牛筋草原生質(zhì)體分離條件的研究

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中圖分類號：Q943.1 文獻(xiàn)標(biāo)志碼：A 文章編號：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號 （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

近年來，單細(xì)胞測序技術(shù)（scRNA-Seq）在生物醫(yī)學(xué)領(lǐng)域得到廣泛應(yīng)用，但植物單細(xì)胞測序進(jìn)展緩慢，難點之一是植物原生質(zhì)體的分離與穩(wěn)定培養(yǎng)。（剩余10962字）