Isolation and purification of mesophyll protoplasts from the leaves of Camellia sinensis
Liu Yan-Li1, Jin Xiao-Fang1, Ma Lin-Long1, Cao Dan1, Gong Zi-Ming1, Wei Chao-Ling2
1. Institute of Fruit and Tea, Hubei Academy of Agricultural Science, Wuhan 430209, China;
2. State Key Laboratory of Tea Plant Biology and Utilization, Anhui Agricultural University, Hefei 230036, China
In the present study, the effect of mannitol concentration, combination of different enzyme (cellulase and macerozyme) solutions, and enzymolysis time on the separation of mesophyll protoplasts and purification of protoplasts from two high density separating solutions were studied in the leaves of tea seedlings. The results demonstrated that high quality mesophyll protoplasts were obtained from tea leaves subjected to enzymatic digestion in a solution containing 0.6 mol/L mannitol, 1.5% cellulase, and 2.5% macerozyme for 16-18 h in the dark. In addition, protoplasts gathered into a band on the layer between 10% mannitol and 25% iodixanol.
刘艳丽, 金孝芳, 马林龙, 曹丹, 龚自明, 韦朝领. 茶树叶肉原生质体的分离与纯化[J]. 植物科学学报, 2017, 35(6): 908-911.
Liu Yan-Li, Jin Xiao-Fang, Ma Lin-Long, Cao Dan, Gong Zi-Ming, Wei Chao-Ling. Isolation and purification of mesophyll protoplasts from the leaves of Camellia sinensis. Plant Science Journal, 2017, 35(6): 908-911.
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