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An auxin-responsive transcription factor AbLBD1 promotes the development of lateral roots and reduces the biosynthesis of tropane alkaloids in Atropa belladonna

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Abstract

Atropa belladonna produces anticholinergic tropane alkaloids (TAs), such as hyoscyamine and scopolamine, which are synthesized in its lateral roots. Yet, the regulation of lateral root growth and TA biosynthesis remain poorly understood. In this study, an auxin-responsive transcription factor (AbLBD1), encoding a lateral organ boundaries domain protein (LBD), was functionally characterized from A. belladonna. Overexpression of AbLBD1 effectively promoted the initiation of lateral roots and adventitious roots of A. belladonna. The root biomass of AbLBD1-overexpressing lines was 1.48 to 2.45 fold higher that of the control. However, the contents of hyoscyamine and scopolamine were markedly reduced in AbLBD1-overexpressing lines. Gene expression analysis showed that the expression levels of TAs biosynthesis genes, namely AbPMT, AbPYKS, AbCYP82M3, and AbH6H, were greatly decreased in AbLBD1-overpressing lines. In conclusion, AbLBD1 plays a positive role in regulating lateral root development but a negative one in regulating TAs biosynthesis in A. belladonna.

Key message

This study proved for the first time that AbLBD1 was a key auxin response regulator in root, which promotes initiation of lateral root and inhibits the biosynthesis of tropane alkaloids.

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Abbreviations

TA:

Tropane alkaloid

LBD:

Lateral organ boundaries domain protein

GST:

Glandular secretory trichome

TF:

Transcription factor

ARF:

Auxin response factor

CRL:

Crown rootless protein

RTCS:

Rootless concerning crown and seminal roots protein

RTCL:

Rootless concerning crown and seminal roots like protein

PGK:

Phosphoglycerate kinase

PMT:

Putrescine N-methyltransferase

PYKS:

Polyketide synthase

CYP82M3:

Tropinone synthase

H6H:

Hyoscyamine 6 beta-hydroxylase

IAA:

Indole-3-acetic acid

IBA:

Indole-3-butanoic acid

ZT:

Zeatin

qPCR:

Real-time quantitative PCR

HPLC:

High performance liquid chromatography

MS:

Murashige & Skoog

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Acknowledgements

This work was financially supported by the National Natural Science Foundation of China (31770335 and U1902212) and the National Transgenic Major Project of China (2019ZX08010-004). The authors declare no competing financial interests.

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Contributions

ZL and JZ conceived research, analyzed data and wrote the manuscript. JZ, JW and XL conducted the experiments. CY conducted plant cultivation and harvesting. XL, JQ and MC analyzed the alkaloids. All authors reviewed the manuscript.

Corresponding author

Correspondence to Zhihua Liao.

Additional information

Communicated by K X Tang.

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Zeng, J., Wang, J., Liu, X. et al. An auxin-responsive transcription factor AbLBD1 promotes the development of lateral roots and reduces the biosynthesis of tropane alkaloids in Atropa belladonna. Plant Cell Tiss Organ Cult 142, 559–569 (2020). https://doi.org/10.1007/s11240-020-01882-7

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  • DOI: https://doi.org/10.1007/s11240-020-01882-7

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