Expression of stilbene synthase gene in transgenic tomato using salicylic acid-inducible Cre/loxP recombination system with self-excision of selectable marker

G., B.; Duan, Xiaoyu; Niu, Jianxin; Ma, Castello; Hao, Qingnan; Zhang, L. X.; Zhang, H. P.

doi:10.1007/s10529-008-9843-x

Cited by 22 publications

(6 citation statements)

References 16 publications

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“…The pPUCM-X1-RNAi was digested with HindIII following the blunting with Klenow enzyme, and then it was digested with SacI. The small fragments were inserted into the p2300-121 expressive vector (Ma et al 2009) at SacI-SmaI sites. The resulting plasmid containing the expression cassettes of the P35s-Lycβ-Tnos in series was designated as p2300-121-X1-RNAi (Fig.…”

Section: Construction Of Vectorsmentioning

confidence: 99%

Regulation of Carotenoid Content in Tomato by Silencing of Lycopene β/ε-Cyclase Genes

Bing-gang

et al. 2010

Plant Mol Biol Rep

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To conduct RNAi interference of Lyc-β and Lyc-ε genes, two plant expression vectors were constructed by inserting the intron fragments of the gusA gene into the two target gene fragments, which were designed in anti-sense directions. After the Agrobacterium tumefaciens-mediated transformation, 13 transgenic tomato plants (seven and six for Lyc-β and Lyc-ε, respectively) were obtained, which was further validated by PCR. Real-time PCR revealed that the messenger RNA abundance of Lyc-β gene and Lyc-ε gene in transgenic tomato plants was significantly reduced to 8.95% and 13.16%, respectively, of the level of the wild-type plant. Subsequent high-performance liquid chromatography analysis found that transgenic tomato plant had significantly increased lycopene content, with the highest value of 13.8 μg/g leaf dry weight, which was about 4.2-fold that of wild-type plant. Moreover, Lyc-β and Lyc-ε interference gene effects were observed on downstream products as well. β-Carotene and lutein contents decreased in Lyc-β RNAi lines, ranging from 40.7 to 117.3 μg/g and 4.9 to 23.5 μg/g leaf dry weight, respectively. In Lyc-ε RNAi lines, β-carotene content increased, ranging from 195.8 to 290.2 μg/g, while lutein content decreased, ranging from 3.7 to 11.3 μg/g. For total carotenoids, Lyc-β RNAi lines resulted in 2.9-fold decrease, while Lyc-ε RNAi lines yielded 1.7-fold increase in contents when compared to wild-type control. This study demonstrated that RNAi gene technology is an effective method for enhancing lycopene content in plants.

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Section: Construction Of Vectorsmentioning

confidence: 99%

Regulation of Carotenoid Content in Tomato by Silencing of Lycopene β/ε-Cyclase Genes

Bing-gang

et al. 2010

Plant Mol Biol Rep

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“…For example, Medicago sativa (alfalfa) transformed with AhRS produced 15 µg/g fresh weight (FW) of resveratrol [15], Oryza sativa L. (rice) transformed with AhRS1 produced 0.697 µg/g FW [16], and Rehmannia glutinosa transformed with AhRS3 produced 2.0 µg/g FW [17]. Furthermore, Lactuca sativa L. (lettuce) transformed with a stilbene synthase gene of Parthenocissus henryana accumulated 56.40 µg/g FW [18], Solanum lycopersicum (tomato) transformed with a grapevine stilbene synthase gene (vst1) accumulated 8.7 µg/g FW [19], Vitis vinifera L. (grapevine) transformed with a novel stilbene synthase gene from Chinese wild Vitis pseudoreticulata produced 2.586 µg/g FW [20], and Ziziphus jujuba Mill. (Huping jujube) transformed with a resveratrol synthase gene from Polygonum cuspidatum produced 0.45 µg/g FW [21].…”

Section: Discussionmentioning

confidence: 99%

Resveratrol Biosynthesis in Hairy Root Cultures of Tan and Purple Seed Coat Peanuts

