File:图片gus.png

AvrXa7 Drives TAL Effector-Specific Host Gene Induction. (A) Promoter fragments used in the transient expression assay. The consensus effector binding elements are underlined. The nonconsensus changes introduced in the mutant versions are in lowercase letters. Os-8N3pBB13 contains the insertion/deletion of xa13 allele in IRBB13, and sequence is not shown. The first 10 (Os-11N3) or 12 (Os-8N3) bases upstream of the EBE and between the EBE region and the ATG are not shown. Each fragment was fused to the ATG of the uidA coding sequence. (B) GUS assay on N. benthamiana using 35S-avrXa7 (left sites) or 35S-pthXo1 (right sites). Sites are stained with X-gluc. Coinoculations with 35SpthXo3 and combinations with promoter fragment Os-8N3pBB13 (7) are not shown. The T-DNA vector containing 35S-avrXa7 or 35S-pthXo1 alone was infiltrated at sites indicated with an asterisk. (C) Average GUS activity was calculated from triplicate coinoculations of the promoter fragments in (A) with 35S-pthXo1, 35S-avrXa7, or 35S-pthXo3. The numbers for each treatment with the indicated plant-expressed effector corresponds to the promoter fragments in (A). Average GUS activity was measured from three excised leaf disks with hybrid promoters as indicated in (A) coinfiltrated with 35S-pthXo1 (black columns), 35S-avrXa7 (gray columns), or 35S-pthXo3 (white). Activity on MUG substrate is expressed as pmol of 4-methylumbelliferone (4-MU) per mg of protein.