Temporal expression patterns of Pasteuria spp. sporulation genes

Abstract

Endospore-forming bacterium in the genus Pasteuria spp. infect multiple agriculturally significant plant parasitic nematodes and has potential as a potent biological control. Success as a biological control requires not only spore attachment to the cuticle, but sporulation and reproduction within the nematode host. Tracking and identifying Pasteuria spp. development is then critical to demonstrating efficacy as a biocontrol. Microscopic observations suggest Pasteuria spp. follows the model bacterium, Bacillus subtilis, sporulation. Here, we identified B. subtilis homologs of sporulation regulators in Pasteuria spp. and characterized the temporal expression of these genes throughout the bacterium’s ∼30-d lifecycle in Meloidogyne arenaria as a means of tracking sporulation development. Detectable levels of transcripts of Spo0F were present as early as 5 d after the nematodes were exposes to Pasteuria spp. and were relatively constant throughout the 30-d lifecycle. Transcripts to Sigma-F were significantly higher in the middle of the lifecycle, while the transcripts of Sigma-G were detectable between 15 and 25 d, nearing the end of the lifecycle. These three markers can be used to track the process of sporulation in the nematode and augment microscopic observations. Tracking sporulation of Pasteuria spp. is important to fully realize its potential as a biological control method as it can more readily identify successful parasitism, define host ranges, and inform in vitro growth progress.

Keywords

• biological control
• pasteuria spp
• meloidogyne spp
• rt-qpcr