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).