Assessment of conditions affecting Agrobacterium-mediated soybean transformation using the cotyledonary node explant

Kelbert, A. J.; Spaner, Dean; Briggs, K. G.; King, John L.

doi:10.1023/b:euph.0000030670.36730.a4

Cited by 217 publications

(189 citation statements)

References 22 publications

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“…Danyang indicated that a half-strength N6 is the most efficient basal medium (data not shown). This is consistent with previous reports that the use of low-salt media during Agrobacterium infection improves T-DNA transformation (Cheng et al, 1997;Paz et al, 2004), although the mechanism of transformation enhancement is still unknown.…”

Section: Resultssupporting

confidence: 92%

“…For instance, the use of L-cysteine enhances Agrobacteriummediated transformation of maize (Frame et al, 2006). Moreover, DTT in combination with L-cysteine in soybean has been shown to enhance T-DNA transfer (Paz et al, 2004). We thus tested the effects of three antioxidants, 1 mM MES, 200 mg/L L-cysteine, and 100 mg/L DTT, in a half-strength N6 solution (Chu et al, 1975) (Table 1).…”

Section: Resultsmentioning

confidence: 99%

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Development of an Efficient Agrobacterium-Mediated Transformation System and Production of Herbicide-Resistant Transgenic Plants in Garlic (Allium sativum L.)

Ahn

Yoon

Jeon

2013

Molecules and Cells

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The genetic improvement of garlic plants (Allium sativum L.) with agronomical beneficial traits is rarely achieved due to the lack of an applicable transformation system. Here, we developed an efficient Agrobacterium-mediated transformation procedure with Danyang, an elite Korean garlic cultivar. Examination of sGFP (synthetic green fluorescence protein) expression revealed that treatment with 2-(N-morpholino) ethanesulfonic acid (MES), L-cysteine and/or dithiothreitol (DTT) gives the highest efficiency in transient gene transfer during Agrobacterium co-cultivation with calli derived from the roots of in vitro plantlets. To increase stable transformation efficiency, a two-step selection was employed on the basis of hygromycin resistance and sGFP expression. Of the hygromycin-resistant calli initially produced, only sGFP-expressing calli were subcultured for selection of transgenic calli. Transgenic plantlets produced from these calli were grown to maturity. The transformation efficiency increased up to 10.6% via our optimized procedure. DNA and RNA gel-blot analysis indicated that transgenic garlic plants stably integrated and expressed the phosphinothricin acetyltransferase (PAT) gene. A herbicide spraying assay demonstrated that transgenic plants of garlic conferred herbicide resistance, whilst nontransgenic plants and weeds died. These results indicate that our transformation system can be efficiently utilized to produce transgenic garlic plants with agronomic benefits.

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Section: Resultssupporting

confidence: 92%

Section: Resultsmentioning

confidence: 99%

Development of an Efficient Agrobacterium-Mediated Transformation System and Production of Herbicide-Resistant Transgenic Plants in Garlic (Allium sativum L.)

Ahn

Yoon

Jeon

2013

Molecules and Cells

View full text Add to dashboard Cite

show abstract

“…Although the transformation improvement mechanism is not clear, the use of low salt contents through the infection and cocultivation stages has been shown to improve the transfer of T-DNA (Fry et al 1987, Cheng et al 1997, Armstrong and Rout 2001, Zhang et al 2002, Paz et al 2004, Vega et al 2008). …”

Section: Rav Souza Et Almentioning

confidence: 99%

“…Cysteine, dithiothreitol (DTT), polyvinyl pyrrolidone, and ascorbic acid (Perl et al 1996, Olhoft and Somers 2001, Paz et al 2004, Frame et al 2006, Mishra et al 2014) are antioxidants that have been used to increase the potential of Agrobacterium to infect tissues by inhibiting the antimicrobial exudates produced by plant cells during the infection and cocultivation stages and/or by preventing the occurrence of necrosis in the plant tissue (Gupta 2010).…”

Section: Rav Souza Et Almentioning

confidence: 99%