关键词: 油莎豆, 原生质体制备, 组织筛选, 酶解条件, 优化, 种质创新

Abstract: In order to obtain Cyperus esculentus protoplasts efficiently，the tissue parts including young shoots，young roots，young leaves，young leaf sheaths，tillering nodes and stolons were screened for protoplast preparation by single factor test method，and the enzymatic hydrolysis conditions such as enzyme combination dosage，enzymatic hydrolysis time，osmotic pressure stabilizer dosage and pH value of the enzyme solution were also optimized.The results showed that the best tissue part for protoplast preparation of Cyperus esculentus was young tillering node；and the optimal enzymatic hydrolysis conditions were as follows：the enzyme combination dosage was 2.0% cellulase+0.9% macerozyme，the enzymatic hydrolysis time was 6 h，the osmotic pressure stabilizer（mannitol）dosage was 11%，and the pH value of the enzyme solution was 6.0.Under these conditions，0.2 g of Cyperus esculentus tillering nodes were added to 4 mL of enzyme solution，and then the protoplast yield could reach 22.91×10⁵ per gram and the protoplast activity could reach 90.72%.In this study，an efficient method for protoplast preparation of Cyperus esculentus was obtained，which could lay a foundation for subsequent protoplast fusion，germplasm innovation and hybrid breeding of Cyperus esculentus.

Key words: Cyperus esculentus, Protoplast preparation, Tissue screening, Enzymatic hydrolysis conditions, Optimization, Germplasm innovation