Biparental immune priming in the pipefish Syngnathus typhle
Table S2. Results from ANOSIM analysis of 29 immune genes to test for F0-parental and F1-offspring treatment effects in one-week-old F1-juveniles. Multivariate ANOSIM analysis to assess differences in the gene expression profiles per parental treatment group applying pairwise comparison to relative gene expression data (−∆Ct-values) based on a Bray-Curtis distance matrix and 999 permutations. Significant p-values are presented in boldface, “ns” indicates no statistically significant difference. Shown are overall R and significance values over two main treatments (parents and offspring) and their interaction followed by pairwise comparisons between parental control (Control), paternal Vibrio (PatV+) and Tenacibaculum (PatT+), maternal Vibrio (MatV+) and Tenacibaculum (MatT+), as well as biparental (Bi-(MatV+PatT+) and Bi-(MatT+PatV+)) treatment groups. ANOSIM analysis was conducted for all target genes (44) and grouped into the following functional gene categories:
genes of the innate immune system (13), of the innate and adaptive immune system (5), of the adaptive immune system (8) and complement component genes (3).
ANOSIM
Immunegenes [29]
Innate [13]
Innate &
Adaptive [5]
Adaptive [8]
Complement [3]
F0-parents (7 treatments)
Overall R ns ns ns ns ns
Overall significance values 0.001 0.001 0.001 0.001 0.001
Bi-(MatV+PatT+) x Bi-(MatT+PatV+) 0.004 0.042 0.004 0.002 ns
Bi-(MatV+PatT+) x MatV+ 0.010 0.034 0.001 0.048 ns
Bi-(MatV+PatT+) x MatT+ 0.001 0.006 0.001 0.001 ns
Bi-(MatV+PatT+) x PatV+ 0.001 0.001 0.001 0.001 0.002
Bi-(MatV+PatT+) x PatT+ ns ns ns ns ns
Bi-(MatV+PatT+) x Control 0.005 0.005 0.003 0.015 ns
Bi-(MatT+PatV+) x MatV+ 0.001 0.003 0.001 0.001 0.001
Bi-(MatT+PatV+) x MatT+ 0.001 0.017 0.001 0.002 0.009
Bi-(MatT+PatV+) x PatV+ 0.001 0.001 0.001 0.001 0.001
Bi-(MatT+PatV+) x PatT+ 0.001 0.001 0.002 0.005 0.042
Bi-(MatT+PatV+) x Control 0.001 0.003 0.001 0.008 ns
MatV+ x MatT+ 0.001 0.002 0.001 0.001 ns
MatV+ x PatV+ 0.001 0.001 0.004 0.006 ns
MatV+ x PatT+ 0.001 0.001 0.010 0.006 0.001
MatV+ x Control 0.002 0.001 0.003 0.007 ns
MatT+ x PatV+ 0.001 0.002 0.001 0.001 0.028
MatT+ x PatT+ 0.005 0.005 0.001 0.006 0.035
MatT+ x Control 0.001 0.001 0.001 0.001 ns
PatV+ x PatT+ 0.001 0.001 0.001 0.001 0.003
PatV+ x Control 0.001 0.001 0.001 0.001 0.008
