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LECTURE 4 and 5:  NATURAL SELECTION: THEORETICAL CONSIDERATIONS

I.  Darwin's deduction of natural selection

     A.  Darwin's logic

     B.  Darwin's Crucial assumption

          1.  Most of the assumptions used by Darwin in his logical deduction of evolution by natural selection are
               self-evident or well established.

          2.  He had, however, two novel insights, associated with assumption of step 7:

                a.  Some individuals possess certain traits that make them are better equipped for competition to obtain
                    resources; they possess these traits because of their genetic makeup; and their increased competitiveness
                    causes them to produce more offspring. (PRINCIPLE OF NATURAL SELECTION)

                b.  Because these traits are hereditary and individuals possessing them produce more offspring, the traits
                     will be more common in the next generation. (PRINCIPLE OF EVOLUTIONARY CHANGE)

          3.  Both of these assumptions are clearly open to experimental verification or refutation (see below).

     C.  Darwin's logic is not circular.

     D.  Darwinian Fitness

          1.  Although Darwin's deduction of natural selection is phrased in terms of differential success among genotypes
               in the competition to obtain resources, which suggests that the driving force behind natural selection is
               differential survival, Darwin clearly recongized that natural selection could also arise due to differential
               reproductive success among individuals.

          2.  His novel insight should thus be rephrased in a more general fashion:  Some individuals have higher FITNESS
               because of their genetic makeup; or , more formally, individuals of some genotypes have a higher fitness, on
               average, than individuals of other genotypes.

               a.  Individual fitness may be defined, at this point, as the number of offspring produced by an individual.

               b.  Average fitness of individuals of a given genotype is influenced both by the probability that such an
                    individual will survive to reproduce and by the number of offspring that it produces if it survives.

               c.  In particular, for an annual organism with one generation per year, for a particular genotype,

                         fitness = W = (probability of survival) x (average number of seeds produced)

     E.  Natural Selection

          1.  Definition:  Natural selection occurs when genotypes differ in average fitness.

          2.  Natural selection IS NOT the same thing as Evolution.

               a.  Natural selection is one cause of Evolutionary change, as we shall soon see.
               b.  Processes other than natural selection can cause Evolutionary change, as we shall also soon see.

II.  Testing Darwin's Crucial Assumption about Natural Selection

     A.   Numerous examples exist of genotypes differing in fitness, indicating that natural selection is ubiquitous in
            nature (e.g. see Endler, J. A.  1986.  Natural Selection in the Wild.  Princeton University Press, Princeton, NJ).

     B.  Example: Leaf-shape polymorphism in Ipomoea hederaceae.

          1.  This example not only demonstrates that genotypes differ in fitness, but also illustrates how fitnesses are
               calculated experimentally.

          2.  This example also shows that which allele is favored, or which genotype has the highest fitness, is not absolute,
               but depends on the environment in which a population exists.

III.  Formalizing Darwin's argument about natural selection causing evolutionary change.

     A.   Applicability of Darwin's logic to Mendelian inheritance

          1.  Although Darwin's argument seems rather impeccable, he derived it long before anything was known about
               the true nature of heredity.

          2.  Consequently, it behooves us to enquire whether the laws of Mendel are compatible with that argument and,
               if so, to determine whether we can derive a theoretical framework that will allow us to predict changes in
               gene frequency.

          3.  We will concentrate on developing such a theoretical framework for predicting evolution at a single locus with
               two alleles, since this is relatively straightforward.  Incorporating multiple loci into the framework is
               conceptually straightforward but analytically too complicated for this course.

     B.  Goal:  predict change in gene frequency from one generation to next

          1.  Information needed for prediction

               a.  initial gene frequency
               b.  fitnesses of the genotypes at the locus of interest

          2.  Possible methods used for prediction

               a.  Simulation
               b.  Derivation of a dynamical equation

IV.  Predicting gene frequency change through simulation

     A.  Features of simulation

          1.  Initial state is a population of zygotes, each of whose genotype is specified and kept track of by the computer

          2.  The values of fitness for each genotype are used to determine whether an individual survives or not.

               a.  For each individual, determine from its genotype what its probability of survival is.
               b.  The computer determines, based on that probability and selection of a random number, whether that
                    individual survives or not.  The computer keeps track of individuals that survive.

