Mate Quality Affects Offspring Sex Ratio in Blue Tits Erik Svensson; Jan-Ake Nil Proceedings: Biological Sciences, Vol. 263, No. 1368 (Mar. 22, 1996), 357-361. Stable URL: bttp//links jstor.org/sici?sic!=0962-8452%28 1996032204 292630434 1368%3C357%3AMQAOSR%3E2,0,.CO%3B2-K Proceedings: Biological Sciences is currently published by The Royal Society ‘Your use of the ISTOR archive indicates your acceptance of JSTOR’s Terms and Conditions of Use, available at hhup:/www.jstororg/about/terms.hml. JSTOR’s Terms and Conditions of Use provides, in part, that unless you have obtained prior permission, you may not download an entire issue of a journal or multiple copies of articles, and you may use content in the JSTOR archive only for your personal, non-commercial use. Please contact the publisher regarding any further use of this work. Publisher contact information may be obtained at hup:/www jstor-org/journals/rst.huml Each copy of any part of a JSTOR transmission must contain the same copyright notice that appears on the sereen or printed page of such transmission, STOR is an independent not-for-profit organization dedicated to creating and preserving a digital archive of scholarly journals, For more information regarding JSTOR, please contact support @jstor.org. bupslwww jstor.org/ ei Jun 18 04:55:11 2004Mate quality affects offspring sex ERIK SVENSSON anv JAN-AKE NILS! ratio in blue tits SON Department of Animal Ecology, Ecology Building, 5-223 62 Lund, Sweden SUMMARY Questions concerning the occurrence of aday in evolutionary biology, but evidence from birds is searce adaptively determine the sex of their young because of the sex ratio manipulation have a long research tradition Ithas been suggested that birds are unable to constraints of their sex determination systems, Here we report results from a study of variation in offspring sex ratio in the blue tit, Paras eaerulews, a small passerine bird, Our data suggest that females are able to manipulate their primary sex ratios away from the equal sex ratio predicted from simple chromosomal segregation at meiosis. Contrary to the situation in some other birds, offspring sex ratio variation in this species was not influenced by timing of breeding (or brood size. We show that females mated to males with high survival prospects bias the hatching sex ratio of their broods in favour of sons. Although it has previously been proposed that females should produce an excess of sons when mated to high-quality males, our data provide the first evidence that the phenotypic quality of fathers contributes to the sex ratio variation in a natural population. 1, INTRODUCTION Sex ratio evolution has attracted much interest among. evolutionary biologists during the past decades (Fisher 1930; Charnov 1982), Studies of animals with chromo- somal sex determination systems are_ particularly interesting because they highlight the interplay be- tween genetic constraints and evolutionary adaptation (Williams 1979; Bull & Charnov 1988). Birds, like most other vertebrates, have chromosomal sex de- termination systems, which led Williams (1979) 10 suggest that sex ratio variation among outcrossed vertebrates is mainly a result of automatic, mendelian segregation of sex chromosomes at meiosis, with litle fr no scope for adaptive parental manipulation of sex ratios (see also Fiala 1981; Harmsen & Cooke 1983; but see Charnov 1982 for'a critique). During recent years, however, there has been an accumulation of ‘empirical evidence that animals with chromosomal sex. determination systems are actually capable of adjusting their offspring sex ratios in an adaptive way (for examples, see Clutton-Brock ef al. 1984; Bortolotti 1986; Madsen & Shine 1992) Many factors have been shown to affect sex ratios in nature, including the reproctuctive value of sons and daughters (Fisher 1930), local mate competition (Hamilton 1967; Madsen & Shine 1992), timing of breeding within the season (Dijkstra ¢ al. 1990; Zijlstra tal, 1992) and maternal body condition (Trivers & Willard 1973; Clutton-Brock etal. 1984). So far, many studies on sex ratio variation in birds and mammals have involved species with strong sexual size di- morphism and small brood or litter sizes, like pol- yeynous mammals and birds of prey (for examples, see Newton & Marquiss 1979; Clutton-Brock tal. 1984; Bortolotti 1986; Bedinarz & Hayden 1991). Here we report the result from a study on sex ratio variation in the blue tit Parus caeralews, a small passerine bird. This Pro. R. Sue. Land (1996) 269, 357-861 357 Printed i Great Brita bird differs from many of the previously studied species in that it is sexually monomorphic and lays large clutches. In such a species, one would expect females to adjust the sex ratio in relation to sex-specific fitness benefits of sons and daughters, rather than in relation to sex-specific costs arising from a high level of sexual size dimorphism, We investigated whether females changed the sex ratios oftheir broods depending on time in the breeding season, as has recently been shown for some other bird species (Dijkstra et al, 1990; Olsen & Cockburn 1991 Zijlstra eal. 1992). In many territorial birds, the finess of juvenile males is more negatively affected by date of hatching than the fitness of juvenile females (Smith & Areese 1989; Nilsson 1989; Dijkstra et al. 1990) Consequently, females might be expected to bias the sex ratio of their offipring towards males in carly broods and females in late broods (Dijkstra eal. 1990) We experimentally tested this hypothesis by delaying breeding for some pairs, and investigated whether experimental females produced more female-biased broods than early control females. It has previously also been suggested that mate properties might influence the sex ratio (Burley 1981). Consequently, we also investigated whether parental characteristies (age and survival prospects) influenced brood sex ratios in this species, 2. METHODS (a) Study site, general methods and experiments We studied a nestbox-breeding blue tit population outside Lund, southern Sweden during the breeding seasons of 1995, and 1994. The blue tit isa small (about 11g), hole-breeding passerine bird in which both parents feed the young. ‘The ‘weight difference between males and female is about 0.5 (i about 4% size dimorphism, data from breeding season of 1993). Clutch sizes range from 6 to 19 eggs. Blue tits are (© 1096 The Royal Society358. E, Svensson and J.