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Schlagwörter:
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Zusammenfassung:
Environmental cues can have profound effects on organismal development and phenotype, often regulated by small RNAs. Using long-term environmental induction approaches, we aim to study the influence of shifts in microbial diet on mouth-form plasticity in Pristionchus pacificus. Utilizing the rapidly propagating isogenic populations of P. pacificus with its direct binary mouth-form phenotype, directed evolution for 101 generations on a Novosphingobium diet revealed immediate and systemic diet- induced plasticity. Periodic diet-reversals to OP50 revealed memories that are not limited to the first- generation offspring but maintained transgenerationally. We combined these long-term induction experiments with unbiased forward genetic screens by performing an EMS mutagenesis in generation F14 on Novosphingobium to find mutants defective in transgenerational inheritance. From a screen of 9,900 ‘F2’ progeny of mutagenized animals, we found 165 potential candidates for transgenerational inheritance defective (tid) after food reversal. Whole genome sequencing revealed that multiple candidate genes were hit several times independently (i.e. >3 alleles). Indeed, generating clean CRISPR mutants in the first four candidate genes revealed a tid phenotype after diet reversal. Providing novel insight into transgenerational inheritance, we will describe novel molecular mechanisms involving sets of epigenetic factors identified through the analysis of these tid mutants.
