A Dual Repeat Cis-Element Determines Expression of GERANYL DIPHOSPHATE SYNTHASE for Monoterpene Production in Phalaenopsis Orchids
is a scented orchid emitting large amount of monoterpenes. GERANYL DIPHOSPHATE SYNTHASE (PbGDPS) is the key enzyme for monoterpene biosynthesis, and shows concomitant expression with the emission of monoterpenes during flower development in . Here, we identified a dual repeat -element in the promote...
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Published in | Frontiers in Plant Science Vol. 9; p. 765 |
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Main Authors | , , , , , , , , |
Format | Journal Article |
Language | English |
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05.06.2018
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Abstract | is a scented orchid emitting large amount of monoterpenes. GERANYL DIPHOSPHATE SYNTHASE (PbGDPS) is the key enzyme for monoterpene biosynthesis, and shows concomitant expression with the emission of monoterpenes during flower development in
. Here, we identified a dual repeat
-element in the
promoter that is critical for monoterpene biosynthesis in
orchids. A strong correlation between the dual repeat and the monoterpene production was revealed by examination of the
promoter fragments over 12
species. Serial-deletion of the 2-kb
promoter fragments demonstrated that the integrity of the dual repeat was crucial for its promoter activities. By screening the
transcription factors (TFs) cDNA library using yeast one-hybrid assay, AtbZIP18, a member of group I of bZIP TFs, was identified to be able to bind the dual repeat. We then identified
in the transcriptome of
, showing 83% identity in the DNA binding region with that of AtbZIP18, and the expression level of
was higher in the scented orchids. In addition, PbbZIP4 transactivated the
promoter fragment containing the dual repeat in dual luciferase assay. Furthermore, transient ectopic expression of
induced a 10-fold production of monoterpenoids in the scentless orchid. In conclusion, these results indicate that the dual repeat is a real TF-bound
-element significant for
gene expression, and thus subsequent monoterpene biosynthesis in the scented
orchids. |
---|---|
AbstractList | Phalaenopsis bellina is a scented orchid emitting large amount of monoterpenes. GERANYL DIPHOSPHATE SYNTHASE (PbGDPS) is the key enzyme for monoterpene biosynthesis, and shows concomitant expression with the emission of monoterpenes during flower development in P. bellina. Here, we identified a dual repeat cis-element in the GDPS promoter that is critical for monoterpene biosynthesis in Phalaenopsis orchids. A strong correlation between the dual repeat and the monoterpene production was revealed by examination of the GDPS promoter fragments over 12 Phalaenopsis species. Serial-deletion of the 2-kb GDPS promoter fragments demonstrated that the integrity of the dual repeat was crucial for its promoter activities. By screening the Arabidopsis transcription factors (TFs) cDNA library using yeast one-hybrid assay, AtbZIP18, a member of group I of bZIP TFs, was identified to be able to bind the dual repeat. We then identified PbbZIP4 in the transcriptome of P. bellina, showing 83% identity in the DNA binding region with that of AtbZIP18, and the expression level of PbbZIP4 was higher in the scented orchids. In addition, PbbZIP4 transactivated the GDPS promoter fragment containing the dual repeat in dual luciferase assay. Furthermore, transient ectopic expression of PbbZIP4 induced a 10-fold production of monoterpenoids in the scentless orchid. In conclusion, these results indicate that the dual repeat is a real TF-bound cis-element significant for GDPS gene expression, and thus subsequent monoterpene biosynthesis in the scented Phalaenopsis orchids.Phalaenopsis bellina is a scented orchid emitting large amount of monoterpenes. GERANYL DIPHOSPHATE SYNTHASE (PbGDPS) is the key enzyme for monoterpene biosynthesis, and shows concomitant expression with the emission of monoterpenes during flower development in P. bellina. Here, we identified a dual repeat cis-element in the GDPS promoter that is critical for monoterpene biosynthesis in Phalaenopsis orchids. A strong correlation between the dual repeat and the monoterpene production was revealed by examination of the GDPS promoter fragments over 12 Phalaenopsis species. Serial-deletion of the 2-kb GDPS promoter fragments demonstrated that the integrity of the dual repeat