Mendel and Inheritance MUPGRET Workshop December 4, 2004

Genetic variation In the beginning geneticists studied differences they could see in plants. These differences are called morphological differences. Individual variants are referred to as phenotypes, ex. tall vs. short plants or red vs. white flowers.

Trait A broad term encompassing a distribution of phenotypic variation. Example: Trait: Disease resistance Phenotype: resistant vs. susceptible Morphological differences associated with the trait might include fungal infection, fungal growth, sporulation, etc.

Mendel Monk at the St. Thomas monastery in the Czech Republic. Performed several experiments between 1856 and 1863 that were the basis for what we know about heredity today. Used garden peas for his research. Published his work in 1866.

Mendel Results are remarkably accurate and some have said they were too good to be unbiased. His papers were largely ignored for more than 30 years until other researchers appreciated its significance.

Garden Pea Pisum sativum Diploid Differed in seed shape, seed color, flower color, pod shape, plant height, etc. Each phenotype Mendel studied was controlled by a single gene.

Terms Wild-type is the phenotype that would normally be expected. Mutant is the phenotype that deviates from the norm, is unexpected but heritable. This definition does not imply that all mutants are bad; in fact, many beneficial mutations have been selected by plant breeders.

Advantages of plants Can make controlled hybrids. Less costly and time consuming to maintain than animals. Can store their seed for long periods of time. One plant can produce tens to hundreds of progeny.

Advantages of plants Can make inbreds in many plant species without severe effects that are typically seen in animals. Generation time is often much less than for animals. Fast plants (Brassica sp.) Arabidopsis

Allele One of two to many alternative forms of the same gene (eg., round allele vs. wrinkled allele; yellow vs. green). Alleles have different DNA sequences that cause the different appearances we see.

Principle of Segregation (Mendel’s First Law) Parental Lines Round Wrinkled X All round F 1 progeny Self-pollinate Round 5474 Wrinkled Round : 1 Wrinkled

Important Observations F 1 progeny are heterozygous but express only one phenotype, the dominant one. In the F 2 generation plants with both phenotypes are observed  some plants have recovered the recessive phenotype. In the F 2 generation there are approximately three times as many of one phenotype as the other.

Mendel’s Results Parent Cross F 1 Phenotype F 2 data Round x wrinkled Round 5474 : 1850 Yellow x green Yellow 6022 : 2001 Purple x white Purple 705 : 224 Inflated x constricted pod Inflated 882 : 299 Green x yellow pod Green 428 : 152 Axial x terminal flower Axial 651 : 207 Long x short stem Long 787 : 277

3 : 1 Ratio The 3 : 1 ratio is the key to interpreting Mendel’s data and the foundation for the the principle of segregation.

The Principle of Segregation Genes come in pairs and each cell has two copies. Each pair of genes can be identical (homozygous) or different (heterozygous). Each reproductive cell (gamete) contains only one copy of the gene.

Mendel’s Principle of Segregation In the formation of gametes, the paired hereditary determinants separate (segregate) in such a way that each gamete is equally likely to contain either member of the pair. One male and one female gamete combine to generate a new individual with two copies of the gene.

Round vs. Wrinkled Parental Lines Round Wrinkled X All round F 1 progeny Self-pollinate Round 5474 Wrinkled Round : 1 Wrinkled

Round vs. wrinkled The SBEI causes the round vs. wrinkled phenotype. SBEI = starch-branching enzyme Wrinkled peas result from absence of the branched form of starch called amylopectin. When dried round peas shrink uniformly and wrinkled do not.

Round vs. wrinkled The non-mutant or wild-type round allele is designated W. The mutant, wrinkled allele is designated w. Seeds that are Ww have half the SBEI of wild-type WW seeds but this is enough to make the seeds shrink uniformly. W is dominant over w.

Round vs. wrinkled An extra DNA sequence is present in the wrinkled allele that produces a non- functional SBEI and blocks the starch synthesis pathway at this step resulting in a lack of amylopectin.

A Molecular View ParentsF1F1 F 2 Progeny WW ww Ww¼WW ¼Ww ¼wW ¼ww 1: 2 : 1 Genotype = 3: 1 Phenotype

Dihybrid crosses reveal Mendel’s law of independent assortment A dihybrid is an individual that is heterozygous at two genes Mendel designed experiments to determine if two genes segregate independently of one another in dihybrids First constructed true-breeding lines for both traits, crossed them to produce dihybrid offspring, and examined the F2 for parental or recombinant types (new combinations not present in the parents).

