Novel AroA from Pseudomonas putida Confers Tobacco Plant with High Tolerance to Glyphosate

Yan, Hai-Qin; Chang, Suhua; Tian, Zhe-Xian; Zhang, Le; Sun, Yicheng; Li, Yan; Wang, Jing; Wang, Yiping

doi:10.1371/journal.pone.0019732

Cited by 18 publications

(15 citation statements)

References 23 publications

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“…In our study, the untransformed MH86 was sensitive to glyphosate, and were dead 7 days after 420 g/ha glyphosate treatment; while transgenic MH86 rice plants expressing aroA J. sp exhibited robust tolerance to glyphosate at a dosage of 3360 g/ha, which is four-fold that of the recommended dosage ( Fig. 5e ) The tolerant level was comparable with that resulted from the expression of several class II aroA genes in other plants 20 22 39 .…”

Section: Discussionsupporting

confidence: 54%

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

Cui

Zhao

et al. 2016

Sci Rep

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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 aroAJ. sp). The purified aroAJ. sp enzyme has a Km value of 30 μM for PEP and 83 μM for S3P, and a significantly higher Ki value for glyphosate (373 μM) than aroAE. coli. AroAJ. sp is characterized as a novel and naturally occurring class I aroA enzyme with glyphosate tolerance. Furthermore, we show that aroAJ. 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.

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Section: Discussionsupporting

confidence: 54%

“…Several class II enzymes have been used to generate transgenetic plants 18 19 20 21 22 . Among them, only the one from Agrobacterium tumefaciens CP4 has been used for the production of transgenic glyphosate-tolerant crops.…”

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confidence: 99%

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

Cui

Zhao

et al. 2016

Sci Rep

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“…Class II EPSPS enzymes have been used mostly to develop glyphosate‐tolerant agriculturally and economically important plants (Chhapekar et al ., 2015; Kahrizi et al ., 2007; Yan et al ., 2011; Ye et al ., 2001; Zhao et al ., 2011). In contrast, class I EPSPS enzymes are inhibited by micromolar glyphosate concentrations and have low affinity for PEP substrates (Cao et al ., 2004; He et al ., 2001; Pollegioni et al ., 2011).…”

Section: Introductionmentioning

confidence: 99%

Overexpression of improved EPSPS gene results in field level glyphosate tolerance and higher grain yield in rice

Achary

Sheri

Manna

et al. 2020

Plant Biotechnology Journal

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Glyphosate is a popular, systemic, broad-spectrum herbicide used in modern agriculture. Being a structural analog of phosphoenolpyruvate (PEP), it inhibits 5-enolpyruvylshikimate 3-phosphate synthase (EPSPS) which is responsible for the biosynthesis of aromatic amino acids and various aromatic secondary metabolites. Taking a lead from glyphosate-resistant weeds, two mutant variants of the rice EPSPS gene were developed by amino acid substitution (T173I + P177S; TIPS-OsEPSPS and G172A + T173I + P177S; GATIPS-OsEPSPS). These mutated EPSPS genes were overexpressed in rice under the control of either native EPSPS or constitutive promoters (maize ubiquitin [ZmUbi] promoter). The overexpression of TIPS-OsEPSPS under the control of the ZmUbi promoter resulted in higher tolerance to glyphosate (up to threefold of the recommended dose) without affecting the fitness and related agronomic traits of plants in both controlled and field conditions. Furthermore, such rice lines produced 17%-19% more grains compared to the wild type (WT) in the absence of glyphosate application and the phenylalanine and tryptophan contents in the transgenic seeds were found to be significantly higher in comparison with WT seeds. Our results also revealed that the native promoter guided expression of modified EPSPS genes did not significantly improve the glyphosate tolerance. The present study describing the introduction of a crop-specific TIPS mutation in class I aroA gene of rice and its overexpression have potential to substantially improve the yield and field level glyphosate tolerance in rice. This is the first report to observe that the EPSPS has role to play in improving grain yield of rice.

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“…Originally, all crops were sensitive to glyphosate; however, transgenic glyphosate-resistant crops including soybean, corn, cotton, potato and rape seed have become widespread and have led to glyphosate becoming one of the most popular herbicides available. Glyphosate reversibly inhibits 5-enolpyruvylshikimate-3phosphate synthase (EPSPS) activity, after which EPSPS cannot be imported into chloroplasts, blocking the downstream synthesis of aromatic amino acids and secondary metabolites from chorismate, and enabling efficient weed control (Peñaloza-Vazquez et al, 1995;Yan et al, 2011).…”

Section: Introductionmentioning

confidence: 99%

Identification of regulated genes conferring resistance to high concentrations of glyphosate in a new strain ofEnterobacter

Fei

Gai

Zhao

2013

FEMS Microbiol Lett

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Glyphosate is a widely used herbicide that inhibits 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS) activity. Most plants and microbes are sensitive to glyphosate. However, transgenic-resistant crops that contain a modified epsps obtained from the resistant microbes have been commercially successful and therefore, new resistance genes and their adaptive regulatory mechanisms are of great interest. In this study, a soil-borne, glyphosate-resistant bacterium was selected and identified as Enterobacter. The EPSPS in this strain was found to have been altered to a resistant one. A total of 42 differentially expressed genes (DEGs) in the glyphosate were screened using microarray techniques. Under treatment, argF, sdhA, ivbL, rrfA-H were downregulated, whereas the transcripts of speA, osmY, pflB, ahpC, fusA, deoA, uxaC, rpoD and a few ribosomal protein genes were upregulated. Data were verified by quantitative real-time PCR on selected genes. All transcriptional changes appeared to protect the bacteria from glyphosate and associated osmotic, acidic and oxidative stresses. Many DEGs may have the potential to confer resistance to glyphosate alone, and some may be closely related to the shikimate pathway, reflecting the complex gene interaction network for glyphosate resistance.

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Novel AroA from Pseudomonas putida Confers Tobacco Plant with High Tolerance to Glyphosate

Cited by 18 publications

References 23 publications

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

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

Overexpression of improved EPSPS gene results in field level glyphosate tolerance and higher grain yield in rice

Identification of regulated genes conferring resistance to high concentrations of glyphosate in a new strain ofEnterobacter

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