基于全基因组的栀子花香成分合成<i>TPS</i>基因系统分析

许文杰; 娄千; 陈启桢; 胡开治; 曹敏; 刘燕琴; 韩蓉蓉; 宋经元

doi:10.11913/PSJ.2095-0837.23017

基于全基因组的栀子花香成分合成TPS基因系统分析

1.
中国医学科学院北京协和医学院药用植物研究所，国家中医药管理局中药资源保护重点研究室，北京 100193
2.
重庆市药物种植研究所，重庆 408435

基金项目: 国家自然科学基金项目（82304674）。

详细信息

作者简介:
许文杰（1990−），女，博士，助理研究员，研究方向为植物次生代谢产物合成（E-mail：wjxu@implad.ac.cn）

通讯作者:
宋经元: E-mail：jysong@implad.ac.cn

中图分类号: Q943.2
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出版历程
- 收稿日期: 2023-03-16
- 录用日期: 2023-05-20
- 网络出版日期: 2023-03-13
- 刊出日期: 2024-01-31

Systematic analysis of TPS genes for the synthesis of floral aroma components of Gardenia jasminoides J. Ellis based on the whole genome

1.
Key Laboratory of Chinese Medicine Resources Conservation, State Administration of Traditional Chinese Medicine of the People’s Republic of China, Institute of Medicinal Plant Development, Chinese Academy of Medical Science and Peking Union Medical College, Beijing 100193, China
2.
Chongqing Institute of Medicinal Plant Cultivation, Chongqing 408435, China

摘要

摘要:
栀子（Gardenia jasminoides J. Ellis）花色洁白且香气怡人，不仅具有极高的观赏价值，还是重要天然香料来源。萜类是栀子花独特香气的主要组成成分，但合成该类挥发性产物的关键萜类合酶（TPS）尚未鉴定。本研究基于栀子高质量基因组，通过转录组数据、系统发育树和保守结构域分析等，全面挖掘栀子萜类花香成分合成相关的TPS基因。结果显示，栀子基因组共注释到44个GjTPS基因，不均匀分布于11条染色体，其中9个GjTPS参与串联复制事件。所有GjTPS被划分为5个亚家族，其中27个属于被子植物特有的TPS-a、TPS-b和TPS-g分支。结合栀子5个器官的转录组数据及实时荧光定量PCR分析，筛选出5个候选GjTPS基因，其在盛开期的花器官中特异高表达，且含有DDXXD和NSE/DTE 活性基序。其中，GjTPS1、GjTPS2、GjTPS3和GjTPS27为TPS-b分支酶，推测为芳樟醇或罗勒烯等栀子主要花香成分合成酶，而TPS-a分支的GjTPS18可能是金合欢烯合酶。
- 栀子 /
- 花香 /
- 萜类合酶 /
- 基因组 /
- 转录组
Abstract:
Gardenia jasminoides J. Ellis flowers are white in color and have a pleasant aroma. These flowers have high ornamental value and are an important source of natural spices. Terpenes are the main components of the unique aroma of G. jasminoides flowers, but the key terpene synthases (TPSs) for the biosynthesis of such volatile products have not yet been identified. Here, based on the high-quality genome of G. jasminoides, this study comprehensively explored the TPS genes related to floral scent biosynthesis through transcriptomic, phylogenetic, and conserved domain analysis. In total, 44 GjTPS genes were identified in G. jasminoides, unevenly distributed on 11 chromosomes, and nine GjTPS genes participated in tandem duplication events. All GjTPSs were clustered into five subfamilies, 27 of which belonged to the angiosperm-specific clades, including TPS-a, TPS-b, and TPS-g. Combining transcriptomic data and quantitative real-time PCR (qRT-PCR) analyses of five organs, five candidate GjTPS genes, with high expression in blooming flowers and containing the active motif DDXXD and NSE/DTE, were screened. Among them, GjTPS1, GjTPS2, GjTPS3, and GjTPS27 were grouped in the TPS-b clade, speculated to be the TPSs responsible for the biosynthesis of the main floral aroma components of G. jasminoides, such as linalool and ocimene, while GjTPS18 from the TPS-a clade may be a farnesene synthase.
- Gardenia jasminoides /
- Floral scent /
- Terpene synthase /
- Genome /
- Transcriptome
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图 1 栀子转录组测序样本基因表达相关性（A）及萜类花香成分合成途径（B）

圆的半径与相关系数成正比，蓝色和红色分别代表负相关和正相关。

Figure 1. Gene expression correlations of RNA-seq samples (A) and biosynthetic pathway of volatile terpenes in Gardenia jasminoides flowers (B)

Circle radius is proportional to correlation coefficient. Blue and red indicate negative and positive correlations of gene expression between pairs of samples, respectively.

