NAME

Zmapqtl - Composite interval mapping module

SYNOPSIS

Zmapqtl [ -o output ] [ -i input ] [ -m mapfile ] [ -l lrfile ] [ -S srfile ] [ -t trait ] [ -M Model ] [ -c chrom ] [ -d walk ] [ -n nbp ] [ -w window ] [ -r perms ] [ -r boots ]

DESCRIPTION

Zmapqtl uses composite interval mapping to map quantitative trait loci to a map of molecular markers. It requires a molecular map that could be a random one produced by Rmap, or a real one in the same format as the output of Rmap. The sample could be a randomly generated one from Rcross or a real one in the same format as the output of Rcross. In addition, the program requires the results of the stepwise linear regression analysis of SRmapqtl for composite interval mapping.

OPTIONS

See QTLcart(1) for more information on the global options -h for help, -A for automatic, -V for non-Verbose -W path for a working directory, -R file to specify a resource file, -e to specify the log file, -s to specify a seed for the random number generator and -X stem to specify a filename stem. The options below are specific to this program.

If you use this program without specifying any options, then you will get into a menu that allows you to set them interactively.

-o

This requires a filename for output. Zmapqtl will append the file if it exists, and create a new file if it does not. If not used, then Zmapqtl will use qtlcart.z.

-i

This requires an input filename. This file must exist. It should be in the same format as the output of Rcross. The default file is qtlcart.cro.

-m

Zmapqtl requires a genetic linkage map. This option requires the name of a file containing the map. It should be in the same format that Rmap outputs. The default file is qtlcart.map.

-t

Use this to specify which trait Zmapqtl will analyze. If this number is greater than the number of traits, then all traits will be analyzed. The default is to analyze trait 1 only.

-l

Allows the user to specify the name of the file containing results from LRmapqtl. Zmapqtl reads those results and uses the information to choose cofactors for some of the analysis methods.

-S

Allows the user to specify the name of the file containing results from SRmapqtl. Zmapqtl reads the results and uses the information to choose cofactors for composite interval mapping model 6.

-M

Zmapqtl assumes the specified model (see below) in the analysis. Model 3 is default.

-c

The user can specify a specific chromosome for Zmapqtl to analyze. If zero, then all will be analyzed.

-d

Zmapqtl walks along the chromosome at a rate that can be specified with this option. The default is to do an analysis every 2 centiMorgans along the chromosome.

-n

Use this to indicate how many background parameters Zmapqtl uses in composite interval mapping. This is used only with model 6, and gives an upper bound. If fewer than this number of markers are ranked in the SRmapqtl.out file, then less than the specified number of markers will be used.

-w

Zmapqtl blocks out a region of this many centiMorgans on either side of the markers flanking the test position when picking background markers. It is 10 by default and is only used in models 5 and 6. We refer to it as the window size.

-r

Zmapqtl can do a permutation test to determine the threshold for rejecting the null hypothesis of no QTL at a site. By default, this option sets the number of permutations equal to 0, which means no permutation test is run. You can set it to a number < 10000 to do the test. See Churchill and Doerge (1994) for more details. The results are in an interim file. Use Eqtl to summarize them when enough repetitions have been done. You need to run Zmapqtl without permutations or bootstraps at least once before you can do the permutation tests. This option only allows for interval mapping (Model 3) or composite interval mapping (Model 6).

-b

When used with argument 1, Zmapqtl will do a single bootstrap. You need to run Prune to actually create the bootstrapped data set: This option merely analyzes it and stores summary statistics in an interim file qtlcart.z3b by default, for model 3. You should also run Zmapqtl without bootstraps or permutation tests before doing a bootstrap analysis.

When used with an argument 2, Zmapqtl will do a jackknife analysis. Again, Zmapqtl should be run without this argument prior to doing a jackknife.

INPUT FORMAT

The input format of the molecular map should be the same as that of the output format from the program Rmap. The input format of the individual data should be the same as the output format of the program Rcross.

