Plant vaccination: stimulation of defense system by caffeine production in planta.

Plants produce up to 100,000 secondary metabolites. One of their biological functions is self-denfese, and it is referred as chemical defense, directly and/or indirectly counteracting biotic and abiotic stresses. Alkaloids constitute 12% of total secondary metabolites, and some of them exhibit detrimental effects on living organisms. Caffeine (1,3,7-trimethylxanthine) is a member of purine alkaloids, and its exogenous application to plants at relatively high concentrations (0.01-0.1%) effectively repelled herbivores and pathogenic microbes. This allowed the construction of transgenic crops that endogenously produce caffeine to tolerate stresses. Experimentally, tobacco and chrysanthemum were successfully transformed with three distinct N-methyltranferases involved in the caffeine biosynthesis pathway. They produced 0.4-5 mug caffeine/g tissue (5 x 10(-4)%), this being three magnitudes lower than values found in caffeine-producing plants and in vitro experiments. Nevertheless, they exhibited strong repellence against pest insects, and high resistance to virus and bacterial infection. They also exhibited accelerated self-defense, as estimated by constitutive expression of defense-related genes, and by elevated production of salicylic acid, a critical signaling molecule for defense response. Since caffeine content was low in transgenic lines, observed effects might not be direct, but rather indirect. We presume that, as endogenously produced caffeine could be toxic, the host plants activated its own self-defense system, which commonly occurs regarding other stresses. Eventually the host became on standby to cope with a broad range of biotic stresses. The procedure resembles mammalian vaccination, in which antigen-antibody system is critical. We propose that plants can also be vaccinated as far as proper "antigenic" chemicals are expressed in planta.