Identification of Peach NAC Gene Family and Role of PpNAC050 in Promoting Fruit Fructose Accumulation

doi:10.16420/j.issn.0513-353x.2023-0085

Acta Horticulturae Sinica ›› 2024, Vol. 51 ›› Issue (9): 1983-1996.doi: 10.16420/j.issn.0513-353x.2023-0085

• Genetic & Breeding · Germplasm Resources · Molecular Biology • Next Articles

Identification of Peach NAC Gene Family and Role of PpNAC050 in Promoting Fruit Fructose Accumulation

LIU Jianhao¹^,², JING Yanfu², LIU Yuexin¹^,², XU Yaoguang², YU Yang², GE Xiuxiu¹^,^**(), XIE Hua¹^,²^,^**()

¹ College of Bioscience and Resources Environment，Beijing University of Agriculture，Beijing 102206，China
² Biotechnology Research Institute，Beijing Academy of Agriculture and Forestry Sciences，Beijing 100097，China

Received:2024-05-17 Revised:2024-07-16 Online:2024-09-25 Published:2024-09-19
Contact: GE Xiuxiu, XIE Hua

Abstract

Abstract:

The plant-specific NAC（NAM，ATAF1/2，CUC2）transcription factors play important roles in regulating fruit quality. To explore the role of the NAC in the regulation of soluble sugars in peach （Prunus persica）fruit quality，a total of 117 potential peach NAC members（PpNAC）were identified with an uneven distribution across all eight chromosomes as well as a scaffold. Phylogenetic analyses indicated that these PpNAC were classified into 13 subgroups. The NAC gene PpNAC050 that is highly expressed during later fruit development were further characterized，and it was in nucleus with transcriptional activation. Transient overexpression of PpNAC050 in peach fruits significantly increased fructose content and glucose content in peach fruit flesh. EMSA and dual-luciferase assays showed that PpNAC050，being a transcriptional repressor，directly bound to the promoters of PpERDL6-1（PpERDL16），a vacuolar membrane monosaccharide transporter gene regulating fructose content，suggesting that PpNAC050 positively regulated fructose accumulation by repressing PpERDL6-1 gene expression.

Key words: peach, fructose, NAC gene family, PpNAC050, PpERDL6-1

LIU Jianhao, JING Yanfu, LIU Yuexin, XU Yaoguang, YU Yang, GE Xiuxiu, XIE Hua. Identification of Peach NAC Gene Family and Role of PpNAC050 in Promoting Fruit Fructose Accumulation[J]. Acta Horticulturae Sinica, 2024, 51(9): 1983-1996.

Figures/Tables 7

Table 1 Primer sequences

基因 Gene	引物序列（5′-3′） Primer sequence	PCR产物大小/bp PCR product size
PpNAC-BD-F	ATCTCAGAGGAGGACCTGCATATGATGAGTATGGCCAATGCCATG	1 638
PpNAC-BD-R	CGGCCGCTGCAGGTCGACGGATCCAGAACTTTGTGGGAAAAGAGTCGC
PpNAC-F	ATGAGTATGGCCAATGCCATG	1 641
PpNAC-R	TCAAGAACTTTGTGGGAAAAGAGTCGC
PpNAC-1300-F	TAAATACTAGTGGATCCGGTACCATGAGTATGGCCAATGCCATG	1 638
PpNAC-1300-R	TCGCCCTTGCTCACCATGGTACCAGAACTTTGTGGGAAAAGAGTCGC
PpNAC-62SK-F	GGCCGCTCTAGAACTAGTGGATCCATGAGTATGG CCAATGCCAT GT	1 638
PpNAC-62SK-R	ACCGGGCCCCCCCTCGAGGTCGACAGAACTTT GTGGGAAAAG AGTC
q-NAC-F	TGAGTATGGCCAATGCCATGT	181
q-NAC-R	TTGGGCAGCAAATCAAGCAC
PpNAC-AD-F	CCATGGAGGCCAGTGAATTCATGAGTATGGCCAATGCCATGTAT	1 638
PpNAC-AD-R	TGCCCACCCGGGTGGAATTC AGAACTTTGTGGGAAAAGAGTCG
PpERDL6-1-NAC-F	GACATGTGTATAAATACCTCAGAACACGAGCATATATTGCATTGCATCCC	50
PpERDL6-1-NAC-R	GGGATGCAATGCAATATATGCTCGTGTTCTGAGGTATTTATACACATGTC
4T-PpNAC-F	CCGCGTGGATCCCCGGAATTCATGAGTATGGCCAATGCCATG	1 638
4T-PpNAC-R	GTCACGATGCGGCCGCTCGAGAGAACTTTGTGGGAAAAGAGTCGC
PpERDL6-1-800-lucF	GGTCGACGGTATCGATAAGCTT GTCACGTCTG ATGTCCCAGT	1 971
PpERDL6-1-800-lucR	GCAGGAATTCGATATCAAGCTT GTTTTTTTCTC ACTACTCTAA
PpERDL6-1-q-F	TCACTATGGTCGGAGCAACG	183
PpERDL6-1-q-R	ATGTAAGTCAGCACGCCAGA

Fig. 1 Chromosomal localization of NAC family genes in peach

Fig. 2 Phylogenetic analysis of NAC family members in peach（Prupe）and Arabidopsis（AT） a：ANAC001/ONAC003；b：TIP；c：OSNAC8；d：NAC2；e：ANAC011；f：NAM/NAC1；g：OSNAC7；h：ONAC022/TERN；j：ANAC063/SENU5；k：ANAC104；l：ATAF/AtNAC3；m：NAP；n：ANAC063.

Fig. 3 Expression（A），fragment amplification（B）and transcriptional activation detection（C）of PpNAC050 S1：First rapid growth phase；S2：Lag phase；S3：Second rapid growth phase. Different lowercase letters indicate in Fig. A significant differences at the 0.05 level in different fruit development periods.

Fig. 4 The subcellular localization of PpNAC050 A：Schematic diagram the recombinant plasmid pCAMBIA1300-PpNAC050-GFP； B：The GFP fluorescence from PpNAC050-GFP fusion protein was localized to the nucleus of the cells.

Fig. 5 The expression level and fructose，glucose and sucrosecontent in peach fruits transiently overexpressing PpNAC050 CK-1，CK-2和CK-3：Empty pGreen0029-62SK；TOE-1，TOE-2 and TOE-3：Overexpression. ns：Non-significant. * P < 0.05，** P < 0.01（Student’s t-test）.

Fig. 6 PpNAC050 suppresses the expression of sugar transporter PpERDL6-1 A：The expression levels of sugar transporter PpERDL6-1 in transiently overexpressing PpNAC050 fruits；B and C：Yeast one-hybrid and EMSA assay showing PpNAC050 binds to the PpERDL6-1 promoter fragment，respectively. D：The effects of PpNAC050 on the PpERDL6-1 sugar transporter promoter with dual-luciferase reporter assay in tobacco. Error bars represent the standard deviation of three biological replicates. ** P < 0.01，Student’s t-test.

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Identification of Peach NAC Gene Family and Role of PpNAC050 in Promoting Fruit Fructose Accumulation

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