Table 1

PCR primers

Fig. 1

Construction of genome editing vectorNote: Design four primers containing TD1 and TD2 fragments, use pCBC-TD1TD2 vector as a template to synthesize sgRNA fragments, and clone into pSKE401 vector through the Bsa Ιrestriction sites on TD1 and TD2 to obtain a dual-target genome editing vector G2-TD1TD2. LB and RB are the left and right borders of Agrobacterium T-DNA, respectively; U6-26p, U6-29p are the promoters of Arabidopsis U6 gene, U6-26t is the transcription terminator; 35Sp-Cas9-NosT and 35Sp -Kana-PolyA are Cas9 and Kana gene expression boxes, respectively

Table 2

Amino acid composition of BnGRP1 protein

Fig. 2

Alignment of genomic sequence, CDS sequence and homologous sequence of BnGRP1 and the selection of CRISPR target sitesNote: T1 and T2 indicate the target sequences of genome editing; PAM sequences are underlined with blue boxes

Fig. 3

Agrobacterium mediated genetic transformation of G2-TD1TD2 in Brassica napus Westar and PCR identification of regenerated seedlings of genetic transformationNote: A: The callus formed by dedifferentiation of explants; B: The adventitious buds formed by callus redifferentiation; C: Regenerated seedlings in rooting medium; D: Regenerated rape seedlings transplanted into nutrient soil; E: PCR identification of regenerated seedlings of genetic transformation

Fig. 4

Variation analysis of BnGRP1 gene sequences in different transgenic plantsNote: The boxes marked the sequence variation sites, and boxes of the same color are marked the sequence variation in the same plant

Fig. 5

Protein sequences of Bngrp1 in different transgenic plantsNote: Red letter show the variation of protein sequences