Over-Expression of a Novel Cry1ie Gene Confers Resistance to Cry1ac-Resistantcotton Bollworm

Over-expression of a novel Cry1Ie gene confers resistance to Cry1Ac-resistantcotton bollworm in transgenic lines of maize

Yuwen Zhang1,2#, Yunjun Liu2#, Yuan Ren1, Yan Liu2, Gemei Liang3, Fuping Song3, Shuxiong Bai3, Jianhua Wang1*, Guoying Wang2*

Supplementary material

Supplemental Fig. 1 Southern blot analysis of IE1103 and IE012

(A) T-DNA cassette of plasmid p3301UbiIe. LB and RB, left and right border of T-DNA; 35S ployA, 35S terminator; Bar, phosphinothricin acetyltransferase gene; 35S, CaMV 35S promoter; Ubiquitin, maize ubiquitin promoter; Cry1Ie, Cry1Ie gene; Nos, Nos terminator. The probe fragment for Southern blot is noted by the line.

(B)Southern blot analysis. M, Fermentas 1kb DNA marker; 1, plasmid p3301UbiIe digested with HindIII; 2, non-transgenic maize Z31 genomic DNA digested with HindIII; 3, IE034 genomic DNA digested with HindIII; 4, IE1103 genomic DNA digested with HindIII; 5, IE012 genomic DNA digested with HindIII.

Supplemental Fig.2 Southern blot analysis of the transgenic plants containing Cry1Ac

(A)T-DNA cassette of plasmid p3301UbiAc. LB and RB, left and right border of T-DNA; 35S ployA, 35S terminator; Bar, phosphinothricin acetyltransferase gene; 35S, CaMV 35S promoter; Ubiquitin, maize ubiquitin promoter; Cry1Ac, Cry1Ac gene; Nos, Nos terminator. The probe fragment for Southern blot is noted by the line.

(B)Southern blot analysis. 1, plasmid p3301UbiAc digested with HindIII; 2, non-transgenic maize Z31 genomic DNA digestedwithHindIII ; 3, transgenic maize AC400 genomic DNA digested withHindIII; 4, transgenic maize AC146 genomic DNA digested by HindIII; 5, non-transgenic maize Z31 genomic DNA digested by KpnI;6, transgenic maize AC400 genomic DNA digested by KpnI ; 7, transgenic maize AC146 genomic DNA digested by KpnI.

Supplemental Fig.3 PCR identification of T3 (A) and T4 (B) generation plants of IE034

(A) M, DL2000 plus DNA marker; 1-18, different T3 generation transgenic plants; 19, non-transgenic maize Z31 as negative control; 20, water as blank control; 21, plasmid p3301UbiIe as positive control.

(B) M, DL2000 plus DNA marker; 1, water as blank control; 2, plasmid p3301UbiIe as positive control; 3,non-transgenic maize Z31 as negative control; 4-13, different T4generation transgenic plants.

Supplemental Fig. 4The leaf damage of transgenic plants after infested with Asian corn borer in the field.