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Identification of GH1 gene family fgt members in Stevia rebaudiana and their expression when grown in darkness

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Abstract

Stevia rebaudiana Bertoni is an important economic crop that is well known for its secondary metabolites, steviol glycosides (SGs), found in leaves. Because the enzymes of deglycosylation (glycoside hydrolases) play important roles in SGs biosynthetic processes, our study is focused on the functions of β-glucosidases in SGs catabolism in stevia. We cloned and characterized 19 stevia GH1 genes based on transcriptomic sequences. The 19 genes were divided into five putative subfamilies in Arabidopsis. Conserved motifs in the SrGH1 proteins were analysed using the online motif-based sequence analysis tool, MEME. Most of the identified proteins contain the conserved ‘TFNEP’ motif (contains the catalytic acid/base) and ‘ITENG’ motif (contains the catalytic nucleophile). Furthermore, the steviol glycoside content and expression of these 19 genes were characterized under constant darkness. The dark treatment lowered the steviol glycoside content significantly, while SrBGLU16 responded to darkness and was markedly upregulated. This study is the first transcriptome-wide analysis of the GH1 family in Stevia rebaudiana. The sequences of 19 SrGH1 members and their expression when grown in darkness were characterized. Among the 19 genes, SrBGLU16 was markedly upregulated by darkness. Thus, we identified SrBGLU16 for further investigation as a possible steviol glycoside beta-glucosidase.

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Funding

This study was funded by the Natural Science Foundation of Jiangsu Province (BK20160600), the Young Science Foundation Project of the National Natural Science Foundation of China (31601371 and 31901597).

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Authors and Affiliations

  1. Institute of Botany, Jiangsu Province and Chinese Academy of Sciences, Nan**g, 210014, China

    Yongheng Yang, Ting Zhang, **aoyang Xu, Yuming Sun, Yongxia Zhang, Menglan Hou, Suzhen Huang, Haiyan Yuan & Haiying Tong

  2. The Jiangsu Provincial Platform for Conservation and Utilization of Agricultural Germplasm, Nan**g, 210014, China

    Yongheng Yang, Ting Zhang, **aoyang Xu, Yuming Sun, Yongxia Zhang, Menglan Hou, Suzhen Huang, Haiyan Yuan & Haiying Tong

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  1. Yongheng Yang
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  2. Ting Zhang
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  3. **aoyang Xu
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  4. Yuming Sun
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  7. Suzhen Huang
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Contributions

Yongheng Yang designed the experiments. Yongheng Yang and Menglan Hou carried out the sequence analysis and gene clone. Menglan Hou carried out the HPLC analysis. **aoyang Xu carried out the RT-PCR. Yongxia Zhang took part in sequence analysis. Yongheng Yang drafted the manuscript. Ting Zhang and Yuming Sun participated and advised in manuscript development. Haiyan Yuan, Haiying Tong and Suzhen Huang advised in manuscript correction. All authors read and approved the final submission.

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Correspondence to Haiyan Yuan or Haiying Tong.

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Yang, Y., Zhang, T., Xu, X. et al. Identification of GH1 gene family fgt members in Stevia rebaudiana and their expression when grown in darkness. Mol Biol Rep 47, 8739–8746 (2020). https://doi.org/10.1007/s11033-020-05920-7

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  • DOI: https://doi.org/10.1007/s11033-020-05920-7

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