Genetic Transformation of Wheat Mediated by Agrobacterium tumefaciens

Cheng, Ming‐Jen; Fry, Joyce; Pang, Sheng‐Zhi; Hong-bin, Zhou; Hironaka, Catherine M.; Duncan, David R.; Conner, Timothy W.; Wan, Yuhang

doi:10.1104/pp.115.3.971

Cited by 590 publications

(384 citation statements)

References 28 publications

(40 reference statements)

Supporting

Mentioning

352

Contrasting

Unclassified

Order By: Relevance

“…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%

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

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.

show abstract

Section: Resultssupporting

confidence: 92%

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

“…Nonionic surfactants improved the efficiency of transient Agrobacterium mediated transformation in wheat (Cheng et al 1997;Wu et al 2003) and Arabidopsis (Chung et al 2000;Kim et al 2009). In the present study also similar results on the effects of a surfactant, Tween 20 on transient GUS expression were obtained.…”

Section: Optimization Of Transformation Protocolmentioning

confidence: 99%

Genetic transformation of Indian isolate of Lemna minor mediated by Agrobacterium tumefaciens and recovery of transgenic plants

Chhabra

Chaudhary

Sainger

et al. 2011

Physiol Mol Biol Plants

View full text Add to dashboard Cite

Transgenic plants of an Indian isolate of Lemna minor have been developed for the first time using Agrobacterium tumefaciens and hard nodular cell masses 'nodular calli' developed on the BAP -pretreated daughter frond explants in B 5 medium containing sucrose (1.0 %) with 2,4-D (5.0 μM) and 2-iP (50.0 μM) or 2,4-D (50.0 μM) and TDZ (5.0 μM) under light conditions. These calli were co-cultured with A. tumefaciens strain EHA105 harboring a binary vector that contained genes for β-glucuronidase with intron and neomycin phosphortransferase. Transformed cells selected on kanamycin selection medium were regenerated into fronds whose transgenic nature was confirmed by histochemical assay for GUS activity, PCR analysis and Southern hybridization. The frequency of transformation obtained was 3.8 % and a period of 11-13 weeks was required from initiation of cultures from explants to fully grown transgenic fronds. The pretreatment of daughter fronds with BAP, use of non-ionic surfactant, presence of acetosyringone in co-cultivation medium, co-culture duration of 3 d and 16 h photoperiod during culture were found crucial for callus induction, frond regeneration and transformation of L. minor. This transformation system can be used for the production of pharmaceutically important protein and in bioremediation.

show abstract

“…The addition of 0.03% Pluronic F68 to the inoculation medium (Cheng, et al, 1997) dramatically increased transient GUS expression by up to 100-fold (Table 4). Figure 5A shows that GUS staining in immature embryos varied from single spots to large patches of blue on the edge and scutelum, although GUS expression in immature embryos was not consistently observed in the same treatment and only about 40% of the experiments showed some GUS expression.…”

Section: Effect Of Pluronic F-68 On Gus Expressionmentioning

confidence: 99%

“…Cultures were collected with a scoop, suspended in inoculation medium (IM , Table 1), adjusted to the required optical density (λ = 600 nm) using a Spectronic 21D spectrophotometer (Milton Roy, USA) and used for inoculation within 30 min. In most of the experiments 0.03% (w/v) Pluronic F-68 was added to IM medium to try to improve transformation efficiency (Cheng et al 1997).…”

Section: Transformationmentioning

confidence: 99%

See 1 more Smart Citation

Agrobacterium-mediated transformation of sorghum: factors that affect transformation efficiency

et al. 2004

View full text Add to dashboard Cite

The results presented in this work support the hypothesis that Agrobacterium-mediated transformation of sorghum is feasible, analogous to what has been demonstrated for other cereals such as rice, maize, barley and wheat. The four factors that we found most influenced transformation were: the sensitivity of immature sorghum embryos to Agrobacterium infection, the growth conditions of the donor plant, type of explant and co-cultivation medium. A major problem during the development of our protocol was a necrotic response which developed in explants after co-cultivation. Immature sorghum embryos proved to be very sensitive to Agrobacterium infection and we found that the level of embryo death after co-cultivation was the limiting step in improving transformation efficiency. The addition of coconut water to the co-cultivation medium, the use of vigorous and actively growing immature embryos and the removal of excess bacteria significantly improved the survival rate of sorghum embryos and was critical for successful transformation. Hygromycin phosphotransferase (hpt) proved to be a good selectable marker for sorghum. We also found that β-glucuronidase (GUS) activity was low in most of the transgenic plant tissues tested, although it was very high in immature inflorescences. Although promising, the overall transformation efficiency of the protocol is still low and further optimization will require particular attention to be given to the number of Agrobacterium in the inoculum and the selection of sorghum genotypes and explants less sensitive to Agrobacterium infection.

show abstract

Genetic Transformation of Wheat Mediated by Agrobacterium tumefaciens

Cited by 590 publications

References 28 publications

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

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

Genetic transformation of Indian isolate of Lemna minor mediated by Agrobacterium tumefaciens and recovery of transgenic plants

Agrobacterium-mediated transformation of sorghum: factors that affect transformation efficiency

Contact Info

Product

Resources

About