Identification and preliminary functional characterization of SWEET sugar transporters involved in fruit development of melon (Cucumis melo L.)

doi:3969/j.issn.1004-1524.2023.02.08

Acta Agriculturae Zhejiangensis ›› 2023, Vol. 35 ›› Issue (2): 308-318.DOI: 3969/j.issn.1004-1524.2023.02.08

• Horticultural Science • Previous Articles Next Articles

Identification and preliminary functional characterization of SWEET sugar transporters involved in fruit development of melon (Cucumis melo L.)

LI Pingfang¹^,²(), YAO Xiefeng²^,^*(), XU Jinhua², ZHU Lingli², YANG Xingping²^,^*()

1. Department of Agriculture, Teaching and Researching Center, Zhejiang Open University, Hangzhou 310012, China
2. Jiangsu Key Laboratory for Horticultural Crop Genetic Improvement, Institute of Vegetable Crops, Jiangsu Academy of Agricultural Sciences, Nanjing 210014, China

Received:2022-05-20 Online:2023-02-25 Published:2023-03-14
Contact: YAO Xiefeng,YANG Xingping

Abstract

Abstract:

Melon (Cucumis melo L.) is one of the most important summer fruits in China and worldwide. The quality and yield of melon fruits are largely determined by the types and amounts of carbohydrates contained in the fruits. SWEET(sugars will eventually be exported transporters) are a new class of proteins with the ability to transport glucose and other oligosaccharides, and recent studies have shown that SWEET sugar transporters may play a role in regulating the fruit development. In this study, 18 SWEETs genes were identified from the melon genome, and further through RT-PCR combined with quantitative real-time PCR (qPCR) methods, three SWEETs genes with high expression level were identified during the whole or certain period of fruit development stages. Subcellular localization showed that two SWEETs genes(CmSWEET3,CmSWEET7a) were localized on the cell membrane. Further cloned into a yeast vector system, it was found that CmSWEET7a has the function of transporting glucose and fructose in vitro. This study laid the foundation for revealing the regulatory role of SWEET in the development process of melon fruit.

Key words: melon, fruit, sugar transporter, SWEET

CLC Number:

S652

LI Pingfang, YAO Xiefeng, XU Jinhua, ZHU Lingli, YANG Xingping. Identification and preliminary functional characterization of SWEET sugar transporters involved in fruit development of melon (Cucumis melo L.)[J]. Acta Agriculturae Zhejiangensis, 2023, 35(2): 308-318.

Figures/Tables 8

References 38

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引物名称	上游引物序列	下游引物序列
Primer name	Forward primer sequence(5'-3')	Reverse primer sequence(5'-3')
CmSWEET1	TCTTGGAAATGCAACTGCTC	GGCAATTCAACATCGTCATC
CmSWEET2	AAAGTGTGGAGTTTATGCCATTT	CCCAACATAGTTCCTATTCCATT
CmSWEET3	CTCCGCAAGTTCCTTATGGT	CAATGGGCTTCCCACTAGAT
CmSWEET5d	GGCCTCTTTCTGTCACCAGT	GCATTCCATAGAACACCCAA
CmSWEET7a	GAGGGAAGGTAAAGGGAAGG	TGTTGCGAGTCCTATCAAGC
CmSWEET7c	ATGGAAGAAGGGATCAGTGG	CCGTACAATGTCCACACCAT
CmSWEET10	TTCTTCAGCCAAACATGGAC	CGGGTGCAATAGATCAGATG
CmSWEET12a	ACGCGGATGAGTTCTTTCTT	GAGCCTTCTTGGGAGCATAG
CmSWEET12c	TCGATACTCATCACCCTGGA	AAGTGCCACCACATATGGAA
CmSWEET12d	TCGAAAGAGAGTTTCTTCCTTACA	GCAGCAGGGTTATGGGTATT
CmSWEET15	CTCCTTTGGTTGTGTCATCG	AATGAAGTGAATGGCCACAA
CmSWEET17c	AACGGAGATGATGGAGGAAG	TTGGCTTTGGAAGGCTTAAC
Actin7	TGCCCAGAAGTTCTATTCCAGC	CATAGTTGAACCACCACTGAGGAC
TOPO-SWEET7a	CAC CAT GGT TTC TTT GGT GC	TCAAGCTCCATGGACCTCAG
TOPO-SWEET3	CACCATGAGATCTCTCTACACCAT	CTAGGAGTTATTTTGATGTGGA
TOPO-SWEET17c	CACCATGGCTGCTAGTTTGAGCT	TCAAAATTTTCCATCTTCCACC
NEV-SWEET7a:	GCGGCCGC AAGCTTGTAAAAGAA AT GGT TTC TTT GGT GC	GCGGCCGCTCAAGCTCCATGGACCTC
NEV-SWEET3:	GCGGCCGC AAGCTTGTAAAAGAA ATGAGATCTCTCTACACC	GCGGCCGCCTAGGAGTTATTTTGATG

