Life Sciences at Indiana State University …

from Molecules to Ecosystems Testing Adaptive Models Of Sex Allocation

in a Polymorphic Species
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Tiffany Ko - Department of Electrical Engineering, Princeton University, Princeton, NJ 08544

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Marisa Korody, Elaina Tuttle, Rusty Gonser - Department of Life Sciences, Indiana State University, Terre Haute, IN 47802

Introduction Abstract Statistical Analysis

Fluorescence Intensity (a.u.)

*Calculated proportion of males in each clutch
The first theories concerning evolution and sex ratio were introduced by Darwin (1871) in *Arcsin square root transformation
his ground-breaking book The Descent of Man and Selection in Relation to Sex. One of his Sex ratio is one of the best reflections of evolution because it determines the *ANOVA
most prevalent theories is that a facultative, or changing, sex ratio is a direct product of natural outcome of reproduction, a key evolutionary process. Two hypotheses explaining *Year, pair type, hatch date
selection. the evolution of sex ratio, the Fisherian and the Trivers-Willard hypotheses, *Post hoc Tukey-Kramer HSD, α=0.05
were tested in the white-throated sparrow (Zonotrichia albicollis), a species * JMP 4.0.4 statistical analysis program
Two major classes of adaptive hypotheses were tested in this study: that exhibits a genetic polymorphism causing differences in plumage (white Fragment Length (bp)
Fisher’s Hypothesis or tan) that correlate with distinct behavioral strategies. These birds mate
* Homeostasis - sex ratio should be maintained at ideal equlibrium ratio of 0.5 dissasortatively, white almost exclusively with tan. Nestlings captured during the
- any biased sex ratio at birth will be in favor of the rarer sex 2000-2004 breeding seasons were sexed via analysis of the CHD gene. White
male/tan female pairs produced female-biased broods when chances of nesting Results
Trivers-Willard Hypothesis success was low. In contrast, tan male/white female pairs produced male- * Sex ratio of WxT clutches do not vary significantly
* Better conditioned parents should invest in the sex that has the best chance of biased broods more often, probably as a result of their increased expenditure in from year to year (F4,36=0.67, p=0.62)
producing the most grand-offspring. parental effort. The Trivers-Willard hypothesis is a better model as the facultative * Sex ratio of TxW clutches vary significantly
changes in sex ratio seem to be linked to differences in parental effort patterns from year to year (F4,49=3.61, p=0.01)
The White-Throated Sparrow exhibited by the two disassortative pair types. We suggest a combination of * Overall average sex ratio of the population did
(Zonotrichia albicollis) ecological and evolutionary theories best explains the evolution of sex ratio in not differ significantly from 0.5.
The white-throated sparrow , is a passerine which inhabits northeastern United States and this and other species.
Canada. They are a unique avian species in that there is a separate genetic polymorphism that
is connected with morphological and behavioral distinctions. These are caused by a pericentric
inversion on the second chromosome, 2m. A physical distinction occurs in plumages; without the
presence of the inversion (homozygous 2/2), the bird has a tan stripe on its head, but with the Methods Sample Collection
Approximately 50% of the world’s species of birds, including * TxW clutch ratios do not differ significantly in early
inversion (heterozygous 2/2m), the stripe is white.
the white-throated sparrow, are not sexually dimorphic. Blood and late clutches (F1,52=1.30, p=0.25)
White-throated sparrows mate samples were collected for sexing through molecular DNA * WxT clutches produce a significantly lower

disassortatively. Naturally, tan analysis. proportion of males in late clutches
females mate nearly exclusively - All blood sample collection was done by Tuttle and field (F1,38=4.96, p=0.03)
with white males and vice versa. teams.

* Cranberry Lake Biological Station, Aidirondack Mountains, NY

Tan White
*
*
Indentified by a serial number and band colors
This study encompasses data collected in 2000-2004. Discussion Fisher’s Hypothesis:
* Overall, not significantly different from 0.5
Parental Promiscuous
* 282 chicks were analyzed for a composite total of 92 clutches. * Clutch sex ratios of TxW≠WxT
The inversion causes variation in behavior where tan morphs tend to take up a more parental role Molecular Analysis Trivers-Willard Hypothesis:
in caring for offspring while whites are much more socially aggressive and sexually promiscuous. A DNA-based sex identification method was used utilizing two CHD (chromo-helicase-
White males often seek extra-pair copulations (EPCs) with neighboring females that are not their * The trends in clutch sex ratio of TxW pairs
DNA-binding) genes, CHD-W and CHD-Z, located on the avian sex chromosomes W and Z, and WxT pairs differ.
social mates. In an earlier study tan males were found not only to be more attractive than white respectively.
males, but also tan males and their white female mates (TxW) were found to nurture their offspring
White male/tan female parents showed evidence that they are able to facultatively shift the
and contribute parental investment much more than white male/tan female parents (WxT). 1) Blood was separated through centrifugation and red blood cells (RBCs) were stored. sex ratio of their clutch in response to environmental cues such as predators. The later breeding
~~~~ 2) DNA was extracted using the Promega DNA IQ System. season harbors many more predators and a greater risk of depredation in nests. Because TxW
The objective of this study was to test the Fisher and the Trivers-Willard hypotheses in white- 3) P8 and P2 primers were used to amplify the needed sequence through PCR. pairs are better parents, they need not shift their sex ratio; they are able to fend off predators.
throated sparrows. 4) PCR product loaded into the ABI Prism 310 Genetic Analyzer, capillary electrophoresis. On the other hand, with more predators, WxT pairs have a disadvantage as they are “poorer”
Predictions parents. Therefore, in order to increase their chances of having some grand-offspring, they
Fisher’s Hypothesis: produce more daughters, instead of more expensive sons. This strongly supports the Trivers-
Clutch sex ratios will be facultative in order to maintain the ideally stable Willard hypothesis. In additional support, the data also show that TxW parents facultatively shift
proportion of 0.5. Regardless of parent pair type, trends will be the same their sex ratio from year to year.
through out.
Future Directions
Trivers-Willard Hypothesis: In the future we hope to look further into parental investments in nestling sex ratio, since
Clutch sex ratios will differ between TxW parents and WxT parents. parental condition and consequent amount of investment is very distinguished throughout the
Trends should correlated to difference in pair type based upon parental data. Another interesting aspect to look at would be the effects of laying order on the sex of the
effort. nestlings. We hope to continue this study in order to further examine cyclic patterns.

Selected References: - Darwin, C. 1871. The Descent of Man and Selection in Relation to Sex. London: John Murray. Acknowledgements: - Research Funding: NSF Grant DEB-0217963
- Ellegren, H. 1996. First gene on the avian W chromosome (CHD) provides a tag for universal sexing of non-ratite birds. Proceedings of the Royal - Facilities: Department of Life Sciences, Indiana State University
Society of London B 263: 1635-1641. - Travel Funding: School of Engineering and Applied Science,
- Fisher, R. A. 1930. The Genetical Theory of Natural Selection. Oxford: Clarendon. Princeton University
- Trivers R. L. and Willard D. E. 1973. Natural selection of the parental ability to vary the sex ratio of offspring. Science 179: 90-92. - Tuttle lab group and field research team