Transformation of tomatoes with osmotin and chitinase genes and their resistance to<i>Fusarium</i>wilt

Ouyang, Bo; Chen, Y.H.; Li, H.X.; Qian, Chunmei; Huang, Shengqin; Ye, Z.B.

doi:10.1080/14620316.2005.11511971

Cited by 54 publications

(33 citation statements)

References 16 publications

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“…Transformation of the tomato cultivar Leeger 87-5 with A. tumefaciens LBA4404 (pBLNSC) was carried out following the procedures as described in the previous study (Ouyang et al 2005;Zhang et al 2006). Briefly, surface-sterilized tomato seeds were sown on half-strength MS medium containing 20 g/l sucrose and 7.5 g/l agar.…”

Section: Genetic Transformation and Sa Treatmentsmentioning

confidence: 99%

Salicylic-Acid-Induced Self-excision of the Marker Gene nptII from Transgenic Tomato Using the Cre–loxP System

Bing-gang

Duan

et al. 2008

Plant Mol Biol Rep

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The presence of antibiotic-resistant genes in genetically engineered crops together with the target gene has generated a number of environmental and consumer concerns. In order to alleviate public concerns over the safety of food derived from transgenic crops, marker gene elimination is desirable. Marker-free transgenic tomato plants were obtained by using a salicylic-acid-regulated Cre-loxP-mediated site-specific DNA recombination system in which the selectable marker neomycin phosphotransferase nptII and cre genes were flanked by two directly oriented loxP sites. Upon induction by salicylic acid, the cre gene produced a recombinase that eliminated sequences encoding nptII and cre genes, sandwiched by two loxP sites from the tomato genome. Regenerant plants with the Cre-loxP system were obtained by selection on kanamycin media and polymerase chain reaction (PCR) screening. Transgenic plants were screened for excision by PCR using nptII, cre, and PR-1a promoter primers following treatment with salicylic acid. The footprint of the excision was determined by sequencing the T-DNA borders after a perfect recombination event. The excision efficiency was 38.7%. A new plant transformation vector, pBLNSC (Genbank accession number EU327497), was developed, containing six cloning sites and the self-excision system. This provided an effective approach to eliminate the selectable marker gene from transgenic tomato, thus expediting public acceptance of genetically modified tomato.Keywords Cre-loxP recombination system . Marker-free . Self-excision . Solanum lycopersicum . Transgenic tomato Abbreviations 35S cauliflower mosaic virus 35S Cef cefotaxime Plant Mol Biol Rep (

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Section: Genetic Transformation and Sa Treatmentsmentioning

confidence: 99%

Salicylic-Acid-Induced Self-excision of the Marker Gene nptII from Transgenic Tomato Using the Cre–loxP System

Bing-gang

Duan

et al. 2008

Plant Mol Biol Rep

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“…Transformation of the tomato cultivar Leeger 87-5 with Agrobacterium tumefaciens LBA4404 was performed following the procedures by Ouyang et al (2005) and Zhang et al (2006). Adventitious shoots (1 cm length) were excised and transferred to a MS medium containing 500 lM SA, 200 mg cefotaxime/ l and 2 mg indole-3-butyric acid/l.…”

Section: Genetic Transformation and Sa Treatmentsmentioning

confidence: 99%

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

Duan

Niu

et al. 2008

Biotechnol Lett

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A plant transformation vector, pCLKSCLA25 (EU327498), was developed to contain eight cloning sites and the inducible self-excision system which provided an effective approach to eliminate the selectable marker gene(s) from transgenic plants. Upon induction by salicylic acid, the cre gene produced a recombinase that eliminated sequences encoding the selectable marker neomycin phosphotransferase and cre itself. The excision efficiency was 41% in transgenic tomato regenarants. The stilbene synthase gene (vst1) from Vitis vinifera L. was cloned into pCLKSCLA25. The expression of vst1 gene contributed to the accumulation of trans-reveratrol from 3.4 to 8.7 mug/g fresh wt in different marker-free transgenic tomato lines.

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“…5 with A. tumefaciens LBA4404 (p35C) was carried out following the procedures as described in the previous study (Ouyang et al 2005). Briefly, surface-sterilized tomato seeds were sown on halfstrength MS medium containing 20 g sucrose l -1 and 7.5 g agar l -1 .…”

Section: Transgenic Plant Generation and Induction By B-estradiolmentioning

confidence: 99%

“…Isolation of genomic DNA from leaf tissues was performed essentially as described previously (Ouyang et al 2005). The PCR primers used for screening DNA excision in transgenic plants are listed in Table 1 and their respective positions are shown in Fig.…”

Section: Molecular Techniquesmentioning

confidence: 99%

Chemical-induced autoexcision of selectable markers in elite tomato plants transformed with a gene conferring resistance to lepidopteran insects

Zhang

Ouyang

et al. 2006

Biotechnol Lett

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Marker-free transgenic tomato plants harboring a synthetic Bacillus thuringiensis endotoxin gene, cryIAc, were obtained by using a chemically regulated, Cre/loxP-mediated site-specific DNA recombination system, in which the selectable marker neomycin phosphotransferase gene flanked by two directly oriented loxP sites was located between the cauliflower mosaic virus 35S promoter and a promoterless cryIAc. Upon induction by 2 microM beta-estradiol, sequences encoding the selectable marker and cre sandwiched by two loxP sites were excised from the tomato genome, leading to activation of the downstream endotoxin gene cryIAc with high expression levels as shown by Northern blot and ELISA assay (250-790 ng g(-1) fresh wt) in T(1) generation. For transgenic line with single transgenic loci, 15% of T(1) progenies were revealed marker-free. This autoexcision strategy provides an effective approach to eliminate a selectable marker gene from transgenic tomato, thus expediting the public acceptance of genetically modified crop.

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Transformation of tomatoes with osmotin and chitinase genes and their resistance toFusariumwilt

Cited by 54 publications

References 16 publications

Salicylic-Acid-Induced Self-excision of the Marker Gene nptII from Transgenic Tomato Using the Cre–loxP System

Salicylic-Acid-Induced Self-excision of the Marker Gene nptII from Transgenic Tomato Using the Cre–loxP System

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

Chemical-induced autoexcision of selectable markers in elite tomato plants transformed with a gene conferring resistance to lepidopteran insects

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