Virus-induced gene silencing (VIGS) in B. napus
The CDS of BnGSTU12 was cloned into pTRV2 digested with BamHI and XbaI for the construction of recombinant vector, pTRV2-BnGSTU12 . The empty vector pTRV2 and pTRV2-BnGSTU12 together with pTRV1 were introduced into AgrobacteriumGV3101. TheAgrobacterium strain suspended in the solution containing 10mM MES, 10mM MgCl2 and 20μM AS, and then injected into the leaves of four-leaf stage B. napus.Ten days after infiltration, the expression of BnGSTU12 was detected using qRT-PCR.
Measurement ofH2O2content, GST, POD, SOD and CAT enzyme activity
Hydrogen peroxide assay kit was purchased from the Grace Biotechnology Institute (Suzhou, China), and H2O2 content was measured according the to the manual. Peroxidase (POD), superoxide dimutase (SOD), catalase activities (CAT) and GST enzyme activity were measured with the commercial kit (Nanjin Jiancheng Bioengineering Institute).