Deficiency of valencene in mandarin hybrids is associated with a deletion in the promoter region of the valencene synthase gene

Abstract Background Valencene is a major sesquiterpene in citrus oil and biosynthesized by valencene synthase (Cstps1; EC: 4.2.3.73) from the 15-carbon substrate farnesyl diphosphate. It is abundant in juice of some mandarins (e.g. Citrus reticulata Blanco cv. Fortune), however, it is undetectable i...

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Main Authors: Qibin Yu, Ming Huang, Hongge Jia, Yuan Yu, Anne Plotto, Elizabeth A. Baldwin, Jinhe Bai, Nian Wang, Frederick G. Gmitter Jr
Format: Article
Language:English
Published: BMC 2019-03-01
Series:BMC Plant Biology
Subjects:
Online Access:http://link.springer.com/article/10.1186/s12870-019-1701-6
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author Qibin Yu
Ming Huang
Hongge Jia
Yuan Yu
Anne Plotto
Elizabeth A. Baldwin
Jinhe Bai
Nian Wang
Frederick G. Gmitter Jr
author_facet Qibin Yu
Ming Huang
Hongge Jia
Yuan Yu
Anne Plotto
Elizabeth A. Baldwin
Jinhe Bai
Nian Wang
Frederick G. Gmitter Jr
author_sort Qibin Yu
collection DOAJ
description Abstract Background Valencene is a major sesquiterpene in citrus oil and biosynthesized by valencene synthase (Cstps1; EC: 4.2.3.73) from the 15-carbon substrate farnesyl diphosphate. It is abundant in juice of some mandarins (e.g. Citrus reticulata Blanco cv. Fortune), however, it is undetectable in others (e.g. C. reticulata Blanco cv. Murcott), We have discovered that the Murcott mandarin Cstps1 gene expression is severely reduced. A previous genetic mapping study using an F1 population of Fortune × Murcott found that the segregation of valencene production in fruit exhibited a Mendelian inheritance ratio of 1:1. There was only one dominant locus associated with valencene content detected on the mandarin genetic map. The goal of this study was to understand the molecular mechanism underlying the valencene deficiency observed in some citrus hybrids. Results There was a clear relationship between presence or absence of the valencene synthase gene (Cstps1) expression, and presence or absence of valencene among randomly selected mandarin hybrids. Cloning the coding regions of Cstps1 from Fortune and Murcott mandarin, and aligning with previous reported Valencia orange Cstps1 sequence, showed that they both exhibited extremely high similarity with the known Cstps1. By further cloning and analyzing the promoter region of Cstps1 from Valencia, Fortune and Murcott, a 12-nucleotide deletion at approximately − 270 bp from the Cstps1 coding region was only found in Murcott. Three binary vectors, designated as p1380-FortP-GUSin, p1380-MurcP-GUSin and p1380-MurcP(+ 12)-GUSin, were developed for promoter activity analysis. Transient over-expression of Fortune Cstps1 promoter in sweet orange showed notable GUS activity, but the Murcott Cstps1 promoter did not. In addition, by re-inserting the 12-nucleotide fragment, the activity of the Murcott Cstps1 promoter was mostly recovered. Conclusion The deficiency of valencene production in some mandarins is probably due to a 12-nucleotide deletion in the promoter region of the Cstps1, which could be a crucial switch of Cstps1 transcription. Our results further enhanced the understanding of valencene biosynthesis in citrus.