          3.  Zygotes of the next generation are formed by random mating among survivors.

               a.  For first zygote, chose a male parent randomly from the survivors and a female parent randomly from
                    the survivors.
               b.  Determine the genotype of that zygote according to Mendel's laws by chosing randomly an allele from
                    the male parent and an allele from the female parent.
               c.  Repeat this process until a new generation of zygotes is produced.

          4.  Repeat steps 2 and 3 for each generation.

     B.  Results of simulation

          1.  Set WAA = 0.5, WAa = Waa = 0.67, initial p = 0.001 and population size = 500

          2.  A simulation of 100 generations shows that p increases to close to 1.0, i.e. the a allele replaces the A allele.

          3.  A set of 10 simulations shows a similar pattern, though the exact trajectory of gene frequency change
               differs for all 10 simulations.

          4.  The mean trajectory of the 10 simulations exhibits a steady, monotonic increase in gene frequency of the a allele.

     C.  Conclusion: simulation indicates that, at least in the case examined, if fitnesses differ for the three genotypes
           (i.e. natural selection occurs), this will be accompanied by evolutionary change (change in gene frequency).

V.  Predicting gene frequency change with a deterministic equation.

     A.  The equation for gene frequency change at a single locus with two alleles subject to natural selection is

             p' = p[pWAA+ qWAa] / [p2WAA+ 2pqWAa +  q2Waa]                                 (I) ,

            where p is the frequency of the A allele, q = 1 - p = frequency of the a allele,  and Wij= fitness of genotype ij.

     B.  This equation may also be written as

             Dp = p' - p = [pq/W..] [p(WAA -  WAa ) + q (WAa -  Waa )                           (II),

            where W.. = p2WAA+ 2pqWAa +  q2Waa  .

     C.  Using the equation.

          1.  Plug in initial value of p and values for the fitnesses, Wij , and calculate new gene frequency, p'.

          2.  Iterate by plugging in the new value of p to calculate subsequent value of p.

     D.  Comparison of predictions using simulations and predictions using deterministic equation.

          1.  Using the same values of the Wij  , the deterministic equation yields a predicted trajectory of gene frequency
               change very similar to the mean trajectory obtained in the simulations.

          2.  This result suggests that deterministic equation is an accurate predictor of evolutionary change for one locus.

VI.  Derivation of Deterministic Equation for Gene Frequency Change

            [If sub/superscripts garbled, try downloading this Alternate version]

VII.  Properties of Evolution Revealed by Deterministic Equation(see accompanying Handout)

     A.  When fitnesses are all equal, no evolutionary change is expected.

          1.  To see this, substitute the constant W for the Wij in equation (I) and simplify to get p' = p , i.e. no
               evolutionary change.

          2.  Consistent with Hardy-Weinberg results: if there is no natural selection, expect no change in gene frequencies.

     B.  We need only know relative ranking the Wij to predict long-term evolutionary change.

          1.  To see this, first recognize that evolutionary equilibrium is achieved when  Dp = 0.

          2.  All possible relationships among the  may be grouped into three categories:

               a.   WAA <,= WAa <,=  Waa  , with one inequality strict.  (Note that this case pertains to

                   WAA > WAa > Waaalso, since this is obtained by just relabeling the alleles).

               b.   WAA < WAa >  Waa

               c.   WAA > WAa <  Waa

      3.  For category a,  p will always decrease until the a allele is fixed.  This can be seen from Eq. (II), as follows:

               WAA <,= WAa <,=  Waa  implies WAA - WAa < 0 and WAa - Waa < 0 , so that  Dp < 0, i.e. the

           frequency of the A allele always decreases.  (Handout)

      4.  For category b, a polymorphism will be maintained.  To see this, note that when WAA < WAa >  Waa  is true,
           a solution to  is given by

           p* = (WAa - Waa ) / [ (WAa - WAA)  +  (WAa - Waa ) ]

           (plug this value of p into Eq. (II)  to verify that it yields Dp = 0).

           a.  That this equilibrium is STABLE (i.e. if the population is perturbed away from this equilibrium, it will return to
              p*) , can be shown mathematically.
           b.  Because the proof is too complicated for this course, however, we rely instead on the results of numerically
                iterating Eq. (I)  for various starting values of p to demonstrate the stability of the equilibrium  (Handout).
 

