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Chinese Journal of Tropical Crops ›› 2020, Vol. 41 ›› Issue (10): 2113-2119.DOI: 10.3969/j.issn.1000-2561.2020.10.017

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Cloning and Expression Analysis of EfGRAS Gene of Relative Sugarcane Wild Species Erianthus fluvus

SHEN Xianyue1,GU Shujie1,XIE Linyan1,QIAN Zhenfeng1,ZENG Dan1,HE Lilian1,2,*(),LI Fusheng1,2,*()   

  1. 1. College of Agronomy and Biotechnology, Yunnan Agricultural University, Kunming, Yunnan 650201, China
    2. Sugarcane Research Institute, Yunnan Agricultural University, Kunming, Yunnan 650201, China
  • Received:2020-07-01 Revised:2020-08-24 Online:2020-10-25 Published:2020-11-26
  • Contact: HE Lilian,LI Fusheng


In order to make deeper research into the drought resistance of sugarcane varieties, the genes of GRAS which playing an important role in the regulation of drought resistance mechanisms in sugarcane varieties for expression analysis were studied. In this experiment, we firstly retrieved the GRAS transcription factor gene sequence information from the transcriptome database, and used RACE-PCR and qRT-PCR techniques to clone a GRAS gene from ‘Sucrose 99-2’ and namde it EfGRAS (GenBank registration No. MT499789), then the bioinformatics analysis and expression analysis under drought stress were conducted on the EfGRAS gene. The results showed that the EfGRAS gene could encode 547 amino acids with a CDS 1644 bp in length, the molecular formula of the protein was C2645H4149N747O816S21, the relative molecular mass was 60.14 kDa, the instability coefficient was 54.85, the fat coefficient was 84.37, and the GRAVY value was ?0.293, which was a class unstable and hydrophilic protein. The subcellular location indicated that it was located in the nucleus. The main secondary structure of EfGRAS protein was α-helix and random coils, which were closely related to sorghum; qRT-PCR analysis showed that after drought stress, the gene expression in ‘Sugarcane 99-2’ was up-regulated by about 57.6 times compared to the control group, and the gene expression in ‘DZ 01-58’ was up-regulated by about 27.4 times compared to the control group. Compared with the control group, the gene expression level in ‘YC 89-9’ had a lower fold of up-regulation. The results would provide a theoretical basis for the further study on the function of EfGRAS in sugarcane.

Key words: Erianthus fluvus, drought stress, EfGRAS gene, clone, expression analysis

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