牛筋草原生质体分离条件的研究

打开文本图片集

中图分类号：Q943.1 文献标志码：A 文章编号：1003-935X（2025）02-0031-08

Isolation Conditions of Protoplasts in Eleusine indica

CUI Xinyu 1，2 ， JIANG Hao 1，2 ， TIAN Xingshan 1 ， ZHANG Chun1 （1.Plant Protection Research Institute，Guangdong Academy of Agricultural Sciences/Key Laboratory of New Techniques for Plant Protection in Guangdong，Guangzhou 51O640，China; 2.School of Life Sciences，South China Normal University，Guangzhou 51O631，China）

Abstract：The isolation and cultivation of high-viability plant protoplastsserveas the foundation for studying plant physiology，biochemistry，andmolecularbiologyatthecellularlevel.However，duetothepresenceofacellwall surrounding plant protoplasts，thereare numerouschallnges in their isolationandculture.Inthisstudy，usingleaves andsheaths of the globall invasive weed Eleusine indicaas materials，single-factor experiments were conducted to optimize the conditions for protoplast preparation from E .indica，including material quantity，enzyme solution composition，enzymatic hydrolysis time，and centrifugation speed.High-activity protoplastsof E .indica were successfully obtained.Specifically，2 gof leaves from E .indicaplants at 5-6 leaf-stage were enzymatically hydrolyzed for 3 hours in a 10mL enzyme solution containing 1.0% cellulase， 0.5% macerozyme，and 0.5% pectinase.The protoplasts were then collected by centrifugation at 2 500r/min .The maximum protoplast yield reached 2.075× （204号 （204 107 protoplasts/ g ， with over 90% viability.Furthermore，the yield of protoplasts prepared from leaves was higher than that from sheaths. This study established an optimized method for preparing protoplasts from E .indica leaves，laying the groundwork for studying the herbicide resistance mechanisms of E .indica at the single-cell level and providing a reference for the preparation of protoplasts from other grass weeds.

Key words： Eleusine indica ; protoplast; enzymatic hydrolysis ;single - cell sequencing

近年来，单细胞测序技术（scRNA-Seq）在生物医学领域得到广泛应用，但植物单细胞测序进展缓慢，难点之一是植物原生质体的分离与稳定培养。（剩余10962字）