      5.  For category c, there is again an equilibrium at

            p* = (WAa - Waa ) / [ (WAa - WAA)  +  (WAa - Waa ) ]

           but this equilibrium is unstable.

           a.  Again, we rely on numerical iteration of the Eq. (I) from various starting p to show this.  (Handout)
           b.  These iterations show that the system will evolve to either fixation or elimination of the  allele, depending
                on whether the initial gene frequency is greater or lower than the unstable equilibrium frequency.

VIII.  Conclusions

    A.  Evolution is a predictive science: Eq. (I) predicts the long-term course of evolutionary change at a single locus
          with two alleles.

    B.  If we know precise values of fitnesses for the different genotypes and are willing to assume that these values do
          not change over time, then Eq. (I) predicts the precise trajectory of evolutionary change.

    C.  In most cases, because fitness is difficult to measure precisely, the best we will be able to do is estimate the
          relative rankings of the fitnesses of the different genotypes.

    D.  However, even though we may not be able to predict the exact trajectory of evolutionary change in these
          cases, the above results indicate that knowing just the relative rankings of fitness does allow us to predict
          the evolutionary equilibrium that will eventually be reached.  Such a prediction of course necessitates       
          assuming that the relative rankings do not change, but in many cases the biology of the situation will make

          this assumption reasonable.

IX.  Testing the Predictive Equation

     A.  We have seen that numerous experiments have demonstrated that genotypes in natural populations frequently differ
           in fitness, i.e. that Natural Selection is ubiquitous in nature.

     B.  We have also developed a theoretical framework for understanding how natural selection causes changes in gene
          frequencies (i.e. causes evolution) for a single locus with two alleles.

     C.  Several questions arise from these two observations:

          1.  Is the equation for gene frequency change we have derived quantitatively correct, i.e. given that we can
               measure fitnesses accurately, does it actually predict the correct evolutionary trajectory a population will follow?

          2.  Are the qualitative predictionsof the equation acurate, i.e., given that we can rank fitness of different genotypes,
               even if we can't measure them with great accuracy, do the rankings predict the evolutionary outcome in
               natural populations?

          3.  Given that we have confidence, from answers to questions 1 and 2, that we understand how natural selection
               causes evolutionary change, what other types of evidence can be used to infer that populations have undergone
               divergent evolution due to natural selection?

     D.  In this lecture, we address each of these questions.

X.  Laboratory Experiments: Demonstrating Quantitative agreement with equation for gene
      frequency change.

     A.  Laboratory experiments as microcosms

          1.  Ideally, to assess whether the predictive equation for gene frequency change accurately and quantitatively
               describes evolution by natural selection, we would like to measure fitnesses in a genetically variable natural
               population, follow gene frequency change over time, and determine whether the actual trajectory of gene
               frequency change matched that predicted by the equation.

          2.  Several things make this extremely difficult:

               a.  Fitness is very difficult to measure accurately in the field.
               b.  Fitnesses do not remain constant, as assumed by the predictive equation.
                    --Environmental fluctuations cause generation to generation changes in the value of a genotype's fitness,
                       even if such fluctuations are not drastic enough to change the fitness ranking of different genotypes.
                    --Therefore, it is generally not possible to measure fitnesses once in the field and use those values to predict
                       gene frequency change over long periods of time.

          3.  One way around these difficulties is to assess the quantitative accuracy of the predictive equation in laboratory
               populations, in which the environment can be held constant and in which accurate measures of fitness can be obtained.

          4.  This approach is legitimate because we the predictive equation is completely general: it should apply in all
               environments, including artificial laboratory environments.

     B. Example: Drosophila melanogaster and the lt locus (B. Wallace.  1963.  American Naturalist 97: 65-66).

          1.  Characteristics of the lt locus

               a.  lt locus carried on CyL (CurlyLobed) chromosome.  This chromosome carries the Curly (Cy) and Lobed (L)
                   genes that give visible phenotypic effects.
               b.  Two alleles: lt+ and lt
               c.  lt+/ lt+ individuals: viable, bright red eyes
               d.  lt+/ lt    individuals:  viable, orange-yellow eyes
               e.  lt / lt      individuals:  inviable
               f.  These characteristics indicate that the lt allele is a RECESSIVE LETHAL, i.e. when homozygous it is lethal,
                    but it has no apparent effects on viability when it is heterozygous (i.e. its lethal effects are recessive; note,
                    however, its effects on eye color are not recessive).

