Host-root exudates increase gene expression of asparagine synthetase in the roots of a hemiparasitic plant Triphysaria versicolor (Scrophulariaceae).
Triphysaria is a facultative root parasite in the Scrophulariaceae family. Similar to other related parasites, the development of the parasitic life cycle is initiated by molecular signals released from appropriate host roots. Using a differential display, we isolated cDNAs preferentially abundant in T. versicolor roots exposed to Trifolium repens (white clover) root exudates in vitro. Sequence analysis indicated that one of the differentially expressed cDNAs had significant homology to the nitrogen-assimilating enzyme, asparagine synthetase (AS). T. versicolor AS cDNA clones were isolated and placed into three distinct classes on the basis of nucleotide sequence variations. All three classes encoded identical AS proteins. AS was expressed in both roots and shoots of in-vitro-cultured T. versicolor. Steady-state levels of AS mRNA increased in T. versicolor roots several-fold when seedlings were exposed to exudate obtained from hydroponically grown Arabidopsis thaliana roots. Therefore, AS transcript levels increased in response to exudates from two different hosts (Trifolium and Arabidopsis). The T. versicolor AS message levels increased to a similar magnitude when seedlings were incubated in the dark. Interestingly, AS levels were unaffected by treatment with the Striga haustoria inducer 2,6-dimethoxybenzoquinone. The potential role of AS in root parasitism is discussed.