Identification and characterization of microRNAs expressed in the African malaria vector Anopheles funestus life stages using high throughput sequencing

Background: Over the past several years, thousands of microRNAs (miRNAs) have been identified in the genomes of various insects through cloning and sequencing or even by computational prediction. However, the number of miRNAs identified in anopheline species is low and little is known about their role. The mosquito Anopheles funestus is one of the dominant malaria vectors in Africa, which infects and kills millions of people every year. Therefore, small RNA molecules isolated from the four life stages (eggs, larvae, pupae and unfed adult females) of An. funestus were sequenced using next generation sequencing technology. Results: High throughput sequencing of four replicates in combination with computational analysis identified 107 mature miRNA sequences expressed in the An. funestus mosquito. These include 20 novel miRNAs without sequence identity in any organism and eight miRNAs not previously reported in the Anopheles genus but are known in non-anopheles mosquitoes. Finally, the changes in the expression of miRNAs during the mosquito development were determined and the analysis showed that many miRNAs have stage-specific expression, and are co-transcribed and co-regulated during development. Conclusions: This study presents the first direct experimental evidence of miRNAs in An. funestus and the first profiling study of miRNA associated with the maturation in this mosquito. Overall, the results indicate that miRNAs play important roles during the growth and development. Silencing such molecules in a specific life stage could decrease the vector population and therefore interrupt malaria transmission.IS

Plant Epigenetic Mechanisms in Response to Biotic Stress

The environment changes faster than the ability of genetic recombination to generate natural genetic diversity. In this context, epigenetic regulation of gene expression has the potential to provide organisms with an alternative mechanism for phenotypic variation by controlling the extent of plasticity that can be achieved in response to environmental changes. There is now substantial evidence suggesting roles for epigenetic regulation of several different aspects of the plant response to biotic stress. At the basic level of gene expression, posttranscriptional gene silencing mediated by small RNAs and chromatin remodelling controlling transcriptional gene silencing are essential for the induced resistance responses activated during pest and pathogen attack. Beyond this, there is also evidence that histone modifications and DNA methylation are associated with immune memory, or defence priming, such as systemic acquired resistance (SAR). In addition, recent evidence indicates that epigenetic modifications can also generate longer-term defence priming responses that can be inherited across generations. In this chapter, we will discuss the roles of epigenetics in these different modes of biotic stress resistance, and suggest ways in which we may in the future be able to exploit epigenetic systems for crop protection