          2.   Genotype of an individual may be ascertained by mating to a CyL/Pm individual, where Pm is an alternate
                version of the CyL chromosome that lacks the Cy and L markers, but contains the Pm marker:

               a.  Cross CyL lt /Pm with individual to be scored and discard offspring that show the Pm  phenotype because
                    these will not be informative about eye color.  Examine eyecolor of remaining flies.
               b.  If individual to be scored is CyL lt+ /CyL lt+, then all surviving offspring will be   lt+/ lt+  and hence will have
                    bright red eyes.
               c.  By contrast, if individual to be scored is CyL lt+ /CyL lt, then half of surviving offspring will be  lt+/ lt+
                    and have bright red eyes, while half of the offspring will be  lt+/ lt and will have orange-yellow eyes.
               d.  Thus, if get half red, half orange-yellow eyed flies, the tested individual was lt+/ lt, while if get all red eyes, the
                    tested individual was  lt+/ lt+ .

          3.  Experimental procedure

               a.  Start a laboratory population consisting entirely of heterozygous lt+/ lt individuals.
               b.  Each generation, remove a sample of flies and test cross each to CyL lt /Pm and determine the genotype
                    of each fly of the sample by scoring eye color of its offspring.
               c.  From these test crosses, determine the gene frequency of lt in the population.
               d.  Transfer the remaining flies to a new population cage to found the next generation.
               e.  Repeat steps b-d over a number of generations.

          4.  Theoretical expectations from a recessive lethal
 
               a.  Wlt+/lt+ = Wlt+/lt = 1,  Wlt/lt = 0 .

               b.  Plugging these values into our predictive equation yields

                    p' = [p(p x Wlt/lt  +  q x Wlt+/lt )/(p2 x Wlt/lt  +  2pq x Wlt+/lt    + q2 x Wlt+/lt+ )]

                        =  [p(p x 0  +  q x 1)/(p2 x 0  +  2pq x 1  +   q2 x 1)] 

                        = p/(1-p)

          5.  Comparing theoretical expectations with observed gene frequencies.

               a.  Match is good (see Graph)

               b.  Conclude:  Predictive equation accurately and quantitatively describes the trajectory of gene frequency
                    change in this case.

     C.  Example: Drosophila pseudoobscura and inversion polymorphisms (T. Dobzhansky.  1954.  Proc. 9th Int. Cong.
           Genetics, in Caryologia, 435-449).

          1.  Natural history

              a.  D. pseudoobscura, a native of western North America, exhibits polymorphisms for chromosomal inversions.
              b.  There are two "alleles", ST and CH
 `            c.  The frequencies of these alleles undergo regular seasonal oscillations (see Graph), which can only be
                   accounted for by fluctuating natural selection.
              d.  This result implies that the three genotypes ST/ST, ST/CH and CH/CH differ in fitness, which should be
                   detectable and measurable under laboratory conditions.
              e.  Given these differences in fitness, it should be possible to use this system to test the quantitative accuracy
                   of the predictive equation for gene frequency change.

          2.  Measuring Fitness (Handout).

              a.  Dobzhansky estimate the relative fitnesses of ST/ST, ST/CH and CH/CH to be 0.90, 1, and 0.41 respectively.
              b.  These fitnesses imply that a polymorphism should be actively maintained in the laboratory environment.

          3.  Experiment

              a.  Established four replicate population cages with initial ST "allele" frequency of 0.2.
              b.  Allowed population to reproduce naturally for approximately 15 generations.
              c.  Approximately every two generations, removed a random sample of flies and determined their genotype,
                   from which the "allele" frequency could be calculated.

          4.  Results (see Graph)

              a.  Excellent match between trajectory from predictive equation and observed "allele" frequencies.
              b.  Evolution led to establishment of a stable polymorphism, as expected with heterozygote superiority.
 


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