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Diana Miculescu
Introduction
Through many experiments with pea plants, Gregor Mendel observed patterns in
breeding parents as well as hybrids and dihybrids. From his results, Mendel observed
3:1 ratios and termed the trait observed in the larger proportion as the dominant
characteristic and the smaller proportion as the recessive characteristic (Yoshio, 2013).
His results led him to propose theories about the method in which the progeny inherits
traits from the parental generation. These theories would later be called the laws of
segregation and independent assortment. The law of segregation states that the
variations of a trait, or alleles, segregate during meiosis so that the offspring obtains a
single allele from each parent (MacLeod, Tweedie, Tait, et al., 2015).
After performing crosses of plants that differed in two traits, called dihybrid
crosses, a modification of the 3:1 ratio was observed. For example, in a dihybrid cross
of a plant the parental generation (P0) contains one parent that is homozygous
dominant for both traits while the other parent is homozygous recessive for both traits
(or one parent homozygous dominant for one trait and recessive for the other). The
cross of the dihybrid leads to a hybrid generation (F1) in which offspring inherit one
dominant allele and one recessive allele for each trait. Once the hybrid generation self-
fertilizes, the offspring (F2 generation) results in a phenotypic ratio of 9:3:3:1. The “9”
part of the ratio refers to the plants that display the dominant phenotypes for both traits.
The “3” part of the ratio refers to the plants that display one of the dominant phenotypes
for one gene and the recessive phenotype for the other gene. Lastly, the “1” part of the
ratio refers to the plants that display both recessive phenotypes for both genes. This
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9:3:3:1 ratio is characteristic of dihybrid crosses. The findings of the dihybrid crosses led
ensures the transmission of traits to offspring occur independently of one another, now
Mendel’s findings in his experiments with pea plants led to the development of
theories of inheritance which would later be combined with the chromosomal theory of
inheritance (Vicedo, 1990). Mendel’s accuracy in his proposals is significant given that
discovered.
observe the Mendelian laws in action, we will use the dihybrid offspring of the rapid-
cycling Brassica rapa that are homozygous for the two traits of stem and leaf color. The
purple color (dominant) in the stem of the plants is caused by anthocyanin which
controlled by the gene anl (Burdzinski & Wendell, 2007). Plants inheriting anl/anl for
suppressed (Burdzinski & Wendell, 2007). The leaf color of light green is controlled by
the gene ygr. Plants inheriting ygr/ygr for leaf color will be light green (recessive), while
those inheriting YGR/ygr or YGY/YGR will be dark green (dominant) in leaf color
proportions with a chi-square test. This will help determine if the deviations of the
observed count for each phenotype from the expected ratio of a dihybrid cross (9:3:3:1)
are significantly different and if the deviations were due to chance. If the possibility of
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chance occurrence in percentage for 3 degrees of freedom lies above 0.05 (upper-tail),
then there is a high chance that the deviations of observed ratios from expected ratios
are due to chance. However, if under 0.05, then the deviations are significantly different
and there is little chance that the deviations in observed ratios are due to chance.
Reservoirs were filled with water, blue algae-control squares were placed in the
water and a saturated water mat was placed on the lid. One wick was placed in each
cell of a quad and each cell was slightly filled with moist soil. Two to three fertilizer
pellets were placed in each cell, more moistened soil was placed in each cell, and then
shallow depressions were made in the soil. Two to three seeds of the offspring (F1
generation: purple stem, dark green leaf anl/ANL, YGR/ygr) of the parent generation (P1
generation: non-purple stem, dark green leaf anl/anl, YGY/YGY and P2 generation:
purple stem, light green leaf ANL/ANL, ygr/ygr) were placed in each depression and soil
was sprinkled on top to cover the seeds. Each cell was watered until the wicks were
saturated. The quads were placed under the lighting system provided in the laboratory.
The plants were watered as needed and the reservoirs were refilled as needed for 2
weeks.