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图 2 栀子与其他物种TPS基因系统发育树

Figure 2. Phylogenetic tree of TPS genes in Gardenia jasminoides and other species

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图 3 栀子TPS基因编码产物保守基序（A）及基因结构（B）

Figure 3. Conserved motifs (A) and gene structures (B) of GjTPS

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图 4 栀子TPS基因家族共线性（A）及表达热图（B）

红色字体标注的GjTPS基因与咖啡TPS存在共线性。

Figure 4. Collinear analysis (A) and gene expression heatmap (B) of GjTPSs

GjTPSs in red are collinear with TPSs of Coffea canephora.

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图 5 栀子萜类花香成分合成候选TPS蛋白局部比对

Figure 5. Partial amino acid alignment of candidate TPSs for volatile terpene biosynthesis in Gardenia jasminoides flowers

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图 6 栀子萜类花香成分合成候选TPS基因相对表达水平

Figure 6. Relative expression levels of candidate TPS genes related to volatile terpene biosynthesis in Gardenia jasminoides flowers

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表 1 不同物种的TPS蛋白信息

Table 1 Protein information of TPSs collected from multiple species

蛋白名称 Protein name	蛋白编号 Protein ID	蛋白名称 Protein name	蛋白编号 Protein ID
FhrTPS1^[8]		AtTPS02	P0CJ43.1
FhrTPS2^[8]		AtTPS03	A4FVP2.1
FhrTPS4^[8]		AtTPS06	Q84UU9.2
FhrTPS6^[8]		AtTPS10	Q9ZUH4.1
FhaTPS7^[8]		AtTPS11	Q4KSH9.2
FhaTPS8^[8]		AtTPS12	Q9T0J9.2
SlTPS3	G1JUH1.1	AtTPS13	Q9T0K1.2
SlTPS5	Q1XBU5.1	AtTPS14	Q84UV0.2
SlTPS9	O64961.1	AtTPS20	A0A178U9Y5.1
SlTPS12	D5KXD2.1	AtTPS21	Q84UU4.2
SlTPS14	G5CV54.1	AtTPS23	P0DI77.1
SlTPS17	G5CV52.1	AtTPS24	Q9LRZ6.1
SlTPS24	NP_001307929.1	AtTPS27	P0DI76.1
SlTPS31	G5CV46.1	LadGERDS	U3LVL5.1
SlTPS40	NP_001234008.2	LadCADS	U3LW50.1
AgPIN1	O24475.1	LadCARS	U3LVZ7.1
AgLIM1	O22340.1	LaBERS	Q2XSC4.1
CaTPS1	XP_027109762.1	LaLINS	Q2XSC5.1
CaTPS2	R4YXW8.2	LaLIMS	Q2XSC6.1
CaTPS3	XP_027093045.1
注：At：拟南芥；Ca：小果咖啡；Lad：薰衣草栽培品种‘Diva’；La：薰衣草；Fhr：香雪兰杂交品种‘Red River’；Fha：香雪兰杂交品种‘Ambiance’；Sl：番茄；Ag：大冷杉。下同。
Notes: At: Arabidopsis thaliana (L.) Heynh.; Ca: Coffea arabica L.; Lad: Lavandula angustifolia ‘Diva’; La: Lavandula angustifolia Mill.; Fhr: Freesia x hybrida ‘Red River’; Fha: Freesia x hybrida ‘Ambiance’; Sl: Solanum lycopersicum L.; Ag: Abies grandis (Douglas ex D. Don) Lindl.. Same below.

下载: 导出CSV

表 2 GjTPS基因表达定量引物

Table 2 Primers of GjTPS for qRT-PCR

基因 Gene	正向引物（5 ' →3 '） Forward primer	反向引物（5 ' →3 '） Reverse primer
GjACTIN	TCCTCTTCCAGCCTTCTATC	GCTCATACGGTCAGCAATAC
GjTPS1	CTTAGGCAACATGGGTACAA	CTCCTTCTGATCCAAGATTCG
GjTPS2	ACTTGGTCTGGCCAATTAC	ATGCCCATGTTGCCTAAG
GjTPS3	GTCTTGGTGAAGAAGGAGAAG	CGTTGTACCATTCCACTCTG
GjTPS18	TCAGCCTTCCAAGTTAAGCG	GTAAAGTTGAATTATGTGAGGGAA
GjTPS19	TTCAGTCTTGCAAGTCAAGCG	GCATTAGTAAAGTTGTATTATGTGAAGA
GjTPS27	GGAGTCAACATCATCCCATAC	GCATGGACTAGAAATACTGGAG

下载: 导出CSV

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