Cloning and Functional Verification of Linalool Synthase Gene PsTPS14 in Tree Peony‘High Noon’

doi:10.16420/j.issn.0513-353x.2023-0383

Acta Horticulturae Sinica ›› 2024, Vol. 51 ›› Issue (6): 1273-1283.doi: 10.16420/j.issn.0513-353x.2023-0383

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

Cloning and Functional Verification of Linalool Synthase Gene PsTPS14 in Tree Peony‘High Noon’

WANG Peiyun¹, LI Ziang¹, BAI Yang¹, YANG Ping¹, YIN Chengpeng¹, LI Chuanrong², ZHANG Xinwen²^,^*(), SONG Xiuhua¹^,^*()

¹ College of Horticulture Science and Engineering，Shandong Agricultural University，Tai’an，Shandong 271018，China
² Forestry College，Shandong Agricultural University，Tai’an，Shandong 271018，China

Received:2024-01-05 Revised:2024-04-08 Online:2024-12-18 Published:2024-06-22
Contact: ZHANG Xinwen, SONG Xiuhua

Abstract

Abstract:

The linalool synthase gene PsTPS14 was cloned from the aromatic tree peony‘High Noon’based on the transcriptome sequencing. The complete CDS sequence of this gene was 903 bp in length encoding 300 amino acids. The analysis of the amino acids sequence showed that the protein has one conservative domain（Terpene_cyclase_plant_C1）. The expression level of PsTPS14 was the highest in half opening stage of petals in tree peony‘High Noon’，consistent with the release pattern of floral volatiles. Subcellular localization analysis showed that PsTPS14 was mainly localized in the chloroplast. The overexpression vector of PsTPS14 was injected into tobacco leaves and the petals of cut flowers of ‘Fengdan’，a light scent tree peony cultivar，by using the transient expression method. The release of linalool in tobacco leaves and‘Fengdan’petals could be detected by GC-MS method. At the same time，the expression level of PsTPS14 was higher in tobacco leaves and‘Fengdan’petals，and the content and activity of linalool synthase were significantly increased. This study showed that PsTPS14 regulated the synthesis of linalool in plants.

Key words: tree peony, linalool, linalool synthase gene, gene cloning, functional verification

WANG Peiyun, LI Ziang, BAI Yang, YANG Ping, YIN Chengpeng, LI Chuanrong, ZHANG Xinwen, SONG Xiuhua. Cloning and Functional Verification of Linalool Synthase Gene PsTPS14 in Tree Peony‘High Noon’[J]. Acta Horticulturae Sinica, 2024, 51(6): 1273-1283.

Figures/Tables 10

Table 1 Sequences of primers used in this study

基因名 Gene name	作用 Function	引物序列（5′-3′） Primer sequence
TPS14	基因克隆 Gene clone	F：ATGGCCTATTCCAGTGGCTCC；R：AGTGTTTAGAATTAAACCACTTTGCTT
TPS14	qRT-PCR	F：GCTCCTCTACTTTCTGGCGTTGTC；R：GCTGTGATCTTGGGCAAGGTTCC
ubiquintin	内参基因 Internal reference gene	F：GACCTATACCAAGCCGAAG；R：CGTTCCAGCACCACAATC

Fig. 1 PCR amplification products of PsTPS14 gene from tree peony

Fig. 2 Phylogenetic tree analysis of PsTPS14 proteins and homologous proteins

Fig. 3 The expression level of PsTPS14 and relative content of linalool in different flowering stages of tree peony‘High Noon’ S1：Squaring stage；S2：Wind bell stage；S3：Ballon stage；S4：Initial opening stage；S5：Half opening stage；S6：Full opening stage；S7：Declining stage. Different lowercase letters indicate significant differences at 0.05 level. The same below.

Fig. 4 The expression level of PsTPS14 and relative content of linalool in different organs of tree peony ‘High Noon’during its full opening stage

Fig. 5 Subcellular localization of PsTPS14 in tree peony

Fig. 6 GC-MS analysis of volatile compounds in tobacco leaves undergoing heterologous transient transformation of PsTPS14

Fig.7 Relative expression level，linalool synthase content and activity of PsTPS14 in tobacco leaves undergoing heterologous transient transformation of PsTPS14

Fig. 8 GC-MS analysis of volatile compounds in Paeonia ostia‘Fengdan’undergoing homologous transient transformation of PsTPS14

Fig. 9 Relative expression levels of PsTPS14 and PSY，content and activity of linalool synthase in Paeonia ostia ‘Fengdan’undergoing homologous transient transformation of PsTPS14

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Cloning and Functional Verification of Linalool Synthase Gene PsTPS14 in Tree Peony‘High Noon’

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