Functional Plant Biology Functional Plant Biology Society
Plant function and evolutionary biology
RESEARCH ARTICLE

Phylogenetic analysis and functional characterisation of strictosidine synthase-like genes in Arabidopsis thaliana

Natalie A. J. Kibble A B , M. Mehdi Sohani A , Neil Shirley A B , Caitlin Byrt A B , Ute Roessner C , Antony Bacic C , Otto Schmidt A and Carolyn J. Schultz A D
+ Author Affiliations
- Author Affiliations

A School of Agriculture Food and Wine, The University of Adelaide, Glen Osmond, SA 5064, Australia.

B Australian Centre for Plant Functional Genomics, The University of Adelaide, Glen Osmond, SA 5064, Australia.

C School of Botany, The University of Melbourne, Parkville, Vic. 3010, Australia.

D Corresponding author. Email: carolyn.schultz@adelaide.edu.au

Functional Plant Biology 36(12) 1098-1109 https://doi.org/10.1071/FP09104
Submitted: 8 May 2009  Accepted: 8 September 2009   Published: 3 December 2009

Abstract

Monoterpenoid indole alkaloids (MIA) are a diverse class of secondary metabolites important for plant protection and are drugs for treating human diseases. Arabidopsis thaliana (L.) is not known to produce MIAs, yet its genome has 15 genes with similarity to the periwinkle (Catharanthus roseus (L.) G. Don) strictosidine synthase (STR) gene. Phylogenetic analysis of strictosidine synthase-like (SSL) proteins reveals four well supported classes of SSLs in Arabidopsis. To determine if Arabidopsis produces active strictosidine synthase, Arabidopsis protein extracts were assayed for enzymatic activity and cDNAs were expressed in Escherichia coli. Arabidopsis protein extracts from leaves and hairy roots do not make strictosidine at levels comparable to C. roseus, but they metabolise one substrate, secologanin, a precursor of strictosidine in other plant species, and produce an ‘unknown’ compound proposed to be a dimer of secologanic acid. Recombinant Arabidopsis proteins expressed in E. coli were not active STRs. Quantitative PCR analysis was performed on class A Ssls and showed they are upregulated by salt, ultraviolet light and salicylic acid treatment. RNAi mutants of Arabidopsis with reduced expression of all four class A Ssls, suggest that class A SSL proteins can modify secologanin. Gene expression and metabolomics data suggests that class A Ssl genes may have a role in plant protection.

Additional keywords: gene duplication, monoterpenoid indole alkaloid, secologanin.


Acknowledgements

This project was supported by the Australian Centre for Plant Functional Genomics, funded by the Australian Research Council and the Grains Research and Development Corporation. The authors thank Kris Ford (ESI–MS/MS), Robert Asenstorfer, Max Tate (MS interpretation), Jelle Lahnstein (RP-HPLC) and Mark Livermore (E. coli expression) for their technical assistance, Ute Baumann for helpful discussions on phylogenetic analysis, and Robert Verpoorte and Magdi El-Sayed for their kind donation of the strictosidine standard.


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