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spelling doaj.art-48af3b67e73b4cc4b1d29edd75fd9cba2022-12-22T02:40:18ZengBMCBMC Plant Biology1471-22292019-03-0119111110.1186/s12870-019-1701-6Deficiency of valencene in mandarin hybrids is associated with a deletion in the promoter region of the valencene synthase geneQibin Yu0Ming Huang1Hongge Jia2Yuan Yu3Anne Plotto4Elizabeth A. Baldwin5Jinhe Bai6Nian Wang7Frederick G. Gmitter Jr8University of Florida, Institute of Food and Agricultural Sciences, Citrus Research and Education CenterUniversity of Florida, Institute of Food and Agricultural Sciences, Citrus Research and Education CenterUniversity of Florida, Institute of Food and Agricultural Sciences, Citrus Research and Education CenterUniversity of Florida, Institute of Food and Agricultural Sciences, Citrus Research and Education CenterUSDA-ARS Horticultural Research LaboratoryUSDA-ARS Horticultural Research LaboratoryUSDA-ARS Horticultural Research LaboratoryUniversity of Florida, Institute of Food and Agricultural Sciences, Citrus Research and Education CenterUniversity of Florida, Institute of Food and Agricultural Sciences, Citrus Research and Education CenterAbstract Background Valencene is a major sesquiterpene in citrus oil and biosynthesized by valencene synthase (Cstps1; EC: 4.2.3.73) from the 15-carbon substrate farnesyl diphosphate. It is abundant in juice of some mandarins (e.g. Citrus reticulata Blanco cv. Fortune), however, it is undetectable in others (e.g. C. reticulata Blanco cv. Murcott), We have discovered that the Murcott mandarin Cstps1 gene expression is severely reduced. A previous genetic mapping study using an F1 population of Fortune × Murcott found that the segregation of valencene production in fruit exhibited a Mendelian inheritance ratio of 1:1. There was only one dominant locus associated with valencene content detected on the mandarin genetic map. The goal of this study was to understand the molecular mechanism underlying the valencene deficiency observed in some citrus hybrids. Results There was a clear relationship between presence or absence of the valencene synthase gene (Cstps1) expression, and presence or absence of valencene among randomly selected mandarin hybrids. Cloning the coding regions of Cstps1 from Fortune and Murcott mandarin, and aligning with previous reported Valencia orange Cstps1 sequence, showed that they both exhibited extremely high similarity with the known Cstps1. By further cloning and analyzing the promoter region of Cstps1 from Valencia, Fortune and Murcott, a 12-nucleotide deletion at approximately − 270 bp from the Cstps1 coding region was only found in Murcott. Three binary vectors, designated as p1380-FortP-GUSin, p1380-MurcP-GUSin and p1380-MurcP(+ 12)-GUSin, were developed for promoter activity analysis. Transient over-expression of Fortune Cstps1 promoter in sweet orange showed notable GUS activity, but the Murcott Cstps1 promoter did not. In addition, by re-inserting the 12-nucleotide fragment, the activity of the Murcott Cstps1 promoter was mostly recovered. Conclusion The deficiency of valencene production in some mandarins is probably due to a 12-nucleotide deletion in the promoter region of the Cstps1, which could be a crucial switch of Cstps1 transcription. Our results further enhanced the understanding of valencene biosynthesis in citrus.http://link.springer.com/article/10.1186/s12870-019-1701-6CloningQTL mappingSesquiterpenecDNA sequencingTransient expressionCitrus
spellingShingle Qibin Yu
Ming Huang
Hongge Jia
Yuan Yu
Anne Plotto
Elizabeth A. Baldwin
Jinhe Bai
Nian Wang
Frederick G. Gmitter Jr
Deficiency of valencene in mandarin hybrids is associated with a deletion in the promoter region of the valencene synthase gene
BMC Plant Biology
Cloning
QTL mapping
Sesquiterpene
cDNA sequencing
Transient expression
Citrus
title Deficiency of valencene in mandarin hybrids is associated with a deletion in the promoter region of the valencene synthase gene
title_full Deficiency of valencene in mandarin hybrids is associated with a deletion in the promoter region of the valencene synthase gene
title_fullStr Deficiency of valencene in mandarin hybrids is associated with a deletion in the promoter region of the valencene synthase gene
title_full_unstemmed Deficiency of valencene in mandarin hybrids is associated with a deletion in the promoter region of the valencene synthase gene
title_short Deficiency of valencene in mandarin hybrids is associated with a deletion in the promoter region of the valencene synthase gene
title_sort deficiency of valencene in mandarin hybrids is associated with a deletion in the promoter region of the valencene synthase gene
topic Cloning
QTL mapping
Sesquiterpene
cDNA sequencing
Transient expression
Citrus
url http://link.springer.com/article/10.1186/s12870-019-1701-6
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