Table 1 Functions over-represented in A. vulgare ovaries in response to Wolbachia infection. Biological process GO accession A S A/S AO ~ SO cell fate determination GO:0001709 0.02 0.05 0.40 level 3 immune effector process GO:0002252 0.07 0.16 0.44 (n= 99) regulation of immune system process GO:0002682 0.04 0.14 0.29 generation of a signal involved in cell-cell signaling GO:0003001 0.04 0.05 0.80 Apoptosis inhibitor muscle contraction
GO:0006936 0.02 0.07 0.29 chromosome segregation GO:0007059 0.18 0.23 TPCA-1 0.78 ensheathment of neurons GO:0007272 0.00 0.02 0.00 circadian rhythm GO:0007623 0.07 0.09 0.78 cell recognition GO:0008037 0.02 0.07 0.29 reproductive behavior GO:0019098 0.04 0.05 0.80 membrane docking GO:0022406 0.04 0.05 0.80
viral reproductive process GO:0022415 0.02 0.05 0.40 cellular pigmentation GO:0033059 0.04 0.05 0.80 leukocyte activation GO:0045321 0.05 0.09 0.56 regulation of response to stimulus GO:0048583 0.12 0.18 0.67 coagulation GO:0050817 0.09 0.11 0.82 regulation of body fluid levels GO:0050878 0.04 0.05 0.80 endocrine process GO:0050886 0.11 0.14 0.79 cellular response to stimulus GO:0051716 0.05 0.07 0.71 In the same manner, two in vitro SSHs were performed by subtracting common transcripts between symbiotic and asymbiotic ovaries (SSH-S), and reciprocally (SSH-A). These selleck kinase inhibitor SSHs were contaminated by a high proportion of mitochondrial ESTs (~40%) that were removed for further analyses. To reveal the functions over-represented, we compared each SSH to SO library by the FatiGO web tool. One biological process (vesicle transport along microtubule) and one molecular function (microtubule motor activity) were over-represented in asymbiotic ovaries (Table 2). Most of the 223 unigenes that are associated to these two
GO terms belong to the kinesin-like protein family. In these two libraries, the BLAST analyses allowed the identification of 1 immune gene in SSH-S and 6 immune Casein kinase 1 genes in SSH-A libraries respectively (Additional File 4: Immune unigenes present in SO, AO, SSH-S, SSH-A, SSH-C, and SSH-NC libraries). Table 2 Functional enrichment analysis: list of GO terms that were over-represented in the lists of unigenes obtained by SSH experiments on ovaries (FatiGO web tool). P-value of Fisher’s exact unilateral tests. Adjusted p-value for multiple test correction. Test # unigenes Ontology domain Level Term GO ID p-value Adj. p-value SSH-A versus SO 223 Biological process 9 vesicle transport along microtubule GO:0047496 1.35E-04 5.97E-02 Molecular function 3 microtubule motor activity GO:0003777 1.13E-03 9.85E-02 SSH-S versus SO 44 no significant term In order to identify genes expressed in response to pathogenic bacteria, we performed SSH libraries between S. typhimurium-challenged and unchallenged asymbiotic A. vulgare females (SSH-C) and reciprocally (SSH-NC).