was crucial for its promoter activities. By screening the Arabidopsis transcription factors (TFs) cDNA library using yeast one-hybrid assay, AtbZIP18, a member of group I of bZIP TFs, was identified to be able to bind the dual repeat. We then identified PbbZIP4 in the transcriptome of P. bellina, showing 83% identity in the DNA binding region with that of AtbZIP18, and the expression level of PbbZIP4 was higher in the scented orchids. In addition, PbbZIP4 transactivated the GDPS promoter fragment containing the dual repeat in dual luciferase assay. Furthermore, transient ectopic expression of PbbZIP4 induced a 10-fold production of monoterpenoids in the scentless orchid. In conclusion, these results indicate that the dual repeat is a real TF-bound cis-element significant for GDPS gene expression, and thus subsequent monoterpene biosynthesis in the scented Phalaenopsis orchids. Phalaenopsis bellina is a scented orchid emitting large amount of monoterpenes. GERANYL DIPHOSPHATE SYNTHASE (PbGDPS) is the key enzyme for monoterpene biosynthesis, and shows concomitant expression with the emission of monoterpenes during flower development in P. bellina . Here, we identified a dual repeat cis -element in the GDPS promoter that is critical for monoterpene biosynthesis in Phalaenopsis orchids. A strong correlation between the dual repeat and the monoterpene production was revealed by examination of the GDPS promoter fragments over 12 Phalaenopsis species. Serial-deletion of the 2-kb GDPS promoter fragments demonstrated that the integrity of the dual repeat was crucial for its promoter activities. By screening the Arabidopsis transcription factors (TFs) cDNA library using yeast one-hybrid assay, AtbZIP18, a member of group I of bZIP TFs, was identified to be able to bind the dual repeat. We then identified PbbZIP4 in the transcriptome of P. bellina , showing 83% identity in the DNA binding region with that of AtbZIP18, and the expression level of PbbZIP4 was higher in the scented orchids. In addition, PbbZIP4 transactivated the GDPS promoter fragment containing the dual repeat in dual luciferase assay. Furthermore, transient ectopic expression of PbbZIP4 induced a 10-fold production of monoterpenoids in the scentless orchid. In conclusion, these results indicate that the dual repeat is a real TF-bound cis -element significant for GDPS gene expression, and thus subsequent monoterpene biosynthesis in the scented Phalaenopsis orchids. Phalaenopsis bellina is a scented orchid emitting large amount of monoterpenes. GERANYL DIPHOSPHATE SYNTHASE (PbGDPS) is the key enzyme for monoterpene biosynthesis, and shows concomitant expression with the emission of monoterpenes during flower development in P. bellina. Here, we identified a dual repeat cis-element in the GDPS promoter that is critical for monoterpene biosynthesis in Phalaenopsis orchids. A strong correlation between the dual repeat and the monoterpene production was revealed by examination of the GDPS promoter fragments over 12 Phalaenopsis species. Serial-deletion of the 2-kb GDPS promoter fragments demonstrated that the integrity of the dual repeat was crucial for its promoter activities. By screening the Arabidopsis transcription factors (TFs) cDNA library using yeast one-hybrid assay, AtbZIP18, a member of group I of bZIP TFs, was identified to be able to bind the dual repeat. We then identified PbbZIP4 in the transcriptome of P. bellina, showing 83% identity in the DNA binding region with that of AtbZIP18, and the expression level of PbbZIP4 was higher in the scented orchids. In addition, PbbZIP4 transactivated the GDPS promoter fragment containing the dual repeat in dual luciferase assay. Furthermore, transient ectopic expression of PbbZIP4 induced a 10-fold production of monoterpenoids in the scentless orchid. In conclusion, these results indicate that the dual repeat is a real TF-bound cis-element significant for GDPS gene expression, and thus subsequent monoterpene biosynthesis in the scented Phalaenopsis orchids. is a scented orchid emitting large amount of monoterpenes. GERANYL DIPHOSPHATE SYNTHASE (PbGDPS) is the key enzyme for monoterpene biosynthesis, and shows concomitant expression with the emission of monoterpenes during flower development in . Here, we identified a dual repeat -element in the promoter that is critical for monoterpene biosynthesis in orchids. A strong correlation between the dual repeat and the monoterpene production was revealed by examination of