Study : A combined small RNA and transcriptome sequencing analysis illuminates the response of resistance to brown planthopper in rice [RNA-seq]
A combined small RNA and transcriptome sequencing analysis illuminates the response of resistance to brown planthopper in rice [RNA-seq]
Here, a combined small RNA and transcriptome sequencing analysis was performed on the BPH6-transgenic and Nipponbare plants sheath samples at the different feeding stage by BPH, and differentially expressed microRNAs (DEMs) and genes (DEGs) were identified and further analyzed for a possible involvement in BPH6 mediated resistance mechanism. A total of 217 DEMs and 7,874 DEGs were identified in the BPH6-transgenic and Nipponbare plants after BPH feeding. At the miRNA level, 29 miRNAs, including the members of miR160, miR166, miR169, miR1861, miR319 and miR390 family and other miRNAs, appeared opposite expression at early or late feeding stages between the two varieties, and 9 miRNAs specially expressed in the BPH6-transgenic plans, suggesting these miRNAs might play an important role in BPH6-mediated resistance to BPH. For the transcriptome, 949 DEGs appeared opposite expression at early or late feeding stages of two rice genotypes were identified and enriched in metabolic process, cellular development, cell wall organization, cellular component movement and hormone transport, and in involved in certain kinds of primary and secondary metabolite synthesis by GO and KEGG enrichment analysis. 24 genes were further selected to be considered as BPH resistance related gene candidates. Finally, integrated analysis of DEMs and DEGs showed that 34 miRNAs corresponding to 42 target genes were selected as BPH resistance related miRNA-mRNA interaction candidates, 18 pairs of which were identified by qRT-PCR, two pairs of which were further confirmed by protoplast fluorescence micrographs and western blotting. Overall design: 18 libraries were constructed, amplified and sequenced with the BPH6-transgenic and Nipponbare sheath before and after BPH feeding.