Abstract
Agrobacterium tumefaciens and A. rhizogenes are the causal agents of gall or hairy root disease, but normally the bacteria do not cause disease in wheat. However, both bacteria grew without inhibition when exposed to intact or wounded wheat roots or embryos, and they colonized wheat root surfaces to levels similar to dicotyledonous plants. A. tumefaciens and A. rhizogenes induced 23% cell death after a 1-h exposure to wheat embryo cells grown in 7.4 mM O2, while the extent of cell death at 2.1 mM 02 was 8%. Contact with A. tumefaciens or A. rhizogenes caused cultured wheat embryo and root cells to rapidly produce H202, which decreased when embryos and roots were cultured at 2.1 mM O2. Browning and autofluorescence, and an increase in ferulic acid in cell walls, were observed in wheat embryo and root epidermal cells exposed to Agrobacterium, but . neither lignin nor callose was detected. Agrobacterium appeared to induce resistance-like responses in wheat that may limit transformation efficiency. The inability to regenerate wheat plants using tissue culture has been a limitation to high efficiency transformation. Regeneration via somatic embryogenesis was improved significantly by simulating the in ovulo environment to which the immature wheat embryos are exposed. Triticum embryo culture medium (TEC) improved callus formation, somatic embryo formation, and regeneration from somatic embryos while reducing precocious germination when compared to growth on Murashige and Skoog medium. Regeneration frequencies were improved when embryos were cultured at the O2 concentration found in the wheat ovule (2.1 mM O2) rather than atmospheric 02concentration (7.4 mM O2). Agrobacterium-mediated transformation of wheat was limited by tissue necrosis following co-cultivation, and by poor plant regeneration. Reduction of necrosis and increased plant regeneration were accomplished by amending the culture medium with antioxidant compounds and by reducing the O2 tension in which the wheat embryos were cu1tured. Twelve days past anthesis (DPA), wheat embryos were co-cultivated with Agrobacterium tumefaciens strains WAg 11 or EHA 101, incubated on TEC medium containing antioxidant compounds (catalase, cysteine and ascorbic acid), and cultured at 2.1 mM O2 concentrations. Transformation was documented in 6.0% ofregeneratedA. tumefaciens WAg 11 exposed wheat plants using the firefly luciferase (luc) reporter system.
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Claims 15
Claim · model-written summary
Both bacteria grew without inhibition on exposure to wheat roots or embryos.
- Taxon
- Wheat
Triticum aestivumt0 - Outcome
- grew without inhibitione0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“both bacteria grew without inhibition when exposed to intact or wounded wheat roots or embryos”
t0“Wheat (Triticum Aestivum L.)”
Claim · model-written summary
The bacteria colonized wheat root surfaces to levels similar to dicotyledonous plants.
- Taxon
- wheatTriticum aestivume0
- Outcome
- colonized wheat root surfacese0
- Comparator
- dicotyledonous plantse0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“they colonized wheat root surfaces to levels similar to dicotyledonous plants”
Claim · model-written summary
Agrobacterium exposure caused 23% embryo cell death at 7.4 mM O2 versus 8% at 2.1 mM O2.
- Taxon
- wheatTriticum aestivume0
- Treatment
- A. tumefaciens and A. rhizogenese0
- Interactor
- A. tumefaciens
Agrobacterium tumefaciense0pathogen - Second interactor
- A. rhizogenese0pathogen
- Outcome
- cell deathe0
- Direction
- inducede0
- Value
- 23%e0
- Qualifier
- 8%e0
- Comparator
- 2.1 mM 02e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“A. tumefaciens and A. rhizogenes induced 23% cell death after a 1-h exposure to wheat embryo cells grown in 7.4 mM O2, while the extent of cell death at 2.1 mM 02 was 8%”
Claim · model-written summary
Contact with either bacterium rapidly induced H202 production in wheat embryo and root cells.
- Taxon
- WheatTriticum aestivumt0
- Treatment
- Contact with A. tumefaciens or A. rhizogenese0
- Interactor
- A. tumefaciensAgrobacterium tumefaciense0pathogen
- Second interactor
- A. rhizogenese0pathogen
- Outcome
- produce H202e0
- Direction
- producee0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Contact with A. tumefaciens or A. rhizogenes caused cultured wheat embryo and root cells to rapidly produce H202”
t0“Wheat (Triticum Aestivum L.)”
Claim · model-written summary
H202 production decreased when wheat embryos and roots were cultured at 2.1 mM O2.
- Taxon
- WheatTriticum aestivumt0
- Treatment
- 2.1 mM O2e0
- Outcome
- H202e1
- Direction
- decreasede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“which decreased when embryos and roots were cultured at 2.1 mM O2”
e1“wheat embryo and root cells to rapidly produce H202”
t0“Wheat (Triticum Aestivum L.)”
