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Plant-parasitic cyst nematodes induce the formation of specialized fe eding cells in infected roots, which involves plant developmental processes that have been shown to be i fluenced by microRNAs (miRNAs) and other small RNAs. This observation provided t h foundation to investigate the potential involvement of small RNAs in plant-cyst nematode interactions. First, we examined the susceptibilities of Arabidopsis DICER-like ( dcl) and RNA-dependent RNA polymerase (rdr) mutants to the sugar beet cyst nematode Het rodera schachtii. The examined mutants exhibited a trend of decreased susceptibility suggesting a role of small RNAs mediating gene regulation processes during the plant – nematode interaction. Second, we generated two s mall RNA libraries from aseptic Arabidopsis roots harvested at 4 and 7 days after infect io with surface-sterilized H. schachtii. Sequences of known miRNAs as well as novel siRNAs were identified. Foll owing this discovery, we used real-time RT-PCR to quantify a total of 15 Arabidopsis tr an cripts that are known targets of six of the different miRNA families found in our study (miR 160, miR164, miR167, miR171, miR396 and miR398) in inoculated and non-inoculated Arabidopsis roots. Our analy ses showed mostly negative correlations between miRNA accumulation and ta rget gene mRNA abundance suggesting regulatory roles of these miRNAs during parasitism. Also, we identified a total of 125 non-miRNA siRNAs. Some of these siRNAs perfectly complement protein -cod g mRNAs or match transposon or retrotransposon sequences in sense or antisense orientati ons. We further quantified a group of siRNAs in H. schachtii-inoculated roots. The examined siRNAs exhibited distinct expression patterns in infected and non-infected roots, providing add itional evidence for the implication of small RNAs in cyst nematode parasitism. These dat a lay the foundation for detailed analyses of the functions of small RNAs during phytonematode parasitism.

A look into the molecular mechanisms of parasitism including the functional characterization of &lt;i&gt;Heterodera schachtii&lt;/i&gt; secreted effector proteins and the response of plant small RNA to nematode infection