TY - JOUR
T1 - The Arabidopsis DWARF27 gene encodes an all-trans-/9-cis-β-carotene isomerase and is induced by auxin, abscisic acid and phosphate deficiency
AU - Abuauf, Haneen W.
AU - Haider, Imran
AU - Jia, Kunpeng
AU - Ablazov, Abdugaffor
AU - Mi, Jianing
AU - Blilou, Ikram
AU - Al-Babili, Salim
N1 - KAUST Repository Item: Exported on 2022-03-16
Acknowledgements: This work was supported by base line funding and a Competitive Research Grant, Round 4 (CRG4) from the King Abdullah University of Science and Technology (KAUST).
PY - 2018/9/11
Y1 - 2018/9/11
N2 - Strigolactones (SLs) are carotenoid-derived plant hormones that influence various aspects of plant growth and development in response to environmental conditions, especially nutrients deficiency. SLs are synthesized via a strict stereo-specific core pathway that leads to the intermediate carlactone, requiring the iron-containing polypeptide DWARF27 (D27) and the carotenoid cleavage dioxygenases 7 (CCD7) and 8 (CCD8). It has been shown that the rice OsD27 is a β-carotene isomerase catalyzing the interconversion of all-trans- into 9-cis-β -carotene. However, data about the enzymatic activity of D27 from other species are missing. Here, we investigated the activity and substrate specificity of the Arabidopsis AtD27 by testing a broad range of carotenoid substrates. Both in vivo and in vitro assays show that AtD27 catalyzes the reverse isomerization of all-trans-/9-cis-β-carotene. AtD27 did not isomerize 13-cis- or 15-cis-β-carotene, indicating high specificity for the C9-C10 double bond. The isomerization reaction was inhibited in the presence of silver acetate, pointing to the involvement of an iron-sulfur cluster. We further investigated the expression of AtD27, using Arabidopsis transgenic lines expressing β-glucuronidase (GUS) under the control of AtD27 native promoter. AtD27 is ubiquitously expressed throughout the plant with the highest expression in immature flowers. In lateral roots, AtD27 expression was induced by treatment with auxin and ABA, while the application of SL analogs did not show an effect. Lower ABA levels in atd27 mutant indicated an interference with the ABA pathway. Quantitative real-time RT-PCR showed that transcript levels of AtD27 and other SL biosynthetic genes in roots are induced upon phosphate starvation. Taken together, our study on AtD27 confirms the postulated enzymatic function of this enzyme, shows its strict substrate- and regio-specificity and indicates an important role in response to multiple plant hormones and phosphate deficiency.
AB - Strigolactones (SLs) are carotenoid-derived plant hormones that influence various aspects of plant growth and development in response to environmental conditions, especially nutrients deficiency. SLs are synthesized via a strict stereo-specific core pathway that leads to the intermediate carlactone, requiring the iron-containing polypeptide DWARF27 (D27) and the carotenoid cleavage dioxygenases 7 (CCD7) and 8 (CCD8). It has been shown that the rice OsD27 is a β-carotene isomerase catalyzing the interconversion of all-trans- into 9-cis-β -carotene. However, data about the enzymatic activity of D27 from other species are missing. Here, we investigated the activity and substrate specificity of the Arabidopsis AtD27 by testing a broad range of carotenoid substrates. Both in vivo and in vitro assays show that AtD27 catalyzes the reverse isomerization of all-trans-/9-cis-β-carotene. AtD27 did not isomerize 13-cis- or 15-cis-β-carotene, indicating high specificity for the C9-C10 double bond. The isomerization reaction was inhibited in the presence of silver acetate, pointing to the involvement of an iron-sulfur cluster. We further investigated the expression of AtD27, using Arabidopsis transgenic lines expressing β-glucuronidase (GUS) under the control of AtD27 native promoter. AtD27 is ubiquitously expressed throughout the plant with the highest expression in immature flowers. In lateral roots, AtD27 expression was induced by treatment with auxin and ABA, while the application of SL analogs did not show an effect. Lower ABA levels in atd27 mutant indicated an interference with the ABA pathway. Quantitative real-time RT-PCR showed that transcript levels of AtD27 and other SL biosynthetic genes in roots are induced upon phosphate starvation. Taken together, our study on AtD27 confirms the postulated enzymatic function of this enzyme, shows its strict substrate- and regio-specificity and indicates an important role in response to multiple plant hormones and phosphate deficiency.
UR - http://hdl.handle.net/10754/628729
UR - https://www.sciencedirect.com/science/article/pii/S0168945218306022
UR - http://www.scopus.com/inward/record.url?scp=85053336526&partnerID=8YFLogxK
U2 - 10.1016/j.plantsci.2018.06.024
DO - 10.1016/j.plantsci.2018.06.024
M3 - Article
C2 - 30466598
AN - SCOPUS:85053336526
SN - 0168-9452
VL - 277
SP - 33
EP - 42
JO - Plant Science
JF - Plant Science
ER -