Cloning, Expression Analysis and Functional Verification of the PrGSTF12 Gene from Paeonia rockii

doi:10.7525/j.issn.1673-5102.2026.05.006

Abstract

Abstract:

Glutathione S-transferase（GST） genes play a crucial role in plant anthocyanin accumulation. To clarify the roles of GSTs in anthocyanin transport and accumulation in the petal blotches of Paeonia rockii， color indices， flavonoid components and contents were determined in petals at different developmental stages. The results indicated that peonidin and cyanidin were the major pigments responsible for the formation of petal blotches. Subsequently， a differentially expressed GST gene， PrGSTF12， was identified and cloned from the petal transcriptome of P. rockii. Subcellular localization， expression analysis， and virus-induced gene silencing（VIGS） were performed to investigate its biological function. Phylogenetic analysis revealed that PrGSTF12 had a close genetic relationship with known GSTs involved in anthocyanin transport. Subcellular localization assays indicated that PrGSTF12 was targeted to the tonoplast， suggesting its function in this cellular compartment. Quantitative real-time PCR analysis showed that the expression level of PrGSTF12 was significantly higher in the basal blotch site than that in the non-blotch site of petals， and its expression level was positively correlated with local anthocyanin accumulation. The VIGS assay demonstrated that silencing PrGSTF12 resulted in noticeable fading of the petal basal blotches， accompanied by a significant reduction in anthocyanin content. Meanwhile， the expression levels of anthocyanin synthesis-related genes， including PrCHS， PrCHI， PrF3H， PrF3'H， PrDFR and PrANS， were markedly decreased. In summary， these results demonstrate that PrGSTF12 plays important roles in anthocyanin transport and accumulation in the petal basal blotches of P. rockii and contributes to basal blotch formation.

Key words: Paeonia rockii, petal blotch, PrGSTF12 gene, gene cloning, functional verification

CLC Number:

S685.11

Sitong CHEN, Sha LUO, Qianru ZHANG, Jiahui LI, Chen TAO, Qianqian SHI. Cloning, Expression Analysis and Functional Verification of the PrGSTF12 Gene from Paeonia rockii[J]. Bulletin of Botanical Research, 2026, 46(5): 838-850.

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引物名称 Primer name	引物序列（5′→3′） Primer sequence（5′→3′）
PrGSTF12-F	TATGGTAGTTAAGGTGTATGG
PrGSTF12-R	TTAATTAACAAGATTCATTAATTTC
pC2300-PrGSTF12-KpnⅠ-F	ACGGGGGACGAGCTCGGTACCATGGTAGTTAAGGTG
pC2300-PrGSTF12-SalⅠ-R	CTTGCTCACCATGGTGTCGACATTAACAAGATTCAT

引物名称 Primer name	核苷酸序列（5′→3′） Nucleotide sequence（5′→3′）
PrGSTF12-pTRV2-F	GTGAGTAAGGTTACCGAATTCACCTGTTGGGAACCACTTTG
PrGSTF12-pTRV2-R	CGTGAGCTCGGTACCGGATCCCTGAGACCAGGAAGATGACT
PrCHS-F	CAAAGGGTCACGAGTTCTA
PrCHS-R	CACGCAAGTGTCCATCA
PrCHI-F	CAGCACAGCCTCGGAAAGTTGTT
PrCHI-R	CAACGAAATCCCAATCAT
PrF3H-F	TTGTCATCAGGCGGTAGA
PrE3H-R	TGTTCACGGCAGGGACT
PrF3′H-F	TTTTTGTTGGGCTTGG
PrF3H-R	ATCGGTTCATGGCTTGT
PrDER-F	AATGGGTATCCGCTTTT
PrDER-R	GAAGATTGCTGGGTATGG
PrANS-F	CTGAGTTGCTTGGCGTA
PrANS-R	CAAAGGGTCACGAGTTCTA

时期 Period	部位 Part	色卡编号 RHSCC	明度 L^*	红绿度 a^*	黄蓝度 b^*	色调角 h	彩度 C
S1	无斑部位Non-blotch part	142C	65.90±0.85^c	-18.83±0.15^f	41.70±0.56^a	45.63±0.55^a	114.40±0.40^b
S2	基部色斑部位Basal blotch part	71B	67.03±1.27^c	-10.97±0.40^d	35.77±0.25^b	37.47±0.21^b	107.10±0.72^cd
S2	无斑部位Non-blotch part	145B	72.70±0.66^b	-17.17±0.38^ef	41.30±0.46^a	44.43±0.60^a	112.40±0.10^bc
S3	基部色斑部位Basal blotch part	59C	42.63±1.10^d	14.97±0.75^b	11.10±0.56^d	18.60±0.20^d	36.20±4.05^e
S3	无斑部位Non-blotch part	145D	73.43±1.21^b	-15.73±0.21^e	36.80±0.60^b	40.40±1.61^ab	112.73±0.15^bc
S4	基部色斑部位Basal blotch part	N80A	45.03±3.45^d	30.47±1.92^a	-4.53±0.64^e	31.50±2.36^abcd	351.30±1.15^a
S4	无斑部位Non-blotch part	155C	80.77±0.40^a	-8.07±0.21^c	25.77±1.90^c	26.20±0.72^cd	107.60±0.10^d