Highly efficient transient gene expression of three tissues in Osmanthus fragrans mediated by Agrobacterium tumefaciens

Zhong, Shiwei; Dong, Bin; Zhou, Jie; Miao, Yunfeng; Yang, Liyuan; Wang, Yiguang; Xiao, Zhen; Fang, Qiu; Wan, Qianqian; Zhao, Hongbo

doi:10.1016/j.scienta.2022.111725

Cited by 10 publications

(4 citation statements)

References 61 publications

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“…While similar to most woody plants, the incipient transgenic technology, hindered by the complexity of the genome, poses challenges for research on O. fragrans . The highly efficient transient gene expression protocols we previously established for Osmanthus can be a valuable tool for molecular regulation validation [ 82 ]. Furthermore, as biotechnology continues to advance, molecular research on Osmanthus will be expected to make greater progress in the future.…”

Section: Discussionmentioning

confidence: 99%

DNA methylome analysis reveals novel insights into active hypomethylated regulatory mechanisms of temperature-dependent flower opening in Osmanthus fragrans

Zhong,

Zhu,

et al. 2024

Horticulture Research

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Abstracts Short-term ambient low temperature (ALT) stimulation is necessary for Osmanthus fragrans to facilitate continued flower opening after floral bud development reaches maturity. DNA methylation, a vital epigenetic modification, regulates various biological processes in response to temperature fluctuations. However, its role in temperature-driven flower opening remains elusive. In this study, we identified the pivotal timeframe during which O. fragrans promptly detected temperature cues. Using whole-genome bisulfite sequencing (WGBS), we explored global DNA hypomethylation during this phase, with the most significant changes occurring in CHH sequence contexts. Auxin transport inhibitor (TIBA) application revealed that ALT-induced endogenous auxin accumulation promoted peduncle elongation. In our mRNA-seq analysis, we discovered the differentially expressed genes (DEGs) with hypo-differentially methylated regions (hypo-DMRs) were mainly enriched in auxin and temperature response, RNA processing, carbohydrate and lipid metabolism. The transcripts of three DNA demethylase genes (OfROS1a, OfDML3, OfDME) showed upregulation. Furthermore, all DNA methylase genes, except for OfCMT2b, also displayed increased expression, specifically with two of them, OfCMT3a and OfCMT1, being associated with hypo-DMRs. Promoter assays showed that OfROS1a, with promoters containing low temperature and auxin responsive elements, were activated by ALT and exogenous IAA at low concentrations but inhibited at high concentrations. Overexpression OfROS1 reduced endogenous auxin levels but enhanced the expression of genes related to auxin response and spliceosome in petunia. Furthermore, OfROS1 promoted sucrose synthesis in petunia corollas. Our data characterized the rapid response of active DNA hypomethylation to ALT and suggested a possible epiregulation of temperature-dependent flower opening in O. fragrans.

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Section: Discussionmentioning

confidence: 99%

DNA methylome analysis reveals novel insights into active hypomethylated regulatory mechanisms of temperature-dependent flower opening in Osmanthus fragrans

Zhong,

Zhu,

et al. 2024

Horticulture Research

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“…We propose that the five hub genes positively regulate the content changes of key metabolites and we revealed a potential molecular mechanism for the leaf color changes of O. fragrans 'Wucaigui'. The functional verification of genes cannot be achieved without an efficient and stable genetic transformation system [47,48]. Although the transformation systems of model plants, like Arabidopsis and tobacco, have been fully developed, it is more convincing to verify gene functions in their original species [49][50][51].…”

Section: Discussionmentioning

confidence: 99%

Integrated Metabolome and Transcriptome Analyses Provide New Insights into the Leaf Color Changes in Osmanthus fragrans cv. ‘Wucaigui’

