茉莉酸甲酯对茶条槭悬浮细胞没食子酸合成的影响

Effect of Methyl Jasmonate on the Biosynthesis of Gallic Acid in Acer ginnala Suspension Cell

为了研究茶条槭悬浮细胞中没食子酸的合成,该研究进行了茉莉酸甲酯诱导试验。通过添加茉莉酸甲酯,利用HPLC检测诱导后细胞中没食子酸的含量变化情况,同时利用电导仪、酸度计、分光光度计法和激光共聚焦显微镜对培养液和细胞进行电导率、pH值、苯丙氨酸解氨酶(PAL)活性以及细胞形态等进行分析。结果表明:(1)相对于正常培养的细胞,添加100μmol·L^-1的茉莉酸甲酯诱导24 h时,没食子酸的含量达到最高为12.49 mg·g^-1,其含量是对照的2倍左右。(2)茉莉酸甲酯的添加导致细胞培养液的pH值和电导率成波动趋势,细胞膜受损,通透性增大,细胞核分散,出现多个细胞核现象。(3)细胞内可溶性蛋白含量在诱导24 h、72 h和5 d时达到高峰,其含量分别是对照的1.4、1.67、2.07倍左右。(4)苯丙氨酸解氨酶活性在诱导24 h和5 d时分别出现一次高峰,其活性分别是对照的2倍和3.75倍。研究认为,茉莉酸甲酯处理短时间内促进了茶条槭细胞内没食子酸含量的积累,细胞内PAL活性和可溶性蛋白含量有所增加,对细胞液中的pH值和电导率影响不显著。

Our objective was to study the influence of methyl jasmonate(MJ)on biosynthesis of gallic acid(GA)in cell suspension of Acer ginnala.MJ was added into A.ginnala suspension and the cells were sampled at a series of time points after induction.The content of GA was determined by an HPLC method,the conductivity of the media was measured by a conductivity meter,the dissolvable protein content of the cells was determined by spectrophotometry and the cell shape was recorded by confocal microscopy.The results showed that:(1)induced by MJ 100μmol·L^-1 for 24 hours,the production of GA reached the peak(12.49 mg·g^-1)which is about twice of that of control.(2)The pH value and conductivity of the media were dynamic after induction by MJ.The cell membrane was damaged,with increased permeability.Few cells had several nuclei.(3)The soluable protein content of the cell had three peaks at 24 h,72 h and 5 d after induced,respectively,which are 1.4,1.67 and 2.07 times of those of control,respectively.(4)The activity of phenylalanine ammonialyase(PAL)reached the peak at 25 h and 5 d of post-induction,respectively,which were twice and 3.75 times of control,respectively.These results indicate that after short-time induction,the GA content is accumulated,the activity of PAL and the content of soluable proteins also increased,whereas the pH value and conductivity of the media did not alter significantly.