Three experiments were conducted to study the allelopathic stimulation and its underlying molecular mechanism of achyranthes medicinal plants in continuously monoculture system. The stimulators in the rhizospheric soil of continuously monocultured achyranthes plants were extracted by water and organic solvents. Results of the bioassay showed that the rhizospheric soil extracts had a significant promotive effect on the growth of achyranthes in continuous monoculture system, implying that the extracts, especially the water extracts might contain plant activators to stimulate the growth of the medicinal plants. Subtractive hybridization suppression (SSH) was used to investigate gene expression profiles of achyranthes in response to the extract treatments. Ten up-regulated genes from SSH-cDNA library were sequenced and assigned. Results indicated that flavonoids and phytosterol might play an important role in the positively allelopathic stimulation on achyranthes plants in continuous monoculture system. Comparative proteomics were employed to further unveil the molecular mechanism of allelopathic stimulation induced by the extracts. Compared with protein expression profile in control, 25 differentially expressed proteins and their functions were detected and identified in the treated plants. The results suggested that the extracts from continuously monocultured rhizospheric soils under Chinese medicinal achyranthes activated the genes encoding the key enzymes involved in terpenes and flavonoids synthesis, which in turn led to increased de novo synthesis of the stimulators, and hence promoted growth of achyranthes in consecutively monoculture system.
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