影响芸苔属植物(Brassica)组织培养过程中外植体褐化的因素

Factors to Browning Reaction of External Implant during Tissue Culture in Brassica

就油菜转基因过程中外植体褐化现象发生的可能原因进行了探讨。研究表明,转导的农杆菌菌株、菌液浓度、预培养和共培养时间对外植体发生褐化无直接影响,虽存在差异,但不显著。在培养基中加入抗氧化试剂PVP(聚乙烯吡咯啉酮)和Vc(抗坏血酸),PVP比Vc对褐化的抑制作用更大。培养温度对褐化的影响是显著的,当培养温度高于30℃时,外植体的褐化程度非常严重,褐化发生率(褐化外植体数/总外植体数)为100%;而当培养温度为25～26℃时,褐化率大大降低(17.6%),褐化程度也较轻。此外,光照时间和光照强度对褐化也有影响,光照时间过长、光照强度过大都使褐化现象变得严重。

In present paper the causes of browing reaction of enternal implant during transplantation were approached.The results showed the strain or strain conentration of transplanted Agrobacterium,and the time spent in preculture and culture had no direct effect to the browning reaction.Though there appeared differences between them,the differences were not so significant.When PVP and Vc were applied in medium,PVP played bigger effect to control the browning reaction than Vc.And the temperature played an important role to the reaction.When the temperature was over 30℃,very serious browing happened and the browning rate was 100%;When the temperature was in a range of 25～26℃,the reaction decreased remarkably to 17.6% and the browning degree was fairly low.In addition,light duration and strength affected browning.When light duration was too long or the strength too powerfull it would resulted serious browning reaction.