基于诺丽叶片愈伤组织的细胞悬浮系的建立  

Establishment of Cell Suspension Culture System Based on the Leaf Callus in Noni

作　　者：张正雪 蓝增全[1] 吴田 ZHANG Zheng-xue;LAN Zeng-quan;WU Tian(College of Environment Science and Engineering Department, Southwest Forestry University, Kunming 650224;College of Horticulture and Gardening, Southwest Forestry University, Kunming 650224)

机构地区：[1]西南林业大学环境科学与工程学院,昆明650224 [2]西南林业大学园林学院,昆明650224

出　　处：《生物技术通报》2018年第5期142-147,共6页Biotechnology Bulletin

基　　金：云南省应用基础研究计划项目(2016FB049);国家林业局推广项目([2015]27);国家星火计划(2014GA830017)

摘　　要：为通过诺丽叶片的细胞悬浮系来获得其次生代谢物。实验基于诱导的诺丽无菌苗叶片愈伤组织,改变液体培养基的激素组成、接种量以及在细胞悬浮培养过程中进行相关参数的测定以确定继代周期。液体培养基为MS+2.0 mg/L NAA+0.2 mg/L KT,最佳初始接种量为60 g/L;在14-22 d进入直线生长期,后缓慢增长,在第26天细胞数量达到峰值11.8×10~5个/mL后细胞数量下降;细胞活力最好出现在第21天,OD_(485)=1.8;初代细胞形态为不规则杆状细胞,在第6代呈规则小细胞团和圆球体;最佳继代周期22-26 d,诺丽叶片细胞悬浮培养液的细胞存活率为77.9%。实验建立了稳定的诺丽叶片细胞悬浮培养体系。This work aims to get the secondary metabolites from the cell suspension culture system of Morinda citrifolia L.（noni）leaves. In the experiment,the cell suspension system of noni leaf was established based on the callus induced from the sterilized leavesand determination of the subculture cycle by changing the collocation of hormone in liquid medium and inoculation amount of callus,andmeasuring some relevant parameter with the process of the culture. Liquid medium was MS＋2.0 mg/L NAA＋0.2 mg/L KT.,and the initialinoculation amount of callus was 60 g/L. During the 14 th-22 nd d,cells were in the linear growth phase,then grew slowly,and reached apeak of 11.8×10^5/mL at the 26 th day,and then declined. The best cell viability appeared at 21 st day when the OD（485）=1.8. The primary cell wasirregular rod-shaped and showed regular clusters of small cells and spheres when in the sixth generation. The best subculture cycle of noni cellsuspension culture system was 22-26 d,and the cell survival rate in the liquid suspension culture of noni leaves was 77.9%. In conclusion,astable cell suspension culture system of noni leaves is established in the experiment.

关 键 词：海巴戟 愈伤组织 细胞活力 生长曲线 

分 类 号：S667.9[农业科学—果树学]