Functional Characterization of An Apple Pyrophosphate-dependent Phosphofructokinase Gene MdPFPβ in Regulating Soluble Sugar Accumulation

doi:10.16420/j.issn.0513-353x.2022-0423

Acta Horticulturae Sinica ›› 2022, Vol. 49 ›› Issue (10): 2223-2235.doi: 10.16420/j.issn.0513-353x.2022-0423

• Research Papers • Previous Articles Next Articles

Functional Characterization of An Apple Pyrophosphate-dependent Phosphofructokinase Gene MdPFPβ in Regulating Soluble Sugar Accumulation

YU Jianqiang¹^,², GU Kaidi¹, WANG Chuanzeng³^,^**(), HU Dagang¹^,^**()

¹State Key Laboratory of Crop Biology,College of Horticultural Science and Engineering,Shandong Agricultural University,Tai'an,Shandong 271018,China
²College of Horticulture and Plant Protection,Yangzhou University,Yangzhou,Jiangsu 225009,China
³Modern Agriculture Research Institute of Yellow River Delta,Shandong Academy of Agricultural Sciences,Dongying,Shandong 257000,China

Received:2022-04-18 Revised:2022-07-27 Online:2022-10-25 Published:2022-10-31
Contact: WANG Chuanzeng,HU Dagang E-mail:fap_296566@163.com;dazeng123321@163.com

Abstract

Abstract:

To elucidate the biological function of the apple（Malus × domestica Borkh.）phospho- fructokinase gene MdPFPβ（Pyrophosphate-fructose 6-phosphate 1-phosphotransferase subunit beta,gene sequence number MD09G1112800）,its sequence and promoter information were analyzed. The results showed that its open reading frame was 483 bp in length,encoding 161 amino acids. Cis-acting element analysis of its promoter using the PlantCare database revealed that regulatory elements related to abscisic acid（ABA）,low temperature,and drought signaling were present in the promoter sequence of MdPFPβ. Quantitative real-time PCR analysis revealed that MdPFPβ was highly expressed in apple leaves and mature fruits. It was also induced by external application of ABA in apple leaves. Overexpressing MdPFPβ significantly increased soluble sugar content in apple calli compared to that in wild-type under ABA treatment. Moreover,ectopic expression of MdPFPβ promoted photosynthetic performance in tomato leaves,as well as the content of soluble sugar in tomato fruits,indicating MdPFPβ plays a key role in regulating soluble sugar content in fruits.

Key words: apple, MdPFPβ, soluble sugar, abscisic acid

CLC Number:

S661.1

YU Jianqiang, GU Kaidi, WANG Chuanzeng, HU Dagang. Functional Characterization of An Apple Pyrophosphate-dependent Phosphofructokinase Gene MdPFPβ in Regulating Soluble Sugar Accumulation[J]. Acta Horticulturae Sinica, 2022, 49(10): 2223-2235.

Figures/Tables 9

Table 1 Primers used in this study

用途Application	引物名称Primer name	引物序列（5′-3′）Primer sequence
cDNA clone	MdPFPβ-F	ATGTACTCCATGAATGGCAAGG
	MdPFPβ-R 18S-F 18S-R	TGCTTGTGCTCCAAGTTCC ACACGGGGAGGTAGTGACAA CCTCCAATGGATCCTCGTTA
内参基因Reference gene
qRT-PCR			MdPFPβ-For1	TCTCGACGTTCCGCTTCCTCTC
qRT-PCR	MdPFPβ-Rev1	CTGGCCGTAGCATTGCTGAAGG

Table 2 Some important cis-acting regulatory elements in the promoter of MdPFPβ

调控序列	序列	位点功能	位置
Regulatory sequence	Sequence	Function of site	Location
ABRE	ACGTG	参与ABA响应元件cis-acting element involved in the abscisic acid responsiveness	-619,+1 719
ARE	AAACCA	参与厌氧响应元件cis-acting regulatory element essential for the anaerobic induction	+485,-1 210
G-box	TACGTG	参与光响应元件cis-acting regulatory element involved in light responsiveness	-1 739,+1 926
LTR	CCGAAA	参与低温响应元件cis-acting regulatory element essential for the anaerobic induction	+415,-697
MBS	CAACTG	参与干旱胁迫响应元件MYB binding site involved in drought-inducibility	+43
TC-rich repeats	GTTTTCTTAC	参与防御和胁迫响应元件MYB binding site involved in drought-inducibility	+1 269
CGTCA-motif	CGTCA	参与茉莉酸响应元件cis-acting regulatory element involved in the MeJA-responsiveness	-1 099

Fig. 1 Genetic information of MdPFPβ（A）and its expression in various tissues（B,root as control）and under ABA treatment（C,0 h water treatment as control） * α = 0.05;** α = 0.01。

Fig. 2 Phenotype,relative expression level of MdPFPβ gene,fresh weight and sugar content of wild type（WT）and MdPFPβ overexpressing apple calli（MdPFPβ-7/9/13）under ABA treatment

Fig. 3 Phenotypes,relative expression level of MdPFPβ and photosynthetic fluorescence parameters of wild-type（WT） and MdPFPβ transgenic tomato leaves（MdPFPβ-22/32/43）

Fig. 4 Photosynthetic rate,stomata conductance and intercellular CO2 concentration of wild-type（WT）and MdPFPβ transgenic tomato leaves（MdPFPβ-22/32/43）

Fig. 5 Starch content,sucrose content,fructose content and glucose content of wild-type（WT）and MdPFPβ transgenic tomato leaves（MdPFPβ-22/32/43）

Fig. 6 Phenotype,fructose content,glucose content and sucrose content of wild-type（WT）and MdPFPβ transgenic tomato fruits（MdPFPβ-22/32/43）

Fig. 7 PFP,SPS,NI,SS-I and S-AI activities of wild-type（WT）and MdPFPβ transgenic tomato fruits（MdPFPβ-22/32/43）

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Functional Characterization of An Apple Pyrophosphate-dependent Phosphofructokinase Gene MdPFPβ in Regulating Soluble Sugar Accumulation

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