一种测定巴西橡胶树树皮中过氧化氢含量的方法

An Improved Technique for Determination of Hydrogen Peroxide in Bark of Rubber Tree(Hevea brasiliensis Muell. Arg.)

研究植物体内H_2O_2的含量变化有助于确认植物提高非生物胁迫耐性的生物技术策略。Ti(Ⅳ)-PAR-H_2O_2比色法灵敏度高、特异性强、稳定性好,广泛用于植物H_2O_2测定,但不同研究所用的反应条件不一致。在本研究中,系统优化了Ti(Ⅳ)-PAR-H_2O_2比色法的反应条件,并比较了丙酮提取法和TCA提取法对橡胶树萌条韧皮部H_2O_2的提取效果。结果表明,丙酮提取法稳定性差,TCA提取法稳定性好。本研究报道了一种简单、快速的测定橡胶树树皮H_2O_2的方法,为进一步研究橡胶树中H_2O_2的生理功能打下良好基础。

Investigation of the H2O2 level induced by abiotic stress will be valuable for identifying biotechnological strategies to improve abiotic stress tolerance in crop plants.The Ti (Ⅳ)-PAR-H2O2 spectrophotometric method for determination of H2O2 was found to give good sensitivity,specificity and stability,and it has beenwidely used for estimation of H2O2 in plant extracts.However,various reaction conditions of this method were used by different authors.This led us to optimize the suitable condition of this method.For sample preparation,extraction with CCl4/CH3Cl3 (3 :1, V :V)and water in the proportion of 1 :3 :5 (V :V :V)after extracted with acetone were compared with the 5% the TCA extract method without activated charcoal in the bark of rubber trees.The 5%TCA method gives more stability than acetone method.In this paper,we described an easy and rapid protocol for determination of H2O2 in rubber tree bark using the Ti (Ⅳ)-PAR-H2O2 spectrophotometric method.Results could provide a basis for the investigation of the physiological function of H2O2 in rubber trees.