Fructokinase Gene PpyFRK5 Plays an Important Role in Sucrose Accumulation of Pear Fruit

doi:10.16420/j.issn.0513-353x.2021-0445

Acta Horticulturae Sinica ›› 2022, Vol. 49 ›› Issue (7): 1429-1440.doi: 10.16420/j.issn.0513-353x.2021-0445

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Fructokinase Gene PpyFRK5 Plays an Important Role in Sucrose Accumulation of Pear Fruit

TAO Xin¹, ZHU Rongxiang¹, GONG Xin¹, WU Lei¹, ZHANG Shaoling¹, ZHAO Jianrong²^,^*(), ZHANG Huping¹^,^*()

¹College of Horticulture,Nanjing Agricultural University,Nanjing 210095,China
²College of Resource and Environment,Anhui Science and Technology University,Fengyang,Anhui 233100,China

Received:2022-03-10 Revised:2022-04-20 Online:2022-07-25 Published:2022-07-29
Contact: ZHAO Jianrong,ZHANG Huping E-mail:zjr0105@163.com;hpzhang@njau.edu.cn

Abstract

Abstract:

As one of the important members of fructokinase gene family,the gene of PpyFRK5 was cloned utilizing‘Hosui’pear（Pyrus pyrifolia Nakai）fruit as the experimental material in the study. Simultaneously,its expression patterns and biological functions were studied. The results indicated that the protein encoded by PpyFRK5 belongs to the phosphofructokinase B subfamily（pfkB）,and contains the highly conserved pfkB characteristic domain and the ATP binding domain and sugar binding domain unique to fructokinase. The expression level of PpyFRK5 exhibited significant distinctions in different pear organs and fruit development stages,and reached the highest in fruits and seeds at the mature stage. PpyFRK5 was localized in the plasma membrane and cytoplasm. During the whole development of the pear fruit,there was a significant positive correlation between the expression level of PpyFRK5 and the sucrose content,and the sucrose content of pear fruits with transient overexpression of PpyFRK5 was significantly increased,while that of pear fruits with transient silencing of PpyFRK5 was significantly decreased;the promoter region of PpyFRK5 contained multiple cis-acting elements involved in light response,hormone induction,and stress response.

Key words: pear, fructokinase, expression pattern, gene function, sucrose accumulation

CLC Number:

S661.2

TAO Xin, ZHU Rongxiang, GONG Xin, WU Lei, ZHANG Shaoling, ZHAO Jianrong, ZHANG Huping. Fructokinase Gene PpyFRK5 Plays an Important Role in Sucrose Accumulation of Pear Fruit[J]. Acta Horticulturae Sinica, 2022, 49(7): 1429-1440.

Figures/Tables 10

References 31

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用途 Use	名称 Name	序列（5′-3′） Sequence
实时荧光定量PCR	PpyFRK5-F	TTCGTTGGCAAGGTTGGAG
Quantitative real-time PCR	PpyFRK5-R	TCACCGTCGTTCTTCAAGGTC
	Tubulin-F	TGGGCTTTGCTCCTCTTAC
	Tubulin-R	CCTTCGTGCTCATCTTACC
基因克隆、超表达和亚细胞定位 Cloning,overexpression,and subcellular localization	35S:PpyFRK5-GFP-F	GAGAACACGGGGGACTCTAGAATGGCGAATTCAGAGAGTCCAC
	35S:PpyFRK5-GFP-R	GCCCTTGCTCACCATGGATCCTTTGGCTTTGGACTTGGAAATG
病毒诱导的基因沉默	pTRV2-PpyFRK5-F	GAAGGCCTCCATGGGGATCCTGTGGTTCTGTCCCTGTGG
Virus induced gene silencing	pTRV2-PpyFRK5-R	GCCTCGAGACGCGTGAGCTCCATCAGACTGGGTTGGAAGA
启动子扩增 Promoter amplification	PpyFRK5-pro-F	GTCGACGGTATCGATAAGCTTCCTTATATTTGTCGACTGTAGGTCGA
	PpyFRK5-pro-R	CGCTCTAGAACTAGTGGATCCTCTCTCTTAACTCTTGCGATTGAGA

