白檀种子胚愈伤组织诱导过程中褐化影响因子研究

Investigation of Factors Influencing Browning During Callus Induction in Symplocos paniculata Seed Embryos

为探究白檀种子贮藏及愈伤组织诱导过程中种子胚组织褐化的影响因子,以白檀种子为材料,测定其贮藏过程中的生理变化,并研究不同处理对白檀种胚组织培养褐化率、转化率及污染率的影响。结果表明,白檀种子采收后,4℃贮藏14 d多酚氧化酶(PPO)活性与丙二醛(MDA)含量均最低,此时进行组织培养种胚褐化率低。白檀种子组织培养在高pH(6.3)和低pH(4.8)下的褐化率高,在pH值5.8时褐化率最低。组织培养接种时去除胚乳,培养基pH值5.3,使用M_(蔗糖)∶M_(葡萄糖)=1∶1混合,碳源浓度为30 g/L,NAA浓度为0.10 mg/L,能获得大量生长状况良好、分化能力较强的愈伤组织。

To investigate the factors influencing seed embryo tissue browning of Symplocos paniculata Thunb.Miq.during storage and callus induction,the physiological changes of seeds during storage were determined and the effects of different treatments on subsequent tissue culture were inverstigated,including browning rate,induction rate,and pollution rate.Test results showed that the activity of polyphenol oxidase(PPO)and content of malonaldehyde(MDA)were at their lowest levels after storing S.paniculata seeds at 4℃for 14 days,and the browning rate of seed embryo tissue culture during this period was lowest.The high browning rate of S.paniculata seeds embryo tissue culture was found at both high(6.3)and low(4.8)pH levels,while the lowest browning rate was observed at pH of 5.8.During tissue culture inoculation,the endosperm was excised and the culture medium was adjusted to a pH of 5.3 with a sucrose-glucose mixture(1∶1)at a carbon concentration of 30 g/L.The addition of NAA at a concentration of 0.10 mg/L resulted in the production of numerous calli exhibiting robust growth and strong differentiation potential.