The Transcription Factor FvbHLH130 Activates Flowering in Fragaria vesca

doi:10.16420/j.issn.0513-353x.2021-1246

Acta Horticulturae Sinica ›› 2023, Vol. 50 ›› Issue (2): 295-306.doi: 10.16420/j.issn.0513-353x.2021-1246

• Research Papers • Previous Articles Next Articles

The Transcription Factor FvbHLH130 Activates Flowering in Fragaria vesca

SONG Yanhong¹, CHEN Yaduo¹, ZHANG Xiaoyu², SONG Pan¹, LIU Lifeng¹, LI Gang¹, ZHAO Xia¹, ZHOU Houcheng¹^,^*()

1. Zhengzhou Fruit Research Institute，Chinese Academy of Agricultural Sciences，Zhengzhou 450009，China
2. College of Horticulture，Henan Agricultural University，Xuchang，Henan 461101，China

Received:2022-08-23 Revised:2022-11-03 Online:2023-02-25 Published:2023-03-06
Contact: *（E-mail：zhouhoucheng@caas）

Abstract

Abstract:

The basic helix-loop-helix（bHLH）transcription factor family of strawberry was analyzed，and the transcription factor FvbHLH130 was identified in Fragaria vesca. qRT-PCR analysis in different tissues showed that FvbHLH130 was flower-specifically expressed. Overexpression of FvbHLH130 in Arabidopsis thaliana activated flowering in transgenic lines. The full-length coding sequence of FvbHLH130 was 960 bp，encoding a 319-amino-acid protein. The prediction of conserved domain of FvbHLH130 showed that 247-297 amino acids were bHLH binding domain. The promoter of FvbHLH130 contained hormone，stress- and low temperature-response elements，which indicated FvbHLH130 might be induced by biotic and abiotic stress. The amino acids sequence of FvbHLH130 revealed up to 93.70% similarity with RcbHLH130. FvbHLH130 sequence was more closely related to Rosaceae bHLH proteins and was homologous to AtFBH4. Agrobacterium-mediated transformation of Arabidopsis thaliana showed that the FvbHLH130 overexpression lines flowered seven days earlier than the control plants，and activated the expression of flowering-related genes AtAP1，AtFT，AtFUL and AtCO. The yeast two-hybrid results showed that FvbHLH130 interacted with FvARF4 and FvARF6，which may form protein complexes in the regulation of flowering.

Key words: Fragaria vesca, bHLH transcription factor, FvbHLH130, flowering

CLC Number:

S668.4

SONG Yanhong, CHEN Yaduo, ZHANG Xiaoyu, SONG Pan, LIU Lifeng, LI Gang, ZHAO Xia, ZHOU Houcheng. The Transcription Factor FvbHLH130 Activates Flowering in Fragaria vesca[J]. Acta Horticulturae Sinica, 2023, 50(2): 295-306.

Figures/Tables 7

References 60

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用途 Usage	引物名称 Primer name	序列（5′-3′） Sequence
FvbHLH130基因编码区克隆 Cloning the CDS of FvbHLH130	FvbHLH130-CDS-F	ATGGATTCAAATACCCATCTCA
FvbHLH130基因编码区克隆 Cloning the CDS of FvbHLH130	FvbHLH130-CDS-R	TCAAACTATCTGATTTTGAA
FvbHLH130基因启动子克隆 Cloning the promoter of FvbHLH130	FvbHLH130-Pro-F	GACATAATAACCTTGCATTT
FvbHLH130基因启动子克隆 Cloning the promoter of FvbHLH130	FvbHLH130-Pro-R	TATACACAACTCTGCTTGCC
构建FvbHLH130过表达载体 Cloning onstruction of FvbHLH130-overexpression vector	FvbHLH130-121-F	gagaacacgggggactctagaATGGATTCAAATACCCATCTCA
	FvbHLH130-121-R	gcccttgctcaccatggatccAACTATCTGATTTTGAACCGG
转基因阳性苗鉴定 Identification of transgenic positive seedlings	35S	GACGCACAATCCCACTATCC
转基因阳性苗鉴定 Identification of transgenic positive seedlings	GFP-R	TTACTTGTACAGCTCGTCCATG
草莓荧光定量内参基因 Reference genes for qRT-PCR in Fragaria vesca	Fv28S-qF	TAACCGCATCAGGTCTCCAA
草莓荧光定量内参基因 Reference genes for qRT-PCR in Fragaria vesca	Fv28S-qR	CTCGAGCAGTTCTCCGACAG
拟南芥荧光定量内参基因 Reference genes for qRT-PCR in F. vesca	At18S-qF	GAGAAGTTACTCCGCAACCT
拟南芥荧光定量内参基因 Reference genes for qRT-PCR in F. vesca	At18S-qR	GAATGATGCGTCGCCAGCACAAAGG
FvbHLH130不同组织表达模式分析 Expression pattern of FvbHLH130 in different tissues	FvbHLH130-qF	ATCCCCGAAGCATTGCAGAA
	FvbHLH130-qR	GCTCGAACATCGCTCAGACT
拟南芥开花相关基因表达分析 Expression of flowering-related genes in Arabidopsis thaliana	AtAP1-qF	CAATGAGCCCTAAAGAGCTT
	AtAP1-qR	GGGAGGCATATTGTGGCCTT
	AtFT-qF	CCCTGCTACAACTGGAACAA
	AtFT-qR	TGACAATTGTAGAAAACTGCG
	AtFUL-qF	GGAGAAGAAAACGGGTCAGC
	AtFUL-qR	ACTCGTTCGTAGTGGTAGGAC
	AtCO-qF	CTACAACGACAATGGTTCCATTAAC
	AtCO-qR	CAGGGTCAGGTTGTTGC
酵母双杂交载体构建 Construction the vector of yeast two-hybrid system	FvARF4-AD-F	gtaccagattacgctcatatgATGGAATTTGATCTGAACCA
	FvARF4-AD-R	cagctcgagctcgatggatccTCAGACCCTAATTGCTGTTG
	FvARF6-AD-F	gtaccagattacgctcatatgATGAGGCTTTCTTCTTCGTC
	FvARF6-AD-R	cagctcgagctcgatggatccTTAGTAGTCAAGGGAGCCCA
	FvARF8-AD-F	gtaccagattacgctcatatgATGAAGCTTTCCACATCAGG
	FvARF8-AD-R	cagctcgagctcgatggatccTCAATACTCAAGTGAGCCGA

