Abstract
Alternative splicing (AS) is an important mechanism by which eukaryotes regulate transcription and protein diversity. The dynamic changes in AS that occur on a genome-wide scale during interactions between plant roots and pathogens remain unknown. Here, we used the interaction between Arabidopsis and Ralstonia solanacearum as a model to explore the AS changes that take place during the response of roots to infection by means of high-throughput RNA-sequencing. We showed that dynamic changes in AS occur much earlier than changes at the level of transcription during R. solanacearum infection. Comparing genes that are regulated at the transcriptional and AS levels indicated that there are few common genes between differentially spliced genes (DSGs) and differentially expressed genes (DEGs). The functional gene ontology (GO) analysis identified that the enriched GO terms for the DSGs were different from those of the DEGs. The DSGs were over-represented in GO terms associated with post-transcriptional and translational regulations, suggesting that AS may act on RNA stability and during post-translation, thus affecting the output of plant defense molecules. Meanwhile, changes in DSGs were infection stage-specific. Furthermore, the nucleotide binding domain and leucine-rich repeat proteins and receptor-like kinases, key regulators in plant immunity, were shown to undergo dynamic changes in AS in response to R. solanacearum. Taken together, AS, along with transcription, modulates plant root defense to R. solanacearum through transcriptome reprogramming.
Highlights
As plants have a sessile lifestyle, they are vulnerable to adverse conditions in their environment that impact their growth and development
Changes in root morphology over time following infection by R. solanacearum were divided into the following periods: no symptoms (NS) from 0 to 12 hpi, root hair emergence (RH) at 12 to 24 hpi, primary root growth arrest and cell death (PC) at 24 to 48 hpi and lateral root emergence (LR) at 48 to 72 hpi (Figure 1a) [9]
To understand the transcriptional reprogramming that occurs in the root during the early stages of infection with R. solanacearum, RNA-seq analysis was performed on seven-day-old Arabidopsis seedlings at 0, 6, 12, 24, 48 and 72 h after infection of roots by GMI1000, with three biological replicates per time point
Summary
As plants have a sessile lifestyle, they are vulnerable to adverse conditions in their environment that impact their growth and development. Ralstonia solanacearum is a biotrophic soil-borne phytopathogen that causes bacterial wilt disease on many economically important crops, such as tobacco, potato and tomato [1]. The bacteria spread to the aerial parts of the plant and cause wilting symptoms [2]. Genes 2020, 11, 1078 lack of an efficient means to control it, R. solanacearum is considered to be one of most destructive plant pathogenic bacteria [1]. The first tier involves pathogen-associated molecular pattern (PAMPs)-triggered immunity (PTI), which is activated by the recognition of PAMPs in microbes through pattern recognition receptors (PRRs), including receptor-like proteins (RLPs) and receptor-like kinases (RLKs) located on the plant cell surface. The second tier comprises effector-triggered immunity (ETI), through which plants utilize intracellular disease resistance (R) proteins to recognize pathogen effectors and activate the defense response. ETI is often accompanied by the hypersensitive response, whereby local cell death occurs in the infected plant tissue
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