6-苄氨基腺嘌呤和萘乙酸对三裂叶野葛毛状根生长和异黄酮含量的影响

Effects of 6-benzylaminopurine and α-naphthaleneacetic acid on growth and isoflavone contents of Pueraria phaseoloides hairy roots

摘要:

为了探讨植物激素对三裂叶野葛毛状根生长和异黄酮化合物含量的影响，采用不同浓度的6-苄氨基腺嘌呤 (6-Benzylaminopurine, 6-BA) 或6-BA和萘乙酸 (α-Naphthaleneacetic, NAA) 结合处理三裂叶野葛毛状根，观察其对毛状根生长的影响，然后利用分光光度计测定毛状根中异黄酮化合物含量及抗氧化酶活性。结果表明：6-BA抑制三裂叶野葛毛状根的生长，降低三裂叶野葛毛状根的生物量, 且随着6-BA浓度的增加，其抑制效果愈明显;同时降低其总异黄酮化合物的含量。与对照相比，不同浓度6-BA和NAA 2.0 mg/L结合抑制毛状根的生长，降低毛状根中总异黄酮化合物含量。6-BA和NAA结合能显著提高毛状根可溶性蛋白质含量和过氧化物酶活性，但降低其超氧化物歧化酶活性；单独6-BA处理的毛状根培养至30 d时，可以检测到典型的DNA ladder带，而6-BA和NAA结合处理的毛状根培养至20 d时就可以检测到DNA ladder带，表明6-BA 或6-BA和NAA结合都可以促进细胞程序性死亡的发生，且NAA具有协同作用。

关键词:三裂叶野葛，毛状根，异黄酮化合物，6-苄氨基腺嘌呤，萘乙酸，超氧化物歧化酶，过氧化物酶，细胞程序性死亡

Abstract:

In order to study the effect of phytohormone on growth and isoflavones contents of Pueraria phaseoloides hairy roots, we cultured the hairy roots with different concentrations of 6-benzylaminopurine (6-BA) alone or in combination with α-naphthaleneacetic acid (NAA). Then we determined the effects of 6-BA alone or in combination with NAA on the growth and the contents of isoflavones compounds and levels of antioxidase activities of hairy roots by spectrophotometry. The results show that 6-BA inhibited the growth, and decreased biomass and total isoflavones compounds of P. phaseoloides hairy roots. Furthermore, the inhibition was increased with the concentrations of 6-BA. Compared with the controls, different concentrations of 6-BA in combination with NAA 2.0 mg/L could inhibit the growth of hairy roots and decrease the content of total isoflavone compounds, and also significantly enhanced the contents of soluble protein and levels of peroxidase (POD) activities, but decreased the activities of superoxide dismutase (SOD). DNA ladders detected by agarose gel electrophoresis can be observed after hairy roots of P. phaseoloides were cultured with 6-BA alone for 30 days, but can appear on the 20th day after culture with 6-BA in combination with NAA 2.0 mg/L. This result indicates that 6-BA or 6-BA in combination with NAA can both stimulate appearance of programmed cell death (PCD), and NAA may play a synergistic role on PCD.

Key words:Pueraria phaseoloides, hairy roots, isoflavones compounds, 6-benzylaminopurine, α-naphthaleneacetic acid, superoxide dismutase, peroxidase, programmed cell death