PatT+ x Control 0.001 0.001 0.009 0.007 ns
F1-offspring (3 treatments)
ANOSIM-overall R 0.061 0.070 0.074 0.001 0.087
Overall significance values 0.001 0.001 0.001 ns 0.001
F1-V+ x F1-T+ ns ns ns ns ns
F1-V+ x F1-N 0.001 0.001 0.001 ns 0.001
F1-T+ x F1-N 0.001 0.001 0.001 ns 0.001
F0-parents x F1-offspring
ANOSIM-overall R 0.149 0.118 0.130 0.076 0.098
Overall significance values 0.001 0.001 0.001 0.001 0.001
Bi-(MatV+PatT+)/F1-V+ X Bi-(MatV+PatT+)/F1-T+ ns ns ns ns ns
Bi-(MatV+PatT+)/F1-V+ X Bi-(MatT+PatV+)/F1-V+ ns ns ns ns ns
Bi-(MatV+PatT+)/F1-V+ X Bi-(MatT+PatV+)/F1-T+ ns ns 0.008 ns ns
Bi-(MatV+PatT+)/F1-T+ X Bi-(MatT+PatV+)/F1-V+ ns ns ns ns ns
Bi-(MatV+PatT+)/F1-T+ X Bi-(MatT+PatV+)/F1-T+ 0.006 0.032 ns 0.043 0.035
Bi-(MatT+PatV+)/F1-V+ X Bi-(MatT+PatV+)/F1-T+ ns ns ns ns ns
Bi-(MatV+PatT+)/F1-N X Bi-(MatT+PatV+)/F1-V+ 0.001 0.001 0.001 ns 0.004 Bi-(MatV+PatT+)/F1-N X Bi-(MatT+PatV+)/F1-T+ 0.003 0.015 0.001 ns 0.018 Bi-(MatV+PatT+)/F1-V+ X Bi-(MatT+PatV+)/F1-N 0.001 0.001 0.001 ns 0.001 Bi-(MatV+PatT+)/F1-T+ X Bi-(MatV+PatT+)/F1-N 0.002 0.023 0.001 ns 0.001 Bi-(MatV+PatT+)/F1-V+ X Bi-(MatV+PatT+)/F1-N 0.002 0.049 0.001 ns 0.005 Bi-(MatV+PatT+)/F1-T+ X Bi-(MatT+PatV+)/F1-N 0.001 0.003 0.001 ns 0.001 Bi-(MatV+PatT+)/F1-N X Bi-(MatT+PatV+)/F1-N 0.028 0.044 0.047 ns ns Bi-(MatT+PatV+)/F1-V+ X Bi-(MatT+PatV+)/F1-N 0.001 0.001 0.001 ns 0.007
Bi-(MatT+PatV+)/F1-T+ X Bi-(MatT+PatV+)/F1-N 0.027 ns 0.003 ns ns
Bi-(MatV+PatT+)/F1-V+ X MatV+/F1-V+ 0.023 ns 0.004 0.038 ns
Bi-(MatV+PatT+)/F1-V+ X MatV+/F1-T+ ns ns 0.005 0.014 ns
Bi-(MatV+PatT+)/F1-V+ X MatT+/F1-V+ 0.006 ns 0.004 0.002 ns
Bi-(MatV+PatT+)/F1-V+ X MatT+/F1-T+ 0.002 0.013 0.004 0.004 ns
Bi-(MatV+PatT+)/F1-V+ X PatV+/F1-V+ 0.002 0.006 0.001 0.002 ns
Bi-(MatV+PatT+)/F1-V+ X PatV+/F1-T+ 0.009 0.048 0.001 0.025 0.031
Bi-(MatV+PatT+)/F1-V+ X PatT+/F1-V+ ns ns ns ns ns
Bi-(MatV+PatT+)/F1-V+ X PatT+/F1-T+ ns ns ns ns ns
Bi-(MatV+PatT+)/F1-T+ X MatV+/F1-V+ 0.005 0.007 0.010 ns ns
Bi-(MatV+PatT+)/F1-T+ X MatV+/F1-T+ 0.038 0.024 0.029 ns ns
Bi-(MatV+PatT+)/F1-T+ X MatT+/F1-V+ 0.024 ns 0.001 0.002 ns
Bi-(MatV+PatT+)/F1-T+ X MatT+/F1-T+ 0.009 0.033 0.002 ns ns
Bi-(MatV+PatT+)/F1-T+ X PatV+/F1-V+ 0.003 0.004 0.002 ns ns
Bi-(MatV+PatT+)/F1-T+ X PatV+/F1-T+ 0.005 0.006 0.002 ns ns
Bi-(MatV+PatT+)/F1-T+ X PatT+/F1-V+ 0.024 0.018 ns ns ns