EXAMPLES

        % Zmapqtl

Calculates the likelihood ratio test statistics of the dataset in qtlcart.cro using the map in qtlcart.map.

        % nice Zmapqtl -A -V -i corn.cro -m corn.map -M 6 -r 500 &

Calculates the likelihood ratio test statistics of the dataset in corn.cro using the map in corn.map. Model 6 is used for analysis and a permutation test with 500 replications is performed. The program is nice'd as a courtesy to other users, and run in the background so that the user can logout and relax.

MODELS

Different parameters for the -M option allow for the analysis of the data assuming different models. Models 1-3 were described in Zeng (1993, 1994).

Fit all the background markers. This was meant for illustration of the method in the original paper (Zeng, 1993), and should not be used for analysis.
Fit all unlinked background markers. This is another illustrative example and also should not be used.
Fit only the mean (Lander and Botstein (1989) method)
Fit a subset of the other markers, namely those unlinked markers with the highest correlation with the trait on each chromosome. This is an ad hoc model programmed in anticipation of model 6 below.
This model uses a pair of markers from each other chromosome and all linked markers that fall outside a window around the flanking markers. This window extends to 10 cM beyond the markers immediately flanking the test position. The window size can be changed with the -w option. This is another ad hoc model programmed in anticipation of model 6 below.
This model uses a specified number of markers that fall outside a window around the flanking markers. This window extends to 10 cM beyond the markers immediately flanking the test position. The number of markers are set by the -n option. You need to run SRmapqtl to rank the markers before using model 6. You should use this for composite interval mapping when using markers ranked by SRmapqtl.
This model requires that you have already run Zmapqtl and Eqtl. It reads in the estimated QTL from the Eqtl.out file and uses them as virtual markers to control for the genetic background. All identified markers are used that do not fall within the window. This has not been extensively tested: Use it at your own risk.
Like model 7, this requires a prior run of Zmapqtl and Eqtl. Instead of using virtual markers, Zmapqtl uses the closest flanking markers to identified QTL. Again, all of these markers outside the window are used. This is good model. You can iterate this process to see if a stable set of cofactors can be identified. See the script below and the Model8.pl(1) man page for more details.

The default is to fit only the mean, that is to use interval mapping.

PERMUTATION TESTS

Churchill and Doerge (1994) describe a method to calculate the threshold values for quantitative trait mapping that we have implemented in this program. Basically, it does a permutation of the trait values and the genotypes and redoes the analysis. Over the number of replicates, two types of thresholds are defined: ``experimentwise'' and ``comparisonwise''. We calculate the experimentwise thresholds, but only give p values for the comparisonwise values to save on storage space. The p values give the proportion of permuted replicates that have loglikelihood ratios larger than the observed ratios.

If you choose to do permutation tests, you need to run Zmapqtl with the model of choice prior to doing the permutation test. Also, if the program terminates prematurely, you can restart it from where it left off to complete the permutation test.

REFERENCES

Churchill, G. A. and R. W. Doerge (1994) Empirical threshold values for quantitative trait mapping. Genetics 138, 963-971.
Lander, E. S. and D. Botstein (1989) Mapping Mendelian factors underlying quantitative traits using RFLP linkage maps. Genetics 121, 185-199.
Zeng, Zhao-Bang (1993) Theoretical basis for separation of multiple linked gene effects in mapping quantitative trait loci. Proc. Natl. Acad. Sci., USA 90, 10972-10976.
Zeng, Zhao-Bang (1994) Precision mapping of quantitative trait loci. Genetics 136, 1457-1468.

CAVEATS

Model 7 has not been extensively tested. Remember that you will need to run Zmapqtl with some other model (say 3 or 6) and then run Eqtl prior to using model 7.

