Molecular Identification of Transgenic hrpZPsta Gene Soybean Lines and Analysis of Resistance to Phytophthora Root Rot
T7 and T8generation transgenic hrpZPsta gene soybean line JN29-705-21 was used as the test material,and disease resistance analysis was performed by molecular identification and indoor artificial inoculation of phytophthora infestans.The results of routine PCR showed that the target gene hrpZPsta,screening marker Badh,promoter 35S and terminator Nos were detected in T7 and T8 transgenic lines,which indicated that hrpZPsta gene could be inherited stably in high generation transgenic lines.Southern hybridization results showed that the hrpZPsta gene was integrated into the soybean genome as a single copy with different integration sites;the inspection results of qRT-PCR showed that hrpZPsta gene was expressed in root,stem,and leaf of the transformed lines,among which the highest expression was in leaf and the lowest in stem.The average relative expression of root,stem,leaf in the T7 generation lines was 2.800,1.772 and 8.367,respectively;and the average rela-tive expression of root,stem,and leaf in the T8 generation lines was 3.028,1.650 and 9.036,respec-tively.The hypocotyl infection method was used to analyze the disease resistance of the transformed lines.The results showed that the resistance level of the offspring of the transgenic lines to phytoph-thora root rot was positive,but the resistance of the non-transgenic recipient lines was medium,indi-cating that the disease resistance of transgenic lines have improved.
万方数据
