Cloning and expression analysis of sugar transporter gene SpSWEET3 from the desert plant Stipagrostis pennata

doi:10.7522/j.issn.1000-694X.2023.00037

Journal of Desert Research ›› 2023, Vol. 43 ›› Issue (5): 129-138.DOI: 10.7522/j.issn.1000-694X.2023.00037

Cloning and expression analysis of sugar transporter gene SpSWEET3 from the desert plant Stipagrostis pennata

Bohan Sun¹(), Dan Yang¹, Fei Wang¹^,²^,³, Rong Li¹^,²^,³(), Hongbin Li¹^,²^,³()

^1.College of Life Science /, Shihezi University，Shihezi 832003，Xinjiang，China
^2.Xinjiang Production and Construction Corps Key Laboratory of Oasis Town and Mountain-basin System Ecology /, Shihezi University，Shihezi 832003，Xinjiang，China
^3.Key Laboratory of Xinjiang Phytomedicine Resource and Utilization of Ministry of Education, Shihezi University，Shihezi 832003，Xinjiang，China

Received:2023-02-07 Revised:2023-04-12 Online:2023-09-20 Published:2023-09-27
Contact: Rong Li,Hongbin Li

Abstract

Abstract:

Sugar transporter （SWEET） plays an important role in plant growth and development and stress response. The purpose of this study is to clone and analyze the expression characteristics of Stipagrosis pennata sugar transporter SpSWEET3 gene， and lay a foundation for further exploring the mechanism and function of its influence on the development of S. pennata root sand trap. The full-length open reading frame sequence of S. pennata SpSWEET3 gene was cloned by molecular cloning technology； The physical and chemical properties of SpSWEET3 gene were analyzed by bioinformatics； The gene expression characteristics were analyzed by qRT-PCR； The subcellular localization was observed and analyzed by laser confocal microscope. SpSWEET3was cloned from the desert plant S. pennata， which includes a 576-bp full-length open reading frame and encodes a basic strongly hydrophobic protein containing 191 amino acids with a relative molecular mass of 21.619 kD. SpSWEET3 belongs to the PQ-loop superfamily and contains the typical MtN3_slv transmembrane domain that catalyzes intracellular sugar efflux. Subcellular localization analysis revealed localization of the gene to the plasma membrane， implying that it may function as a transmembrane protein. qRT-PCR results showed that the SpSWEET3 gene had a higher cumulative expression in S. pennata root tissues， indicating its important role for sand rhizosheath development. Expression of the gene SpSWEET3 was significantly induced by drought stress with PEG， indicating its importance for drought adaptation. Protein interaction analysis showed that SpSWEET3 may transport sugar by interacting with proteins involved in the maintenance of nucleic acid structural stability， transcription factors， and transmembrane transport. The results of this study provide a foundation for in-depth investigation of the functions of SWEET genes and the mechanisms by which they regulate plant tissue development and adaptation to stress.

Key words: Stipagrostis pennata, SpSWEET3, sugar transporter gene, bioinformatics analysis, subcellular localization, rhizosheath, drought response

CLC Number:

Q943

Bohan Sun, Dan Yang, Fei Wang, Rong Li, Hongbin Li. Cloning and expression analysis of sugar transporter gene SpSWEET3 from the desert plant Stipagrostis pennata[J]. Journal of Desert Research, 2023, 43(5): 129-138.

Figures/Tables 10

Fig.1 Stipagrostis pennata plants and the wild growth environment.

Table 1 Primers used in this study

引物名称	引物序列(5'-3')	引物功能
SpSWEET3-F	ATGATCACTGGCATACGC	基因克隆
SpSWEET3-R	AGATAACATAGCTGTGTATTTACTACT
SpSWEET3-EF	GCTCTAGAATGATCACTGGCATACGC
SpSWEET3-ER	TCCCCCGGGAGATAACATAGCTGTGTATTTACTACT
q-ELF-F	ACTACCATCCCTATGAGCCAACT	基因表达分析
q-ELF-R	ATCACTGCCAGCCTGAAGACA
q-GAPDH-F	AGTCCGTCGCCATCGTCA
q-GAPDH-R	CGTGCCCATGCCTTCTGT
q-APR6-F	TCAATTCCAAGAAATGGCTCG
q-APR6-R	TGGGTTCCACCAGTAACAAGG
q-SpSWEET3-F	TGACATTTAAAAGAGTGATA
q-SpSWEET3-R	AGAACTCACAACTGGAAGGC

Fig.2 Gel electrophoresis diagram of SpSWEET3 gene cloning

Fig.3 Sequence analysis of SpSWEET3 protein

Fig.4 Bioinformatic analysis of SpSWEET3

Fig.5 Gene structure and conserved motif analysis of SpSWEET3

Fig.6 Expression pattern analysis of SpSWEET3

Fig.7 Double enzymatic identification of SpSWEET3recombinant plasmid

Fig.8 Subcellular localization analysis of SpSWEET3

Fig.9 Protein interaction analysis of SpSWEET3

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[2]	QIU Dong, WU Nan, ZHANG Yuan-ming, PAN Hui-xia. Ecological Regulation of Stipagrostis pennata by Rhizosheath Microhabitat [J]. JOURNAL OF DESERT RESEARCH, 2012, 32(6): 1647-1654.

Cloning and expression analysis of sugar transporter gene SpSWEET3 from the desert plant Stipagrostis pennata

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