Isolation, characterization, and evaluation of three Citrus sinensis-derived constitutive gene promoters.

Publisher: Springer Country of Publication: Germany NLM ID: 9880970 Publication Model: Print-Electronic Cited Medium: Internet ISSN: 1432-203X (Electronic) Linking ISSN: 07217714 NLM ISO Abbreviation: Plant Cell Rep Subsets: MEDLINE

Original Publication: Berlin ; New York : Springer, 1981-

Citrus sinensis/*metabolism ; Plants, Genetically Modified/*metabolism; Citrus sinensis/genetics ; Cyclophilins/genetics ; Cyclophilins/metabolism ; Gene Expression Regulation, Plant/genetics ; Gene Expression Regulation, Plant/physiology ; Plant Proteins/genetics ; Plant Proteins/metabolism ; Plants, Genetically Modified/genetics ; Promoter Regions, Genetic/genetics ; Promoter Regions, Genetic/physiology ; RNA, Messenger/genetics

Key Message: Regulatory sequences from the citrus constitutive genes cyclophilin (CsCYP), glyceraldehyde-3-phosphate dehydrogenase C2 (CsGAPC2), and elongation factor 1-alpha (CsEF1) were isolated, fused to the uidA gene, and qualitatively and quantitatively evaluated in transgenic sweet orange plants. The 5' upstream region of a gene (the promoter) is the most important component for the initiation and regulation of gene transcription of both native genes and transgenes in plants. The isolation and characterization of gene regulatory sequences are essential to the development of intragenic or cisgenic genetic manipulation strategies, which imply the use of genetic material from the same species or from closely related species. We describe herein the isolation and evaluation of the promoter sequence from three constitutively expressed citrus genes: cyclophilin (CsCYP), glyceraldehyde-3-phosphate dehydrogenase C2 (CsGAPC2), and elongation factor 1-alpha (CsEF1). The functionality of the promoters was confirmed by a histochemical GUS assay in leaves, stems, and roots of stably transformed citrus plants expressing the promoter-uidA construct. Lower uidA mRNA levels were detected when the transgene was under the control of citrus promoters as compared to the expression under the control of the CaMV35S promoter. The association of the uidA gene with the citrus-derived promoters resulted in mRNA levels of up to 60-41.8% of the value obtained with the construct containing CaMV35S driving the uidA gene. Moreover, a lower inter-individual variability in transgene expression was observed amongst the different transgenic lines, where gene constructs containing citrus-derived promoters were used. In silico analysis of the citrus-derived promoter sequences revealed that their activity may be controlled by several putative cis-regulatory elements. These citrus promoters will expand the availability of regulatory sequences for driving gene expression in citrus gene-modification programs.

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140183/2013-2) Conselho Nacional de Desenvolvimento Científico e Tecnológico; 2013/19700-2 Fundação de Amparo à Pesquisa do Estado de São Paulo

Keywords: Cis-regulatory elements; Cultivar improvement; Gene expression; Sweet orange

0 (Plant Proteins)
0 (RNA, Messenger)
EC 5.2.1.- (Cyclophilins)

Date Created: 20180526 Date Completed: 20181022 Latest Revision: 20220318

20240829

10.1007/s00299-018-2298-1

29796947