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Transcription and enzymatic analysis of beta-glucosidase VvBG1 in grape berry ripening

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Abstract

Although beta-glucosidase was recently demonstrated to be involved at transcriptional level in fleshy fruit development, it remains unclear whether BG possesses biochemical activity. In the present study, real-time reverse transcription polymerase chain reaction analysis showed that the mRNA expression levels of β-glucosidase gene VvBG1 increased rapidly concomitant with berry red-colouring in Muscat Hamburg grapevine (Vitis vinifera), suggesting that the VvBG1 gene might be involved in the berry ripening. In order to explore VvBG1 biochemical activity, a 60-kD recombinant protein, VvBG1, was expressed in Escherichia coli BL21 (DE3) and purified. The results gained from a combination of VvBG1-pulp-homogeneity-incubation test and gas chromatography-mass spectroscopy analysis showed that VvBG1-treated berry samples contained more free ABA than the control, demonstrating that VvBG1 has high glycosyl hydrolase activity that can transform the ABA-glucosyl ester to free ABA. In conclusion, both the high expression and increased hydrolase activity of VvBG1 could suggest an important role of ABA in grape berry ripening.

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Abbreviations

ABA:

Abscisic acid

ABA-GE:

ABA glucosyl ester

NCED:

9-cis-Epoxycarotenoid dioxygenase

BG:

Beta-glucosidase

CYP:

ABA 8′-hydroxylase

GT:

ABA glucosyltransferase

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Acknowledgments

This work was supported financially by the National Key Basic Research ‘973’ Program of China (grant no. 2012CB126306), the National Science Foundation of China (grant no 31272144), and the Project of Construction of Innovative Teams and Teacher Career Development for Universities and Colleges under Beijing Municipality (grant no. IDHT20140509).

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Authors and Affiliations

  1. Beijing Key Laboratory of New Technology in Agricultural Application, College of Plant Science and Technology, Beijing University of Agriculture, No. 7 Beinong Road, Changping District, Beijing, 102206, People’s Republic of China

    Jing-hua Sun, Yin-hang Dong, Chun-li Li & Yuan-yue Shen

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  1. Jing-hua Sun
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  2. Yin-hang Dong
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  3. Chun-li Li
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  4. Yuan-yue Shen
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Correspondence to Yuan-yue Shen.

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Sun, Jh., Dong, Yh., Li, Cl. et al. Transcription and enzymatic analysis of beta-glucosidase VvBG1 in grape berry ripening. Plant Growth Regul 75, 67–73 (2015). https://doi.org/10.1007/s10725-014-9932-x

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  • DOI: https://doi.org/10.1007/s10725-014-9932-x

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