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Acta Agronomica Sinica ›› 2020, Vol. 46 ›› Issue (7): 1120-1127.doi: 10.3724/SP.J.1006.2020.94173

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Cloning and functional analysis of the sweet potato sucrose transporter IbSUT3

WANG Dan-Dan1,**,LIU Hong-Juan1,**,WANG Hong-Xia2,ZHANG Peng2,*(),SHI Chun-Yu1,*()   

  1. 1 College of Agronomy, Shandong Agricultural University, Tai’an 271018, Shandong, China
    2 National Key Laboratory of Plant Molecular Genetics, CAS Center for Excellence in Molecular Plant Sciences, Institute of Plant Physiology and Ecology, Chinese Academy of Sciences, Shanghai 200032, China
  • Received:2019-11-11 Accepted:2020-03-24 Online:2020-07-12 Published:2020-04-03
  • Contact: Peng ZHANG,Chun-Yu SHI E-mail:zhangpeng@cemps.ac.cn;scyu@sdau.edu.cn
  • About author:** Contributed equally to this work
  • Supported by:
    National Natural Science Foundation of China Youth Fund(31701357);Shandong Agriculture Innovation Team(SDAIT-16-01)

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Abstract:

Sucrose, as the main transporter of photosynthate, is transported by sucrose transporters. In this study, we cloned IbSUT3 gene from sweet potato (Ipomoea batatas Lam. GenBank accession number MN233361), by RACE technique. The full-length sequence of the gene is 1607 bp and the complete open reading frame is 1518 bp, encoding 505 amino acids. The predicted protein has the molecular weight of 53.82 kD and isoelectric point of 9.19, containing 12 transmembrane domains. Multiple sequence alignment analysis indicated the protein, belonging to Group I, was significantly different from Group IV in evolution and shared high similarities with SUTs from other plant species. IbSUT3 had the function of transporting sucrose verified by SUSY7/ura3 system. Subcellular localization showed that IbSUT3 protein was located in the tobacco protoplasts plasma membrane. The qRT-PCR results showed that IbSUT3 highly expressed in leaves and induced by drought, high salt, low temperature and exogenous abscisic acid, suggesting that IbSUT3 is involved in response to various abiotic stresses, including abscisic acid.

Key words: sucrose transporters, IbSUT3 gene, subcellular localization, function analysis, adversity stresses

Table 1

Information of primers"

引物Primer 序列 Sequence (5'-3') 目的 Purpose
E3-F AGATAGTAATGGTGGCCTCCATTGC EST序列扩增
E3-R GCGAAGCCAATGAGGAAGACAGC EST sequence amplification
Sp-F1 AGATAGTAATGGTGGCGTCCATTGC 3'端和5'端RACE扩增
Sp-F2 GCGTCCTTTATGTGGCTTTGTGG 3' and 5'-RACE amplification
Sp-R1 TGAGATTGGCGCAGTAAACG
Sp-R2 GCGGCGAAGCCAATGAGGAA
IbSUT3-F CCATTCAGGCTGCGCAACT IbSUT3全长扩增
IbSUT3-R TTAAGAAATTAATTCCAAGGTCCAT IbSUT3 full-length amplification
SUT3Q-F CGGAAAATCCTCCTTGCAAGTG 荧光定量PCR
SUT3Q-R GACGCCACCATTACTATCTTCC Real-time PCR
Actin-F CTGGTGTTATGGTTGGGATGG
Actin-R GGGGTGCCTCGGTAAGAAG
SUT3m-F CATCGATATGGAGAGAGACTCCGTTAACG 酵母功能验证
SUT3m-R CCCCGGGTCAGTGGAAACCACCACCAGAT Yeast function verification
SUT3g-F ACGCGTCGACATGGAGAGAGACTCCGTTAACGGAA 亚细胞定位
SUT3g-R GGACTAGTACGTGGAAACCACCACCAGATAGCTCA Subcellular localization

Fig. 1

PCR amplified products of IbSUT3 A: the EST sequences amplification of IbSUT3; B: the 3'-RACE and 5'-RACE amplification of IbSUT3; C: the full length amplification of IbSUT3."

Fig. 2

Putative transmembrane predication of IbSUT3"

Fig. 3

Multiple alignment of the amino acid sequences of IbSUT3 with other species"

Fig. 4

Phylogenetic analysis between IbSUT3 and SUTs proteins from other plants species Nt: Nicotiana tabacum; St: Solanum tuberosum; Ib: Ipomoea batatas; Md: Malus domestica; Le: Solanum lycopersicum."

Fig. 5

IbSUT3 function analysis A: growth of transformed yeast strain on media of 2% sucrose; B: growth of transformed yeast strain on media of 2% glucose."

Fig. 6

Analysis of IbSUT3 subcellular localization"

Fig. 7

Expression of IbSUT3 in different tissues L: leaf; B: petiol; J: stem; WR: white root; DR: developmental root; MR: mature root. Bars indexed with different lowercase letters are significantly different at the 0.05 probability level."

Fig. 8

Expression profiles of IbSUT3 under stresses Bars indexed with different lowercase letters are significantly different at the 0.05 probability level."

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