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 inFrontiers in Plant Science Vol. 9; p. 765
Main Authors Chuang, Yu-Chen, Hung, Yi-Chu, Hsu, Chi-Yu, Yeh, Chuan-Ming, Mitsuda, Nobutaka, Ohme-Takagi, Masaru, Tsai, Wen-Chieh, Chen, Wen-Huei, Chen, Hong-Hwa
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LanguageEnglish
Published Switzerland Frontiers Media SA 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
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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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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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SSID ssj0000500997
Score 2.2707727
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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SourceType Open Website
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StartPage 765
SubjectTerms cis-element
floral scent
monoterpene
orchid
Phalaenopsis
Plant culture
Plant Science
promoter
SB1-1110
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Title A Dual Repeat Cis-Element Determines Expression of GERANYL DIPHOSPHATE SYNTHASE for Monoterpene Production in Phalaenopsis Orchids
URI https://cir.nii.ac.jp/crid/1872272493011362432
https://www.ncbi.nlm.nih.gov/pubmed/29922327
https://www.proquest.com/docview/2057441435
https://pubmed.ncbi.nlm.nih.gov/PMC5996158
https://doaj.org/article/9cdf317e5cab489899abe2421092f587
Volume 9
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