School of Biotechnology, Jiangsu University of Science and Technology, Zhenjiang 212100, China.
School of Biotechnology, Jiangsu University of Science and Technology, Zhenjiang 212100, China.
J Biotechnol. 2024 May 10;386:64-71. doi: 10.1016/j.jbiotec.2024.03.008. Epub 2024 Mar 20.
With the world's population rapidly increasing, the demand for high-quality protein is on the rise. Edible fungi breeding technology stands as a crucial avenue to obtain strains with high yield, high-quality protein, and robust stress resistance. To address the protein supply gap, Atmospheric and Room Temperature Plasma (ARTP) mutagenesis, and spore hybridization techniques were employed to enhance Pleurotus djamor mycelium protein production. Beginning with the original strain Pleurotus djamor JD-1, ARTP was utilized to mutate spore suspension. The optimal treatment time for Pleurotus djamor spores, determined to achieve optimal mortality, was 240 s. Through primary and secondary screenings, 6 mutant strains out of 39 were selected, exhibiting improved protein yield and growth rates compared to the original strain. Among these mutagenic strains, 240S-4 showcased the highest performance, with a mycelial growth rate of 9.5±0.71 mm/d, a biomass of 21.45±0.54 g/L, a protein content of 28.75±0.92%, and a remarkable protein promotion rate of 128.03±7.29%. Additionally, employing spore hybridization and breeding, 7 single-nuclei strains were selected for pin-two hybridization, resulting in 21 hybrid strains. The biomass and protein content of 9 hybrid strains surpassed those of the original strains. One hybrid strain, H-5, exhibited remarkable mycelial protein production, boasting a mycelial growth rate of 26.5±0.7 mm/d, a biomass of 21.70±0.46 g/L, a protein content of 28.44±0.22%, and a protein promotion rate of 128.02±1.73%. Notably, both strains demonstrated about a 28% higher mycelial protein yield than the original strains, indicating comparable effectiveness between hybrid breeding and mutagenesis breeding. Finally, we analyzed the original and selected strains by molecular biological identification, which further proved the effectiveness of the breeding method. These findings present novel insights and serve as a reference for enhancing edible fungi breeding, offering promising avenues to meet the escalating protein demand.
随着世界人口的快速增长，对高质量蛋白质的需求也在不断上升。食用真菌养殖技术是获得高产、高质量蛋白质和强抗逆性菌株的关键途径。为了解决蛋白质供应缺口问题，采用大气室温等离子体（ARTP）诱变和孢子杂交技术来提高糙皮侧耳菌丝体蛋白质的生产。从原始菌株糙皮侧耳 JD-1 开始，使用 ARTP 对孢子悬浮液进行诱变。确定糙皮侧耳孢子最佳处理时间以达到最佳死亡率为 240s。通过初筛和复筛，从 39 个突变株中筛选出 6 个突变株，与原始菌株相比，这些突变株的蛋白质产量和生长速率都有所提高。在这些诱变菌株中，240S-4 的表现最为出色，菌丝生长速度为 9.5±0.71mm/d，生物量为 21.45±0.54g/L，蛋白质含量为 28.75±0.92%，蛋白质促进率高达 128.03±7.29%。此外，通过孢子杂交和选育，选择了 7 个单核菌株进行双核杂交，得到了 21 个杂交菌株。9 个杂交菌株的生物量和蛋白质含量均超过了原始菌株。其中一个杂交菌株 H-5 的菌丝体蛋白产量显著，其菌丝生长速度为 26.5±0.7mm/d，生物量为 21.70±0.46g/L，蛋白质含量为 28.44±0.22%，蛋白质促进率为 128.02±1.73%。值得注意的是，这两个菌株的菌丝体蛋白产量比原始菌株高出约 28%，表明杂交选育和诱变选育的效果相当。最后，我们通过分子生物学鉴定对原始菌株和选育菌株进行了分析，进一步证明了选育方法的有效性。这些发现为提高食用真菌的选育提供了新的思路和参考，为满足不断增长的蛋白质需求提供了有前景的途径。
