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LECTURE 19:    SPECIATION GENETICS AND HALDANE'S RULE

I.  Haldane's rule

    A.  Statement of Haldane's rule:  When in the F offspring produced by crossing two taxa, one sex is absent,
          rare or sterile, that sex is the heterogametic sex.

        1.  In organisms with X-Y sex determination (e.g. Drosophila, mammals), males are the hetergametic
             sex (i.e. males are XY, females are XX).

        2.  Consequently, Haldane's rule states that in these organisms, whenever there is partial reproductive
             isolation, males should exhibit reduced viability or sterility more so than females.

        3.  By contrast, in organisms with Z-W sex determination (e.g. birds, butterflies), females are the
             heterogametic sex (i.e. males are ZZ, females are ZW).

        4.  Haldane's rule states that in these organisms, whenever there is partial reproductive isolation,
             females should exhibit reduced viability or sterility more so than males.

    B.  Evidence

        1.  A recent compendium of several hundred cases of sex-specific reproductive isolation indicates that
             in most cases, Haldane's rule is obeyed. 

        2.  The near universality of this phenomenon indicates that it is one of the most robust generalizations
             in evolutionary biology.

II. Explaining Haldane's rule:  Y-chromosome effect

         1.  As the figure shows, hybrids obtain one copy of each chromosome from each parental species.

 

        2.  One obvious simple explanation for Haldane's rule is that the inviability or sterility of males is
             due to an incompatibility allele on the Y chromosome.

        3.  However, by crosses, one can make males that that carry all chromosomes from one species,
             but a Y-chromosome from the other species:
 

            a.  If there were incompatibility alleles on the Y chromosome, and these are responsible
                 for the sex-limitation of  inviability or sterility, then such individuals should always
                 be inviable or sterile.
            b.  Many experiments have been done to test this--sometimes such individuals are sterile,
                 but most of the time they are viable and fertile.

        4.  Consequently, it does not appear that the presence of incompatibility alleles on the Y chromosome
             can be a general explanation for Haldane's rule.
 
 

III.  Explaining Haldane's rule: Muller's X-Autosome imbalance theory ("dominance theory")

    A.  Explanation of theory

        1.  Suppose genetic incompatibility between two species, with genotypes AAbb and aaBB
             respectively, results from alleles A and B being combined in hybrids (i.e. A and B together
             is detrimental.)

        2.  This effect by itself will not produce hybrid inviability or sterility that is confined to just the
             heterogametic sex.

        3.  However, let us further assume that the detrimental A-B interaction is buffered by the presence
             of  a and b alleles, so that in the heterozygote AaBb, the detrimental interaction is recessive and
             not expressed.

        4.   In this case, if both A and B are on autosomes, then both sexes will be buffered and neither sex
              will exhibit hybrid inviability or sterility.

        5.   However, if one gene, say A, is on the X chromosome, the result is more interesting.

            a.  Female hybrids are AaBb, the A-B interaction is buffered, and females exhibit no
                 detrimental effects.
            b.  By contrast, male hybrids are AYBb (Y chromosomes are essentially devoid of genes).
            c.  At the A locus, male hybrids are "hemizygous" they contain only one copy of the A gene.
            d.  There is thus no a allele to provide an a-b interaction to buffer the A-B interaction and
                 males are thus inviable or sterile.

        6.  Thus, according to Muller's hypothesis, recessive incompatibility alleles accumulate on all
             chromosomes.

            a.  Given recessivity of detrimental interactions, pairs of incompatibility alleles that are
                 both on autosomes do not contribute to hybrid inviability or infertility.
            b.  But pairs of alleles in which at least one locus is on the X chromosome will contribute
                 to hybrid inviability/sterility in males but not in females.

        7.  This effect depends on incompatibility alleles being on average recessive--it has been shown
             theoretically that if such alleles are dominant on average, then hybrid inviability/sterility will
             tend to occur in females rather than males.

        8.  There is no reason to expect incompatibility alleles to be recessive on average, but we can't rule
             out this possibility either.  Whether Muller's hypothesis is correct is thus an empirical matter
             open to experiment.

    B.  Experimental test of Muller's hypothesis--rationale

        1.  If hybrid breakdown is stronger in the heterogametic sex (males) because there isn't a sex
             chromosome carrying an allele to buffer the A-B interaction, then one should see similar
             breakdown in females in which there isn't a buffering allele.

        2.  To create such females, use attached-X chromosome: two X-chromosomes that are connected and
             segregate together at meiosis.

        3.  The cross that is done, then is

                   AAbb  female   x    a-BB male

                   Using Normal female:

                            XAXA bb    x     XaY BB   =>        XAXaBb       female:   viable and fertile

                                                                                   XAY Bb         male:   inviable or sterile

                   Using Attached-X female:
 
                            XA--XAYbb      x     XaY BB     =>        XA--XAXa Bb    dies (too many X's)

                                                                                            XA--XA Y Bb       female (2 X's)

        4.  If Muller's hypothesis is correct, then the XA--XA YBb females should also be inviable or sterile
             because they carry an A-B interaction that is not buffered by an a.

    C.  Experimental test of Muller's hypothesis--results.

      (From, C. C. Laurie.  1997.  The weaker sex is
heterogametic: 75 years of Haldane's rule.
Genetics 147: 937-951.)
 
 
 
 
 
 
 
 
 
 
 
 
 
 
 

        1.  Two sets of crosses (D. melanogaster female x D. simulans or D.mauritiana male, and
             D. simulans female x D. teissieri male) are consistent with Muller's explanation in that
             the attached-X females are inviable, just as are the regular hybrid males.

