Factors affecting soybean cotyledonary node transformation

Meurer, Curtis A.; Dinkins, Randy D.; Collins, G. B.

doi:10.1007/s002990050553

Cited by 107 publications

(54 citation statements)

References 15 publications

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“…The transformation efficiency in our experiments was found to be 3 %. (The results of the molecular analysis is summarized in Table 1) It is still preferred to use host strains like EHA105 with higher copy number of Vir G gene (Meurer et al 1998) which may help in achieving increased efficiency.…”

Section: Resultsmentioning

confidence: 99%

Tissue culture independent transformation of the forage crop sunnhemp (Crotalaria juncea L.): an easy method towards generation of transgenics

Rao¹,

Agrawal²,

Mahmood

et al. 2011

Physiol Mol Biol Plants

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A transformation system which is free of in vitro plant regeneration following Agrobacterium infection is established for the forage legume, Sunnhemp (Crotalaria juncea L.) where in the entire embryo axis of the germinating seed was used as the target tissue for transformation. After standardization of transformation conditions, the cotyledonary node of the embryo axis was infected with Agrobacterium host LBA 4404 harboring the recombinant vector pCAMBIA 2301. The bivalent 1D gene of the two major foot and mouth disease virus (FMDV) serotypes 'O' and 'A 22 ' and the neomycin phosphotransferase (nptII) gene were used as the markers for optimization of the protocol. The embryo axes were pricked randomly on the cotyledonary node and co-cultivated with Agrobacterium. The germlings were then allowed to grow under standard growth room conditions in to mature fertile plants. 60 T 0 plants were established from 3 separate experiments. Three hundred seeds from the 60 T 0 plants were sown to raise the T 1 generation of which 180 were analyzed for integration of bivalent FMDV gene 1D "O" and "A 22 " and the nptII gene. Eighteen out of these 180 plants amplified both the marker genes. Two independent transgenic lines 24 and 37, showed elevated levels of expression of 12 μg and 8 μg (per gm of fresh leaf) of the bivalent ID antigen "O" and "A 22 " . The results showed that the transformation efficiency was 3 %. To the best of our knowledge, this is the first successful attempt of Agrobacterium tumefaciens mediated transformation of Sunnhemp. The protocol can generate whole plant transformants with relative ease and should be compatible to all genotypes of Sunnhemp.

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Section: Resultsmentioning

confidence: 99%

Tissue culture independent transformation of the forage crop sunnhemp (Crotalaria juncea L.): an easy method towards generation of transgenics

Rao¹,

Agrawal²,

Mahmood

et al. 2011

Physiol Mol Biol Plants

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“…GFP expression was observed by a LED 450 nm blue light source with a Kodak Wratten filter #15. The GFP foci per cotyledon (Foci/Cot) and percentage of response (Resp/Cot) were measured on the abaxial surface of the cotyledon (Meurer et al 1998). The Foci/Cot was the number of GFP expressing foci on each cotyledon that responded with more than one spot of GFP expression, and the percentage of the response was the number of cotyledons with more than one spot of GFP expression among total explants.…”

Section: Visualization Of Gfp Expressionmentioning

confidence: 99%

Relationship of Transformation Efficiency and Metabolites Induced in Korean Soybean Cotyledons Treated with Sonication

Song¹,

Yim²,

Jung³

et al. 2013

Korean Journal of Crop Science

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The interaction between Agrobacterium and soybean has been studied at the transcriptome level but not at the metabolic level. However, it is necessary to investigate the difference in metabolites between susceptible and non-susceptible cultivars for high efficiency transformation. We investigated the difference in metabolites from sonicated soybean cotyledons of Korean cultivars and Bert cultivar. To identify difference in metabolites, sonicated extracts were analysed by Fourier transform ion cyclotron resonance mass spectrometry (FT-ICR/MS). The soybean cultivars were classified by susceptibility using green fluorescent protein expression. We found a difference in metabolites between the high susceptible and low susceptible cultivars. The FT-ICR/MS experimental m/z data of different metabolites were compared with theoretical m/z in KNApSAcK database. The candidate list was made using KNApSAcK and focused on phenolic compounds. These candidate metabolites are speculated to influence factors in the interaction. This list of candidates may be useful to investigate the interaction between Agrobacterium and plants to increase transformation efficiency.

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“…Several researchers reported that sonication treatment significantly decreased the morphogenic potential or shoot proliferation of explants (de Oliveira et al, 2009;Meurer et al, 1998). In this study, we investigated the influence of sonication treatment on SEG from immature cotyledons of P. mume.…”

Section: Somatic Embryogenesis With Gfp Expressionmentioning

confidence: 99%

Improving Infection Efficiency of Agrobacterium to Immature Cotyledon Explants of Japanese Apricot (Prunus mume) by Sonication Treatment

Gao–Takai

Tao

2014

J. Japan. Soc. Hort. Sci.

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The infection efficiency of Agrobacterium to cotyledon explants of Japanese apricot (Prunus mume) was markedly improved by sonication treatment. The use of sGFP(S65T) reporter gene in this study enabled direct observation of transgene expression, thus allowing the evaluation of Agrobacterium infection efficiency. Immature cotyledons of different cultivars and different developmental stages were subjected to sonication treatment of durations ranging from 10 sec to 2 min. When early-stage immature cotyledons of 'Nanko' were subjected to sonication treatment, the transient GFP expression frequency after co-cultivation was nearly 100% and GFP fluorescence was distributed over almost the entire cotyledon surface. In contrast, when the inoculation was carried out according to the standard dipping method, transient GFP expression frequency was less than 10% and GFP fluorescence was observed only in spots. The highest frequency of somatic embryogenesis (SEG) with GFP expression was obtained from 40 sec sonication treatment of May 14 'Nanko' immature cotyledons. However, with younger cotyledons, which have a higher SEG frequency, 20 sec of sonication treatment was found to be sufficient to increase the infection frequency.

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Factors affecting soybean cotyledonary node transformation

Cited by 107 publications

References 15 publications

Tissue culture independent transformation of the forage crop sunnhemp (Crotalaria juncea L.): an easy method towards generation of transgenics

Tissue culture independent transformation of the forage crop sunnhemp (Crotalaria juncea L.): an easy method towards generation of transgenics

Relationship of Transformation Efficiency and Metabolites Induced in Korean Soybean Cotyledons Treated with Sonication

Improving Infection Efficiency of Agrobacterium to Immature Cotyledon Explants of Japanese Apricot (Prunus mume) by Sonication Treatment

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