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A novel mutated acetolactate synthase gene conferring specific resistance to pyrimidinyl carboxy herbicides in rice

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Abstract

Acetolactate synthase (ALS) is the first common enzyme in the biosynthetic pathway of branched-chain amino acids. Mutations of specific amino acids in ALS have been known to confer resistance to ALS-inhibiting herbicides such as sulfonylureas and pyrimidinyl carboxy (PC) herbicides. However, mutations conferring exclusive resistance to PC have not yet been reported to date. We selected PC resistant rice calli, which were derived from anther culture, using one of the PCs, bispyribac-sodium (BS), as a selection agent. Two lines of BS-resistant plants carrying a novel mutation, the 95th Glycine to Alanine (G95A), in ALS were obtained. In vitro ALS activity assay indicated that the recombinant protein of G95A-mutated ALS (ALS-G95A) conferred highly specific resistance to PC herbicides. In order to determine if the ALS-G95A gene could be used as a selection marker for rice transformation, the ALS-G95A gene was connected to ubiquitin promoter and introduced into rice. PC resistant plants containing integrated ALS-G95A gene were obtained after selection with BS as a selection agent. In conclusion, novel G95A mutated ALS gene confers highly specific resistant to PC-herbicides and can be used as a selection marker.

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Abbreviations

ALS:

Acetolactate synthase

BS:

Bispyribac-sodium

PS:

Pyrithiobac-sodium

PM:

Pyriminobac

CS:

Chlorsulfuron

BM:

Bensulfuron-methyl

IQ:

Imazaquin

IP:

Imazapyr

PC:

Pyrimidinyl carboxy herbicides

SU:

Sulfonylureas

IM:

Imidazolinones

PCs:

(BS, PS and PM)

SUs:

(CS and BM)

IMs:

(IQ and IP)

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Acknowledgements

We thank the members of Laboratory of Plant Breeding, Tohoku University, for their advice. This study was partially supported by a Grant-in-Aid from the Ministry of Education, Science, Sports and Culture, Japan. A.O. is the recipient of Research Fellowships of the Japan Society for the promotion of Science for Young Scientists.

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Authors and Affiliations

  1. Laboratory of Environmental Biotechnology, Graduate School of Agricultural Science, Tohoku University, 1-1 Tsutumidori-Amamiyamachi, Aoba-ku, Sendai, Miyagi, 981-8555, Japan

    Ayako Okuzaki & Kinya Toriyama

  2. Life Science Research Institute, Kumiai Chemical Industry Co., Tamai 276, Kakegawa, Shizuoka, 436-0011, Japan

    Tsutomu Shimizu, Koichiro Kaku & Kiyoshi Kawai

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  1. Ayako Okuzaki
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  2. Tsutomu Shimizu
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  3. Koichiro Kaku
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  4. Kiyoshi Kawai
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  5. Kinya Toriyama
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Correspondence to Kinya Toriyama.

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Okuzaki, A., Shimizu, T., Kaku, K. et al. A novel mutated acetolactate synthase gene conferring specific resistance to pyrimidinyl carboxy herbicides in rice. Plant Mol Biol 64, 219–224 (2007). https://doi.org/10.1007/s11103-007-9146-y

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  • DOI: https://doi.org/10.1007/s11103-007-9146-y

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