        2.  However, a cross between D.simulans female and D. mauritiana or D. sechellia male does not
             support Muller's explanation because the attached-X females are viable and fertile, while
             the normal hybrid males are sterile.

        3.  One possible explanation for these different results arises from the recognition that in the
             two cases supporting Muller's explanation, hybrid males were inviable, whereas in the one case
             that does not support explanation, hybrid males were viable but infertile.

        4.  The data thus suggests that Muller's explanation may hold for sex-limited hybrid inviability,
             but not for sex-limited hybrid sterility.

        5.  This makes sense.

            a.  Viability tends to be affected by the same genes in males and females.  Consequently,
                 any incompatibility genes on the X chromosome that are expressed in hybrid males and
                 cause inviability will also be expressed in hybrid attached-X females and cause inviability:
 

            b.  By contrast, it is reasonably well established that different genes affect fertility in males
                 and females.  Consequently, it is possible that there may be incompatibility genes
                 affecting male fertility on the X chromosomes but no incompatibility genes affecting
                 female sterility.

            c.  In such a situation, Haldane's rule would hold, but the attached-X test of Muller's
                 explanation would fail.  (see Figure)

        6.  Possible reasons for more incompatibility genes affecting male fertility than affecting female
             fertility on X chromosome (the "Differential Accumulation Theory"):

            a.  Incompatibility genes affecting male fertility may accumulate more quickly on all
                 chromosomes than inc. genes affecting female fertility.
                --One possible reason is that male traits evolve faster due to sexual selection.
            b.  Such genes affecting male fertility may accumulate more rapidly than those affecting
                 female sterility on X chromosome alone, accumulating at similar rates on autosomes.
                --See below for explanation.
 

IV.  Explaining Haldane's rule for fertility: differential accumulation theory

    A.  Experimental method

        1.  General rationale:

            a.  introgress small pieces of  single chromosomes into the genome of another species
            b.  determine, for each piece, whether it carries incompatibility alleles affecting male
                 fertility, female fertility, or both.
            c.  If there are more pieces that affect male fertility, implies alleles affecting male fertility
                 accumulate (evolve) more rapidly than alleles affecting female fertility.

        2.  Procedure: Backcross 
 
            a.  Insert a small gene segment carrying a marker allele into a chromosome of one species.
            b.  Make this segment homozygous by appropriate crosses.
            c.  Cross with a second species.  The hybrid carries one copy of the chromosome with the
                 insert.
            d.  Backcross repeatedly to second species, selecting individuals with the marker each
                 generation.
            e.  Each generation, recombination shortens the segment from the first species' chromosome
            f.  After, say, 15 generations, the segment from the first species is very small.

        3.  Procedure: Testing effects of introgressed segment on fertility.

            a.  Cross a male and female, each heterozygous for the introgressed segment, to
                 produce individuals that are homozygous for that segment.

        4.  Repeat this procedure for many introgressed segments located in different parts of the genome.

            a.  Each segment that has an effect on viability or fertility indicates the presence of at
                 least one incompatability gene within that segment.
            b.  Hence, can obtain a mininimum estimate of the number of incompatibility genes of
                 various types.
 
     B.  Example:  True et.al.  1996.  Genetics 142: 819-837.

        1.  Backcrossing

            a.  Introduced a marker using P-element (a transposon) tagging into D. mauritiana.
            b. P-element carried mini-white gene affecting eye color, which allows individuals
                carrying the marker to be scored easily.
            c.  P-elements insert essentially randomly all over the genome.
            d.  Obtained single inserts in 185 individuals, all in different autosomal locations.
            e.  Each of these individuals was used to establish a backcross line and the corresponding
                 marker and flanking DNA was introgressed into D. simulans.
            f.  Each line was then tested for the effect of the introgressed segment on viability and
                fertility.

        2.  Results

            a.  Results presented in table, which shows the number of introgression lines in each
                 category:

                                                                   Male     Male      Male
                                                                   fertile     sterile     untested

                                 Female fertile             112        53            ---
                                 Female sterile                6           4             2
                                 Female untested          ---           8           ----

            b.  Of the 175 segments, 112 had no detectable effects on viability or sterility in either sex.
            c.  Of the remaining segments, 53 reduced only male fertility, while 6 reduced only female
                 fertility and 4 reduced both male and female fertility.
            d.  In other words, about 9 times as many segments had incompatibility alleles affecting
                 male fertility.

        3.  Conclusion:  Incompatibility alleles affecting male fertility accumulate much faster than those
             affecting female fertility, supporting the differential accumulation theory.
 
     C.  Differential Accumulation on X chromosome.

        1.  Wu et al. performed a similar experiment introgressing X-chromosome segments
             from D.mauritiana into D. simulans.
 
        2.  Results indicate that there are more than 40 incompatibility genes on the X-chromosome
             contributing to male sterility.

        3.  By contrast, there are apparently no X-chromosome incompatibility genes affecting female
             fertility (this is demonstrated by complete fertility of attached-X females).

        4.  Conclusion:  Results support the differential accumulation theory.

V.  Conclusions

    A.  As a general phenomenon, Haldane's rule seems to be caused by several different phenomena.

        1. In some cases, it seems to be due to the type of recessivity of incompatibility interactions
            envisioned by Muller's theory.

        2.  In other cases, it seems to be due to a generally more rapid accumulation of alleles affecting
             male hybrid inviability/sterility than alleles affecting female hybrid inviability/sterility.

        3.  There is some evidence that there may also be more rapid accumulation of incompatibility
             alleles on the X chromosome than on autosomes, which can account for not only Haldane's
             rule but also the Large-X effect.

    B.  This multiplicity of explanations seems to be the rule in evolutionary biology:  the same phenomenon
          may have different explanations in different groups of organisms.


   


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