Liang Lihui, Su Qihang, Wang Yawei, Du Peichen, Zhao Suzhen, Zhang Huanjie, Gao Xiaofeng
National R & D Center for Edible Fungus Processing Technology, Henan University, Kaifeng 475004, China.
College of Agriculture, Henan University, Kaifeng 475004, China.
J Fungi (Basel). 2025 Aug 14;11(8):591. doi: 10.3390/jof11080591.
, a fungal species of within the Pluteaceae family, is predominantly cultivated in southern China. Polysaccharides, the primary bioactive constituents of , exhibit diverse pharmacological activities. However, current cultivation practices face challenges due to the genetic heterogeneity of strains, leading to inconsistent content and compositional variability of polysaccharides and other functional components. ARTP, denoting atmospheric and room-temperature plasma, is a technology capable of generating plasma jets at ambient pressure with temperatures ranging from 25 to 40 °C. These jets feature high concentrations of highly reactive species, including but not limited to excited-state helium atoms, oxygen atoms, nitrogen atoms, and OH radicals. This study aims to develop high-yielding exopolysaccharide (EPS) strains through integrated ARTP mutagenesis and genome shuffling, thereby overcoming current cultivation bottlenecks. ARTP mutagenesis and genome shuffling significantly boosted EPS production in . ARTP generated nine stable mutants with >20% higher EPS yields. Subsequent genome shuffling (three rounds of protoplast fusion) produced the hybrid strain SL212, which achieved 46.85 g/L of EPS, an 111.67% increase over that of the parent strain under identical conditions. Metabolomics and transcriptomics analyses revealed that differential metabolites and genes were mainly enriched in galactose metabolism, ABC transporter pathways, and the tricarboxylic acid cycle. These pathways enhance monosaccharide biosynthesis and generate ATP, providing both precursors and energy for polysaccharide polymerization, thereby driving EPS overproduction. Preliminary mechanistic analysis identified the key contributing factors driving the elevated polysaccharide biosynthesis.
香菇是光茸菌科的一种真菌，主要在中国南方种植。香菇的主要生物活性成分多糖具有多种药理活性。然而，由于菌株的遗传异质性，目前的栽培实践面临挑战，导致多糖和其他功能成分的含量不一致和组成变异性。ARTP表示常压室温等离子体，是一种能够在环境压力下产生温度范围为25至40°C的等离子体射流的技术。这些射流具有高浓度的高活性物种，包括但不限于激发态氦原子、氧原子、氮原子和OH自由基。本研究旨在通过整合ARTP诱变和基因组改组来开发高产胞外多糖（EPS）菌株，从而克服当前的栽培瓶颈。ARTP诱变和基因组改组显著提高了香菇的EPS产量。ARTP产生了9个稳定的突变体，其EPS产量提高了20%以上。随后的基因组改组（三轮原生质体融合）产生了杂交菌株SL212，在相同条件下，其EPS产量达到46.85 g/L，比亲本菌株增加了111.67%。代谢组学和转录组学分析表明，差异代谢物和基因主要富集在半乳糖代谢、ABC转运蛋白途径和三羧酸循环中。这些途径增强了单糖生物合成并产生ATP，为多糖聚合提供前体和能量，从而推动EPS的过量生产。初步的机理分析确定了驱动多糖生物合成增加的关键因素。
