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Temporal expression patterns of Pasteuria spp. sporulation genes

Ruhiyyih Dyrdahl-Young

Ruhiyyih Dyrdahl-Young

Department of Entomology and Nematology, University of FloridaUSA
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Dyrdahl-Young, Ruhiyyih
, 
Weiming Hu

Weiming Hu

Department of Entomology and Nematology, University of FloridaUSA
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Hu, Weiming
 and 
Peter DiGennaro

Peter DiGennaro

Department of Entomology and Nematology, University of FloridaUSA
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DiGennaro, Peter
  
Jul 29, 2019
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Article Category: Arts & Humanities
Published Online: Jul 29, 2019
Page range: 1 - 8
Received: Dec 12, 2018
DOI: https://doi.org/10.21307/jofnem-2019-039
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© 2019 Ruhiyyih Dyrdahl-Young et al., published by Sciendo
This work is licensed under the Creative Commons Attribution 4.0 International License.

Figure 1:

The Bacillus subtillus homologous sporulation gene Spo0F was identified and sequenced in Pasteuria penetrans. Spo0F nucleotide and translated amino acid sequence (A). The Pasteuria spp. sequences are identical and share a 72% identity with the B. subtilis sequence (B). The sequences and positions of primers used in the qPCR gene expression analysis for this gene are noted.
The Bacillus subtillus homologous sporulation gene Spo0F was identified and sequenced in Pasteuria penetrans. Spo0F nucleotide and translated amino acid sequence (A). The Pasteuria spp. sequences are identical and share a 72% identity with the B. subtilis sequence (B). The sequences and positions of primers used in the qPCR gene expression analysis for this gene are noted.

Figure 2:

The conserved RNA sigma factor Sigma-G was identified in Pasteuria penetrans. The nucleotide sequence is 71% similar the Bacillus subtilis sequence (A). The amino acid sequence (B) has 83% identity with the B. subtilis, Sigma-G.
The conserved RNA sigma factor Sigma-G was identified in Pasteuria penetrans. The nucleotide sequence is 71% similar the Bacillus subtilis sequence (A). The amino acid sequence (B) has 83% identity with the B. subtilis, Sigma-G.

Figure 3:

Nucleotide sequence (A) and amino acid residue (B) of Bacillus subtilis and Pasteuria penetrans sigma factor, Sigma-F.
Nucleotide sequence (A) and amino acid residue (B) of Bacillus subtilis and Pasteuria penetrans sigma factor, Sigma-F.

Figure 4:

To track the lifecycle of the obligate endoparasite Pasteuria penetrans, the timing of the expression of Bacillus subtilis homologous sporulation gene Spo0F (A) and sigma factors Sigma-F (B) and Sigma-G (C) was quantified throughout the 30 d lifecycle using RT-qPCR. The results represent three independent replicates each from individual root systems. Pairwise comparisons using Student’s t-test (α = 0.05) were used to examine differences in the expression levels at all timepoints. *Statistically differences. There were significantly more copies of the transcript Spo0F on day 15 than days 25 or 30. There were also significantly more Sigma-F transcripts on day 20 than on day 25 or 30.
To track the lifecycle of the obligate endoparasite Pasteuria penetrans, the timing of the expression of Bacillus subtilis homologous sporulation gene Spo0F (A) and sigma factors Sigma-F (B) and Sigma-G (C) was quantified throughout the 30 d lifecycle using RT-qPCR. The results represent three independent replicates each from individual root systems. Pairwise comparisons using Student’s t-test (α = 0.05) were used to examine differences in the expression levels at all timepoints. *Statistically differences. There were significantly more copies of the transcript Spo0F on day 15 than days 25 or 30. There were also significantly more Sigma-F transcripts on day 20 than on day 25 or 30.

Figure 5:

Light microscopic images of the development of Pasteuria penetrans. Photographs taken 10 (A), 15 (B), 20 (C), and 25 (D) days after Meloidogyne arenaria exposure to P. penetrans. Arrows indicate microcolonies (A), thali (B), nascent sporogonium (C) and sporogonium (D). Scale bar represents 10 µm, two images are shown for each developmental stage.
Light microscopic images of the development of Pasteuria penetrans. Photographs taken 10 (A), 15 (B), 20 (C), and 25 (D) days after Meloidogyne arenaria exposure to P. penetrans. Arrows indicate microcolonies (A), thali (B), nascent sporogonium (C) and sporogonium (D). Scale bar represents 10 µm, two images are shown for each developmental stage.

Figure 6:

The percentage of each sporulation structures was assessed to track the lifecycle of Pasteuria penetrans in its nematode host. For each of the four time points assessed, five independent infected nematodes samples were excised from inoculated roots and the numbers of microcolonies (A), thali (B), nascent sporogonium (C), sporogonium (D), and mature endospores (E) were counted by two observers. Shown is mean observations expressed as a percentage of the total number of sporulation structures identified. Pairwise comparisons using t-test (α = 0.05) was conducted to compare a given time point to the initial time point (15 d).
The percentage of each sporulation structures was assessed to track the lifecycle of Pasteuria penetrans in its nematode host. For each of the four time points assessed, five independent infected nematodes samples were excised from inoculated roots and the numbers of microcolonies (A), thali (B), nascent sporogonium (C), sporogonium (D), and mature endospores (E) were counted by two observers. Shown is mean observations expressed as a percentage of the total number of sporulation structures identified. Pairwise comparisons using t-test (α = 0.05) was conducted to compare a given time point to the initial time point (15 d).
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