茶树组培外植体灭菌技术优化
Optimisation of Sterilisation Techniques for Tissue Culture Explantation of Tea Plants
王兴雯 1袁连玉 1罗理勇 1曾亮2
作者信息
- 1. 西南大学食品科学学院，重庆400700
- 2. 西部（重庆）科学城种质创制科学中心
- 折叠
摘要
为建立\'福云6号\'茶树外植体高效无菌体系，解决组织培养中污染重、褐化高的技术瓶颈，本研究以带腋芽茎段和幼嫩叶片为材料，系统比较了二氯异氰脲酸钠（DCCNa）与次氯酸钠在不同浓度、处理时间及组合下的灭菌效果，并探讨了培养基中添加0.05%植物组培抗菌剂（PPM）的辅助作用。结果表明，DCCNa在较低浓度（5%）下即表现出优异的灭菌能力，组织毒性显著低于次氯酸钠。优化获得的最佳灭菌方案为：茎段经75%酒精处理60 s结合5% DCCNa处理15 min，污染率与褐变率均降至7.78%，存活率达84.44%；叶片经75%酒精处理30 s结合5% DCCNa处理12 min，污染率仅3.33%，褐变率7.50%，存活率高达89.17%。在最佳表面灭菌基础上，培养基中添加0.05% PPM可进一步降低内生菌污染，茎段存活率升至83.33%，叶片愈伤存活率升至87.78%，并促进组培苗生长。本研究成功建立了以DCCNa为核心的高效、低毒灭菌体系，为\'福云6号\'快速繁殖与遗传转化提供了关键技术支撑。
Abstract
To establish a highly efficient aseptic system for \'Fuyun No. 6\' tea plant explants and to address the technical bottlenecks of high contamination and browning rates in tissue culture, this study used stem segments with axillary buds and young leaves as materials to systematically compare the sterilisation efficacy of sodium dichloroisocyanurate (DCCNa) and sodium hypochlorite at different concentrations, treatment durations and combinations, and to investigate the auxiliary effect of adding 0.05% Plant Tissue Culture Antimicrobial (PPM) to the culture medium. The results indicated that DCCNa exhibited excellent sterilisation efficacy even at a relatively low concentration (5%), whilst its tissue toxicity was significantly lower than that of sodium hypochlorite. The optimised sterilisation protocol was as follows: stem segments treated with 75% ethanol for 60 seconds followed by 5% DCCNa for 15 minutes, resulting in contamination and browning rates both reduced to 7.78%, with a survival rate of 84.44%; for leaves, treatment with 75% ethanol for 30 seconds combined with 5% DCCNa for 12 minutes resulted in a contamination rate of only 3.33%, a browning rate of 7.50%, and a survival rate as high as 89.17%. Building upon the optimal surface sterilisation protocol, the addition of 0.05% PPM to the culture medium further reduced endogenous bacterial contamination, with stem segment survival rates rising to 83.33% and leaf callus survival rates increasing to 87.78%, whilst also promoting the growth of tissue-cultured seedlings. This study successfully established a highly efficient, low-toxicity sterilisation system centred on DCCNa, providing crucial technical support for the rapid propagation and genetic transformation of \'Fuyun No. 6\'.关键词
茶树组培/福云6号/二氯异氰脲酸钠/外植体灭菌Key words
tea tissue culture/Fuyun No. 6/sodium dichloroisocyanurate/explant sterilisation引用本文复制引用
王兴雯,袁连玉,罗理勇,曾亮.茶树组培外植体灭菌技术优化[EB/OL].(2026-05-09)[2026-05-11].http://www.paper.edu.cn/releasepaper/content/202605-21.学科分类
农业科学研究/农作物/园艺
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