Efficient biolistic transformation of maize (Zea mays L.) and wheat (Triticum aestivum L.) using the phosphomannose isomerase gene, pmi, as the selectable marker

Wright, Martha; Dawson, John F.; Dunder, Erik; Suttie, J.; Reed, J. N.; Kramer, Catherine; Chang, Yung Fu; Novitzky, R.; Wang, H.; Artim-Moore, L.

doi:10.1007/s002990100318

Cited by 165 publications

(99 citation statements)

References 27 publications

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“…pUbiT10rga2-1A, which contains a 3-kb T10rga2-1A full-length cDNA with 65 bp upstream and 203 bp downstream of the coding sequence was generated by using a similar strategy. A total of 350 immature embryos of the Bobwhite accession SH 98 56 (18) were cotransformed with pUBiT10rga1, pUbiT10rga2-1A, and a plasmid containing the selectable phosphomannose isomerase marker (19) by using the PDS-1000͞He biolistic particle delivery System (Bio-Rad). Regeneration and selection of the transformed plants were performed as described (18,19).…”

Section: Pcr Amplification Of the T10rga1 And T10rga2-1a Genes And Rtmentioning

confidence: 99%

“…A total of 350 immature embryos of the Bobwhite accession SH 98 56 (18) were cotransformed with pUBiT10rga1, pUbiT10rga2-1A, and a plasmid containing the selectable phosphomannose isomerase marker (19) by using the PDS-1000͞He biolistic particle delivery System (Bio-Rad). Regeneration and selection of the transformed plants were performed as described (18,19). Eight independent T0 transgenic lines containing either the two transgenes or only one of the transgenes were obtained.…”

Section: Pcr Amplification Of the T10rga1 And T10rga2-1a Genes And Rtmentioning

confidence: 99%

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Map-based isolation of the leaf rust disease resistance gene Lr10 from the hexaploid wheat ( Triticum aestivum L.) genome

Feuillet

Travella

Stein

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

454

300

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More than 50 leaf rust resistance (Lr) genes against the fungal pathogen Puccinia triticina have been identified in the wheat gene pool, and a large number of them have been extensively used in breeding. Of the 50 Lr genes, all are known only from their phenotype and͞or map position except for Lr21, which was cloned recently. For many years, the problems of molecular work in the large (1.6 ؋ 10 10 bp), highly repetitive (80%), and hexaploid bread wheat (Triticum aestivum L.) genome have hampered map-based cloning. Here, we report the isolation of the Lr gene Lr10 from hexaploid wheat by using a combination of subgenome map-based cloning and haplotype studies in the genus Triticum. Lr10 is a single-copy gene on chromosome 1AS. It encodes a CC-NBS-LRR type of protein with an N-terminal domain, which is under diversifying selection. When overexpressed in transgenic wheat plants, Lr10 confers enhanced resistance to leaf rust. Lr10 has similarities to RPM1 in Arabidopsis thaliana and to resistance gene analogs in rice and barley, but is not closely related to other wheat Lr genes based on Southern analysis. We conclude that map-based cloning of genes of agronomic importance in hexaploid wheat is now feasible, opening perspectives for molecular bread wheat improvement trough transgenic strategies and diagnostic allele detection.

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Section: Pcr Amplification Of the T10rga1 And T10rga2-1a Genes And Rtmentioning

confidence: 99%

Section: Pcr Amplification Of the T10rga1 And T10rga2-1a Genes And Rtmentioning

confidence: 99%

Map-based isolation of the leaf rust disease resistance gene Lr10 from the hexaploid wheat ( Triticum aestivum L.) genome

Feuillet

Travella

Stein

et al. 2003

Proc. Natl. Acad. Sci. U.S.A.

454

300

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show abstract

“…This result is in accordance with previously published efficiency data for biolistic transformation of wheat. It was reported to be around 1.6% in the majority of the transformation experiments published (Li et al 2012) although it reached 12-20% in some case (Ogawa et al 2008;Wright et al 2001) or even exceeded 70% (Pellegrineschi et al 2002), where whole circular plasmids were used. The transgene integration efficiencies were found to be the same (Fu et al 2000) for the minimal cassette and for the whole plasmid transformation.…”

Section: Discussionmentioning

confidence: 96%

Generating Marker-Free Transgenic Wheat Using Minimal Gene Cassette and Cold-Inducible Cre/Lox System

