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Transformation of Azospirillum brasilense Cd with an ACC Deaminase Gene from Enterobacter cloacae UW4 Fused to the Tet r Gene Promoter Improves Its Fitness and Plant Growth Promoting Ability

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Abstract

It has been reported that PGPB, containing ACC deaminase, can cleave the plant ethylene precursor ACC and thereby lower ethylene concentration in a developing or stressed plant, protecting it against the deleterious effects of stress ethylene and facilitating the formation of longer roots. In a previous work we have demonstrated expression of the ACC deaminase gene (acdS) from Enterobacter cloacae UW4 under the control of the lac promoter in Azospirillum brasilense Cd. With the inference that a construct including the ACC deaminase gene under the control of a constitutive promoter weaker than the lac promoter might impose less metabolic load on Azospirillum and improve its fitness, it was decided to clone acdS under the control of a tetracycline resistance gene promoter. The ACC deaminase structural gene was fused to the Tet r gene promoter by overlap extension using PCR, cloned in pRK415, and transferred into A. brasilense Cd. The resulting transformants showed lower ACC deaminase activity than those with the lac promoter controlled acdS gene. However, acdS under the control of the Tet r gene promoter imposed lesser metabolic load on Azospirillum brasilense Cd. The result was significantly increased IAA synthesis and greater bacterial growth rate, as well as increased ability to survive on the surface of tomato leaves and to promote the growth of tomato seedlings.

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Acknowledgements

This work was supported by a grant from the Natural Sciences and Engineering Research Council of Canada to Bernard R. Glick. Gina Holguin participated in this work thanks to the support of the director of CIBNOR, Dr. Mario Martinez García, who supported her stay at the University of Waterloo. We thank Mr. Taylor Morey for English editing, and Ms. Olivia Arjona for gas chromatography analysis.

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Authors and Affiliations

  1. CIBNOR, Calle Mar Bermejo No. 195, Col. Playa Palo de Sta. Rita, Apartado Postal No. 128, La Paz, BCS, 23090, México

    G. Holguin

  2. Department of Biology, University of Waterloo, Waterloo ON N2L 3G1, Canada

    B. R. Glick

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  1. G. Holguin
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Correspondence to G. Holguin or B. R. Glick.

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Holguin, G., Glick, B. Transformation of Azospirillum brasilense Cd with an ACC Deaminase Gene from Enterobacter cloacae UW4 Fused to the Tet r Gene Promoter Improves Its Fitness and Plant Growth Promoting Ability . Microb Ecol 46, 122–133 (2003). https://doi.org/10.1007/s00248-002-1036-x

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