引物名称	上游引物序列	下游引物序列
Primer name	Forward primer sequence(5'-3')	Reverse primer sequence(5'-3')
CmSWEET1	TCTTGGAAATGCAACTGCTC	GGCAATTCAACATCGTCATC
CmSWEET2	AAAGTGTGGAGTTTATGCCATTT	CCCAACATAGTTCCTATTCCATT
CmSWEET3	CTCCGCAAGTTCCTTATGGT	CAATGGGCTTCCCACTAGAT
CmSWEET5d	GGCCTCTTTCTGTCACCAGT	GCATTCCATAGAACACCCAA
CmSWEET7a	GAGGGAAGGTAAAGGGAAGG	TGTTGCGAGTCCTATCAAGC
CmSWEET7c	ATGGAAGAAGGGATCAGTGG	CCGTACAATGTCCACACCAT
CmSWEET10	TTCTTCAGCCAAACATGGAC	CGGGTGCAATAGATCAGATG
CmSWEET12a	ACGCGGATGAGTTCTTTCTT	GAGCCTTCTTGGGAGCATAG
CmSWEET12c	TCGATACTCATCACCCTGGA	AAGTGCCACCACATATGGAA
CmSWEET12d	TCGAAAGAGAGTTTCTTCCTTACA	GCAGCAGGGTTATGGGTATT
CmSWEET15	CTCCTTTGGTTGTGTCATCG	AATGAAGTGAATGGCCACAA
CmSWEET17c	AACGGAGATGATGGAGGAAG	TTGGCTTTGGAAGGCTTAAC
Actin7	TGCCCAGAAGTTCTATTCCAGC	CATAGTTGAACCACCACTGAGGAC
TOPO-SWEET7a	CAC CAT GGT TTC TTT GGT GC	TCAAGCTCCATGGACCTCAG
TOPO-SWEET3	CACCATGAGATCTCTCTACACCAT	CTAGGAGTTATTTTGATGTGGA
TOPO-SWEET17c	CACCATGGCTGCTAGTTTGAGCT	TCAAAATTTTCCATCTTCCACC
NEV-SWEET7a:	GCGGCCGC AAGCTTGTAAAAGAA AT GGT TTC TTT GGT GC	GCGGCCGCTCAAGCTCCATGGACCTC
NEV-SWEET3:	GCGGCCGC AAGCTTGTAAAAGAA ATGAGATCTCTCTACACC	GCGGCCGCCTAGGAGTTATTTTGATG

