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Salicylic acid involved in chilling-induced accumulation of calycosin-7-O-β-d-glucoside in Astragalus membranaceus adventitious roots

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Abstract

Calycosin and calycosin-7-O-β-d-glucoside (CG) are major isoflavonoids in Astragalus membranaceus and have multiple beneficial activities. Adventitious roots (ARs) are becoming attractive resources to obtain biologically active compounds. Salicylic acid (SA) is an important endogenous phytohormone, which is involved in the regulation of biotic and abiotic stresses. However, little is known about the potential role of SA on isoflavonoid accumulations under chilling stress. In the present study, calycosin was found to accumulate mostly in its glucosyl conjugate (CG) form in A. membranaceus ARs (AMARs). Compared to control conditions (25 °C), chilling (5 °C) induced the accumulation of CG, which was confirmed by increased expression levels of gene-encoding enzymes in the CG biosynthetic pathway. Furthermore, chilling triggered the accumulation of SA prior to CG accumulation. In addition, the inhibition of SA biosynthesis with paclobutrazol (PAC) in chilling-exposed AMARs suppressed the accumulation of CG and gene expressions, while exogenous addition of SA to PAC-treated AMARs restored CG content and gene expressions. These results indicated that in AMARs, SA involved in chilling-induced CG accumulation by regulating the expression levels of genes in the CG biosynthetic pathway.

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Abbreviations

4CL:

4-Coumaroyl:CoA-ligase

ARs:

Adventitious roots

AMARs:

Astragalus membranaceus adventitious roots

BA:

Benzoic acid

BA2H:

Benzoic acid 2-hydroxylase

C4H:

Cinnamate-4-hydroxylase

CG:

Calycosin-7-O-β-d-glucoside

CHI:

Chalcone isomerase

CHR:

Chalcone reductase

CHS:

Chalcone synthase

ELISA:

Enzyme-linked immunosorbent assay

HPLC:

High-performance liquid chromatography

I3′H:

Isoflavone 3′-hydroxylase

IBA:

Indole-3-butyric acid

IC:

Isochorismate

IFS:

Isoflavone synthase

IOMT:

Isoflavone O-methyltransferase

MS:

Murashige and Skoog

PAC:

Paclobutrazol

PAL:

Phenylalanine ammonia-lyase

pH:

Hydrogenion concentration

PBS:

Phosphate buffered saline

qPCR:

Quantitative real-time PCR

ROS:

Reactive oxygen species

rpm:

Revolutions per min

SA:

Salicylic acid

UCGT:

UDP-glucose: calycosin-7-O-β-d-glucosyltransferase

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Acknowledgements

This study was funded by the National Natural Science Foundation of China (21462044 and 30860036).

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Correspondence to Songquan Wu or Xueli Quan.

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Communicated by R. Baczek-Kwinta.

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Liu, J., Lan, X., Lv, S. et al. Salicylic acid involved in chilling-induced accumulation of calycosin-7-O-β-d-glucoside in Astragalus membranaceus adventitious roots. Acta Physiol Plant 41, 120 (2019). https://doi.org/10.1007/s11738-019-2909-7

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Keywords

  • Astragalus membranaceus
  • Adventitious roots
  • Calycosin-7-O-β-d-glucoside
  • Gene expression
  • Salicylic acid
  • Chilling