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Mutations in ORP1 Conferring Oxathiapiprolin Resistance Confirmed by Genome Editing using CRISPR/Cas9 in Phytophthora capsici and P. sojae.
Phytopathology. 2018 Dec; 108(12):1412-1419.P

Abstract

Oxathiapiprolin is a novel fungicide that was recently registered in a number of countries to control plant-pathogenic oomycetes such as Phytophthora capsici. In our previous study, point mutations G770V and G839W in oxysterol binding protein-related protein 1 (ORP1) were detected in oxathiapiprolin-resistant P. capsici isolates (PcORP1). Here, we used the CRISPR/Cas9 system to verify the effects of these two point mutations on P. capsici phenotypes. Transformants containing heterozygous G770V and G839W mutations in PcORP1 showed high levels of oxathiapiprolin resistance. The G770V transformants showed otherwise similar phenotypes compared with the wild-type isolate BYA5, including sporangia and zoospore production, cyst germination, and pathogenicity. However, two independent transformants with heterozygous G839W mutations in PcORP1 could not produce sporangia. Three transformants with an unexpected point mutation in PcORP1 (ΔN837) showed high oxathiapiprolin resistance, and either similar or significantly reduced fitness compared with BYA5. The same deletion (ΔN837) was confirmed to confer oxathiapiprolin resistance in P. sojae by using CRISPR/Cas9. These homozygous P. sojae mutants also showed either similar or strongly reduced fitness compared with the wild-type parent isolate P6497. These results improve our understanding of oxathiapiprolin resistance in Phytophthora spp., and will be useful for the development of novel oxysterol-binding protein homolog inhibitor fungicides.

Authors+Show Affiliations

First and fifth authors: State Key Laboratory of Crop Stress Biology for Arid Areas, College of Plant Protection, Northwest A&F University, Yangling 712100, China; first, second, third, and fifth authors: Department of Plant Pathology, College of Plant Protection, China Agricultural University, Beijing 100193, China; and first and fourth authors: Center for Genome Research and Biocomputing and Department of Botany and Plant Pathology, Oregon State University, Corvallis 97331.First and fifth authors: State Key Laboratory of Crop Stress Biology for Arid Areas, College of Plant Protection, Northwest A&F University, Yangling 712100, China; first, second, third, and fifth authors: Department of Plant Pathology, College of Plant Protection, China Agricultural University, Beijing 100193, China; and first and fourth authors: Center for Genome Research and Biocomputing and Department of Botany and Plant Pathology, Oregon State University, Corvallis 97331.First and fifth authors: State Key Laboratory of Crop Stress Biology for Arid Areas, College of Plant Protection, Northwest A&F University, Yangling 712100, China; first, second, third, and fifth authors: Department of Plant Pathology, College of Plant Protection, China Agricultural University, Beijing 100193, China; and first and fourth authors: Center for Genome Research and Biocomputing and Department of Botany and Plant Pathology, Oregon State University, Corvallis 97331.First and fifth authors: State Key Laboratory of Crop Stress Biology for Arid Areas, College of Plant Protection, Northwest A&F University, Yangling 712100, China; first, second, third, and fifth authors: Department of Plant Pathology, College of Plant Protection, China Agricultural University, Beijing 100193, China; and first and fourth authors: Center for Genome Research and Biocomputing and Department of Botany and Plant Pathology, Oregon State University, Corvallis 97331.First and fifth authors: State Key Laboratory of Crop Stress Biology for Arid Areas, College of Plant Protection, Northwest A&F University, Yangling 712100, China; first, second, third, and fifth authors: Department of Plant Pathology, College of Plant Protection, China Agricultural University, Beijing 100193, China; and first and fourth authors: Center for Genome Research and Biocomputing and Department of Botany and Plant Pathology, Oregon State University, Corvallis 97331.