用途 Use	名称 Name	序列（5′-3′） Sequence
实时荧光定量PCR	PpyFRK5-F	TTCGTTGGCAAGGTTGGAG
Quantitative real-time PCR	PpyFRK5-R	TCACCGTCGTTCTTCAAGGTC
	Tubulin-F	TGGGCTTTGCTCCTCTTAC
	Tubulin-R	CCTTCGTGCTCATCTTACC
基因克隆、超表达和亚细胞定位 Cloning,overexpression,and subcellular localization	35S:PpyFRK5-GFP-F	GAGAACACGGGGGACTCTAGAATGGCGAATTCAGAGAGTCCAC
	35S:PpyFRK5-GFP-R	GCCCTTGCTCACCATGGATCCTTTGGCTTTGGACTTGGAAATG
病毒诱导的基因沉默	pTRV2-PpyFRK5-F	GAAGGCCTCCATGGGGATCCTGTGGTTCTGTCCCTGTGG
Virus induced gene silencing	pTRV2-PpyFRK5-R	GCCTCGAGACGCGTGAGCTCCATCAGACTGGGTTGGAAGA
启动子扩增 Promoter amplification	PpyFRK5-pro-F	GTCGACGGTATCGATAAGCTTCCTTATATTTGTCGACTGTAGGTCGA
	PpyFRK5-pro-R	CGCTCTAGAACTAGTGGATCCTCTCTCTTAACTCTTGCGATTGAGA

顺式作用元件 cis-element	序列 Sequence	功能 Probable function
ABRE	ACGTG	脱落酸响应元件Involved in the abscisic acid response
ARE	AAACCA	厌氧诱导响应元件Essential for the anaerobic induction
AE-Box	ACAAACAA	光响应元件Part of a module for light response
CGTCA-motif	CGTCA	MeJA响应元件Involved in the MeJA response
Box-4	ATTAAT	光响应元件Involved in light response
CAAT-Box	CCAAT;CAAT	启动子和增强子区域元件Element in the promoter and enhancer regions
G-Box	CACGTT	光响应元件Involved in light response
LTR	CCGAAA	低温响应元件Involved in low-temperature response
MBS	CAACTG	MYB结合位点MYB binding site
TATA-Box	TATA;TATAAA	转录起始位点Approximately -30 from the transcription start
RY-element	CATGCATG	种子特异性调控响应元件Involved in seed-specific regulation
TC-rich repeats	ATTCTCTAAC	防御和胁迫响应元件Involved in defense and stress response
TCT-motif	TCTTAC	光响应元件Part of a light responsive element
TGACG-motif	TGACG	MeJA响应元件Involved in the MeJA response
as-1	TGACG	转录结合位点Transcription factor-binding site
circadian	CAAAGATATC	昼夜节律控制响应元件Involved in circadian control

顺式作用元件 cis-element	序列 Sequence	功能 Probable function
ABRE	ACGTG	脱落酸响应元件Involved in the abscisic acid response
ARE	AAACCA	厌氧诱导响应元件Essential for the anaerobic induction
AE-Box	ACAAACAA	光响应元件Part of a module for light response
CGTCA-motif	CGTCA	MeJA响应元件Involved in the MeJA response
Box-4	ATTAAT	光响应元件Involved in light response
CAAT-Box	CCAAT;CAAT	启动子和增强子区域元件Element in the promoter and enhancer regions
G-Box	CACGTT	光响应元件Involved in light response
LTR	CCGAAA	低温响应元件Involved in low-temperature response
MBS	CAACTG	MYB结合位点MYB binding site
TATA-Box	TATA;TATAAA	转录起始位点Approximately -30 from the transcription start
RY-element	CATGCATG	种子特异性调控响应元件Involved in seed-specific regulation
TC-rich repeats	ATTCTCTAAC	防御和胁迫响应元件Involved in defense and stress response
TCT-motif	TCTTAC	光响应元件Part of a light responsive element
TGACG-motif	TGACG	MeJA响应元件Involved in the MeJA response
as-1	TGACG	转录结合位点Transcription factor-binding site
circadian	CAAAGATATC	昼夜节律控制响应元件Involved in circadian control

Fructokinase Gene PpyFRK5 Plays an Important Role in Sucrose Accumulation of Pear Fruit

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