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As glyphosate is a broad spectrum herbicide extensively used in agriculture worldwide, identification of new  aroA  genes with high level of glyphosate tolerance is essential for the development and breeding of transgenic glyphosate-tolerant crops. In this study, an  aroA  gene was cloned from a  Janibacter  sp. strain isolated from marine sediment (designated as  aroA   J. sp  ). The purified aroA  J. sp   enzyme has a  K   m   value of 30 μM for PEP and 83 μM for S3P, and a significantly higher  K   i   value for glyphosate (373 μM) than aroA  E. coli  . AroA  J. sp   is characterized as a novel and naturally occurring class I aroA enzyme with glyphosate tolerance. Furthermore, we show that aroA  J. sp   can be used as an effective selectable marker in both  japonica  and  indica  rice cultivar. Transgenic rice lines were tested by herbicide bioassay and it was confirmed that they could tolerate up to 3360 g/ha glyphosate, a dosage four-fold that of the recommended agricultural application level. To our knowledge, it is the first report of a naturally occurring novel class I  aroA  gene which can be efficiently utilized to study and develop transgenic glyphosate-tolerant crops, and can facilitate a more economical and simplified weed control system.

A Novel Naturally Occurring Class I 5-Enolpyruvylshikimate-3-Phosphate Synthase from Janibacter sp. Confers High Glyphosate Tolerance to Rice