et al. 2014

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Abstract:Abstract The precise elimination of selectable marker genes is highly desirable, when their function is no longer needed, because their presence raised worldwide public concerns against the release of genetically modified plants. This is the first report of simultaneous application of the minimal gene cassette and cold-inducible Cre/lox recombination system in wheat. The bar selection and cre-recombinase genes were eliminated from T0 and T1 transgenic lines with 44% and 51% efficiency. This approach provides a new, reasonably effective technique to produce selection genefree transgenic wheat lines either immediately after tissue culture, or from the subsequent transgenic generation. The advantage of this method is that it does not require any additional cold treatment during the plant regeneration/growing because the transgene elimination is ensured by the vernalisation. Application of this method prevents gene flow by pollen and seed, because the selection and recombinase genes are eliminated before pollen development, therefore reducing the risk of GM plants. Powered by Editorial Manager® and ProduXion Manager® from Aries Systems CorporationGenerating marker-free transgenic wheat using minimal gene cassette and cold inducible Cre/lox system AbstractThe precise elimination of selectable marker genes is highly desirable, when their function is no longer needed, because their presence raised worldwide public concerns against the release of genetically modified plants. This is the first report of simultaneous application of the minimal gene cassette and cold-inducible Cre/lox recombination system in wheat. The bar selection and cre-recombinase genes were eliminated from T 0 and T 1 transgenic lines with 44% and 51% efficiency. This approach provides a new, reasonably effective technique to produce selection gene-free transgenic wheat lines either immediately after tissue culture, or from the subsequent transgenic generation. The advantage of this method is that it does not require any additional cold treatment during the plant regeneration/growing because the transgene elimination is ensured by the vernalisation. Application of this method prevents gene flow by pollen and seed, because the selection and recombinase genes are eliminated before pollen development, therefore reducing the risk of GM plants.

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“…An amount of 5 µg of total RNA was taken for first strand cDNA synthesis using oligo-dT (20) by using M-MuLV reverse transcriptase (MBI Fermentas, Germany), following manufacturer's instructions. After the first strand cDNA synthesis, Polymerase chain reaction was performed for the amplification of specific chitinase gene sequence using gene specifc primers.…”

Section: Rt-pcr Of Chitinase and Bar Genesmentioning

confidence: 99%

“…The phenomenon of multiple copy integration is commonly encountered in particle-bombardment-mediated genetic transformation of plants [15][16][17][18]. In earlier studies, a variable number of incorporated transgene copies ranging between 1 and 20 were reported in diverse crop plants [19][20][21][22][23][24][25][26][27][28][29][30].…”

Section: Figure: 7 Extraction Of Total Rna and Rt-pcr Analysismentioning

confidence: 99%

In vitro Regeneration and Optimization of Conditions for transformation methods in Pearl millet, Pennisetum glaucum (L.)

Naravula¹,

Maheshwari²,

Naidu³

et al. 2016

IJCBS

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Pearl millet is a dual-purpose crop used for grain and fodder and is grown primarily in Asia and Africa, where it occupies some 27 million ha. It is capable of growing on some of the poorest soils in dry, hot regions of Africa and Asia, where, as a poor man's source of dietary energy, it sustains a large proportion of the populace. It is also grown in other countries where, under relatively more favorable conditions, it provides grain for bullocks, dairy cows, and poultry. Downy mildew caused by Sclerospora graminicola (Sacc.) J. Schroet. is the most widespread and destructive disease of pearl millet causing severe economic losses. New genes can be introduced into this plant through Agrobacterium mediated and bombardment genetic transformation for its genetic improvement, which is dependent on the availability of suitable in vitro techniques. An efficient regeneration system has been developed for in vitro culture of pearl millet (Pennisetum glaucum L.) from the immature inflorescence. High frequency callus and shoot regeneration was obtained on Murashige and Skoog nutrient agar medium supplemented with 2mg/l 2,4-D and 0.2 mg/l NAA, 2 mg/l Kinetin and 30 g/l sucrose. On transfer to soil, the regenerated plantlets survived and appeared to be morphologically similar to the normal seed-grown plants. Histological analysis revealed the de novo origin of shoots from embryogenic callus in in vitro cultured pearl millet. Parameters affecting transformation were optimized by assaying phosphinothricin resistance to transformed calli and basta test for these leaves of plantlets after transferring to pots. These tissues appear to be susceptible to Agrobacterium infection and Particle gun flow mediated transformation carrying pCAMBIA2300 with osmotin and chitinase double construct and pPUR with bar genes, as well as shoot multiplication. The embryogenic callus was found competent to take up the DNA, which was monitored by transient bar gene with GV2600 at 0.6 O.D for Agrobacterium infection and 1µg/µl plasmid DNA from E.coli for cobombardment was found to be compatible in giving transgenics.

show abstract

Efficient biolistic transformation of maize (Zea mays L.) and wheat (Triticum aestivum L.) using the phosphomannose isomerase gene, pmi, as the selectable marker

Cited by 165 publications

References 27 publications

Map-based isolation of the leaf rust disease resistance gene Lr10 from the hexaploid wheat ( Triticum aestivum L.) genome

Map-based isolation of the leaf rust disease resistance gene Lr10 from the hexaploid wheat ( Triticum aestivum L.) genome

Generating Marker-Free Transgenic Wheat Using Minimal Gene Cassette and Cold-Inducible Cre/Lox System

In vitro Regeneration and Optimization of Conditions for transformation methods in Pearl millet, Pennisetum glaucum (L.)

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