In his book On the Origin of Species, Charles Darwin (1859) proposed that all organisms on earth evolved from a single proto-organism by descent with modification. He also proposed that the primary force of evolution is natural selection. Most biologists accepted the first proposition almost immediately, but the second proposal was controversial and was criticized by such prominent biologists as Thomas Huxley, Moritz Wagner, and William Bateson.
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However, Morgan's mutation-selection theory or mutationism gradually became unpopular as the neo-Darwinism advocated by Fisher (1930), Wright (1931), Haldane (1932), Dobzhansky (1937, 1951), and others gained support from many investigators in the 1940s. In neo-Darwinism, natural selection is assumed to play a much more important role than mutation, sometimes creating new characters in the presence of genetic recombination. Although there are several reasons for this change (see Nei 1987, chap. 14), two are particularly important. First, most geneticists at that time believed that the amount of genetic variability contained in natural populations is so large that any genetic change can occur by natural selection without waiting for new mutations. Second, mathematical geneticists showed that the gene frequency change by mutation is much smaller than the change by natural selection. Neo-Darwinism reached its pinnacle in the 1950s and 1960s, and at this time almost every morphological or physiological character was thought to have evolved by natural selection (Dobzhansky 1951; Mayr 1963).
This situation again started to change as molecular data on evolution accumulated in the 1960s. Studying the GC content of the genomes of various organisms, early molecular evolutionists such as Sueoka (1962) and Freese (1962) indicated the possibility that the basic process of evolution at the nucleotide level is determined by mutation. Comparative study of amino acid sequences of hemoglobins, cytochrome c, and fibrinopeptides from various organisms also suggested that most amino acid substitutions in a protein do not change the protein function appreciably and are therefore selectively neutral or nearly neutral, as mentioned below. However, this interpretation was immediately challenged by eminent neo-Darwinians such as Simpson (1964) and Mayr (1965), and this initiated a heated controversy over selectionism versus neutralism.
An even more intense controversy on this subject was generated when protein electrophoresis revealed that the extent of genetic variation within populations is much higher than previously thought. At that time, most evolutionists believed that the high degree of genetic variation can be maintained only by some form of balancing selection (Mayr 1963; Ford 1964). However, a number of authors argued that this variation can also be explained by neutral mutations. From the beginning of the 1980s, the study of molecular evolution was conducted mainly at the DNA level, but the controversy is still continuing. This long-standing controversy over selectionism versus neutralism indicates that understanding of the mechanism of evolution is fundamental in biology and that the resolution of the problem is extremely complicated. However, some of the controversies were caused by misconceptions of the problems, misinterpretations of empirical observations, faulty statistical analysis, and others.
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One important progress in the study of evolution in this era was that the neutral theory generated many predictions about the allele frequency distribution within populations and the relationships between genetic variation within and between species, so that one could test the applicability of the neutral theory to actual data by using various statistical methods. In other words, we could use the neutral theory as a null hypothesis for studying molecular evolution (Kimura and Ohta 1971). This type of statistical study of evolution was almost never done before the neutral theory was proposed. The results of these studies are summarized by Lewontin (1974), Nei (1975, 1987), Wills (1981), and Kimura (1983). Although the interpretations of the results by these and other authors were not necessarily the same, it was clear by the early 1980s that the extent and pattern of protein polymorphism within species roughly agree with what would be expected from neutral theory (e.g., Yamazaki and Maruyama 1972; Nei, Fuerst, and Chakraborty 1976; Chakraborty, Fuerst, and Nei 1978; Skibinski and Ward 1981; Nei and Graur 1984).
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In 1981 even stronger support of the neutral theory came from studies of the evolutionary rate of pseudogenes (Li, Gojobori, and Nei 1981; Miyata and Yasunaga 1981). Pseudogenes are nonfunctional genes because they contain nonsense or frameshift mutations, and therefore according to the neutral theory the rate of nucleotide substitution is expected to be high and is more or less equal to the total mutation rate. By contrast, if neo-Darwinism is right, one would expect that virtually no nucleotide substitution occurs because they are functionless and there is no way for positive selection to operate. When Li, Gojobori, and Nei (1981) studied the rate of nucleotide substitution for three globin pseudogenes from the human, mouse, and rabbit, the average rate was about 5 × 10−9 per site per year and was much higher than the rates for the first, second, and third codon position rates of the functional genes (table 1). An independent study by Miyata and Yasunaga (1981) about a mouse globin pseudogene also showed a high rate of substitution. These observations clearly supported the neutral theory rather than the neo-Darwinian evolution.
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At present, we do not know the relative importance of chance and natural selection. However, the chance factor working here is a new feature of random evolution, which is qualitatively different from the neutral evolution of genes by random genetic drift. This view of evolution is based on a large amount of molecular data, and in this sense it is different from Morgan's mutationism, which was largely speculative. For this reason I have called it neomutationism (Nei 1983, 1984) or the neoclassical theory of evolution (Nei 1987, chap. 14). Whatever it is called, however, recent molecular data supports the theory of mutation-driven evolution rather than neo-Darwinism.