Zhang,

Qiu,

Wang

et al. 2024

Forests

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Osmanthus fragrans, belonging to the family Oleaceae, is listed as one of the most important traditional ornamental plant species in China. A new cultivar O. fragrans ‘Wucaigui’ has a very diversified form in terms of leaf colors, in which the leaf color changes from red to yellow-green and finally to dark green. To understand the mechanisms involved in leaf color changes, metabolome and transcriptome studies were performed on leaves at different developmental stages. A total of 79 metabolites, two chlorophyll, 26 carotenoids, and 51 anthocyanins, were detected in the 6 different developmental stages. An orthogonal partial least squares discriminant analysis identified key metabolites at different developmental stages, including lutein, pelargonidin-3-O-(6-O-p-coumaroyl)-glucoside, neoxanthin, and α-carotene. A total of 48,837 genes were obtained by transcriptome sequencing, including 3295 novel genes. Using a weighted gene co-expression network analysis to study the correlations between key metabolites and differentially expressed genes, we determined the characteristic modules having the highest correlations with key metabolites and selected associated candidate genes. Five genes (OfSHOU4L, OfATL1B, OfUGE5 OfEIF1AX, and OfUGE3) were finally identified as hub genes using real-time fluorescence quantitative PCR. In addition, we proposed a model based on the changes in key metabolite contents and the network regulatory map during the changes in O. fragrans ‘Wucaigui’ leaf color. The positive regulation of OfUGE3 led to an increase in the lutein content, which resulted in the leaves changing from grayish brown to moderate brown; during the change from moderate brown to dark greenish-yellow, the positive regulation of three genes (OfHOU4L, OfATL1B, and OfUGE5) increased the content of pelargonidin-3-O-(6-O-p-coumaroyl)-glucoside and the red color of the leaves gradually faded to dark greenish-yellow and then to strong yellow-green; the positive regulation of OfEIF1AX increased the content of neoxanthin; the stages in which the color changed from strong yellow-green to yellow-green and then to moderate olive-green were positively regulated by OfUGE3, which resulted in higher α-carotene content. These findings provided new insights into the mechanisms underlying the processes involved in O. fragrans ‘Wucaigui’ leaf color changes at the metabolic and transcriptional levels. This work seeks to contribute to the development of artificial regulate and control technology in the breeding and production of O. fragrans and other ornamental plants.

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“…This line of research requires the identification of genes encoding enzymes involved in the synthesis of flower color and fragrance substances and the determination of how the expression of these genes is regulated, which may require the use of transgenic model plants [83]. Meanwhile, existing research shows that calli and others have carried out tissue culture of O. fragrans [84,85], but due to the low efficiency of callus tissue proliferation and the difficulty of differentiation of indefinite buds, the tissue culture and regeneration system of O. fragrans has not been successfully established [86]. The lack of complete sequencing of the O. fragrans genome may limit the cloning of O. fragrans genes and the development of gene transformation requires a stable O. fragrans tissue culture technology.…”

Section: Prospectmentioning

confidence: 99%

Recent Advances in Flower Color and Fragrance of Osmanthus fragrans

Qian

Shan

Zhao

et al. 2023

Forests

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Osmanthus fragrans is an evergreen shrub or tree of the Oleaceae family with a long history of cultivation in Asian countries and is one of the ten traditionally famous flowers in China, with important cultural and economic value. The unique floral color and fragrance of O. fragrans are formed by a variety of endogenous metabolites that distinguish it from other flowers and exhibit extraordinary ornamental value. However, many studies on the flower color and fragrance of this plant have been mainly based on bioactive extracts and physiological characteristics, leading to a notable lack of molecular machinery and systematic research. In this review, recent advances in bioactive ingredients associated with the underlying regulatory mechanisms, as well as the prospect for industrial utilization, are comprehensively presented and critically evaluated. In particular, the isolated components and essential genes required for flower color and fragrance are also well summarized, which will provide a scientific basis for molecular breeding for ornamental applications and facilitate the discovery of novel natural products for the future industrial development of O. fragrans. In prospect, we plan to use genetic research and high-throughput omics to analyze the genes related to the flower color and fragrance of O. fragrans, and at the same time, we will hybridize and breed excellent O. fragrans varieties that are resistant to low temperature.

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Highly efficient transient gene expression of three tissues in Osmanthus fragrans mediated by Agrobacterium tumefaciens

Cited by 10 publications

References 61 publications

DNA methylome analysis reveals novel insights into active hypomethylated regulatory mechanisms of temperature-dependent flower opening in Osmanthus fragrans

DNA methylome analysis reveals novel insights into active hypomethylated regulatory mechanisms of temperature-dependent flower opening in Osmanthus fragrans

Integrated Metabolome and Transcriptome Analyses Provide New Insights into the Leaf Color Changes in Osmanthus fragrans cv. ‘Wucaigui’

Recent Advances in Flower Color and Fragrance of Osmanthus fragrans

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