Model 8 requires a prior run of Zmapqtl and Eqtl as well. There is a shell script called Model8.csh in the scripts subdirectory that allows you to easily iterate using model 8. Try using it to see whether your set of cofactors (or estimated QTL) becomes stable after a few iterations. A low threshold may lead to adding and dropping putative QTL over consecutive analyses using Model 8. Here is the shell script:

        #!/usr/bin/csh
        #
        #  Run Model 8 iteration
        #           Copyright 2001 Christopher J. Basten
        #   Usage:
        #   Model8  bindir  stem   siglevel  iterations  max_nbp
        #     bindir is the binary subdirectory
        #     stem is the filename stem
        #     siglevel  is the significance level to declare a QTL
        #     iterations is the number of iterations
        #     max_nbp is the maximal number of background parameters.
        #
        if ( $1 == '-h' ) then
        echo "    Usage:  Model8.csh bindir  stem   siglevel  iterations  max_nbp"
        echo "Where"
        echo "        bindir  = QTL Cart. binary directory"
        echo "          stem  = filename stem"
        echo "      siglevel  = Significance level to declare a QTL"
        echo "    iterations  = number of iterations"
        echo "       max_nbp  = maximal number of background parameters"
        echo " "
        echo "Now exiting"
        exit
        endif
        set bindir=$1
        set stem=$2
        set siglevel=$3
        set iterations=$4
        set maxnbp=$5
        $bindir/Qstats -X $stem -A -V
        $bindir/Zmapqtl -A -V -M 3
        $bindir/Eqtl -A -V -S $siglevel
        #
        #  Save the original files
        #
        /usr/bin/mv $stem.eqt $stem.eqt.0
        /usr/bin/mv $stem.z $stem.z.0
        /usr/bin/cp $stem.sr $stem.sr.0
        #
        #  Use model 8 iteratively with cofactors from previous run.
        #
        set i=1
        while ( $i < $iterations )
        echo "Doing iteration $i"
        $bindir/Zmapqtl -A -V -M 8 -n $maxnbp
        /usr/bin/rm $stem.sr 
        $bindir/Eqtl -A -V -S $siglevel
        /usr/bin/cp $stem.sr $stem.sr.$i
        /usr/bin/mv $stem.eqt $stem.eqt.$i
        /usr/bin/mv $stem.z $stem.z.$i
        @ i++
        end
        /usr/bin/rm $stem.sr 
        echo "Finished"

The above script has been translated into Perl to make it more useful. The Model8.pl script can take command line parameters and is self-documenting.

BUGS

It is likely that we will abandon the internal permutation tests in Zmapqtl. It is more efficient to use Prune and a batch file to do the same job. This paradigm will allow users to do permutation tests with any of the programs. Of course, you will need access to a UNIX platform to do this.

SEE ALSO

Emap(1), Rmap(1), Rqtl(1), Rcross(1), Qstats(1), LRmapqtl(1), BTmapqtl(1), SRmapqtl(1), JZmapqtl(1), Eqtl(1), Prune(1), Preplot(1), MImapqtl(1), MultiRegress(1), Examples(1) SSupdate.pl(1), Prepraw.pl(1), EWThreshold.pl(1), GetMaxLR.pl(1), Permute.pl(1), Vert.pl(1), CWTupdate.pl(1), Ztrim.pl(1), SRcompare.pl(1), Ttransform.pl(1), TestExamples.pl(1), Model8.pl(1), Dobasics.pl(1), Bootstrap.pl(1)

CONTACT INFO

In general, it is best to contact us via email (zeng@statgen.ncsu.edu)

        Christopher J. Basten, B. S. Weir and Z.-B. Zeng
        Bioinformatics Research Center, North Carolina State University
        1523 Partners II Building/840 Main Campus Drive
        Raleigh, NC 27695-7566     USA
        Phone: (919)515-1934

Please report all bugs via email to qtlcart-bug@statgen.ncsu.edu.

The QTL Cartographer web site ( http://statgen.ncsu.edu/qtlcart ) has links to the manual, man pages, ftp server and supplemental materials.