Expression of tobacco tocopherol cyclase in rice regulates antioxidative defense and drought tolerance

Woo, Hee Jong; Sohn, Soo-In; Shin, Kong Sik; Kim, Jae Kwang; Kim, Beom Gi; Lim, Myung Ho

doi:10.1007/s11240-014-0530-4

Cited by 11 publications

(4 citation statements)

References 37 publications

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“…[39]. Similarly, Woo et al [40] reported the high content of total tocopherol, and the antioxidant status activity in Nt.TC-transgenic rice compared to control under drought stress. In addition, vte1 mutants of Arabidopsis showed high contents of ascorbate and glutathione which known as antioxidants to compensate tocopherol de ciency.…”

Section: Biochemical Activity Of the Ros-scavenging Enzymesmentioning

confidence: 81%

Constitutive Overexpressing At.TC Improves Drought-mediated Oxidative Tolerance in Transgenic Brassica Napus L.

Sobhanverdi

Majidi

Abbasi

et al. 2021

Preprint

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Background: Environmental stresses are the most important factors limiting crops production in worldwide. Tocopherol, belonging to family of vitamin E compounds, is an amphipathic antioxidants involved in oxidative responses. In the current study, we generated transgenic canola plants overexpressing Arabidopsis VTE1 gene (At.TC) through Agrobacterium tumefaciens system. Methods and results: The putative transgenic plants were successfully regenerated and acclimated in greenhouse conditions. The transcriptional activity of the At.TC gene was evaluated by RT-PCR. In addition, the relative gene expression analysis by qRT-PCR confirmed an increased expression pattern of the transformed gene in canola transgenic lines, with the highest level in R. Line1. Given the results, the transgenic plants, particularly H. Line1 and R. Line2 showed a lower lipid peroxidation compared to WTs under FC 30%. Moreover, two ROS scavenging enzymes including CAT and PPO were up-regulated in transgenic lines; however, no significant pattern was observed for Ascorbate Peroxidase. Also, the amount of leaf tocopherol was significantly more in all T1 lines under drought stress (FC 30%). Conclusion: Taken together, here we successfully developed transgenic lines overexpressing At.TC gene constituently throughout the plant. The results confirmed that the generated transgenic plants are resistant to drought stress, thereby paving the way toward introducing canola plants to deal with the climate change and water shortage.

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Section: Biochemical Activity Of the Ros-scavenging Enzymesmentioning

confidence: 81%

Constitutive Overexpressing At.TC Improves Drought-mediated Oxidative Tolerance in Transgenic Brassica Napus L.

Sobhanverdi

Majidi

Abbasi

et al. 2021

Preprint

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“…Tocopherol cyclase (TC/VTE1) catalyzes the conversion of 2,3-dimethyl-5-phytyl-1,4-benzoquinone (DMPBQ) to γ-tocopherol ( S2 Fig ). Previous studies have reported the characterization of NtTC , a tocopherol cyclase ortholog originating from tobacco, and have demonstrated a relative increase in tocopherol content in the leaves of transgenic rice plants upon overexpression of NtTC [ 27 ]. To enhance the tocopherol contents of rice seed, the plasmid binary pCMF-TC was constructed as shown in Fig 1 and introduced into A .…”

Section: Resultsmentioning

confidence: 99%

“…The resulting binary expression vector, designated as pCMF, was verified by DNA sequencing and restriction enzyme analysis. The full-length NtTC gene (Genbank accession number KJ645980) was obtained from pMJ102TC [ 27 ] by PCR with the following primers incorporating Bam HI and Hin dIII sites: forward primer, 5′-TTGGATCCATGGAGAACATATACGATTTTTCCACCATCTCTTC-3′ and reverse primer, 5′-TTAAGCTTACAGGCCAGGAGGTTTGAGAAATGGAG-3′. The gene was then inserted into MCS of the pCMF vector with Bam HI and Hin dIII, yielding pCMF-TC.…”

Section: Methodsmentioning

confidence: 99%

“…In our present study, we describe the construction of a spontaneous self-excision binary vector using the oxidative-stress inducible mFLP / FRT system for rice transformation. In a recent report, Woo et al [ 27 ] showed that overexpression of NtTC could increase the tocopherol contents in leaves of rice plants. Therefore, for efficient generation of marker-free transgenic rice plants via the spontaneous auto-excision method with enhancement of the tocopherol content of rice seeds, we generated marker-free T 1 transgenic rice plants overexpressing NtTC by Agrobacterium -mediated transformation using hygromycin as a selective agent, followed by segregation of selectable marker genes and analysis of tocopherol content.…”

Section: Introductionmentioning

confidence: 99%

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Development of Selectable Marker-Free Transgenic Rice Plants with Enhanced Seed Tocopherol Content through FLP/FRT-Mediated Spontaneous Auto-Excision

et al. 2015

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Development of marker-free transgenic plants is a technical alternative for avoiding concerns about the safety of selectable marker genes used in genetically modified (GM) crops. Here, we describe the construction of a spontaneous self-excision binary vector using an oxidative stress-inducible modified FLP/FRT system and its successful application to produce marker-free transgenic rice plants with enhanced seed tocopherol content. To generate selectable marker-free transgenic rice plants, we constructed a binary vector using the hpt selectable marker gene and the rice codon-optimized FLP (mFLP) gene under the control of an oxidative stress-inducible promoter between two FRT sites, along with multiple cloning sites for convenient cloning of genes of interest. Using this pCMF binary vector with the NtTC gene, marker-free T1 transgenic rice plants expressing NtTC were produced by Agrobacterium-mediated stable transformation using hygromycin as a selective agent, followed by segregation of selectable marker genes. Furthermore, α-, γ-, and total tocopherol levels were significantly increased in seeds of the marker-free transgenic TC line compared with those of wild-type plants. Thus, this spontaneous auto-excision system, incorporating an oxidative stress-inducible mFLP/FRT system to eliminate the selectable marker gene, can be easily adopted and used to efficiently generate marker-free transgenic rice plants. Moreover, nutritional enhancement of rice seeds through elevation of tocopherol content coupled with this marker-free strategy may improve human health and public acceptance of GM rice.

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ROS Modulation in Crop Plants Under Drought Stress

Verma

Srivastava

Tiwari

et al. 2019

Reactive Oxygen, Nitrogen and Sulfur Species in Plants

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Expression of tobacco tocopherol cyclase in rice regulates antioxidative defense and drought tolerance

Cited by 11 publications

References 37 publications

Constitutive Overexpressing At.TC Improves Drought-mediated Oxidative Tolerance in Transgenic Brassica Napus L.

Constitutive Overexpressing At.TC Improves Drought-mediated Oxidative Tolerance in Transgenic Brassica Napus L.

Development of Selectable Marker-Free Transgenic Rice Plants with Enhanced Seed Tocopherol Content through FLP/FRT-Mediated Spontaneous Auto-Excision

ROS Modulation in Crop Plants Under Drought Stress

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