Abstract
Somatic embryogenesis in Arabidopsis is achieved by culturing bending-cotyledon embryos on a 2,4-D-containing induction medium for 14 d followed by a transfer on to a hormone-free development medium. Several genes orthologous to Arabidopsis SHOOTMERISTEMLESS (STM), CLAVATA 1 (CLV1), and ZWILLE (ZLL) were isolated from Brassica oleracea (Bo), B. rapa (Br), and B. napus (Bn), and ectopically expressed in Arabidopsis to assess their effects on somatic embryogenesis. Ectopic expression of BoSTM, BrSTM, and BnSTM increased the number of somatic embryos, whereas a different effect was observed in lines overexpressing BnCLV1 in which somatic embryo formation was severely repressed. The introduction of BnZLL did not have any effects on Arabidopsis somatic embryogenesis. The increased embryo-forming capacity observed in lines overexpressing Brassica STM was associated with a lower requirement for the inductive signal 2,4-D, and a higher expression of WUSCHEL (WUS) which demarcates the formation of embryogenic cells. This was in contrast to the 35S::BnCLV1 lines which showed the highest requirement for exogenous 2,4-D and a reduced WUS expression. Microarray studies were conducted to monitor global changes in transcript levels during Arabidopsis somatic embryogenesis between the wild-type (WT) line and a BoSTM-overexpressing line, which showed the most pronounced enhancement of somatic embryo yield. The introduction of BoSTM affected the expression of many genes involved in hormone perception and signalling, as well as genes encoding DNA methyltransferases and enzymes of glutathione metabolism. Pharmacological experiments performed to confirm some of the microarray results showed that Arabidopsis somatic embryogenesis is encouraged by a global hypomethylation of the DNA during the induction phase and by a switch of the glutathione pool towards an oxidized state during the subsequent development phase. Both events occurred in the 35S::BoSTM line, but not in the WT line. Altered expression of Brassica STM also had profound effects on B. napus microspore-derived embryogenesis. The yield of microspore-derived embryos increased in lines overexpressing BnSTM and significantly decreased in antisense lines down-regulating BnSTM.
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Claims 12
Claim · model-written summary
Ectopic BoSTM, BrSTM and BnSTM expression increased somatic embryo number in Arabidopsis.
- Taxon
- Arabidopsis
Arabidopsise1 - Treatment
- Ectopic expression of BoSTM, BrSTM, and BnSTMe0
- Gene
- BoSTM, BrSTM, and BnSTMe0
- Outcome
- number of somatic embryose0
- Direction
- increasede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Ectopic expression of BoSTM, BrSTM, and BnSTM increased the number of somatic embryos”
e1“Somatic embryogenesis in Arabidopsis”
Claim · model-written summary
BnCLV1 overexpression severely repressed somatic embryo formation.
- Taxon
- ArabidopsisArabidopsise1
- Treatment
- overexpressing BnCLV1e0
- Gene
- BnCLV1e0
- Outcome
- somatic embryo formatione0
- Direction
- severely repressede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“lines overexpressing BnCLV1 in which somatic embryo formation was severely repressed”
e1“Somatic embryogenesis in Arabidopsis”
Claim · model-written summary
BnZLL introduction had no effect on Arabidopsis somatic embryogenesis.
- Taxon
- ArabidopsisArabidopsise0
- Treatment
- introduction of BnZLLe0
- Gene
- BnZLLe0
- Outcome
- Arabidopsis somatic embryogenesise0
- Direction
- did not have any effectse0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The introduction of BnZLL did not have any effects on Arabidopsis somatic embryogenesis”
Claim · model-written summary
Brassica STM overexpression lowered the 2,4-D requirement for embryo formation.
- Taxon
- ArabidopsisArabidopsise1
- Gene
- Brassica STMe0
- Outcome
- requirement for the inductive signal 2,4-De0
- Direction
- lowere0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“increased embryo-forming capacity observed in lines overexpressing Brassica STM was associated with a lower requirement for the inductive signal 2,4-D”
e1“Somatic embryogenesis in Arabidopsis”
Claim · model-written summary
Brassica STM overexpression was associated with higher WUS expression.
