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Diagnostic kit for rice blight resistance
Thursday, 2019/12/12 | 08:35:45

Joon-Seob EomDangping Luo, Genelou Atienza-GrandeJungil YangChonghui JiVan Thi LuuJosé C. Huguet-TapiaSi Nian CharBo LiuHanna NguyenSarah Maria SchmidtBoris SzurekCasiana Vera CruzFrank F. WhiteRicardo OlivaBing Yang and Wolf B. Frommer

Nat Biotechnol. 2019; 37(11): 1372–1379. Published online 2019 Oct 28. doi: 10.1038/s41587-019-0268-y

Abstract

Blight-resistant rice lines are the most effective solution for bacterial blight, caused by Xanthomonas oryzae pv. oryzae (Xoo). Key resistance mechanisms involve SWEET genes as susceptibility factors. Bacterial transcription activator-like (TAL) effectors bind to effector-binding elements (EBEs) in SWEET gene promoters and induce SWEET genes. EBE variants that cannot be recognized by TAL effectors abrogate induction, causing resistance. Here we describe a diagnostic kit to enable analysis of bacterial blight in the field and identification of suitable resistant lines. Specifically, we include a SWEET promoter database, RT–PCR primers for detecting SWEET induction, engineered reporter rice lines to visualize SWEET protein accumulation and knock-out rice lines to identify virulence mechanisms in bacterial isolates. We also developed CRISPR–Cas9 genome-edited Kitaake rice to evaluate the efficacy of EBE mutations in resistance, software to predict the optimal resistance gene set for a specific geographic region, and two resistant ‘mega’ rice lines that will empower farmers to plant lines that are most likely to resist rice blight.

 

See https://www.ncbi.nlm.nih.gov/pmc/articles/PMC6831515/

Fig. 1

SWEET11SWEET13SWEET14 EBE pentagon and PCR detection of SWEET induction.

a, Arrows indicate which TAL effectors can overcome a particular resistance mechanism by activating any of the other SWEET genes or by activating the same SWEET gene via targeting another EBE in the same promoter. For example, xa13-based resistance (resulting from a variant in the SWEET11 promoter that is not recognized by PthXo1) can be overcome by other TAL effectors (e.g., PthXo2 and PthXo3) that target the EBEs in another SWEET gene promoter or, in the case of SWEET14, target different EBEs in the same promoter. b, Example of SWEET gene induction as detected by RT–PCR using the SWEETup primer set. RT–PCR products are shown for the SWEET11SWEET13 and SWEET14 genes in Kitaake infected by Xoo strain ME2 lacking a SWEET-targeting TAL effector and ME2 transformed with plasmids encoding PthXo1 (targeting SWEET11), PthXo2 or PthXo2B (both targeting SWEET13) and PthXo3, TalC, TalF or AvrXa7 (all targeting SWEET14). Actin served as a control. Leaves were infected using leaf-clipping assays; scissors dipped in water served as an additional negative control. The experiment was repeated twice independently with similar results.

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