5.0 mM CaCl2 和 5.0 mM MgCl2 的培养基中进行消化。在 25°C 下消化 15 至 120 分钟即可形成原生质体。原生质体再生的最佳条件是将原生质体放在一层含有 0.3 M 蔗糖、5.0 mM CaCl2 和 5.0 mM MgCl2 的复杂生长培养基上，然后在其上覆盖一层含有 0.5 M 蔗糖、5.0 mM MgCl2 和 5.0 mM CaCl2 的 0.8%低熔点琼脂糖。CpI1 菌株的最大再生效率为 36.9%，ACN1 菌株为 1.3%，EAN1pec 菌株为 27%，EuI1c 菌株为 20%。