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Beadle and Tatum irradiated the mold with X-rays to induce changes to a sequence of nucleic acids, called mutations . They mated the irradiated mold spores and attempted to grow them on both a complete medium and a minimal medium. They looked for mutants that grew on a complete medium, supplemented with vitamins and amino acids, but did not grow on the minimal medium lacking these supplements. Such molds theoretically contained mutations in the genes that encoded biosynthetic pathways. Upon finding such mutants, they systematically tested each to determine which vitamin or amino acid it was unable to produce ( [link] ) and published this work in 1941. G.W. Beadle, E.L. Tatum. “Genetic Control of Biochemical Reactions in Neurospora.” Proceedings of the National Academy of Sciences 27 no. 11 (1941):499–506.

Diagram of Beadle and Tatum’s experiment. Wild type spores are exposed to X-rays to form mutagenized spores. The wild type and mutagenized spores are then crossed. The mutants are then grown on complete (with amino acids) and minimal media (without amino acids). Mutants that grow only on complete medium are identified. Spores that cannot grow on a minimal medium are tested on a minimal medium with a single amino acid added. Spores that grow inonly one of these tubes have a mutation in the pathway that produces that particular amino acid.
Beadle and Tatum ’s experiment involved the mating of irradiated and nonirradiated mold spores. These spores were grown on both complete medium and a minimal medium to determine which amino acid or vitamin the mutant was unable to produce on its own.

Subsequent work by Beadle, Tatum, and colleagues showed that they could isolate different classes of mutants that required a particular supplement, like the amino acid arginine ( [link] ). With some knowledge of the arginine biosynthesis pathway, they identified three classes of arginine mutants by supplementing the minimal medium with intermediates (citrulline or ornithine) in the pathway. The three mutants differed in their abilities to grow in each of the media, which led the group of scientists to propose, in 1945, that each type of mutant had a defect in a different gene in the arginine biosynthesis pathway. This led to the so-called one gene–one enzyme hypothesis , which suggested that each gene encodes one enzyme.

Subsequent knowledge about the processes of transcription and translation led scientists to revise this to the “one gene–one polypeptide” hypothesis. Although there are some genes that do not encode polypeptides (but rather encode for transfer RNAs [tRNAs] or ribosomal RNAs [rRNAs], which we will discuss later), the one gene–one enzyme hypothesis is true in many cases, especially in microbes. Beadle and Tatum’s discovery of the link between genes and corresponding characteristics earned them the 1958 Nobel Prize in Physiology and Medicine and has since become the basis for modern molecular genetics.

The table at the top is labeled Beadle and Tatum Experiments and shows the growth pattern of 4 different spores. The wild type spore grew on minimal medium (MM), MM + Ornithing, MM + Citruline and MM + Arginine. Mutant 1 did not grow on MM but did grow on MM + Ornithing, MM + Citruline and MM + Arginine. Mutant 2 did not grow on MM or  MM + Ornithing but did grow on  MM + Citruline and MM + Arginine. Mutant 3 did not grow on MM, MM + Ornithing, or MM + Citruline but did grow on MM + Arginine.  Underneath the table is a diagram that explains these results. The top diagram shows a pathway where gene 1 produces enzyme 1 and enzyme 1 produces ornithine. Gene 2 produces enzyme 2 which converts ornithine to citruline. Gene 3 produces enzyme 3 which converts citruline to arginine. Mutant 1 had a mutation in gene 1 that destroyed the function of enzyme 1, so one of the amino acids are produced. Mutant 2 had a mutation in gene 2 that destroyed the function of enzyme 2. So, Ornithine is still produced but citruline and arginine are not. Mutant 3 had a mutation in gene 3 that destroyed the function of enzyme 3. So, ornithine and citruline are produced but arginine is not.
Three classes of arginine mutants were identified, each differing in their ability to grow in the presence of intermediates in the arginine biosynthesis pathway. From this, Beadle and Tatum concluded that each mutant was defective in a different gene encoding a different enzyme in the arginine biosynthesis pathway, leading to them to their one gene–one enzyme hypothesis.
  • What organism did Morgan and his colleagues use to develop the Chromosomal Theory of Inheritance? What traits did they track?
  • What did Hämmerling prove with his experiments on Acetabularia ?

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Source:  OpenStax, Microbiology. OpenStax CNX. Nov 01, 2016 Download for free at http://cnx.org/content/col12087/1.4
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