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Identification of a cytochrome P450 hydroxylase, CYP81E22, as a causative gene for the high sensitivity of soybean to herbicide bentazon

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Abstract

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A frame shift invoked by a single-base deletion in the gene encoding a cytochrome P450 hydroxylase, CYP81E22, causes the loss of bentazon detoxification function in soybean.

Abstract

Bentazon is an effective herbicide in soybean cultivation applied at post-emergence stages for control of several broadleaf weeds. However, some soybean cultivars are highly sensitive to bentazon and are killed upon application. In this study, the gene related to the high sensitivity of soybean cultivars to bentazon was mapped to chromosome 16, and its location was narrowed down to a 257-kb region where three cytochrome P450 genes were located. In these genes, a single-base deletion of cytosine was detected in the coding region of Glyma.16G149300, CYP81E22, at + 1465 bp downstream from the translation start codon, leading to a frame shift in the open reading frame and creating a premature stop codon. This stop codon resulted in the loss of more than half of the P450, and consequently, the remaining molecule failed to form a functioning protein. This single-base deletion was common among the highly sensitive cultivars screened from the soybean mini-core collection and other previously reported highly sensitive cultivars. Furthermore, we screened plant lines from the targeting-induced local lesions in genomes library of the soybean cultivar Enrei based on a modelled 3D structure of CYP81E22. The lines with mutations in Glyma.16G149300 were highly sensitive to bentazon, which provides strong evidence that Glyma.16G149300 is the gene responsible for high sensitivity to bentazon.

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Acknowledgements

We thank Hitoshi Sato, Osamu Fujii, Yuko Sato, Akihiro Takahashi, Kosei Awatsu, Hisaho Takagai, and Kenei Takahashi (NARO) for the field management of yield trials and Fumiyo Ito for technical assistance in the experiments.

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Authors and Affiliations

  1. Tohoku Agricultural Research Center, National Agriculture and Food Research Organization (NARO), 297 Uenodai, Kariwano, Daisen, Akita, 019-2112, Japan

    Shin Kato & Akio Kikuchi

  2. Institute of Crop Science, NARO, 2-1-2 Kannondai, Tsukuba, Ibaraki, 305-8518, Japan

    Yuko Yokota, Yukiko Fujisawa, Takashi Sayama, Akito Kaga & Masao Ishimoto

  3. Advanced Analysis Center, NARO, 2-1-2 Kannondai, Tsukuba, Ibaraki, 305-8602, Japan

    Rintaro Suzuki

  4. Western Region Agricultural Research Center, NARO, 1-3-1 Senyu-cho, Zentsuji, Kagawa, 765-8508, Japan

    Takashi Sayama & Kunihiko Komatsu

  5. Faculty of Agriculture, Saga University, Saga, 840-8502, Japan

    Toyoaki Anai

  6. Kyushu Okinawa Agricultural Research Center, NARO, 2421 Suya, Koshi, Kumamoto, 861-1192, Japan

    Nobuhiko Oki

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  1. Shin Kato
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  2. Yuko Yokota
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  3. Rintaro Suzuki
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  4. Yukiko Fujisawa
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  5. Takashi Sayama
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  7. Toyoaki Anai
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  8. Kunihiko Komatsu
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  9. Nobuhiko Oki
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  10. Akio Kikuchi
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  11. Masao Ishimoto
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Contributions

SK and MI planned and designed the overall research. SK, YY, TS, KK, and NO performed the positional cloning. SK, YY, and AK evaluated the reactions of plant materials to bentazon. SK, YY, RS, AK, and TA screened the mutant lines with the mutation in the candidate gene. SK, YY, YF, and MI determined and compared the sequences of the candidate genes.

Corresponding author

Correspondence to Masao Ishimoto.

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Communicated by Brian Diers.

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Kato, S., Yokota, Y., Suzuki, R. et al. Identification of a cytochrome P450 hydroxylase, CYP81E22, as a causative gene for the high sensitivity of soybean to herbicide bentazon. Theor Appl Genet 133, 2105–2115 (2020). https://doi.org/10.1007/s00122-020-03580-6

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