Rapid mutation of Spirulina platensis by a new mutagenesis system of atmospheric and room temperature plasmas (ARTP) and generation of a mutant library with diverse phenotypes.
Rapid mutation of Spirulina platensis by a new mutagenesis system of atmospheric and room temperature plasmas (ARTP) and generation of a mutant library with diverse phenotypes.
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DOI：
10.1371/journal.pone.0077046
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发表时间：
2013
期刊：
影响因子：
3.7
通讯作者：
Xing X
中科院分区：
文献类型：
--
作者：
Fang M;Jin L;Zhang C;Tan Y;Jiang P;Ge N;Heping Li;Xing X
在本文中，我们旨在通过产生碳水化合物含量和生长速度增加的突变体来提高钝顶螺旋藻的碳水化合物生产力。 ARTP 被用作一种新的诱变工具来生成具有不同表型的钝顶葡萄球菌突变体文库。建立了通过用氦驱动的 ARTP 处理 60 秒来快速突变 S.platensis 的方案以及使用 96 孔微孔板和酶标仪进行突变体的高通量筛选方法。然后构建了62个突变体的突变体库，并选出了理想的突变体。诱变后的突变体特征在传代9次左右趋于稳定，总突变频率和比生长率正突变频率分别达到45%和25%。突变体库中的突变体在细胞生长速率、碳水化合物含量和絮凝强度方面表现出不同的表型。细胞碳水化合物含量较野生株增加20%以上的阳性突变频率为32.3%。与野生株相比，代表性突变体3-A10和3-B2的碳水化合物含量分别增加了40.3%和78.0%，而突变体4-B3的比生长率则增加了10.5%。代表性突变体的碳水化合物含量在不同的传代培养过程中保持稳定，表明遗传稳定性较高。 ARTP 被证明是一种有效的非转基因诱变工具，可用于生成多细胞微藻突变体库。
In this paper, we aimed to improve the carbohydrate productivity of Spirulina platensis by generating mutants with increased carbohydrate content and growth rate. ARTP was used as a new mutagenesis tool to generate a mutant library of S. platensis with diverse phenotypes. Protocol for rapid mutation of S. platensis by 60 s treatment with helium driven ARTP and high throughput screening method of the mutants using the 96-well microplate and microplate reader was established. A mutant library of 62 mutants was then constructed and ideal mutants were selected out. The characteristics of the mutants after the mutagenesis inclined to be stable after around 9th subculture, where the total mutation frequency and positive mutation frequency in terms of specific growth rate reached 45% and 25%, respectively. The mutants in mutant library showed diverse phenotypes in terms of cell growth rate, carbohydrate content and flocculation intensity. The positive mutation frequency in terms of cellular carbohydrate content with the increase by more than 20% percent than the wild strain was 32.3%. Compared with the wild strain, the representative mutants 3-A10 and 3-B2 showed 40.3% and 78.0% increase in carbohydrate content, respectively, while the mutant 4-B3 showed 10.5% increase in specific growth rate. The carbohydrate contents of the representative mutants were stable during different subcultures, indicating high genetic stability. ARTP was demonstrated to be an effective and non-GMO mutagenesis tool to generate the mutant library for multicellular microalgae.
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