the promoter fragments over 12 species. Serial-deletion of the 2-kb promoter fragments demonstrated that the integrity of the dual repeat was crucial for its promoter activities. By screening the transcription factors (TFs) cDNA library using yeast one-hybrid assay, AtbZIP18, a member of group I of bZIP TFs, was identified to be able to bind the dual repeat. We then identified in the transcriptome of , showing 83% identity in the DNA binding region with that of AtbZIP18, and the expression level of was higher in the scented orchids. In addition, PbbZIP4 transactivated the promoter fragment containing the dual repeat in dual luciferase assay. Furthermore, transient ectopic expression of induced a 10-fold production of monoterpenoids in the scentless orchid. In conclusion, these results indicate that the dual repeat is a real TF-bound -element significant for gene expression, and thus subsequent monoterpene biosynthesis in the scented orchids. |
Author | Nobutaka Mitsuda Hong-Hwa Chen Chi-Yu Hsu Masaru Ohme-Takagi Wen-Huei Chen Yu-Chen Chuang Yi-Chu Hung Chuan-Ming Yeh Wen-Chieh Tsai |
AuthorAffiliation | 5 Orchid Research and Development Center, National Cheng Kung University , Tainan , Taiwan 2 Division of Strategic Research and Development, Graduate School of Science and Engineering, Saitama University , Saitama , Japan 1 Department of Life Sciences, National Cheng Kung University , Tainan , Taiwan 4 Institute of Tropical Plant Sciences, National Cheng Kung University , Tainan , Taiwan 3 Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology , Tsukuba , Japan |
AuthorAffiliation_xml | – name: 1 Department of Life Sciences, National Cheng Kung University , Tainan , Taiwan – name: 3 Bioproduction Research Institute, National Institute of Advanced Industrial Science and Technology , Tsukuba , Japan – name: 4 Institute of Tropical Plant Sciences, National Cheng Kung University , Tainan , Taiwan – name: 5 Orchid Research and Development Center, National Cheng Kung University , Tainan , Taiwan – name: 2 Division of Strategic Research and Development, Graduate School of Science and Engineering, Saitama University , Saitama , Japan |
Author_xml | – sequence: 1 givenname: Yu-Chen surname: Chuang fullname: Chuang, Yu-Chen – sequence: 2 givenname: Yi-Chu surname: Hung fullname: Hung, Yi-Chu – sequence: 3 givenname: Chi-Yu surname: Hsu fullname: Hsu, Chi-Yu – sequence: 4 givenname: Chuan-Ming surname: Yeh fullname: Yeh, Chuan-Ming – sequence: 5 givenname: Nobutaka surname: Mitsuda fullname: Mitsuda, Nobutaka – sequence: 6 givenname: Masaru surname: Ohme-Takagi fullname: Ohme-Takagi, Masaru – sequence: 7 givenname: Wen-Chieh surname: Tsai fullname: Tsai, Wen-Chieh – sequence: 8 givenname: Wen-Huei surname: Chen fullname: Chen, Wen-Huei – sequence: 9 givenname: Hong-Hwa surname: Chen fullname: Chen, Hong-Hwa |
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Copyright | Copyright © 2018 Chuang, Hung, Hsu, Yeh, Mitsuda, Ohme-Takagi, Tsai, Chen and Chen. 2018 Chuang, Hung, Hsu, Yeh, Mitsuda, Ohme-Takagi, Tsai, Chen and Chen |
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Keywords | transcription factor monoterpene Phalaenopsis yeast-one hybrid screening cis-element floral scent promoter orchid |
Language | English |
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Notes | ObjectType-Article-1 SourceType-Scholarly Journals-1 ObjectType-Feature-2 content type line 23 Edited by: Swee-Suak Ko, Academia Sinica, Taiwan This article was submitted to Plant Breeding, a section of the journal Frontiers in Plant Science Reviewed by: Dinesh A. Nagegowda, Central Institute of Medicinal and Aromatic Plants (CIMAP), India; Tariq Akhtar, University of Guelph, Canada |
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Snippet | is a scented orchid emitting large amount of monoterpenes. GERANYL DIPHOSPHATE SYNTHASE (PbGDPS) is the key enzyme for monoterpene biosynthesis, and shows... Phalaenopsis bellina is a scented orchid emitting large amount of monoterpenes. GERANYL DIPHOSPHATE SYNTHASE (PbGDPS) is the key enzyme for monoterpene... Phalaenopsis bellina is a scented orchid emitting large amount of monoterpenes. GERANYL DIPHOSPHATE SYNTHASE (PbGDPS) is the key enzyme for monoterpene... |
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Title | A Dual Repeat Cis-Element Determines Expression of GERANYL DIPHOSPHATE SYNTHASE for Monoterpene Production in Phalaenopsis Orchids |
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