Bi-(MatV+PatT+)/F1-T+ X PatT+/F1-T+ ns ns ns ns ns
Bi-(MatT+PatV+)/F1-T+ X PatV+/F1-V+ 0.001 0.023 0.001 0.001 ns
Bi-(MatT+PatV+)/F1-T+ X PatV+/F1-T+ 0.001 ns 0.001 0.001 0.001
Bi-(MatT+PatV+)/F1-V+ X MatV+/F1-V+ 0.001 0.002 0.003 0.005 0.007
Bi-(MatT+PatV+)/F1-V+ X MatV+/F1-T+ 0.008 0.019 0.046 0.028 0.006
Bi-(MatT+PatV+)/F1-T+ X PatT+/F1-V+ 0.002 0.003 0.003 0.029 ns
Bi-(MatT+PatV+)/F1-T+ X PatT+/F1-T+ 0.002 0.005 0.014 ns ns
Bi-(MatT+PatV+)/F1-V+ X MatT+/F1-V+ 0.005 ns 0.001 0.037 ns
Bi-(MatT+PatV+)/F1-V+ X MatT+/F1-T+ 0.006 ns 0.001 ns 0.021
Bi-(MatT+PatV+)/F1-V+ X PatV+/F1-V+ 0.004 ns 0.007 0.003 ns
Bi-(MatT+PatV+)/F1-V+ X PatV+/F1-T+ 0.005 ns 0.011 0.015 0.005
Bi-(MatT+PatV+)/F1-V+ X PatT+/F1-V+ 0.041 0.027 ns ns ns
Bi-(MatT+PatV+)/F1-V+ X PatT+/F1-T+ ns ns ns ns ns
Bi-(MatT+PatV+)/F1-T+ X MatV+/F1-V+ 0.001 0.002 0.001 0.001 0.001
Bi-(MatT+PatV+)/F1-T+ X MatV+/F1-T+ 0.001 ns 0.001 0.002 0.002
Bi-(MatT+PatV+)/F1-T+ X MatT+/F1-V+ 0.002 0.007 0.001 0.002 0.018
Bi-(MatT+PatV+)/F1-T+ X MatT+/F1-T+ 0.001 0.018 0.001 ns 0.002
Bi-(MatT+PatV+)/F1-T+ X MatT+/F1-N 0.001 0.005 0.001 0.032 ns
Bi-(MatT+PatV+)/F1-T+ X PatV+/F1-N 0.002 ns 0.016 0.001 0.014
Bi-(MatT+PatV+)/F1-T+ X PatT+/F1-N 0.004 0.011 0.005 0.007 ns
Bi-(MatT+PatV+)/F1-T+ X MatV+/F1-N 0.014 ns 0.005 0.006 ns
Bi-(MatV+PatT+)/F1-V+ X MatV+/F1-N 0.018 ns 0.002 ns ns
Bi-(MatV+PatT+)/F1-V+ X MatT+/F1-N 0.001 0.002 0.001 0.001 0.001
Bi-(MatV+PatT+)/F1-V+ X PatV+/F1-N 0.002 0.003 0.003 0.001 0.015
Bi-(MatV+PatT+)/F1-V+ X PatT+/F1-N 0.005 0.021 0.039 0.029 0.015
Bi-(MatV+PatT+)/F1-T+ X MatV+/F1-N 0.009 0.022 0.003 ns 0.029
Bi-(MatV+PatT+)/F1-T+ X MatT+/F1-N 0.001 0.003 0.001 0.007 0.001
Bi-(MatV+PatT+)/F1-T+ X PatV+/F1-N 0.004 0.003 0.010 0.040 0.039
Bi-(MatV+PatT+)/F1-T+ X PatT+/F1-N 0.002 0.003 0.014 ns 0.003
Bi-(MatT+PatV+)/F1-V+ X MatV+/F1-N 0.002 0.001 0.001 ns ns
Bi-(MatT+PatV+)/F1-V+ X MatT+/F1-N 0.001 0.001 0.001 ns 0.008
Bi-(MatT+PatV+)/F1-V+ X PatV+/F1-N 0.001 0.001 0.001 0.008 0.013
Bi-(MatT+PatV+)/F1-V+ X PatT+/F1-N 0.001 0.003 0.002 ns 0.028
Bi-(MatV+PatT+)/F1-N X MatV+/F1-N ns ns 0.011 ns ns
Bi-(MatV+PatT+)/F1-N X MatT+/F1-N 0.010 0.034 0.001 ns ns
Bi-(MatV+PatT+)/F1-N X PatV+/F1-N 0.001 0.001 0.001 0.003 0.001
Bi-(MatV+PatT+)/F1-N X PatT+/F1-N ns ns trend 0.020 ns
Bi-(MatV+PatT+)/F1-N X MatV+/F1-V+ 0.001 0.001 0.001 0.006 0.001
