Designer TAL Effectors Induce Disease Susceptibility and Resistance to Xanthomonas oryzae pv. Oryzae in Rice  Ting Li, Sheng Huang, Junhui Zhou, Bing Yang  Molecular Plant  Volume 6, Issue 3, Pages 781-789 (May 2013) DOI: 10.1093/mp/sst034 Copyright © 2013 The Authors. All rights reserved. Terms and Conditions

Figure 1 Structure of a Typical Design TAL Effector (dTALE). The dTALEs consist of a scaffold of the repeat-less AvrXa10 and the custom-engineered TAL effector repeat domains. Each dTALE contains an N-terminal type III secretion signal (T3S signal), three functional nuclear localization motifs (NLS), and an acidic trans-activating domain (AD), and a varying number of repeats as each represented by the 12th and 13th amino acids (Variable repeats). Four types of 34-amino-acid repeats, each recognizing one of four nucleotides, are used to assemble the repeat domain of each dTALE. Molecular Plant 2013 6, 781-789DOI: (10.1093/mp/sst034) Copyright © 2013 The Authors. All rights reserved. Terms and Conditions

Figure 2 dTALE-xa27 Induced the Susceptible Allele of Xa27 and Triggered Resistance Responses in Rice IR24. APromoter sequence for dTALE-xa27 (xa27 EBE, shaded in tan) in IR24 and the TAL effector AvrXa27 binding element of Xa27 (Xa27 EBE, shaded in gray) in IRBB27. BDisease reactions of IR24 leaves to PXO99 strains containing or lacking dTALE-xa27. Hypersensitive reaction (HR) is indicated by tissue browning within 48 h compared to susceptible reaction (yellow coloration) at the inoculation sites. CPopulations of PXO99 and derivative in leaves of IR24. Populations are expressed as colony forming unit (cfu) per inoculation spot 4 d post inoculation. Lower-case letters above columns indicate significant differences at the <0.01 level using Tukey statistic and ANOVA analysis. Bars indicate 1 SD. DGene induction of xa27 in IR24. RNA from leaves treated with water, PXO99, and dTALE-xa27-containing PXO99 was used for quantitative RT–PCR with gene-specific primers and rice TFIIA gamma subunit gene in chromosome 5 used as an internal control. Molecular Plant 2013 6, 781-789DOI: (10.1093/mp/sst034) Copyright © 2013 The Authors. All rights reserved. Terms and Conditions

Figure 3 Four dTALEs Targeted Four Xa2-Like Genes in Nipponbare for Gene Activation and Disease Reaction. AFour dTALEs and their corresponding EBEs in the respective promoters of Xa27 homologs. BDisease reactions of Nipponbare leaves in response to PXO99 strains containing different dTALEs. (C–F) Gene induction of individual Xa27 homologs in response to different Xoo strains as detected by RT–PCR with gene-specific primers. The rice TFIIA gamma subunit gene was used as an internal control. Molecular Plant 2013 6, 781-789DOI: (10.1093/mp/sst034) Copyright © 2013 The Authors. All rights reserved. Terms and Conditions

Figure 4 dTALE11 Induced and Overcame Resistant Gene xa13. APromoter sequences of the allelic Os8N3 and xa13 with TAL effector binding elements (EBEs). PthXo1 recognizes Os8N3 (also named OsSWEET11 or Xa13) in susceptible rice IR24 but not xa13 in the IR24-derived near isogenic resistant line IRBB13. dTALE11 was designed to activate both alleles by recognizing the identical promoter sequences (underlined and overlapping with the TATA-box) in IR24 and IRBB13. BdTALE11 with RVDs and their corresponding target nucleotides (EBE). (C, D) Gene induction detected with RT–PCR. Leaf RNA of IR24 and IRBB13 from different treatments was subjected to quantitative RT–PCR with gene-specific primers. PCR for TFIIAγ5 was used as an internal control. ME2 is a pthXo1 knockout PXO99. EDisease severities (lesion lengths) caused by different Xoo strains in IR24 (gray bars) and IRBB13 (dark bars). Lesions were measured 10 d post inoculation. Error bars indicate ±SD. Molecular Plant 2013 6, 781-789DOI: (10.1093/mp/sst034) Copyright © 2013 The Authors. All rights reserved. Terms and Conditions

Figure 5 OsSWEET12 Induction Conferred Disease Susceptibility to Xoo. ATwo dTALE-binding elements (underlined) in the promoter of OsSWEET12. BRepeat units (RVDs) of two dTALEs and their EBEs. COsSWEET12 induction as detected by RT–PCR. Leaf RNA of rice Nipponbare treated with different Xoo strains was subjected to RT–PCR with gene-specific primers. RT–PCR with TFIIAγ5 was used as an internal control. DLesion lengths caused by Xoo in Nipponbare leaves. Lesions in 4-week-old plants were measured 10 d post inoculation. Error bars indicate ±SD. Molecular Plant 2013 6, 781-789DOI: (10.1093/mp/sst034) Copyright © 2013 The Authors. All rights reserved. Terms and Conditions