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DAB and Evans Blue Staining and Measurement of Enzymatic Activity Related Parameters

YC Yongrun Cao
YZ Yue Zhang
YC Yuyu Chen
NY Ning Yu
SL Shah Liaqat
WW Weixun Wu
DC Daibo Chen
SC Shihua Cheng
XW Xinghua Wei
LC Liyong Cao
YZ Yingxin Zhang
QL Qunen Liu
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Top leaves were collected at heading stage and immediately submerged in 10% SDS. After 10 min, the leaves were subsequently submerged in a 1 mg/mL solution of DAB or 0.5 mg/mL solution of Evans Blue incubated for 8 h in the dark at room temperature, respectively. Samples were then decolored in 95% boiling ethanol for 30 min and soaked for 48 h in 95% ethanol until all of the chlorophyll had been removed. The cleared leaves were then photographed.

Malondialdehyde (MDA) content, Peroxidase (POD), Super Oxide Dismutase (SOD), Catalase (CAT) activity, Soluble Protein (SP) and H202 content of wild type and ltn-212 top leaves were measured from heading stage plants and using kits from Nanjing Jiancheng Bioengineering Research Institute.

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