After 2 weeks, pollen was transferred back and forth from the F1 generation
Twenty days after the cross pollination, the plants were removed from the
reservoirs and allowed to dry out. Once the plants dried out, the seeds (F2 generation)
were harvested from the seeds pods. The harvested seeds were placed on a wet filter
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paper in a petri dish and placed under the lighting system for a week to sprout (until
Results
Of the seeds in the petri dish, the results of the seeds with observable and
distinguishable stem and leaf colors were recorded (regardless if they were alive or
partly dried out). The results observed for Group 1 were 19 plants with purple stems and
dark green leaves, 6 plants with purple stems and light green leaves, 5 plants with non-
purple stems and dark green leaves, and 2 plants with non-purple stems and light green
leaves (Figure 1, Table 1). The class results were combined to yield 26 plants with
purple stems and dark green leaves, 9 plants with purple stems and light green leaves,
14 plants with non-purple stems and dark green leaves, and 4 plants with non-purple
stems and light green leaves (Table 2). The 2 value was calculated to be 2.3793 with 3
Table 1. Phenotypic and genotypic results of stem and leaf colors of Brassica rapa (F2
generation) from Figure 1
Phenotype purple stems, dark purple stems, light non-purple stems, non-purple stems,
green leaves green leaves dark green leaves light green leaves
Genotype* -/ANL, YGR/- -/ANL, ygr/ygr anl/anl, YGR/- anl/anl, ygr/ygr
Group 1 19 6 5 2
Class combined 26 9 14 4
*(-) in genotype indicates either dominant or recessive allele may be present with no phenotypic
difference
approximately 30 were expected to have purple stems and green leaves, 10 to have
purple steams and light green leaves, 10 to have non-purple stems and dark green
leaves, and 3 to have non-purple stems and light green leaves (Table 2). Of the
observed (class combined), the number of non-purple stems and dark green leaves was
considerably more than expected and of the purple stems and dark green leaves less
plants were observed than expected (Table 2). All plants that were phenotypically non-
purple stemmed must have a genotype of anl/anl. All plants displaying some pigment of
purple must at least one dominant allele of ANL. All plants that were phenotypically light
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green leafed must have a genotype of ygr/ygr, and all plants displaying dark green
chi-square value was calculated (Table 2). With a 2 value calculated to be 2.3793 with
3 degrees of freedom, the probability that the deviation of the observed values from the
expected values was a chance occurrence is .5026. A probability greater than 0.05 is
not considered statistically significant; therefore, the deviation of the observed values
from the expected values must have occurred by chance. For a total of only 53 plants,
the 2 value calculated may not be reliable. The method in the characteristics of the
plants in the petri dish were interpreted by other groups are unknown. It may be that not
all plants were recorded, if say, they were not “alive” or not. This could be reason why
the observed data did not turn out as expected of the 9:3:3:1 ratio of a dihybrid cross.
While the 2 value calculated may not be reliable due to the low count of results,
these results do support that specific traits could be passed from one generation to the
next without phenotypically displaying that trait. This is important beyond the application
of crossing plants, since some traits that cause disorders can be carried by a parent and
passed on to the offspring without any knowledge of carrying that trait. The expansion of
knowledge in the field of genetics has greatly increased awareness of such genetic
disorders. And with the awareness of such genetic disorders, methods to decrease the
References
Burdzinski C., and Wendell D.. Mapping the anthocyaninless (anl) locus in rapid-cycling
Brassica rapa (RBr) to linkage group R9. BMC Genet. 2007;8:64. Accessed at
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC2048511/
Miko, I.. Gregor Mendel and the principles of inheritance. Nature Education
mendel-and-the-principles-of-inheritance-593
Slankster, E., Chase J., Jones L., and Wendell D.. DNA-based genetic markers for rapid
cycling Brassica rapa (Fast Plants type) designed for the teaching laboratory.
http://www.ncbi.nlm.nih.gov/pmc/articles/PMC3365483/
Yoshio T. Gregor Mendel and the Seven Genes (2). Interdisciplinary Bio Central. March
2015.
T. brucei crosses: Proof that the genetic system is Mendelian and involves