Improved glyphosate resistance of 5-enolpyruvylshikimate-3-phosphate synthase from Vitis vinifera in transgenic Arabidopsis and rice by DNA shuffling

Tian, Yong‐Sheng; Xu, Jing; Xing, Xiangzhuo; Wang, Zhao; Fu, Xin; Peng, Ri‐He; Yao, Qizheng

doi:10.1007/s11032-015-0327-0

Cited by 17 publications

(21 citation statements)

References 40 publications

(44 reference statements)

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“…It was thought that since EPSP enzyme was not found in animals, so glyphosate is not toxic to them. However, the results of previous studies showed that it was toxic to fish species (10,14). Loro et al (15) reported that glyphosate-based herbicide was toxic to Rhamdia quelen and Leporinus obtusidens.…”

Section: Introductionmentioning

confidence: 99%

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Acute toxicity effect of glyphosate on survival rate of common carp, Cyprinus carpio

Vajargah

Yalsuyi

Sattari

et al. 2018

Environ Health Eng Manag

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Background: Herbicides are usually used to control weeds and some of them like glyphosate are nonselective herbicides. Aquatic environments are usually the last destinations of agricultural pesticides, which disrupt the metabolic processes of organisms. Thus, the aim of this study was to evaluate acute toxicity of glyphosate on the survival rate of common carp, Cyprinus carpio. Methods: A total of 135 common carp averaged 7 ± 0.8 g in weight were exposed to 0, 25, 50, 75, 100, 125, 150, 175 and 200 mL.L-1 glyphosate (15 fish in each treatment, with triplicates) for 96 hours. The aquariums capacity was 98 L in volume (80 × 35 × 35 cm) and physicochemical parameters were the same for all groups (pH 7.4-8, temperature = 26 ± 1ºC, DO = 7 mg.L-1 and total hardness of 190 mg CaCO 3). LC 10 , LC 20 , LC 30 , LC 40 , LC 50 , LC 60 , LC 70 , LC 80, LC 90 and LC 95 of glyphosate were calculated at 24, 48, 72 and 96 hours after adding glyphosate using probit test. Results: Mortality was observed in all treatments which exposed to higher than 50 mL.L-1 after 96 hours. The results showed that 96-hour LC 50 of glyphosate for common carp was 92.71 mL.L-1. The fish exposed to different concentrations of glyphosate showed clinical signs such as increased mucus secretion, skin darkening and death with mouth open. Conclusion: Glyphosate disrupts the synthesis of amino acids in plants by inhibiting enzymatic activity of 5-enolpyruvylshikimate-3-phosphate synthase (EPSP). This enzyme is absent in animals. However, glyphosate is toxic for common carp.

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Section: Introductionmentioning

confidence: 99%

“…Two-thirds of the total volume of glyphosate has been sprayed in the recent decades (9). Glyphosate inhibits the 5-enolpyruvylshikimate-3-phosphate synthase (EPSP) activity in plants and impairs the synthesis of the aromatic amino acids (10). EPSP is found only in plants and some bacteria (11).…”

Section: Introductionmentioning

confidence: 99%

Acute toxicity effect of glyphosate on survival rate of common carp, Cyprinus carpio

Vajargah

Yalsuyi

Sattari

et al. 2018

Environ Health Eng Manag

View full text Add to dashboard Cite

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“…Therefore, initial libraries of EPSPS variants, generated by DNA shuffling (Tian et al, 2013) or error-prone PCR (Mao et al, 2017), are typically tested in E. coli by selecting for colony growth in the presence of glyphosate at inhibitory concentrations. Improved protein variants are then characterized and verified by generating transgenic plants, such as rice (Oryza sativa; Tian et al, 2013Tian et al, , 2015 and Arabidopsis (Arabidopsis thaliana; Tian et al, 2015;Mao et al, 2017).…”

Section: Engineering Enzymes For Glyphosate Tolerance and Their Applimentioning

confidence: 99%

Applications of Protein Engineering and Directed Evolution in Plant Research

Engqvist

Rabe

2019

Plant Physiol.

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“…In vitro mutated plant EPSPS genes have also been used to confer resistance to glyphosate in different crop plants [ 18 , 20 ]. Tian et al [ 21 , 22 ] have used in vitro -directed shuffling of the EPSPS genes from Malus domestica and Vitis vinefera to produce glyphosate-resistant EPSPS.…”

Section: Introductionmentioning

confidence: 99%

An intragenic approach to confer glyphosate resistance in chile (Capsicum annuum) by introducing an in vitro mutagenized chile EPSPS gene encoding for a glyphosate resistant EPSPS protein

et al. 2018

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Chile pepper (Capsicum annuum) is an important high valued crop worldwide, and when grown on a large scale has problems with weeds. One important herbicide used is glyphosate. Glyphosate inactivates the enzyme 5-enolpyruvylshikimate-3-phosphate synthase (EPSPS), a key enzyme in the synthesis of aromatic amino acids. A transgenic approach towards making glyphosate resistant plants, entails introducing copies of a gene encoding for glyphosate-resistant EPSPS enzyme into the plant. The main objective of our work was to use an intragenic approach to confer resistance to glyphosate in chile which would require using only chile genes for transformation including the selectable marker. Tobacco was used as the transgenic system to identify different gene constructs that would allow for the development of the intragenic system for chile, since chile transformation is inefficient. An EPSPS gene was isolated from chile and mutagenized to introduce substitutions that are known to make the encoded enzyme resistant to glyphosate. The promoter for EPSPS gene was isolated from chile and the mutagenized chile EPSPS cDNA was engineered behind both the CaMV35S promoter and the EPSPS promoter. The leaves from the transformants were checked for resistance to glyphosate using a cut leaf assay. In tobacco, though both gene constructs exhibited some degree of resistance to glyphosate, the construct with the CaMV35S promoter was more effective and as such chile was transformed with this gene construct. The chile transformants showed resistance to low concentrations of glyphosate. Furthermore, preliminary studies showed that the mutated EPSPS gene driven by the CaMV35S promoter could be used as a selectable marker for transformation. We have shown that an intragenic approach can be used to confer glyphosate-resistance in chile. However, we need a stronger chile promoter and a mutated chile gene that encodes for a more glyphosate resistant EPSPS protein.

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Improved glyphosate resistance of 5-enolpyruvylshikimate-3-phosphate synthase from Vitis vinifera in transgenic Arabidopsis and rice by DNA shuffling

Cited by 17 publications

References 40 publications

Acute toxicity effect of glyphosate on survival rate of common carp, Cyprinus carpio

Acute toxicity effect of glyphosate on survival rate of common carp, Cyprinus carpio

Applications of Protein Engineering and Directed Evolution in Plant Research

An intragenic approach to confer glyphosate resistance in chile (Capsicum annuum) by introducing an in vitro mutagenized chile EPSPS gene encoding for a glyphosate resistant EPSPS protein

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