Dataset Information

Identification and Characterization of Transcription Factors Involved in Geraniol Biosynthesis in Rosa chinensis.

ABSTRACT: Fragrance is an important characteristic of rose flowers and is largely determined by the terpenes. Rose has a unique NUDX1 (NUDIX HYDROLASES 1)-dependent monoterpene geraniol biosynthesis pathway, but little is known about its transcriptional regulation. In this study, we characterized two China rose (Rosa chinensis) materials from the 'Old Blush' variety with contrasting aromas. We profiled the volatile metabolome of both materials, and the results revealed that geraniol was the main component that distinguishes the aroma of these two materials. We performed a comparative transcriptome analysis of the two rose materials, from which we identified the hydrolase RcNUDX1 as a key factor affecting geraniol content, as well as 17 transcription factor genes co-expressed with RcNUDX1. We also determined that the transcription factor RcWRKY70 binds to four W-box motifs in the promoter of RcNUDX1, repressing RcNUDX1 expression, based on yeast one-hybrid and transient dual-luciferase assays. These results provide important information concerning the transcriptional regulatory framework underlying the control of geraniol production in rose.

SUBMITTER: Yu J

PROVIDER: S-EPMC9739587 | biostudies-literature | 2022 Nov

REPOSITORIES: biostudies-literature

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Identification and Characterization of Transcription Factors Involved in Geraniol Biosynthesis in <i>Rosa chinensis</i>.

Yu Jiayao J Liu Xiaoyu X Peng Yifang Y Li Qi Q Han Yu Y

International journal of molecular sciences 20221124 23

Fragrance is an important characteristic of rose flowers and is largely determined by the terpenes. Rose has a unique NUDX1 (NUDIX HYDROLASES 1)-dependent monoterpene geraniol biosynthesis pathway, but little is known about its transcriptional regulation. In this study, we characterized two China rose (<i>Rosa chinensis</i>) materials from the 'Old Blush' variety with contrasting aromas. We profiled the volatile metabolome of both materials, and the results revealed that geraniol was the main co ...[more]

PMID: 36499007

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Project description:BackgroundRose (Rosa chinensis) is a traditional famous flower with valuable ornamental characteristics. However, drought stress restricts its growth and development, leading to an abnormal phenotype. One of the main transcription factor (TF) protein groups in the plant kingdom are the APETALA2/ethylene-responsive factor (AP2/ERF) proteins and are potentially involved in the growth and stress responses of various plants.ResultsOur investigation mainly focused on exploring the genome of rose and thereby we discovered 135 apparent AP2/ERF TFs. Phylogenic analyses revealed that RcAP2/ERF genes are categorized into DREB, Soloist, AP2, and ERF subfamilies, and are further classified these into 17 groups, with the same as Malus domestica and Arabidopsis thaliana. The analysis of the gene structure revealed that the introns ranged from 0 to 9 in number. Pattern examination demonstrated that the RcAP2/ERF predominantly consists of typical AP2 domains, of which the 2nd motif is the most ubiquitous. Distributions of cis-acting elements indicated that members of the AP2/ERF family are frequently involved in growth and development, phytohormone and stress response in rose species. Also, the distribution mapping of the rose chromosomes indicated that AP2/ERF class genes are dispersed among all seven chromosomes. Additionally, we isolated a novel DREB A2 subgroup gene and named it RcDREB2B. Subsequently, the RcDREB2B transcript accumulation was repressed under the mild and severe drought stress in the root samples of rose. RcDREB2B was targeted to the nucleus and exhibited transactivation in yeast cells. The overexpression of RcDREB2B was found to promote sensitivity to a higher salt concentration, ABA, and PEG at the germination and post-germination stages. Twelve putative osmotic and ABA-related genes were impaired in RcDREB2B-overexpressing plants.ConclusionsThe results provide comprehensive information regarding the gene structure, phylogenic, and distribution of the rose AP2/ERF family and bring insight into the complex transcriptional gene regulation of RcAP2/ERF. Findings in this study would also contribute to further understanding of the RcDREB2B gene in rose.

Project description:BackgroundIncreased defense against a variety of pathogens in plants is achieved through activation of a mechanism known as systemic acquired resistance (SAR). The broad-spectrum resistance brought about by SAR is mediated through salicylic acid (SA). An important step in SA biosynthesis in Arabidopsis is the conversion of chorismate to isochorismate through the action of isochorismate synthase, encoded by the ICS1 gene. Also AVRPPHB SUSCEPTIBLE 3 (PBS3) plays an important role in SA metabolism, as pbs3 mutants accumulate drastically reduced levels of SA-glucoside, a putative storage form of SA. Bioinformatics analysis previously performed by us identified WRKY28 and WRKY46 as possible regulators of ICS1 and PBS3.ResultsExpression studies with ICS1 promoter::β-glucuronidase (GUS) genes in Arabidopsis thaliana protoplasts cotransfected with 35S::WRKY28 showed that over expression of WRKY28 resulted in a strong increase in GUS expression. Moreover, qRT-PCR analyses indicated that the endogenous ICS1 and PBS3 genes were highly expressed in protoplasts overexpressing WRKY28 or WRKY46, respectively. Electrophoretic mobility shift assays indentified potential WRKY28 binding sites in the ICS1 promoter, positioned -445 and -460 base pairs upstream of the transcription start site. Mutation of these sites in protoplast transactivation assays showed that these binding sites are functionally important for activation of the ICS1 promoter. Chromatin immunoprecipitation assays with haemagglutinin-epitope-tagged WRKY28 showed that the region of the ICS1 promoter containing the binding sites at -445 and -460 was highly enriched in the immunoprecipitated DNA.ConclusionsThe results obtained here confirm results from our multiple microarray co-expression analyses indicating that WRKY28 and WRKY46 are transcriptional activators of ICS1 and PBS3, respectively, and support this in silico screening as a powerful tool for identifying new components of stress signaling pathways.

Dataset Information

Identification and Characterization of Transcription Factors Involved in Geraniol Biosynthesis in Rosa chinensis.

Publications

Identification and Characterization of Transcription Factors Involved in Geraniol Biosynthesis in <i>Rosa chinensis</i>.

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