Development of Transgenic Rice Harbouring Mutated Rice 5-Enolpyruvylshikimate 3-Phosphate Synthase (Os-mEPSPS) and Allium sativum Leaf Agglutinin (ASAL) Genes Conferring Tolerance to Herbicides and Sap-Sucking Insects

Chandrasekhar, Kottakota; Reddy, G. M.; Singh, Jitender; Vani, K. N. K.; Vijayalakshmi, M.; Kaul, Tanushri; Reddy, Malireddy K.

doi:10.1007/s11105-014-0715-3

Cited by 31 publications

(22 citation statements)

References 27 publications

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“…Among these, EPSPS from Agrobacterium tumefaciens CP4 has been used to generate the commercial glyphosate-resistant crops currently available in the market (Funke et al 2006). However, biosafety concerns are being raised in relation to the transformation of the heterologous glyphosate-resistant EPSPS from pathogenic bacteria into crop plants (Kottakota et al 2014). Recently, the discovery of glyphosate-resistant crops with multiple residue mutations in their EPSPS resourced from plants has provided an alternative to the EPSPS of microorganisms for crop engineering (Funke et al 2009;Tian et al 2013).…”

Section: Introductionmentioning

confidence: 99%

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

Tian

Xing

et al. 2015

Mol Breeding

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To date, only AroA variant derived from Agrobacterium tumefaciens CP4 has been used to generate the commercial glyphosate-resistant crops currently available in the market. This single source of the EPSPS gene may have caused the decrease in herbicide tolerance, which has become a major concern of those involved in field management programs. Therefore, it is of interest to increase aroA/EPSPS gene diversity and seek new glyphosate-tolerant genes for developing glyphosate-tolerant crops. In the current study, EPSPS gene from Vitis vinifera (VvEPSPS) was cloned using reverse transcription polymerase chain reaction. However, wild type VvEPSPS cannot be directly used for developing transgenic crops because of its extreme glyphosate sensitivity. Recent studies have demonstrated that DNA shuffling is an effective strategy in producing multi-mutated EPSPS resourced from plants (EPSPS plant ) with improved glyphosate resistance in bacteria and plants. After performing DNA shuffling on VvEPSPS gene, one highly glyphosateresistant mutant with seven amino acid variations was isolated after five rounds of shuffling and screening. The mutant showed seven amino acid changes in the EPSPS gene, namely, Q93R, T113A, P117L, G126A, C160Y, N239H, and V343A. The assay of glyphosate resistance further confirmed the potential of the VvEPSPS mutant in developing glyphosate-resistant crops.

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Section: Introductionmentioning

confidence: 99%

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

Tian

Xing

et al. 2015

Mol Breeding

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“…Although glyphosate tolerant gene aroA has been used directly as selectable marker gene in Arabidopsis and other species 34 35 36 , there are few reports of its application in rice. In most of the reported glyphosate-tolerance rice events, hpt was used as the selectable marker gene 22 37 . Zhao et al used a glyphosate-tolerant aroA variant G6 gene as the selectable marker gene in a japonica rice XiuShui-110 21 .…”

Section: Discussionmentioning

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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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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“…The embryogenic calli, developed from mature rice seeds, were infected and co-cultivated with Agrobacterium culture containing either recombinant pMDC99-OsmEPSPS or recombinant pMDC99-OsmEPSPS-igrA vector separately. The transformed calli were selected on 50 mg/l hygromycin with three rounds of selections and putative transgenic rice plants were generated from secondary calli after regeneration and rooting according to Chandrasekhar et al (2014) .…”

Section: Methodsmentioning

confidence: 99%

Co-expression of P173S Mutant Rice EPSPS and igrA Genes Results in Higher Glyphosate Tolerance in Transgenic Rice

et al. 2018

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Weeds and their devastating effects have been a great threat since the start of agriculture. They compete with crop plants in the field and negatively influence the crop yield quality and quantity along with survival of the plants. Glyphosate is an important broad-spectrum systemic herbicide which has been widely used to combat various weed problems since last two decades. It is very effective even at low concentrations, and possesses low environmental toxicity and soil residual activity. However, the residual concentration of glyphosate inside the plant has been of major concern as it severely affects the important metabolic pathways, and results in poor plant growth and grain yield. In this study, we compared the glyphosate tolerance efficiency of two different transgenic groups over expressing proline/173/serine (P173S) rice EPSPS glyphosate tolerant mutant gene (OsmEPSPS) alone and in combination with the glyphosate detoxifying encoding igrA gene, recently characterized from Pseudomonas. The molecular analysis of all transgenic plant lines showed a stable integration of transgenes and their active expression in foliar tissues. The physiological analysis of glyphosate treated transgenic lines at seed germination and vegetative stages showed a significant difference in glyphosate tolerance between the two transgenic groups. The transgenic plants with OsmEPSPS and igrA genes, representing dual glyphosate tolerance mechanisms, showed an improved root-shoot growth, physiology, overall phenotype and higher level of glyphosate tolerance compared to the OsmEPSPS transgenic plants. This study highlights the advantage of igrA led detoxification mechanism as a crucial component of glyphosate tolerance strategy in combination with glyphosate tolerant OsmEPSPS gene, which offered a better option to tackle in vivo glyphosate accumulation and imparted more robust glyphosate tolerance in rice transgenic plants.

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Development of Transgenic Rice Harbouring Mutated Rice 5-Enolpyruvylshikimate 3-Phosphate Synthase (Os-mEPSPS) and Allium sativum Leaf Agglutinin (ASAL) Genes Conferring Tolerance to Herbicides and Sap-Sucking Insects

Cited by 31 publications

References 27 publications

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

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

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

Co-expression of P173S Mutant Rice EPSPS and igrA Genes Results in Higher Glyphosate Tolerance in Transgenic Rice

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