Mendel and two genes x Round Yellow Wrinkled Green All F1 Round, Yellow Round Yellow 315 Round Green 108 Wrinkled Yellow 101 Wrinkled Green 32

Dihybrid cross produces a predictable ratio of phenotypes genotype phenotype number phenotypic ratio Parent Y_R_ 315 9/16 Recombinant yyR_ 108 3/16 Recombinant Y_rr 101 3/16 Parent yyrr 32 1/16 Ratio of yellow (dominant) to green (recessive)=3:1 (12:4) Ratio of round (dominant) to wrinkled (recessive)=3:1 (12:4)

Ratio for a cross with 2 genes Crosses with two genes are called dihybrid. Dihybrid crosses have genetic ratios of 9:3:3:1.

Mendel and two genes Round Yellow 315 Round Green 108 Wrinkled Yellow 101 Wrinkled Green 32 Round = 423 Wrinkled = 133 Yellow = 416 Green = 140 Each gene has a 3 : 1 ratio.

Summary of Mendel's work Inheritance is particulate - not blending There are two copies of each trait in a germ cell Gametes contain one copy of the trait Alleles (different forms of the trait) segregate randomly Alleles are dominant or recessive - thus the difference between genotype and phenotype Different traits assort independently

Rules of Probability Independent events - probability of two events occurring together What is the probability that both A and B will occur? Solution = determine probability of each and multiply them together. Mutually exclusive events - probability of one or another event occurring. What is the probability of A or B occurring? Solution = determine the probability of each and add them together.

PRODUCT RULE From James Birchler

Mutually exclusive ways! SUM RULE From James Birchler

Dominant All Dominant All Recessive Recessive From James Birchler

Punnett Square method = 16 possible gamete combinations for each parent Thus, a 16  16 Punnett Square with 256 genotypes That’s one big Punnett Square! Loci (Genes) Assort Independently - So we can look at each locus independently to get the answer. Branch diagrams are also convenient tools

Extensions to Mendel Complexities in relating genotype to phenotype

Some Extensions to Mendel’s Analysis Single-gene inheritance In which pairs of alleles show deviations from complete dominance and recessiveness In which different forms of the gene are not limited to two alleles Where one gene may determine more than one trait Multifactorial inheritance in which the phenotype arises from the interaction of one or more genes with the environment, chance, and each other

Dominance is not always complete Crosses between true-breeding strains can produce hybrids with phenotypes different from both parents Incomplete dominance F1 hybrids that differ from both parents express an intermediate phenotype. Neither allele is dominant or recessive to the other Phenotypic ratios are same as genotypic ratios Codominance F1hybrids express phenotype of both parents equally Phenotypic ratios are same as genotypic ratios

Summary of dominance relationships

Incomplete dominance in snapdragons

Codominant blood group alleles

A gene can have more than two alleles Genes may have multiple alleles that segregate in populations Alleles may be unique to every pair of alleles in an individual

Mendelian inheritance in humans Many traits in humans are due to the interaction of multiple genes and do not show a simple Mendelian pattern of inheritance. A few traits represent single-genes. Examples include sickle-cell anemia, cystic fibrosis, Tay-Sachs disease, and Huntington’s disease Because we can not do breeding experiments on humans, we must use pedigrees to study inheritance Pedigrees are an orderly diagram of relevant genetic features extending through multiple generations Pedigrees help us infer if a trait is from a single gene and if the trait is dominant or recessive

Anatomy of a pedigree

A vertical pattern of inheritance indicates a rare dominant trait Huntington’s disease: A rare dominant trait

A horizontal pattern of inheritance indicates a rare recessive trait Cystic fibrosis: a recessive condition

Mutation Found in 'Muscle Man' Toddler Somewhere in Germany is a baby Superman, born in Berlin with bulging arm and leg muscles. Not yet 5, he can hold seven-pound weights with arms extended, something many adults cannot do. He has muscles twice the size of other kids his age and half their body fat. DNA testing showed why: The boy has a genetic mutation that boosts muscle growth (by blocking myostatin). AP news 23 June 2004 Researchers would not disclose the German boy's identity but said he was born to a somewhat muscular mother, a 24-year-old former professional sprinter. Her brother and three other close male relatives all were unusually strong. In the mother, one copy of the gene is mutated and the other is normal; the boy has two mutated copies. One almost definitely came from his father.

Double Muscling in Cattle