The Use of Arabidopsis to Study Interactions between Parasitic Angiosperms and Their Plant Hosts.

Parasitic plants invade host plants in order to rob them of water, minerals and nutrients. The consequences to the infected hosts can be debilitating and some of the world's most pernicious agricultural weeds are parasitic. Parasitic genera of the Scrophulariaceae and Orobanchaceae directly invade roots of neighboring plants via underground structures called haustoria. The mechanisms by which these parasites identify and associate with host plants present unsurpassed opportunities for studying chemical signaling in plant-plant interactions. Seeds of some parasites require specific host factors for efficient germination, thereby insuring the availability of an appropriate host root prior to germination. A second set of signal molecules is required to induce haustorium development and the beginning of heterotrophy. Later stages in parasitism also require the presence of host factors, although these have not yet been well characterized. Arabidopsis is being used as a model host plant to identify genetic loci associated with stimulating parasite germination, haustorium development, and parasite support. Arabidopsis is also being employed to explore how host plants respond to parasite attack. Current methodologies and recent findings in Arabidopsis - parasitic plant interactions will be discussed.

Differential induction of Orobanche seed germination by Arabidopsis thaliana.

Parasitic plants, including the root holoparasites Orobanche spp., cause devastating damage to crops worldwide. Arabidopsis thaliana (L.) is widely used an amenable model for the study of plant biology, including plant-pathogen interactions. Bringing the two plants together in a controlled system will enable the study of the molecular and genetic basis involved in host-parasitic plant interactions and should provide tools for the detection of genes responsible for incompatibility and resistance responses. The objective of this study was to screen Arabidopsis lines for reduced germination of Orobanche seeds. A 96-cell well bioassay was developed to test the potential of lines, ecotypes and mutants of Arabidopsis to induce germination of Orobanche. Screening of 50 A. thaliana ecotypes did not reveal non-inducing ecotypes. Screening of 13000 A. thaliana fast neutron mutated M2 plants detected 94 non-inducing mutant plants of which 34 were rescued, self pollinated, and M3 seeds collected. M3 seedlings from five lines were reduced in their ability to induce germination. In a separate assay, we determined that the reduced germination rates corresponded with reduced distance from the roots at which germination occurred. While further studies are necessary to determine the segregation of low germination phenotypes, these lines might prove useful for studying the genetic basis of variation in germination stimulant production in A. thaliana.

Quinone oxidoreductase message levels are differentially regulated in parasitic and non-parasitic plants exposed to allelopathic quinones.

Allelopathic chemicals released by plants into the rhizosphere have effects on neighboring plants ranging from phytoxicity to inducing organogenesis. The allelopathic activity of naturally occurring quinones and phenols is primarily a function of reactive radicals generated during redox cycling between quinone and hydroquinone states. We isolated cDNAs encoding two distinct quinone oxidoreductases from roots of the parasitic plant Triphysaria treated with the allelopathic quinone 2,6-dimethoxybenzoquinone (DMBQ). TvQR1 is a member of the zeta-crystallin quinone oxidoreductase family that catalyzes one-electron quinone reductions, generating free radical semiquinones. TvQR2 belongs to a family of detoxifying quinone oxidoreductases that catalyze bivalent redox reactions which avoid the radical intermediate. TvQR1 and TvQR2 message levels are rapidly upregulated in Triphysaria roots as a primary response to treatment with various allelopathic quinones. Inhibition of quinone oxidoreductase enzymatic activity with dicumarol prior to quinone treatment resulted in increased transcript levels. While TvQR2 homologs were upregulated by DMBQ in roots of all plants examined, TvQR1 homologs were upregulated only in roots of parasitic plants. Phylogenetic trees constructed of TvQR1 and TvQR2 protein homologs in Archea, Eubacteria and Eukaryotes indicated that both gene families are ancient, yet the families have dissimilar evolutionary histories in angiosperms. We hypothesize that TvQR2-like proteins function to detoxify allelopathic quinones in the rhizosphere, while TvQR1 has specific functions associated with haustorium development in parasitic plants.