用途 Usage	引物名称 Primer name	序列（5′-3′） Sequence
FvbHLH130基因编码区克隆 Cloning the CDS of FvbHLH130	FvbHLH130-CDS-F	ATGGATTCAAATACCCATCTCA
FvbHLH130基因编码区克隆 Cloning the CDS of FvbHLH130	FvbHLH130-CDS-R	TCAAACTATCTGATTTTGAA
FvbHLH130基因启动子克隆 Cloning the promoter of FvbHLH130	FvbHLH130-Pro-F	GACATAATAACCTTGCATTT
FvbHLH130基因启动子克隆 Cloning the promoter of FvbHLH130	FvbHLH130-Pro-R	TATACACAACTCTGCTTGCC
构建FvbHLH130过表达载体 Cloning onstruction of FvbHLH130-overexpression vector	FvbHLH130-121-F	gagaacacgggggactctagaATGGATTCAAATACCCATCTCA
	FvbHLH130-121-R	gcccttgctcaccatggatccAACTATCTGATTTTGAACCGG
转基因阳性苗鉴定 Identification of transgenic positive seedlings	35S	GACGCACAATCCCACTATCC
转基因阳性苗鉴定 Identification of transgenic positive seedlings	GFP-R	TTACTTGTACAGCTCGTCCATG
草莓荧光定量内参基因 Reference genes for qRT-PCR in Fragaria vesca	Fv28S-qF	TAACCGCATCAGGTCTCCAA
草莓荧光定量内参基因 Reference genes for qRT-PCR in Fragaria vesca	Fv28S-qR	CTCGAGCAGTTCTCCGACAG
拟南芥荧光定量内参基因 Reference genes for qRT-PCR in F. vesca	At18S-qF	GAGAAGTTACTCCGCAACCT
拟南芥荧光定量内参基因 Reference genes for qRT-PCR in F. vesca	At18S-qR	GAATGATGCGTCGCCAGCACAAAGG
FvbHLH130不同组织表达模式分析 Expression pattern of FvbHLH130 in different tissues	FvbHLH130-qF	ATCCCCGAAGCATTGCAGAA
	FvbHLH130-qR	GCTCGAACATCGCTCAGACT
拟南芥开花相关基因表达分析 Expression of flowering-related genes in Arabidopsis thaliana	AtAP1-qF	CAATGAGCCCTAAAGAGCTT
	AtAP1-qR	GGGAGGCATATTGTGGCCTT
	AtFT-qF	CCCTGCTACAACTGGAACAA
	AtFT-qR	TGACAATTGTAGAAAACTGCG
	AtFUL-qF	GGAGAAGAAAACGGGTCAGC
	AtFUL-qR	ACTCGTTCGTAGTGGTAGGAC
	AtCO-qF	CTACAACGACAATGGTTCCATTAAC
	AtCO-qR	CAGGGTCAGGTTGTTGC
酵母双杂交载体构建 Construction the vector of yeast two-hybrid system	FvARF4-AD-F	gtaccagattacgctcatatgATGGAATTTGATCTGAACCA
	FvARF4-AD-R	cagctcgagctcgatggatccTCAGACCCTAATTGCTGTTG
	FvARF6-AD-F	gtaccagattacgctcatatgATGAGGCTTTCTTCTTCGTC
	FvARF6-AD-R	cagctcgagctcgatggatccTTAGTAGTCAAGGGAGCCCA
	FvARF8-AD-F	gtaccagattacgctcatatgATGAAGCTTTCCACATCAGG
	FvARF8-AD-R	cagctcgagctcgatggatccTCAATACTCAAGTGAGCCGA

The Transcription Factor FvbHLH130 Activates Flowering in Fragaria vesca

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