To system-wide identify the degradation substrates of SKIP14.A2, 35S::SKIP14.A2 transgenic ZS11 hairy roots were constructed, and the independently expressed mGFP (with the promoter of 2×35S) was used as a screening marker for positive transgenic materials. Hairy roots with GFP signals produced by the same cotyledon are used as "OE" materials, while hairy roots without GFP signals are used as "CK" materials. RT-PCR assay with these two groups showed that the expression of target gene in OE-1/2/3 sample was increased by 29.309, 73.164 and 28.573 times compared with CK1/2/3, respectively. Subsequently, OE and CK groups were carried out for ubiquitinome (4D-fastDIA) analysis.