- Taxon
- ArabidopsisArabidopsise1
- Material
- lines overexpressing Brassica STMe0
- Gene
- WUSCHEL (WUS)e0
- Outcome
- expression of WUSCHEL (WUS)e0
- Direction
- highere0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“lines overexpressing Brassica STM was associated with a lower requirement for the inductive signal 2,4-D, and a higher expression of WUSCHEL (WUS)”
e1“Somatic embryogenesis in Arabidopsis”
Claim · model-written summary
BnCLV1 overexpression lines had the highest exogenous 2,4-D requirement.
- Taxon
- ArabidopsisArabidopsise1
- Material
- 35S::BnCLV1 linese0
- Outcome
- requirement for exogenous 2,4-De0
- Direction
- higheste0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“35S::BnCLV1 lines which showed the highest requirement for exogenous 2,4-D”
e1“Somatic embryogenesis in Arabidopsis”
Claim · model-written summary
BnCLV1 overexpression lines had reduced WUS expression.
- Taxon
- ArabidopsisArabidopsise1
- Material
- 35S::BnCLV1 linese0
- Gene
- WUSe0
- Outcome
- WUS expressione0
- Direction
- reducede0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“35S::BnCLV1 lines which showed the highest requirement for exogenous 2,4-D and a reduced WUS expression”
e1“Somatic embryogenesis in Arabidopsis”
Claim · model-written summary
BoSTM altered expression of genes for hormone perception, DNA methylation and glutathione metabolism.
- Taxon
- ArabidopsisArabidopsise1
- Treatment
- introduction of BoSTMe0
- Gene
- BoSTMe0
- Outcome
- expression of many genes involved in hormone perception and signallinge0
- Direction
- affectede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The introduction of BoSTM affected the expression of many genes involved in hormone perception and signalling, as well as genes encoding DNA methyltransferases and enzymes of glutathione metabolism”
e1“Somatic embryogenesis in Arabidopsis”
Claim · model-written summary
Global DNA hypomethylation during induction encouraged Arabidopsis somatic embryogenesis.
- Taxon
- ArabidopsisArabidopsise0
- Treatment
- global hypomethylation of the DNAe0
- Outcome
- Arabidopsis somatic embryogenesise0
- Direction
- encouragede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Arabidopsis somatic embryogenesis is encouraged by a global hypomethylation of the DNA during the induction phase”
Claim · model-written summary
An oxidized glutathione pool during development encouraged Arabidopsis somatic embryogenesis.
- Taxon
- ArabidopsisArabidopsise0
- Treatment
- a switch of the glutathione pool towards an oxidized statee0
- Outcome
- Arabidopsis somatic embryogenesise0
- Direction
- encouragede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“Arabidopsis somatic embryogenesis is encouraged by a global hypomethylation of the DNA during the induction phase and by a switch of the glutathione pool towards an oxidized state during the subsequent development phase”
Claim · model-written summary
BnSTM overexpression increased microspore embryo yield in B. napus.
- Taxon
- B. napus
_Brassica_napus_-_Habit.jpg?width=60)
Brassica napuse1 - Treatment
- overexpressing BnSTMe0
- Gene
- BnSTMe0
- Outcome
- yield of microspore-derived embryose0
- Direction
- increasede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
_Brassica_napus_-_Habit.jpg?width=500)
Verbatim
e0“The yield of microspore-derived embryos increased in lines overexpressing BnSTM”
e1“B. napus microspore-derived embryogenesis”
Claim · model-written summary
BnSTM antisense downregulation significantly decreased microspore embryo yield in B. napus.
- Taxon
- B. napusBrassica napuse1
- Treatment
- antisense lines down-regulating BnSTMe0
- Gene
- BnSTMe0
- Outcome
- yield of microspore-derived embryose0
- Direction
- decreasede0
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“The yield of microspore-derived embryos increased in lines overexpressing BnSTM and significantly decreased in antisense lines down-regulating BnSTM”
e1“B. napus microspore-derived embryogenesis”
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