Phosphatidylinositol Transporters Gene SbSEC14 C in Sugarbeet: Cloning and Expression Analysis Under Low Temperature Stress

doi:10.11924/j.issn.1000-6850.casb20191100883

Abstract

Abstract:

Phosphatidylinositol transporters (PITPs) are widely present in eukaryotic cells and can regulate the independent transport of phosphatidylinositol (PtdIns) or phosphatidylcholine (PtdCho) monomers between lipid bilayers in vitro. They are involved in many important life processes such as phosphoinositide metabolism, membrane transport, polar growth, signal transduction, stress, cytoplasmic movement, and cell cycle regulation. They play an important role in plant stress response and development regulation. In order to study the sugarbeet phosphatidylinositol transporter gene and its expression under low temperature stress, in this study, using a predicted phosphatidylinositol transporter gene CRS1 from sugarbeet genome database as a template, a beet SEC14 gene with a length of 765 bp, an open reading frame of 596 bp, and a coding of 198 amino acids was obtained by gene cloning. It was named SbSEC14. The analysis of the physicochemical properties showed that the protein was an unstable hydrophilic protein. The secondary and tertiary structure analysis of the protein showed that the protein's α-helix accounted for the highest proportion, 47.47%, and the β-turn angle accounted for the lowest, 5.56%. The conserved structural analysis of the protein showed that the protein had a typical SEC14 domain. The phylogenetic tree analysis of the protein showed that the beet SbSEC14 gene was most closely related to spinach and quinoa's. Real-time fluorescence quantitative results showed that the constitutive expression of SbSEC14 gene appeared in sugarbeet, the highest expression was in sugarbeet leaves, the lowest expression was in sugarbeet roots, and the expression in leaves was about 2.5 times that of roots. Sugarbeet seedlings were treated at 4℃ for 0, 2, 6, 12, and 24 h, the expression level of this gene showed an upward trend when the plants were treated for 0-2 h, the expression level was decreased when the plants were treated for 2-6 h and the expression level tended to be steady for 6-24 h. Therefore, it is predicted that this gene is related to the resistance of sugarbeet to low temperature stress.

Key words: sugarbeet, phosphatidylinositol transporter, SEC14, low temperature, abiotic stress

CLC Number:

S566.3

Zou Fengkang, Jia Hailun, Ding Guangzhou, Chen Li. Phosphatidylinositol Transporters Gene SbSEC14 C in Sugarbeet: Cloning and Expression Analysis Under Low Temperature Stress[J]. Chinese Agricultural Science Bulletin, 2020, 36(32): 39-48.

Figures/Tables 16

References 41

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引物	引物序列(5'-3')	用途
Sbsec14	R:GCTAGTTTTGGAAGTGAAGGAAAG	基因克隆
Sbsec14	F:ATGCTCTCAAGTCATAGATCACCT	基因克隆
GAPDH	R:GTTGGAACACGGAA AGCC	内参基因
GAPDH	F:TGGAGAGGTGGAAGG	内参基因
qSbsec14	R:GAGGAAACCGTGCCTGCCATCT	荧光定量
qSbsec14	F:TATTATCCAGCGAGGGGCTACAA	荧光定量

引物	引物序列(5'-3')	用途
Sbsec14	R:GCTAGTTTTGGAAGTGAAGGAAAG	基因克隆
Sbsec14	F:ATGCTCTCAAGTCATAGATCACCT	基因克隆
GAPDH	R:GTTGGAACACGGAA AGCC	内参基因
GAPDH	F:TGGAGAGGTGGAAGG	内参基因
qSbsec14	R:GAGGAAACCGTGCCTGCCATCT	荧光定量
qSbsec14	F:TATTATCCAGCGAGGGGCTACAA	荧光定量