基因名 Gene name	甜瓜基因组ID Melon genome IDs	拟南芥同源基因 Arabidopsis hits	E值 E-value	开放阅读框 ORF length/bp	多肽Deduced polypeptide
					长度	分子量	等电点	跨膜区域
					Length/aa	MW/ku	pI	TM
CmSWEET1	MELO3C008417.2.1	AtSWEET1	6.00×10^-94	759	252	27.7	9.25	7
CmSWEET3	MELO3C005869.2.1	AtSWEET3	3.00×10^-43	627	208	23.5	9.06	5
CmSWEET5a	MELO3C002256.2.1	AtSWEET5	2.00×10^-72	552	183	21	9.42	5
CmSWEET5b	MELO3C009951.2.1	AtSWEET5	9.00×10^-67	747	248	28.2	8.91	7
CmSWEET5c	MELO3C009950.2.1	AtSWEET5	5.00×10^-63	705	234	26	8.72	7
CmSWEET7a	MELO3C016259.2.1	AtSWEET7	3.00×10^-74	789	262	29.1	9.1	7
CmSWEET7b	MELO3C005758.2.1	AtSWEET7	3.00×10^-39	504	265	29.1	9.66	7
CmSWEET9a	MELO3C008674.2.1	AtSWEET9	4.00×10^-76	798	262	30.1	9.07	7
CmSWEET9b	MELO3C031232.2.1	AtSWEET9	4.00×10^-59	639	212	24.3	9.8	6
CmSWEET10	MELO3C026184.2.1	AtSWEET10	5.00×10^-82	891	296	33	9.23	7
CmSWEET12a	MELO3C026183.2.1	AtSWEET12	1.00×10^-79	894	297	33.3	8.85	7
CmSWEET12b	MELO3C002381.2.1	AtSWEET12	2.00×10^-72	876	291	32.9	6.44	7
CmSWEET12c	MELO3C002380.2.1	AtSWEET12	3.00×10^-38	729	242	27.4	8.97	5
CmSWEET15	MELO3C022341.2.1	AtSWEET15	2.00×10^-74	822	273	30.9	9.19	6
CmSWEET17a	MELO3C000200.2.1	AtSWEET17	2.00×10^-42	582	193	21.6	7.66	4
CmSWEET17b	MELO3C004222.2.1	AtSWEET17	1.00×10^-31	456	151	16.6	8.67	5
CmSWEET17c	MELO3C027076.2.1	AtSWEET17	2.00×10^-27	615	204	22.5	7.8	4
CmSWEET17d	MELO3C031734.2.1	AtSWEET17	4.00×10^-16	501	166	19.5	9.86	2

基因名 Gene name	甜瓜基因组ID Melon genome IDs	拟南芥同源基因 Arabidopsis hits	E值 E-value	开放阅读框 ORF length/bp	多肽Deduced polypeptide
					长度	分子量	等电点	跨膜区域
					Length/aa	MW/ku	pI	TM
CmSWEET1	MELO3C008417.2.1	AtSWEET1	6.00×10^-94	759	252	27.7	9.25	7
CmSWEET3	MELO3C005869.2.1	AtSWEET3	3.00×10^-43	627	208	23.5	9.06	5
CmSWEET5a	MELO3C002256.2.1	AtSWEET5	2.00×10^-72	552	183	21	9.42	5
CmSWEET5b	MELO3C009951.2.1	AtSWEET5	9.00×10^-67	747	248	28.2	8.91	7
CmSWEET5c	MELO3C009950.2.1	AtSWEET5	5.00×10^-63	705	234	26	8.72	7
CmSWEET7a	MELO3C016259.2.1	AtSWEET7	3.00×10^-74	789	262	29.1	9.1	7
CmSWEET7b	MELO3C005758.2.1	AtSWEET7	3.00×10^-39	504	265	29.1	9.66	7
CmSWEET9a	MELO3C008674.2.1	AtSWEET9	4.00×10^-76	798	262	30.1	9.07	7
CmSWEET9b	MELO3C031232.2.1	AtSWEET9	4.00×10^-59	639	212	24.3	9.8	6
CmSWEET10	MELO3C026184.2.1	AtSWEET10	5.00×10^-82	891	296	33	9.23	7
CmSWEET12a	MELO3C026183.2.1	AtSWEET12	1.00×10^-79	894	297	33.3	8.85	7
CmSWEET12b	MELO3C002381.2.1	AtSWEET12	2.00×10^-72	876	291	32.9	6.44	7
CmSWEET12c	MELO3C002380.2.1	AtSWEET12	3.00×10^-38	729	242	27.4	8.97	5
CmSWEET15	MELO3C022341.2.1	AtSWEET15	2.00×10^-74	822	273	30.9	9.19	6
CmSWEET17a	MELO3C000200.2.1	AtSWEET17	2.00×10^-42	582	193	21.6	7.66	4
CmSWEET17b	MELO3C004222.2.1	AtSWEET17	1.00×10^-31	456	151	16.6	8.67	5
CmSWEET17c	MELO3C027076.2.1	AtSWEET17	2.00×10^-27	615	204	22.5	7.8	4
CmSWEET17d	MELO3C031734.2.1	AtSWEET17	4.00×10^-16	501	166	19.5	9.86	2

Identification and preliminary functional characterization of SWEET sugar transporters involved in fruit development of melon (Cucumis melo L.)

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