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Sugar Tech

, Volume 20, Issue 6, pp 669–680 | Cite as

A Small GTP-Binding Gene ScRan from Sugarcane is Involved in Responses to Various Hormone Stresses and Sporisirium scitamineum Challenge

  • Ning Huang
  • Hui Ling
  • Xu Zhang
  • Huaying Mao
  • Yachun Su
  • Weihua Su
  • Feng Liu
  • Liping Xu
  • Rukai Chen
  • Youxiong Que
Research Article
  • 198 Downloads

Abstract

The ras-like nuclear GTP-binding protein (Ran) is known as a molecular switch and plays an important role in eukaryotic metabolism. In this study, a differentially expressed EST sequence from our previous suppression subtractive hybridization libraries of sugarcane under Sporisirium scitamineum infection was used as the probe, and an 836-bp-long sugarcane Ran gene (ScRan) was cloned from sugarcane smut-resistant genotype YC05-179. Bioinformatic analysis revealed that the ScRan gene contained a 666-bp-long complete open-reading frame, encoding a stable 25-kDa acidic protein with four core domains GDGGTGKT (I), DTAG (II), NKVD (III), and EISAK (IV), and without the signal peptide. The inducing expression results of ScRan in Escherichia coli showed that the molecular weight of ScRan was about 20–35 kDa. Sequence alignment and phylogenetic analysis indicated that ScRan protein was conservative in evolution. ScRan protein was located in both the nucleus and the cytoplasm and could interact with sugarcane Sc14-3-3 protein (GenBank Acc. No.: AY222859). qRT-PCR analysis demonstrated that the expression of ScRan was higher in root than all the other sugarcane tissues including bud, leaf sheath, stem pith, and epidermis. The upregulation of ScRan in case of abscisic acid (ABA), salicylic acid (SA), and methyl jasmonic acid (MeJA) stimuli indicated its positive involvement in sugarcane responses to phytohormone ABA, SA, and MeJA signaling. ScRan was positively responded to S. scitamineum infection in the early stage but was inhibited in the later stage. These results would facilitate understanding the important role of the ScRan gene in sugarcane defense against various hormone stresses and S. scitamineum stress.

Keywords

Ras-like nuclear GTP-binding protein Hormone stress Sugarcane Bioinformatics analysis Sporisorium scitamineum 

Abbreviations

Ran

Ras-like nuclear GTP-binding protein

Sugarcane

Saccharum spp. hydrid

MW

Molecular weight

S. scitamineum

Sporisorium scitamineum

SSH

Suppression subtractive hybridization

ABA

Abscisic acid

SA

Salicylic acid

MeJA

Methyl jasmonate

qRT-PCR

Quantitative real-time PCR

IPTG

Isopropyl-β-D-thiogalactopyranoside

ATMT

Agrobacterium tumefacien-mediated transformation

GFP

Green fluorescent protein

YFP

Yellow fluorescent protein

Notes

Acknowledgements

The empty vector pEZYHb was created by Yu-Zhu Zhang and provided by Addgene (http://www.addgene.org/18669/). The vector pMDC84 used for analyzing subcellular location of gene was created by Mark D. Curtis and provided by the Arabidopsis biological resource center (http://www.arabidopsis.org/servlet/TairObject?id=1001376087&type=stock).

Funding

This work was supported by the National Natural Science Foundation of China (31671752 and 31101196), the Natural Science Foundation of Fujian province, China (2015J06006), and China Agriculture Research System (CARS-17). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.

Compliance with Ethical Standards

Conflict of interest

The authors declare that they have no conflict of interest.

Supplementary material

12355_2018_598_MOESM1_ESM.docx (31 kb)
Supplementary material 1 (DOCX 31 kb)

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Copyright information

© Society for Sugar Research & Promotion 2018

Authors and Affiliations

  1. 1.Key Laboratory of Sugarcane Biology and Genetic Breeding, Ministry of AgricultureFujian Agriculture and Forestry UniversityFuzhou CityChina
  2. 2.Key Laboratory of Genetics, Breeding and Multiple Utilization of Crops, Ministry of EducationFujian Agriculture and Forestry UniversityFuzhou CityChina
  3. 3.Guangxi Collaborative Innovation Center of Sugarcane IndustryGuangxi UniversityNanningChina

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