Claim · model-written summary
Agrobacterium exposure produced browning and autofluorescence in wheat embryo and root epidermal cells.
- Taxon
- wheatTriticum aestivume0
- Interactor
- Agrobacterium
.jpg?width=60)
Agrobacteriume0pathogen - Outcome
- Browning and autofluorescencee0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
.jpg?width=500)
Verbatim
e0“Browning and autofluorescence, and an increase in ferulic acid in cell walls, were observed in wheat embryo and root epidermal cells exposed to Agrobacterium”
Claim · model-written summary
Ferulic acid increased in cell walls of Agrobacterium-exposed wheat cells.
- Taxon
- wheatTriticum aestivume0
- Interactor
- AgrobacteriumAgrobacteriume0pathogen
- Outcome
- ferulic acid in cell wallse0
- Direction
- increasee0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“an increase in ferulic acid in cell walls, were observed in wheat embryo and root epidermal cells exposed to Agrobacterium”
Claim · model-written summary
Neither lignin nor callose was detected in exposed wheat cells.
- Taxon
- WheatTriticum aestivumt0
- Outcome
- neither lignin nor callose was detectede0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“neither lignin nor callose was detected”
t0“Wheat (Triticum Aestivum L.)”
Claim · model-written summary
Simulating the in ovulo environment significantly improved wheat regeneration by somatic embryogenesis.
- Taxon
- wheatTriticum aestivume0
- Treatment
- simulating the in ovulo environmente0
- Outcome
- Regeneration via somatic embryogenesise0
- Direction
- improvede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Regeneration via somatic embryogenesis was improved significantly by simulating the in ovulo environment to which the immature wheat embryos are exposed”
Claim · model-written summary
TEC improved callus and somatic embryo formation and regeneration compared with Murashige and Skoog medium.
- Taxon
- WheatTriticum aestivumt0
- Treatment
- Triticum embryo culture medium (TEC)e0
- Outcome
- callus formation, somatic embryo formation, and regeneration from somatic embryose0
- Direction
- improvede0
- Comparator
- Murashige and Skoog mediume0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Triticum embryo culture medium (TEC) improved callus formation, somatic embryo formation, and regeneration from somatic embryos while reducing precocious germination when compared to growth on Murashige and Skoog medium”
t0“Wheat (Triticum Aestivum L.)”
Claim · model-written summary
TEC reduced precocious germination compared with Murashige and Skoog medium.
- Taxon
- WheatTriticum aestivumt0
- Treatment
- Triticum embryo culture medium (TEC)e0
- Outcome
- precocious germinatione0
- Direction
- reducinge0
- Comparator
- Murashige and Skoog mediume0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Triticum embryo culture medium (TEC) improved callus formation, somatic embryo formation, and regeneration from somatic embryos while reducing precocious germination when compared to growth on Murashige and Skoog medium”
t0“Wheat (Triticum Aestivum L.)”
Claim · model-written summary
Wheat regeneration frequencies improved at 2.1 mM O2 rather than 7.4 mM O2.
- Taxon
- wheatTriticum aestivume0
- Treatment
- 2.1 mM O2e0
- Outcome
- Regeneration frequenciese0
- Direction
- improvede0
- Comparator
- 7.4 mM O2e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Regeneration frequencies were improved when embryos were cultured at the O2 concentration found in the wheat ovule (2.1 mM O2) rather than atmospheric 02concentration (7.4 mM O2)”
Claim · model-written summary
Antioxidants and reduced O2 lowered necrosis after Agrobacterium cocultivation.
- Taxon
- wheatTriticum aestivume0
- Treatment
- amending the culture medium with antioxidant compounds and by reducing the O2 tensione0
- Outcome
- necrosise0
- Direction
- Reductione0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Reduction of necrosis and increased plant regeneration were accomplished by amending the culture medium with antioxidant compounds and by reducing the O2 tension in which the wheat embryos were cu1tured”
Claim · model-written summary
Antioxidants and reduced O2 increased wheat plant regeneration.
- Taxon
- wheatTriticum aestivume0
- Treatment
- amending the culture medium with antioxidant compounds and by reducing the O2 tensione0
- Outcome
- plant regeneratione0
- Direction
- increasede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Reduction of necrosis and increased plant regeneration were accomplished by amending the culture medium with antioxidant compounds and by reducing the O2 tension in which the wheat embryos were cu1tured”
Claim · model-written summary
Transformation was documented in 6.0% of regenerated wheat plants exposed to WAg 11.
- Taxon
- wheatTriticum aestivume0
- Interactor
- A. tumefaciens WAg 11Agrobacterium tumefaciense0pathogen
- Outcome
- Transformatione0
- Value
- 6.0%e0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Transformation was documented in 6.0% ofregeneratedA. tumefaciens WAg 11 exposed wheat plants using the firefly luciferase (luc) reporter system”
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