Bi-(MatV+PatT+)/F1-N X MatV+/F1-T+ 0.001 0.003 0.001 0.007 0.001
Bi-(MatV+PatT+)/F1-N X MatT+/F1-V+ 0.001 0.001 0.001 0.004 0.001
Bi-(MatV+PatT+)/F1-N X MatT+/F1-T+ 0.001 0.002 0.001 ns 0.001
Bi-(MatV+PatT+)/F1-N X PatV+/F1-V+ 0.001 0.001 0.001 0.033 0.001
Bi-(MatV+PatT+)/F1-N X PatV+/F1-T+ 0.001 0.001 0.001 ns 0.001
Bi-(MatV+PatT+)/F1-N X PatT+/F1-V+ 0.001 0.009 0.001 0.024 0.004
Bi-(MatV+PatT+)/F1-N X PatT+/F1-T+ 0.001 0.005 0.001 ns 0.001
Bi-(MatT+PatV+)/F1-N X MatV+/F1-V+ 0.001 0.001 0.001 0.001 0.001
Bi-(MatT+PatV+)/F1-N X MatV+/F1-T+ 0.001 0.002 0.001 0.007 0.001
Bi-(MatT+PatV+)/F1-N X MatV+/F1-N 0.003 0.020 0.009 0.044 ns
Bi-(MatT+PatV+)/F1-N X MatT+/F1-V+ 0.001 0.003 0.001 0.001 0.001
Bi-(MatT+PatV+)/F1-N X MatT+/F1-T+ 0.001 0.006 0.001 ns 0.001
Bi-(MatT+PatV+)/F1-N X MatT+/F1-N 0.001 0.029 0.001 0.039 ns
Bi-(MatT+PatV+)/F1-N X PatV+/F1-V+ 0.001 0.001 0.001 0.001 0.001
Bi-(MatT+PatV+)/F1-N X PatV+/F1-T+ 0.001 0.001 0.001 0.001 0.001
Bi-(MatT+PatV+)/F1-N X PatV+/F1-N 0.001 0.005 0.018 0.005 0.003
Bi-(MatT+PatV+)/F1-N X PatT+/F1-V+ 0.001 0.001 0.001 0.021 0.009
Bi-(MatT+PatV+)/F1-N X PatT+/F1-T+ 0.001 0.001 0.001 ns 0.002
Bi-(MatT+PatV+)/F1-N X PatT+/F1-N 0.004 0.008 0.004 0.013 0.049
Bi-(MatT+PatV+)/F1-N X Control/F1-V+ 0.001 0.001 0.001 0.004 0.001
Bi-(MatT+PatV+)/F1-N X Control/F1-T+ 0.001 0.001 0.001 ns 0.001
Bi-(MatT+PatV+)/F1-N X Control/F1-N 0.003 0.013 0.004 ns ns
Bi-(MatV+PatT+)/F1-N X Control/F1-V+ 0.001 0.001 0.001 0.046 0.001
Bi-(MatV+PatT+)/F1-N X Control/F1-T+ 0.001 0.006 0.001 ns 0.001
Bi-(MatV+PatT+)/F1-N X Control/F1-N ns ns 0.009 ns ns
Bi-(MatT+PatV+)/F1-V+ X Control/F1-V+ 0.047 0.020 ns ns ns
Bi-(MatT+PatV+)/F1-V+ X Control/F1-T+ 0.034 0.036 0.014 ns ns
Bi-(MatT+PatV+)/F1-V+ X Control/F1-N 0.003 0.014 0.001 0.012 ns
Bi-(MatT+PatV+)/F1-T+ X Control/F1-V+ 0.001 0.001 0.001 0.008 ns
Bi-(MatT+PatV+)/F1-T+ X Control/F1-T+ 0.001 0.003 0.001 0.042 ns
Bi-(MatT+PatV+)/F1-T+ X Control/F1-N 0.005 0.031 0.003 0.001 ns
Bi-(MatV+PatT+)/F1-V+ X Control/F1-V+ 0.023 0.008 0.019 0.005 ns
Bi-(MatV+PatT+)/F1-V+ X Control/F1-T+ ns ns 0.027 ns ns
Bi-(MatV+PatT+)/F1-V+ X Control/F1-N 0.006 0.046 0.003 0.005 ns
Bi-(MatV+PatT+)/F1-T+ X Control/F1-V+ 0.015 0.001 ns ns ns
Bi-(MatV+PatT+)/F1-T+ X Control/F1-T+ 0.042 0.010 ns ns ns
Bi-(MatV+PatT+)/F1-T+ X Control/F1-N 0.013 ns 0.004 0.040 0.024