Pub Type(s)

Journal Article
Research Support, Non-U.S. Gov't

Language

eng

PubMed ID

29979095

Citation

Miao, Jianqiang, et al. "Mutations in ORP1 Conferring Oxathiapiprolin Resistance Confirmed By Genome Editing Using CRISPR/Cas9 in Phytophthora Capsici and P. Sojae." Phytopathology, vol. 108, no. 12, 2018, pp. 1412-1419.
Miao J, Chi Y, Lin D, et al. Mutations in ORP1 Conferring Oxathiapiprolin Resistance Confirmed by Genome Editing using CRISPR/Cas9 in Phytophthora capsici and P. sojae. Phytopathology. 2018;108(12):1412-1419.
Miao, J., Chi, Y., Lin, D., Tyler, B. M., & Liu, X. (2018). Mutations in ORP1 Conferring Oxathiapiprolin Resistance Confirmed by Genome Editing using CRISPR/Cas9 in Phytophthora capsici and P. sojae. Phytopathology, 108(12), 1412-1419. https://doi.org/10.1094/PHYTO-01-18-0010-R
Miao J, et al. Mutations in ORP1 Conferring Oxathiapiprolin Resistance Confirmed By Genome Editing Using CRISPR/Cas9 in Phytophthora Capsici and P. Sojae. Phytopathology. 2018;108(12):1412-1419. PubMed PMID: 29979095.
* Article titles in AMA citation format should be in sentence-case
TY - JOUR T1 - Mutations in ORP1 Conferring Oxathiapiprolin Resistance Confirmed by Genome Editing using CRISPR/Cas9 in Phytophthora capsici and P. sojae. AU - Miao,Jianqiang, AU - Chi,Yuandong, AU - Lin,Dong, AU - Tyler,Brett M, AU - Liu,Xili, Y1 - 2018/10/31/ PY - 2018/7/7/pubmed PY - 2019/2/15/medline PY - 2018/7/7/entrez SP - 1412 EP - 1419 JF - Phytopathology JO - Phytopathology VL - 108 IS - 12 N2 - Oxathiapiprolin is a novel fungicide that was recently registered in a number of countries to control plant-pathogenic oomycetes such as Phytophthora capsici. In our previous study, point mutations G770V and G839W in oxysterol binding protein-related protein 1 (ORP1) were detected in oxathiapiprolin-resistant P. capsici isolates (PcORP1). Here, we used the CRISPR/Cas9 system to verify the effects of these two point mutations on P. capsici phenotypes. Transformants containing heterozygous G770V and G839W mutations in PcORP1 showed high levels of oxathiapiprolin resistance. The G770V transformants showed otherwise similar phenotypes compared with the wild-type isolate BYA5, including sporangia and zoospore production, cyst germination, and pathogenicity. However, two independent transformants with heterozygous G839W mutations in PcORP1 could not produce sporangia. Three transformants with an unexpected point mutation in PcORP1 (ΔN837) showed high oxathiapiprolin resistance, and either similar or significantly reduced fitness compared with BYA5. The same deletion (ΔN837) was confirmed to confer oxathiapiprolin resistance in P. sojae by using CRISPR/Cas9. These homozygous P. sojae mutants also showed either similar or strongly reduced fitness compared with the wild-type parent isolate P6497. These results improve our understanding of oxathiapiprolin resistance in Phytophthora spp., and will be useful for the development of novel oxysterol-binding protein homolog inhibitor fungicides. SN - 0031-949X UR - https://www.unboundmedicine.com/medline/citation/29979095/Mutations_in_ORP1_Conferring_Oxathiapiprolin_Resistance_Confirmed_by_Genome_Editing_using_CRISPR/Cas9_in_Phytophthora_capsici_and_P__sojae_ L2 - http://apsjournals.apsnet.org/doi/full/10.1094/PHYTO-01-18-0010-R?url_ver=Z39.88-2003&rfr_id=ori:rid:crossref.org&rfr_dat=cr_pub=pubmed DB - PRIME DP - Unbound Medicine ER -