Box-Behnken设计-响应面法优化人参干细胞培养体系  

Establishment of Panax ginseng cambial meristematic cell culture by Box-Behnken

作　　者：金司阳 刘青 卢伊娜 JIN Si-yang;LIU Qing;LU Yi-na(Technology and Innovation Center,Shanghai JAKA Biotech Co.,Ltd.Shanghai 201506,China)

机构地区：[1]上海珈凯生物股份有限公司技术创新中心,上海201506

出　　处：《生物技术》2023年第2期239-247,225,共10页Biotechnology

摘　　要：[目的]研究人参形成层干细胞(Cambial Meristematic Cell,CMCs)培养体系的构建及相关工艺优化。[方法]通过Box-Behnken设计-响应面法优化人参CMCs培养相关工艺。[结果]从人参形成层中分离得到人参形成层细胞CMCs,诱导分离的最佳培养条件为:B5+IAA 6 mg/L+IBA 6 mg/L+蔗糖20 g/L+琼脂粉7 g/L+L-抗坏血酸100 mg/L+柠檬酸150 mg/L,诱导率为50.54%;固体增殖的最佳培养条件为:B5+IAA 2.9 mg/L+IBA 2.6 mg/L+NAA 5.9 mg/L+蔗糖20 g/L+L-抗坏血酸100 mg/L+柠檬酸150 mg/L,增殖量为1.01 g;悬浮培养的最佳培养条件为:B5+IAA 5 mg/L+IBA 5 mg/L+NAA 5 mg/L+蔗糖20 g/L+L-抗坏血酸100 mg/L+柠檬酸150 mg/L,pH 6.2,增殖量为3.05 g;均为24.6℃暗培养。[结论]人参CMCs具备干细胞特征,Box-Behnken设计-响应面法适用于人参CMCs培养工艺的优化。[Objective] To optimize Panax ginseng cambial meristematic cells(CMCs) culture were established and optimized.[Method]The Box-Behnken based optimization was used to isolate and culture of P.ginseng cambial meristematic cell.[Result]P.ginseng CMCs were obtained from P.ginseng cambium.The optimal conditions for induction were B5+IAA 6 mg/L+IBA 6 mg/L+ sucrose 20 g/L+agar powder 7 g/L+L-ascorbic acid 100 mg/L+citric acid 150 mg/L,the induction rate was 50.54%.The solid proliferation was B5+IAA 2.9 mg/L+IBA 2.6 mg/L+NAA 5.9 mg/L+sucrose 20 g/L+L-ascorbic acid 100 mg/L+citric acid 150 mg/L,the proliferation was 1.01 g.And suspension culture was B5+IAA 5 mg/L+IBA 5 mg/L+NAA 5 mg/L+sucrose 20 g/L+L-ascorbic acid 100 mg/L+citric acid 150 mg/L,pH 6.2,the proliferation was 3.05 g.Both culture temperature was 24.6 ℃,dark culture.[Conclusion]P.ginseng CMCs culture was a stable culture system.The Box-Behnken could use in P.ginseng CMCs optimization.

关 键 词：人参 形成层干细胞 植物干细胞 Box-Behnken 

分 类 号：Q945[生物学—植物学]