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.
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Change history
07 July 2018
In the original publication of this article, the word “Sporisorium” was incorrectly published as “Sporisirium” in the title and the abstract section.
07 July 2018
In the original publication of this article, the word ?Sporisorium? was incorrectly published as ?Sporisirium? in the title and the abstract section.
07 July 2018
In the original publication of this article, the word ?Sporisorium? was incorrectly published as ?Sporisirium? in the title and the abstract section.
07 July 2018
In the original publication of this article, the word ?Sporisorium? was incorrectly published as ?Sporisirium? in the title and the abstract section.
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
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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.
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Huang, N., Ling, H., Zhang, X. et al. A Small GTP-Binding Gene ScRan from Sugarcane is Involved in Responses to Various Hormone Stresses and Sporisirium scitamineum Challenge. Sugar Tech 20, 669–680 (2018). https://doi.org/10.1007/s12355-018-0598-y
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DOI: https://doi.org/10.1007/s12355-018-0598-y