叶绿素等的测定方法（Determination of chlorophyll and other methods）.docVIP

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叶绿素等的测定方法（Determination of chlorophyll and other methods）.doc

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叶绿素等的测定方法（Determination of chlorophyll and other methods）

叶绿素等的测定方法（Determination of chlorophyll and other methods） 1 Determination of chlorophyll content: 80% acetone solution is prepared: 4L acetone + 1L distilled water. That leaves about 0.5g on a 50ml centrifuge tube (three repeats), adding 25ml concentration of acetone 80%, placed in the dark about extraction was removed after 36 hours, 4 times diluted respectively in the wavelength of 663nm, 645nm, 652nm and 470nm determination of optical density, with acetone for 80% blank. (refer to Chen Jianxun, editor of Wang Xiaofengs Guide to plant physiology experiment, P35 ~ 36) 95% ethanol solution can also be used at 665, 649 and 470nm to have the maximum absorption peak Ca=13.95D665-6.8649 Cb=24.96D649-7.32D665 Cx.c= (1000D470-2.05Ca-114Cb) /245 2 Determination of antioxidant enzyme activity (2.5G - like) 0.05mol/L phosphoric acid buffer solution (PBS) (pH=7.8) solution is prepared: 65.5506g, Na2HPO4, 12H2O + 2.65285g, NaH2PO4, 2H2O, constant volume to 4L. (refer to Chen Jianxun, editor in chief of Wang Xiaofengs experimental guide to plant physiology, P134). Fresh samples of cryopreservation, leaf about 2g (or root) in 50ml centrifuge tube, adding 20ml concentration of 0.05mol/L phosphate buffer solution (pH=7.8) (preferably in phosphate buffer solution, the cooler from grinding at high temperature to inactivate enzymes), grinding (by grinding mill), frozen the 8000r/min centrifuge centrifuge for 20 minutes, the supernatant for crude enzyme. 2.1 determination of malondialdehyde (MDA): 20%, three chloroacetic acid (TCA) solution preparation: called 200g three chloroacetic acid, with distilled water constant volume to 1000ml. (refer to Chen Jianxun, editor in chief of Wang Xiaofengs experimental guide to plant physiology, P124); 0.5% thiobarbituric acid (TBA) solution is prepared: 5g thiobarbituric acid (TBA), dissolved in 20%, three chloroacetic acid (TCA) solution, and fixed to 1000ml. (refer to Chen Jianxun, editor of Wang Xiaofengs plant physiology laboratory guide, P12