Agrobacterium-mediated transformation of Pisum sativum in vitro and in vivo

Švábová, Lenka; Smýkal, Petr; Griga, Miroslav; Ondřej, Vladan

doi:10.1007/s10535-005-0009-6

Cited by 36 publications

(14 citation statements)

References 52 publications

(41 reference statements)

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“…We have also compared the explant source from garden-grown mature plants with those from in vitro seedling-derived plants in culture; the former were more responsive. For an efficient and reliable transformation system of pea, in vivo seedlings were reportedly preferable (Svábová et al 2005). Tiwari et al (2007) had shown higher transformation rates of mature leaves than younger leaves of Gentiana macrophylla.…”

Section: Discussionmentioning

confidence: 99%

Hairy root cultures of butterfly pea (Clitoria ternatea L.): Agrobacterium × plant factors influencing transformation

Swain

Sahu

Pal

et al. 2011

World J Microbiol Biotechnol

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Transformed rhizoclones were developed from Agrobacterium-treated explants of the medicinally important twinning legume Clitoria ternatea L. Several key factors influencing transformation events were optimized. A4T was the most infectious among the strains employed. Internode segments were more responsive than leaves, outdoor-grown explants preferred to those from in vitro cultures. High frequency transformation, resulting in up to 85.8% rhizogenesis, was attained using pre-pricked internodal explants for immersion (10 min) in Agrobacterium rhizogenes suspension grown overnight with acetosyringone (100 μM) to an OD(660) ≅ 0.6, diluted to a density of 10(9) cells ml(-1), followed by 5-day co-cultivation. Roots were individually cultured in MS0 supplemented with the bacteriostatic antibiotic cefotaxime (500 μg ml(-1)). Rhizoclones were renewed through successive subcultures in MS0 under diffused illumination. The T ( L )-DNA rolB and rolC ORF were detected in rhizoclones through PCR amplification. The T ( R )-DNA gene encoding mannopine synthase (man2) was revealed by positive amplification and opine gene expression substantiated by agropine and mannopine biosynthesis in all selected transformed rhizoclones. The implication of such findings is discussed on the context of utilization of such genetically transformed root cultures towards sustainable production of medicinally useful phytocompounds, besides providing a means for plant conservation.

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Section: Discussionmentioning

confidence: 99%

Hairy root cultures of butterfly pea (Clitoria ternatea L.): Agrobacterium × plant factors influencing transformation

Swain

Sahu

Pal

et al. 2011

World J Microbiol Biotechnol

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“…Although pea is amenable to genetic transformation, this remains a challenge and precludes systematic characterization of gene function [234,235]. However, virus-induced gene silencing (VIGS) has become an important reverse genetics tool for functional genomics and VIGS vectors based on Pea early browning virus (PEBV, genus Tobravirus) are available for legume species and were successfully used to silence pea genes involved in symbiosis with nitrogen-fixing Rhizobium as well as development [236,237].…”

Section: Genomic Analysis Of Peamentioning

confidence: 99%

Pea (Pisum sativum L.) in the Genomic Era

Smýkal

Aubert²,

Burstin³

et al. 2012

Agronomy

201

158

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Pea (Pisum sativum L.) was the original model organism used in Mendel's discovery (1866) of the laws of inheritance, making it the foundation of modern plant genetics. However, subsequent progress in pea genomics has lagged behind many other plant species. Although the size and repetitive nature of the pea genome has so far restricted its sequencing, comprehensive genomic and post genomic resources already exist. These include BAC libraries, several types of molecular marker sets, both transcriptome and proteome datasets and mutant populations for reverse genetics. The availability of the full genome sequences of three legume species has offered significant opportunities for genome wide comparison revealing synteny and co-linearity to pea. A combination of a candidate gene and colinearity approach has successfully led to the identification of genes underlying agronomically important traits including virus resistances and plant architecture. Some of this knowledge has already been applied to marker assisted selection (MAS) programs, increasing precision and shortening the breeding cycle. Yet, complete translation of marker discovery to pea breeding is still to be achieved. Molecular analysis of pea collections has shown that although substantial variation is present within the cultivated genepool, wild material offers the possibility to incorporate novel traits that may have been inadvertently eliminated. Association mapping analysis of diverse pea germplasm promises to identify genetic variation related to desirable agronomic traits, which are historically difficult to breed for in a traditional manner. The availability of high throughput 'omics' methodologies offers great promise for the development of novel, highly accurate selective breeding tools for improved pea genotypes that are sustainable under current and future climates and farming systems.

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“…the so-called vacuum infiltration method), has been reported as an effective procedure to increase contact area between explant and Agrobacterium (Bechtold et al 1993). The vacuumassisted technique has been successfully applied in transformations of Arabidopsis thaliana (Clough and Bent 1998), Petunia hybrida (Tjokrokusumo et al 2000), Agapanthus (Suzuki et al 2001), wheat (Amoah et al 2001), Pinus radiata (Charity et al 2002), Brassica napus (Wang et al 2003), pea (Svabova et al 2005), and Coffea canephora (Kumar et al 2006). In addition, the successful regeneration of transformed plants, either directly from starting explants or indirectly from transformed callus, is an important step in plant genetic transformation.…”

Section: Introductionmentioning

confidence: 99%

Overexpression of farnesyl pyrophosphate synthase (FPS) gene affected artemisinin content and growth of Artemisia annua L.

Banyai

Kirdmanee

Mii

et al. 2010

Plant Cell Tiss Organ Cult

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Transgenic plants of Artemisia annua L., a medicinal plant that produces the compound artemisinin which has an anti-malarial activity, were developed following Agrobacterium tumefaciens-mediated transformation of leaf explants. A. tumefaciens strain EHA105 carrying either pCAMBIA1301 or pCAMBIAFPS was used. Both plasmids harbored the hygromycin phosphotransferase II (hptII) gene as a selectable gene, but the latter plasmid also harbored the gene encoding for farnesyl pyrophosphate synthase (FPS), a key enzyme for artemisinin biosynthesis. Shoot regeneration was observed either directly from leaf sections or via intervening callus when explants were incubated on solidified Murashige and Skoog (MS) (1962) medium containing 0.1 mg l -1 a-naphthaleneacetic acid (NAA), 1 mg l -1 N 6 -benzyladenine (BA), 30 mg l -1 meropenem and 10 mg l -1 hygromycin. Applying vacuum infiltration dramatically increased transformation efficiency up to 7.3 and 19.7% when plasmids with and without FPS gene were used, respectively. All putative transgenic regenerants showed positive bands of hptII gene following Southern blot analysis. Expression of FPS was observed in all transgenic lines, and FPS over-expressed lines exhibited higher artemisinin content and yield, of 2.5-and 3.6-fold, respectively, than that detected in wild-type plants. A relatively high correlation (R 2 = 0.78) was observed between level of expression of FPS and artemisinin content. However, gene silencing was detected in some transgenic lines, especially for those lines containing two copies of the FPS transgene, and with some lines exhibiting reduced growth.

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Agrobacterium-mediated transformation of Pisum sativum in vitro and in vivo

Cited by 36 publications

References 52 publications

Hairy root cultures of butterfly pea (Clitoria ternatea L.): Agrobacterium × plant factors influencing transformation

Hairy root cultures of butterfly pea (Clitoria ternatea L.): Agrobacterium × plant factors influencing transformation

Pea (Pisum sativum L.) in the Genomic Era

Overexpression of farnesyl pyrophosphate synthase (FPS) gene affected artemisinin content and growth of Artemisia annua L.

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