Functional Analysis of SoCHR35 in the Biosynthesis of Ocimene in Syringa oblata

doi:10.16420/j.issn.0513-353x.2024-0817

Acta Horticulturae Sinica ›› 2025, Vol. 52 ›› Issue (10): 2655-2665.doi: 10.16420/j.issn.0513-353x.2024-0817

• Genetic & Breeding·Germplasm Resources·Molecular Biology • Previous Articles Next Articles

Functional Analysis of SoCHR35 in the Biosynthesis of Ocimene in Syringa oblata

WANG Jinxuan¹, NI Ying¹, MENG Xin³, LENG Zhuo¹, MA Bo¹, LENG Pingsheng¹^,², WU Jing¹^,²^,^*(), HU Zenghui¹^,²^,^*()

¹ Beijing Advanced Innovation Center for Tree Breeding by Molecular Design，Ancient Tree Health and Culture Engineering Technology Research Center，National Forestry and Grassland Administration，College of Landscape Architecture，Beijing University of Agriculture，Beijing 102206，China
² Beijing Laboratory for Urban and Rural Ecological Environment，Beijing 102206，China
³ National Botanical Garden，Beijing 100093，China

Received:2025-02-14 Revised:2025-07-21 Online:2025-10-25 Published:2025-10-28
Contact: WU Jing, HU Zenghui

Abstract

Abstract:

In order to investigate the role of SoCHR35（Chromatin remodeling complex ATPase 35），a member of RNA-mediated DNA methylation pathway，in the biosynthesis of lilac floral substance—ocimene，the SoCHR35 gene was cloned and analyzed，including bioinformatics，spatiotemporal expression pattern，functional characterization，and subcellular localization. The results showed that the total length of the CDS sequence of SoCHR35 was 2 073 bp，encoding for 690 amino acids. The protein had the typical SNF2 conserved domains of DEXDc and HELICc，and was clustered with CHR35 of Perilla frutescens and Sesamum indicum. The spatiotemporal expression analysis showed that with the flower development the expression level of SoCHR35 first decreased and then increased，which was negatively correlated with the release of floral scents. After transiently overexpressing SoCHR35 in lilac petals，the release amount of ocimene was significantly reduced，while transiently silencing of SoCHR35 lead to a remarkable increase in the release amount of ocimene. Through subcellular localization，SoCHR35 was found to be located in chloroplasts，which was same to the expression position of SoTPS2，a ocimene synthase gene. Moreover，the transient silencing of SoCHR35 resulted in a significant enhancement of SoTPS2 expression in petals. These results indicated that SoCHR35 negatively regulated the biosynthesis of ocimene in lilac petals to impact the floral scent release，which may result from a direct or indirect regulation of SoTPS2.

Key words: Syringa oblata, ocimene, SoCHR35, DNA methylation, floral scent

WANG Jinxuan, NI Ying, MENG Xin, LENG Zhuo, MA Bo, LENG Pingsheng, WU Jing, HU Zenghui. Functional Analysis of SoCHR35 in the Biosynthesis of Ocimene in Syringa oblata[J]. Acta Horticulturae Sinica, 2025, 52(10): 2655-2665.

Figures/Tables 7

Table 1 Primer sequences used in this study

引物用途 Primer function	引物名称 Primer name	引物序列（5′-3′） Primer sequence
克隆基因 Gene clone	SoCHR35-F	ATGGTTAATGACATTGTTCTACATGT
克隆基因 Gene clone	SoCHR35-R	CCTTTTGAAGACAGCAACCACAT
实时荧光定量PCR qRT-PCR	SoActin-F	TGGAATGTGCTGAGAGATGC
	SoActin-R	TGCTGACCGTATGAGCAAAG
	qSoCHR35-F	AATTTTCCCACATCCACGGC
	qSoCHR35-R	TTCCAGAACCAGGTCATGA
	qSoTPS2-F	GGTTTGCTTCAGGGCACTTG
	qSoTPS2-R	CATCCACCTGAACGGTTCCA
基因过表达 Gene overexpression	PRI101-SoCHR35-F	ATGCCCGTCGACCCCGGGATGGTTAATGACATTGTTCTACATGTTGGC
基因过表达 Gene overexpression	PRI101-SoCHR35-R	TGCTCACCATGGATCCTTACCTTTTGAAGACAGCAACCACATCT TGGGAGATGATACGCTGTT
基因沉默 Gene silencing	TRV2-SoCHR35-F	GTCCAGTCCTGGCCTCGTCGGCCGATAACTATAAGAGGAAGGTTACCGTAATACAAGATC
	TRV2-SoCHR35-R	GACCACAAGTGGCCAGACTGGCCTAAGAAATTCTTCAACTTTGGGTGAACATAAACTGC
	TRV1-F	TTACAGGTTATTTGGGCTAG
	TRV1-R	CCGGGTTCAATTCCTTATC

Fig. 1 Alignment and analysis of SoCHR35 with CHR35 protein sequences of other species The black，red，and blue parts represent homology = 100%，≥ 75%，≥ 50%，respectively. Red boxes are DEXDc conserved motifs，and green boxes are HELICc conserved motifs. Oe：Olea europaea；Si：Sesamum indicum；Ac：Actinidia chinensis；Ap：Andrographis paniculata；Ca：Capsicum annuum；Ce：Coffea eugenioides

Fig. 2 Phylogenetic tree of SoCHR35 from Syringa oblata and homologous proteins of other plant species

Fig. 3 The expression pattern of SoCHR35 in different organs（A）and at different floral developmental stages（B） Different lowercase letters indicate significant differences at the P < 0.05 level

Fig. 4 The effect on floral scent of SoCHR35 transient overexpression A：Gene overexpression efficiency；B：The release amount of floral scents；C：Total ion current（TIC）chromatograms of the floral scent compounds emitted from lilac of 35S and 35S:SoCHR35 t-test，**P < 0.01，***P < 0.001

Fig. 5 The effect on floral scent of SoCHR35 transient silencing A：Gene silencing efficiency；B：The release amount of floral；C：Total ion current（TIC）chromatograms of the floral scent compounds emitted from lilac of TRV and TRV:SoCHR35 t-test，** P < 0.01，**** P < 0.0001

Fig. 6 The subcellular localization of SoCHR35（A）and the expression analysis of downstream SoTPS2（B） t-test，*** P < 0.001

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Functional Analysis of SoCHR35 in the Biosynthesis of Ocimene in Syringa oblata

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