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Journal of Plant Biochemistry and Biotechnology

, Volume 28, Issue 4, pp 460–469 | Cite as

Cloning and expression analysis of the StCUL1 gene in potato

  • Peng-Xiang Pang
  • Li Shi
  • Xiao-Juan Wang
  • Yan-Nan Chang
  • Yong-Ping Luo
  • Jin-Lin Feng
  • Hemu Eri
  • Gang GaoEmail author
Original Article
  • 84 Downloads

Abstract

The Cullin-RING E3 ubiquitin ligase (CRL) complex is the most common E3 ligase, and the SCF complex (CRL1) has the most diverse functions. Cullin1(CUL1) is a scaffolding protein for assembly of the complex. SCF has been shown to participate in the non-biological stress response pathways. In this study, a classic CUL1 protein was identified in Solanum tuberosum, StCUL1. A full-length cDNA of the StCUL1 gene was obtained from ED13 (a potato variety) by Ralstonia solanacearum inoculation using the RACE method. Sequence analysis indicated that the gene comprised 2662 bp, with an open reading frame of 2229 bp encoding 743 amino acids. The expression levels of the StCUL1 gene in potato treated with R. solanacearum and exogenous hormones (such as salicylic acid, jasmonic acid methyl ester and abscisic acid) at different time points were determined by real-time PCR. The results indicated that StCUL1 was induced not only by pathogenic bacteria, but also by exogenous hormones, with sustained high expression. However, there were some differences in the modes of expression. Tissue localization analysis indicated that its expression was tissue specific, and it was mainly in the phloem of the vascular system of stems and leaves.

Keywords

Potato Ralstonia solanacearum Cullin1 StCUL1 Gene expression Tissue localization 

Abbreviations

CRL

Cullin-RING E3 ubiquitin ligase

UPS

Ubiquitin–proteasome system

HECT

Homology to E6-AP Carboxyl Terminus

cdc53

Cell division control protein 53

RBX1

Ring-box 1

SKP1

Suppressor of kinetochore protein 1

SLY1

S-phase kinase-associated protein 1

Max2

MORE AXILLARY BRANCHES 2

Notes

Acknowledgements

We would like to thank the institutes of Vegetables and Flowers and Plant Protection of the Chinese Academy of Agricultural Sciences for potato materials. This work was supported by the project of National Natural Science Foundation of China (31771858).

Compliance with ethical standards

Conflict of interest

No conflict of interest was declared.

Supplementary material

13562_2019_495_MOESM1_ESM.tif (25.6 mb)
Supplementary Figure 1 StCUL1 shares high structural similarity in different species. A Protein sequence alignment of StCUL1 showing the conserved domains. The amino acid sequence of StCUL1 was aligned with Solanum lycopersicum (XP_004229226.1), Capsicum annuum (XP_016542284.1), Nicotiana sylvestris (XP_009780619.1), and Petunia hybrida (BAW00386.1) homologs. The Cullin domain, consisting of 150 (418-567) amino acid residues and the cullin-nedd8 domain, consisting of 68 (669-736) amino acid residues, are represented by a solid line, which is conserved across many species. The comparison software was performed using DNAMAN6.0 software. Black indicates 100% conservatism, dark gray shading indicates 70% conservatism, and light gray shading indicates 50% conservatism. B A phylogenetic tree was generated from a multiple sequence alignment using Neighbour-Joining method with 1000 bootstrap replicates by MEGA 5 software. (TIFF 26184 kb)
13562_2019_495_MOESM2_ESM.tif (5.7 mb)
Supplementary Figure 2 Structure representation of StCUL1 protein predicted using the PSIPRED Web Server (http://bioinf.cs.ucl.ac.uk/psipred/). A Secondary structure showing alpha helix、extended strand、beta-turn and random coil. B Ribbon representation of the 3D structure of StCUL1 modified on SWISS Web Server (https://swissmodel.expasy.org/). (TIFF 5860 kb)
13562_2019_495_MOESM3_ESM.tif (6.3 mb)
Supplementary Figure 3 Cis-acting regulatory elements in the promoter regions of StCUL1. The 1572 bp upstream of the start codon was analyzed based on the PLACE, PlantCARE and potato genome sequence databases from GenBank (https://www.ncbi.nlm.nih.gov/). Different cis-elements showed by colored squares. (TIFF 6424 kb)

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Copyright information

© Society for Plant Biochemistry and Biotechnology 2019

Authors and Affiliations

  1. 1.College of Life ScienceShanxi Normal UniversityLinfenPeople’s Republic of China

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