Cloning and Functional Analysis of Peony PlSVP Gene in Regulating Flowering

doi:10.16420/j.issn.0513-353x.2021-1253

Acta Horticulturae Sinica ›› 2022, Vol. 49 ›› Issue (11): 2367-2376.doi: 10.16420/j.issn.0513-353x.2021-1253

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Cloning and Functional Analysis of Peony PlSVP Gene in Regulating Flowering

JI Fengjiao¹, MA Yan¹, QI Shuai¹, GUO Xianfeng¹^,^*(), CHEN Junqiang²^,^*()

¹Colloge of Forestry，Shandong Agricultural University，Tai’an，Shandong 271018
²Shandong Academy of Forestry，Jinan 250014，China

Received:2022-03-08 Revised:2022-09-13 Online:2022-11-25 Published:2022-11-25
Contact: GUO Xianfeng,CHEN Junqiang E-mail:guoxf@sdau.edu.cn;chenjq2000@126.com

Abstract

Abstract:

Cloning SHORT VEGETATIVE PHASE（SVP）gene from peony and exploring its function in regulating flowering will provide gene resource for molecular breeding of peony. In this study，PlSVP gene was isolated from Paeonia lactiflora‘Da Fugui’by RT-PCR method. The open reading frame of PlSVP gene was 687 bp in length，encoding 228 amino acids. Phylogenetic analysis indicated that PlSVP protein had the closest relationship with the homologue from P. suffruticosa. The qRT-PCR results showed that the expression of PlSVP gene was generally confined to vegetative organs. The highest expression was detected in roots，followed by in leaves，and the expression level in the basal leaves was higher than that in the top leaves. Overexpression of PlSVP gene in Arabidopsis Col-0 indicated that both bolting and flowering of the transgenic plants were delayed，by 7 and 15 d，respectively. And the complementary overexpression study in Arabidopsis svp-41 verified the inhibition effect of PlSVP gene. Further study that the expression of the floral promotors FT，SOC1，LFY and AP1 in transgenic plants was significantly down-regulated. These results suggested that PlSVP gene distinctively and negatively control flowering time，and possibly via inhibiting the expression of FT，SOC1，LFY and AP1.

Key words: herbaceous peony, Paeonia lactiflora, SVP gene, overexpression, complementary function, flowering

CLC Number:

S682.12

JI Fengjiao, MA Yan, QI Shuai, GUO Xianfeng, CHEN Junqiang. Cloning and Functional Analysis of Peony PlSVP Gene in Regulating Flowering[J]. Acta Horticulturae Sinica, 2022, 49(11): 2367-2376.

Figures/Tables 7

Table 1 List of primers

用途 Purpose	引物 Primer	核酸序列（5′-3′） Nucleotide sequence
基因克隆	PlSVP-F	ATGGCGAGAGAGAAGATTCAGAT
Gene cloning	PlSVP-R	TCAACCCGAATATGGCAGTC
载体构建Vector	pBI121-PlSVP-F	CGCGGATCCGCGATGGCGAGAGAGAAGATTCAGAT
construction	pBI121-PlSVP-R	CCGGAATTCCGGTCAACCCGAATATGGCAGTC
	35S	GACGCACAATCCCACTATCC
内参Internal	PlActin-F	ACTGCTGAACGGGAAATT
reference	PlActin-R	ATGGCTGGAACAGGACTT
qRT-PCR	qPlSVP-F	AGAACTGGCGGCTGAGACAA
	qPlSVP-R	TCCTTCCTCGCAAACCATG
内参Internal	Atactin-F	TCTCTATGCCAGTGGTCGTA
reference	Atactin-R	CCTCAGGACAACGGAATC
qRT-PCR	AtFT-F	TGGAACAACCTTTGGCAATG
	AtFT-R	GTCTTCTTCCTCCGCAGC
	AtAP1-F	CAGATCAAGGAGAGGGAA
	AtAP1-R	TTGATACAGACCACCCAT
	AtSOC1-F	CTCCAATATGCAAGATACCA
	AtSOC1-R	TATGCCTTCTCCCAAGAGTT
	AtLFY-F	CCCAAGAAGGGTTATCTGA
	AtLFY-R	AAACGGATGCTCCCTCTG

Fig. 1 Phylogenetic tree based on the amino acid alignment of PlSVP and SVP proteins from other plant species The scale indicates the reference length of the difference values between sequences. The number at each branch of the phylogenetic tree indicates the confidence level of the branch.

Fig. 2 Relative expression of PlSVP gene in different tissues of Paeonia lactiflora‘Da Fugui’ Different letters on bars indicate significant difference（P < 0.05）.

Fig. 3 Bolting and flowering time of Arabidopsis thaliana svp-41：Arabidopsis svp-41 mutants;a：Transgenic wild type Arabidopsis plants;b：Transgenic Arabidopsis svp-41 mutants;Different letters on bars indicate significant difference（P < 0.05）.

Fig. 4 Expression of PlSVP gene in transgenic Arabidopsis a1-a4：Expression of PlSVP gene in different transgenic lines of wild type Arabidopsis plants;b1-b4：Expression of PlSVP gene in different transgenic lines of Arabidopsis svp-41 mutants;** represents significant difference between transgenic lines and the control（P < 0.01）.

Fig. 5 Expression of SVP downstream genes in transgenic Arabidopsis A：Expression of AtSVP downstream gene in PlSVP-transformed wild type Arabidopsis plants;B：Expression of AtSVP downstream gene in PlSVP- transformed Arabidopsis svp-41 mutants;** represents significant difference between transgenic lines and the control（P < 0.01）.

Fig. 6 Schematic diagram of spatial expression of PlSVP gene in Paeonia lactiflora and the function of negatively regulating flowering time in Arabidopsis

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Cloning and Functional Analysis of Peony PlSVP Gene in Regulating Flowering

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