I am new to divergence time dating in MrBayes 3.2, though I have used BEAST often. I am analyzing some data for which I prefer to use MrBayes for reasons not applicable to this question.
I ran an analysis with node constraints and calibrations as follows:
Begin mrbayes;
[Hard constraints here; Identical to those listed in prset topologypr below. Not provided here in Biostars for space considerations.]
lset nst=6 rates=gamma ngammacat=4;
prset topologypr = constraints(Ingroup,AesculusHandeliodendron,AesculusGenus,sectCalothyrsus,AsianClade,node1,node2,node3,sectsParryanaPaviaMacrothyrsusHippocastanum,sectsParryanaPaviaMacrothyrsus,sectPavia,node4,node5,sectAesculus);
prset clockratepr = lognormal(-5,0.6); [mean 0.00001, SD 0.005; centered around small numbers with room for increasing]
prset clockvarpr = tk02; [use with default settings]
prset sampleprob = 0.01;
prset treeagepr = offsetlognormal(72.1,73.53,1.25);
calibrate AesculusGenus = offsetlognormal(66.0,66.1,0.8);
calibrate sectAesculus = uniform(1.75,22.5);
prset nodeagepr = calibrated;
mcmc ngen=20000000 nruns=2 nchains=4 samplefreq=1000 checkpoint=yes checkfreq=1000 temp=0.2 mcmcdiagn=no;
;
end;
I can confirm from a log of what is echoed to screen in MrBayes that these priors were set as intended:
Defining constraint called 'sectAesculus' [others defined above; not included here for space]
Setting Nst to 6
Setting Rates to Gamma
Setting Ngammacat to 4
Successfully set likelihood model parameters
Setting Topologypr to Constraints
Successfully set prior model parameters
Setting Clockratepr to Lognormal(-5.000000,0.600000)
Successfully set prior model parameters
Setting Clockvarpr to TK02
Successfully set prior model parameters
Setting Sampleprob to 0.01000
Successfully set prior model parameters
Setting Treeagepr to Offsetlognormal(72.10,73.53,1.25)
Successfully set prior model parameters
Setting age of constraint node 'AesculusGenus' to Offsetlognormal(66.00,66.10,0.80)
Setting age of constraint node 'sectAesculus' to Uniform(1.75,22.50)
Setting Nodeagepr to Calibrated
Successfully set prior model parameters
Setting number of generations to 20000000
[...and so on]
However, my results in no way reflect these node age calibrations and look more like uncalibrated branch lengths. See here for a screen shot of the MCC Tree displayed in FigTree with node ages on branches.
This is not a case of extremely strong signal in the data leading to results outside of the prior bounds as I do get the expected old ages using these calibration priors in BEAST.
What have I done wrong?
Thanks in advance!