Abstract
Somatic embryogenesis (SE), or embryo development from in vitro cultured vegetative explants, can be induced in Arabidopsis by the synthetic auxin 2,4-dichlorophenoxyacetic acid (2,4-D) or by overexpression of specific transcription factors, such as AT-HOOK MOTIF NUCLEAR LOCALIZED 15 (AHL15). Here, we explored the role of endogenous auxin [indole-3-acetic acid (IAA)] during 2,4-D and AHL15-induced SE. Using the pWOX2:NLS-YFP reporter, we identified three distinct developmental stages for 2,4-D and AHL15-induced SE in Arabidopsis, with these being (i) acquisition of embryo identity; (ii) formation of pro-embryos; and (iii) somatic embryo patterning and development. The acquisition of embryo identity coincided with enhanced expression of the indole-3-pyruvic acid auxin biosynthesis YUCCA genes, resulting in an enhanced pDR5:GFP-reported auxin response in the embryo-forming tissues. Chemical inhibition of the indole-3-pyruvic acid pathway did not affect the acquisition of embryo identity, but significantly reduced or completely inhibited the formation of pro-embryos. Co-application of IAA with auxin biosynthesis inhibitors in the AHL15-induced SE system rescued differentiated somatic embryo formation, confirming that increased IAA levels are important during the last two stages of SE. Our analyses also showed that polar auxin transport, with AUXIN/LIKE-AUX influx and PIN-FORMED1 efflux carriers as important drivers, is required for the transition of embryonic cells to proembryos and, later, for correct cell fate specification and differentiation. Taken together, our results indicate that endogenous IAA biosynthesis and its polar transport are not required for the acquisition of embryo identity, but rather to maintain embryonic cell identity and for the formation of multicellular proembryos and their development into histodifferentiated embryos.
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Claims 8
Claim · model-written summary
Reporter analysis identified three stages of induced somatic embryogenesis: embryo identity, pro-embryos, and patterning and development.
- Taxon
- Arabidopsis
Arabidopsise0 - Treatment
- 2,4-D and AHL15-induced SEe0
- Outcome
- three distinct developmental stagese0
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“three distinct developmental stages for 2,4-D and AHL15-induced SE in Arabidopsis, with these being (i) acquisition of embryo identity; (ii) formation of pro-embryos; and (iii) somatic embryo patterning and development.”
Claim · model-written summary
Acquisition of embryo identity coincided with enhanced YUCCA expression.
- Taxon
- ArabidopsisArabidopsise0
- Organ / stage
- acquisition of embryo identitye1
- Gene
- YUCCA genese1
- Outcome
- enhanced expressione1
- Direction
- enhancede1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“2,4-D and AHL15-induced SE in Arabidopsis”
e1“The acquisition of embryo identity coincided with enhanced expression of the indole-3-pyruvic acid auxin biosynthesis YUCCA genes”
Claim · model-written summary
Acquisition of embryo identity coincided with enhanced auxin response in embryo-forming tissues.
- Taxon
- ArabidopsisArabidopsise0
- Organ / stage
- embryo-forming tissuese1
- Gene
- pDR5:GFPe1
- Outcome
- auxin responsee1
- Direction
- enhancede1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“2,4-D and AHL15-induced SE in Arabidopsis”
e1“resulting in an enhanced pDR5:GFP-reported auxin response in the embryo-forming tissues”
Claim · model-written summary
Inhibiting the indole-3-pyruvic acid pathway did not affect embryo identity acquisition.
- Taxon
- ArabidopsisArabidopsise0
- Treatment
- Chemical inhibition of the indole-3-pyruvic acid pathwaye1
- Outcome
- acquisition of embryo identitye1
- Direction
- did not affecte1
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“2,4-D and AHL15-induced SE in Arabidopsis”
e1“Chemical inhibition of the indole-3-pyruvic acid pathway did not affect the acquisition of embryo identity”
Claim · model-written summary
Inhibiting the indole-3-pyruvic acid pathway significantly reduced or eliminated pro-embryo formation.
- Taxon
- ArabidopsisArabidopsise0
- Treatment
- Chemical inhibition of the indole-3-pyruvic acid pathwaye1
- Outcome
- formation of pro-embryose1
- Direction
- reduced or completely inhibitede1
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“2,4-D and AHL15-induced SE in Arabidopsis”
e1“Chemical inhibition of the indole-3-pyruvic acid pathway did not affect the acquisition of embryo identity, but significantly reduced or completely inhibited the formation of pro-embryos.”
Claim · model-written summary
Adding IAA with biosynthesis inhibitors rescued differentiated embryo formation in the AHL15 system.
- Taxon
- ArabidopsisArabidopsise0
- Treatment
- Co-application of IAA with auxin biosynthesis inhibitorse1
- Outcome
- differentiated somatic embryo formatione1
- Direction
- rescuede1
Not reportedComparator
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“2,4-D and AHL15-induced SE in Arabidopsis”
e1“Co-application of IAA with auxin biosynthesis inhibitors in the AHL15-induced SE system rescued differentiated somatic embryo formation”
Claim · model-written summary
Polar auxin transport was required for transition of embryonic cells to proembryos.
- Taxon
- ArabidopsisArabidopsise0
- Gene
- AUXIN/LIKE-AUX influx and PIN-FORMED1 efflux carrierse1
- Outcome
- transition of embryonic cells to proembryose1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“2,4-D and AHL15-induced SE in Arabidopsis”
e1“polar auxin transport, with AUXIN/LIKE-AUX influx and PIN-FORMED1 efflux carriers as important drivers, is required for the transition of embryonic cells to proembryos”
Claim · model-written summary
Polar auxin transport was required for correct cell fate specification and differentiation.
- Taxon
- ArabidopsisArabidopsise0
- Gene
- AUXIN/LIKE-AUX influx and PIN-FORMED1 efflux carrierse1
- Outcome
- correct cell fate specification and differentiatione1
Organisms pictured · Representative pictures of the matched species, not figures from the paper.
Verbatim
e0“2,4-D and AHL15-induced SE in Arabidopsis”
e1“polar auxin transport, with AUXIN/LIKE-AUX influx and PIN-FORMED1 efflux carriers as important drivers, is required for the transition of embryonic cells to proembryos and, later, for correct cell fate specification and differentiation.”
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