Cai Ruihang, Xiao Yu, Xing Jiajia, Yu Kongjian, Li Xiaola, Chai Yiqiu
Zhejiang Institute of Subtropical Crops, Zhejiang Academy of Agricultural Sciences, Wenzhou, China.
State Key Laboratory of Rice Biology and Ministry of Agriculture Key Laboratory of Molecular Biology of Crop Pathogens and Insects, Institute of Biotechnology, Zhejiang University, Hangzhou, China.
Front Microbiol. 2024 Jun 27;15:1333793. doi: 10.3389/fmicb.2024.1333793. eCollection 2024.
is a well-known edible and medicinal mushroom with a long history of use in China, and it contains a variety of secondary metabolites with interesting bioactive ingredients. However, recent researches have mainly focused on cultivation conditions, secondary metabolite compositions and pharmacological activities of , the lack of an efficient and stable genetic transformation system has largely limited further research on the relationship between secondary metabolites and biosynthetic gene clusters in . In this study, single-factor experiments were used to compare the effects of different osmotic stabilizers, enzyme concentrations and enzyme digestion times on protoplast yield, and we found that the highest yield of 5.53 × 10 protoplasts/mL was obtained with 0.7 M mannitol, 6 mg/mL snail enzyme and 4 h of enzyme digestion time, and the regeneration rate of protoplasts was up to approximately 30% using 0.7 M mannitol as an osmotic stabilizer. On this basis, a PEG-mediated genetic transformation system of was successfully established for the first time, which lays the foundation for further genetic transformation of .
是一种在中国有着悠久使用历史的著名食药用菌，它含有多种具有有趣生物活性成分的次生代谢产物。然而，最近的研究主要集中在其栽培条件、次生代谢产物组成和药理活性方面，缺乏高效稳定的遗传转化系统在很大程度上限制了对其次生代谢产物与生物合成基因簇之间关系的进一步研究。在本研究中，采用单因素实验比较了不同渗透稳定剂、酶浓度和酶解时间对原生质体产量的影响，我们发现使用0.7 M甘露醇、6 mg/mL蜗牛酶和4 h酶解时间可获得最高产量为5.53×10个原生质体/mL，并且以0.7 M甘露醇作为渗透稳定剂时原生质体的再生率高达约30%。在此基础上，首次成功建立了的PEG介导的遗传转化系统，这为的进一步遗传转化奠定了基础。