Park

Yeo

et al. 2021

Agronomy

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Peanut (Arachis hypogaea) is a crop that can produce resveratrol, a compound with various biological properties, such as those that exert antioxidant, anticancer, and anti-inflammatory effects. In this study, trans-resveratrol was detected in the roots, leaves, and stems of tan and purple seed coat peanuts (Arachis hypogaea) cultivated in a growth chamber. Both cultivars showed higher levels of resveratrol in the roots than the other plant parts. Thus, both cultivars were inoculated with Agrobacterium rhizogenes, in vitro, to promote hairy root development, thereby producing enhanced levels of t-resveratrol. After 1 month of culture, hairy roots from the two cultivars showed higher levels of fresh weight than those of seedling roots. Furthermore, both cultivars contained higher t-resveratrol levels than those of their seedling roots (6.88 ± 0.21 mg/g and 28.07 ± 0.46 mg/g, respectively); however, purple seed coat peanut hairy roots contained higher t-resveratrol levels than those of tan seed coat peanut hairy roots, ranging from 70.16 to 166.76 mg/g and from 46.61 to 54.31 mg/g, respectively. The findings of this study indicate that peanut hairy roots could be a good source for t-resveratrol production due to their rapid growth, high biomass, and substantial amount of resveratrol.

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“…In this study, we present the feasibility of a designed seed-specifically regulated autoexcision system for NptII cassette excision, based on a Cre/loxP site-specific recombination system in N. tabacum. In recent studies on marker gene elimination using the Cre/loxP recombination system, Cre recombinase was expressed through various strategies including constitutive (Arumugam et al, 2007), transient (Jia et al, 2006), inducible (Wang et al, 2005;Ma et al, 2009), and genetically regulated (Mlynárová et al, 2006;Li et al, 2007;Moon et al, 2010) systems. Eleven T1 progeny plants for each homogeneous line were analyzed with PCR using E1/gusR and nptIIF/R primers.…”

Section: Discussionmentioning

confidence: 99%

Efficient seed-specifically regulated autoexcision of marker gene (nptII) with inducible expression of interest gene in transgenic Nicotiana tabacum

Hamzeh¹,

Motallebi²,

Zamani³

2016

Turk J Biol

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In this study, we developed the seed-specifically regulated autoexcision system based on Cre/loxP site-specific recombination for coelimination of nptII and cre genes from transgenic plants. To accomplish this, a seed-specific gene promoter, BcNA1, from Brassica napus was used to drive conditional expression of recombinase. NptII and recombinase cassettes were placed between two directly repeated loxP sites. The loxP-flanked DNA was located between the SP-DDEE synthetic pathogen inducible promoter and promoterless β-glucuronidase (Gus) gene, as a model gene of interest. Upon seed-specific expression of cre recombinase, the excision event would eliminate loxP-flanked DNA and bring the gus reporter gene under the control of the inducible promoter. This Cre/loxP system was transformed into Nicotiana tabacum via Agrobacterium-mediated transformation. The regenerated plants were obtained by selection on kanamycin medium and PCR screening. Based on seed germination assay on kanamycin-containing medium, the transgenic lines were subdivided into homogeneous and heterogeneous categories. Phenotypic and molecular analysis of T1 progeny plants indicated that completely NptII-free plants were obtained in homogeneous transgenic lines. Coelimination of NptII and recombinase cassettes were verified with PCR, sequencing, and histochemical Gus staining assay. Full excision efficiency (100%) demonstrates the effectiveness of this autoexcision system for the production of marker gene-free transgenic plants.

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Expression of stilbene synthase gene in transgenic tomato using salicylic acid-inducible Cre/loxP recombination system with self-excision of selectable marker

Cited by 22 publications

References 16 publications

Regulation of Carotenoid Content in Tomato by Silencing of Lycopene β/ε-Cyclase Genes

Regulation of Carotenoid Content in Tomato by Silencing of Lycopene β/ε-Cyclase Genes

Resveratrol Biosynthesis in Hairy Root Cultures of Tan and Purple Seed Coat Peanuts

Efficient seed-specifically regulated autoexcision of marker gene (nptII) with inducible expression of interest gene in transgenic Nicotiana tabacum

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