-A. Nilsson Sex ratios in blue tits socially monogamous in our population (only three recorded cases of polygyny out of more than 500 breeding attempts luring 1992-1995), accordingly we clasified the male eaught uring nestling feeding of a brood as the females’ mate, We ‘timated overwinter survival of adults through ringing and subsequent recaptures in the breeding season of 1994. If an adult was replaced by another bied, and it was not found breeding elsewhere in the study area, it was considered to be dead, Bach year we capture more than 95% of all breeders during nestling feeding and adult dispersal distances sith the study area are very short (B, Svenswon, unpublished data), 40 our estimate is probably very close o true adult survival, Because of the failure to capture one or both parents in one or both ofthe breeding seasons, and hecause ‘of some cases of brood desertion and or predation prior to hatching, the sample sizes in thie study varies between different analyses. To experimentally manipulate the timing of breeding, we removed clutches of experimental pairs one day alter they were completed, which revulted in most of the paity laying replacement clutches, Experimental pairs were chosen randomly throughout the egslaying period to avoid any bias in parental and territory quality. In total, we studied 21 frst, broods and 20 experimentally delayed broods. (b) Blood sampling and flow cytometric analyses During the breeding season of 1993, we took blood samples from 389 nestlings in 41 broods when the young were 9-13= days-old. OF these broods, 34 had suffered no. previous nestling mortality and in each of the remaining broods one ‘young had died before blood sampling. Five broods contained fone unhatched egg each, but only one of these eggs showed jgns of embryonic development. Excluding thie brood from the data set does not affect the results from our analyses Blood samples were taken by tarsus or wing vein puncture ‘The blood (about 15p from each nesting) was suspended in 4 Iysit.staining trisodiumeitrate butler and froze 10 —20 prior to analysis “To determine the sex of the nestlings, flow cytometric analyses ofthe blood samples were done by B. Baldetorp and M. Ferno at the Department of Oncology, Lund University, according to the methods described by Nakamura st al (1990) and De Vita er al. (1994). Briefly, low cytometry ‘quantifies the nuclear DNA-content on a per-cell basis. As Females ate the heterogametic sex in most bird species (Charnov 1982), they have a lower amount of nuclear DNA in their erythtoeytes than have males, a difference which is Possible to quantity spectophotometrically through flow eytometry, For calibration ofthe instrument, reference cells from rainbow trout Oncorynchas mykic and blue tts of known, sex were used. "To determine the borderline between males andl females and to evaluate the accuracy of flow eytometry as a method for sex determination of blue tts, we employed two different methods, one which assumes no prior knowledge ofthe sex of| the individuals (the EM-algorithm) and one which uses a sample of individuals of known sex (see below). First, we fied two normal distributions to the DNA-values from all nestlings, using an EMcalgorithns (Dempster eal. 1977). The ENLalgorithm was used to find maximum likelihood esti= mates of the parameters ina mixture of two Gaussian distributions (sce Dempster al, 1977 and Titeringson 1992 for further details). The intersection point for the two listibutions were chosen as the borderline for classification, fof males and females, The two DNA-value distributions were clearly separated ffom each other, and over 97% of all neslings were outside the overlapping zone, indicating that the accuracy of flow eytometry as a sexing method for blue Proc R Sec, Lad B (196) tits was very high. Second, we also compared the DNA- values of males and females that we identified by plumage characters during thei first winter (A= 37 birds, 28 males ‘and nine females). The repeatability value between diferent measurement sesions for these individuals was 86% and the DNAwvalue distributions were clearly separated. between males and females (99.99%, confidence limit for DNA-ratios| between rainbow trout; blue tt: males, 2.019-2.035; females, 2.043-2.075). The borderlines for sex clasifications de- {ermined from the EM-algorithm and fom the known-sex individuals were similar, and all ofoue results and conchsions| remain unaffected irrespective of the method used, (c) Statistics To achieve normality and be able to perform parametric statistical tess, we transformed brood sex ratios into arcsiny/t-values’ before analysis (Sokal & Rohlf 1995) ‘Throughout this paper, we define brood sex ratio as the proportion of males in a brood. Likelihood rato tess were tused when comparing observed and expected brood sex :atis ftom the binomial distribution, hence the testquantile is the summed ratios between the maximum of the likelihood function under H, and the maximum under Hy (Sokal & Rohif 1995). All probability values in this paper reer to tw tailed tests, All statistical tests were performed with SYSTAT (1992) statistical package, 3. RESULTS AND DISCUSSION Brood sex ratios varied between 0.21 and 1.0 (mean 57, sd.=0.20), and the observed variation differed significantly from the variation expected! from a binomial distribution (figure 1). This suggests that the variation between broods do not automatically fallow from simple chromosomal segregation at meiosis (Williams 1979), but that females in some way can control the sex of the individual eggs. In birds, a variety of mechanisms have been proposed, including maternal control over chromosomal segregation, sex- specific destruction of embryos or differential growth of male and female follicles (Godfray & Harvey 1986). Recent work suggests that ultra-sellish genetic elements in combination with autosomal modifier loci could explain biased sex ratios under some circumstances (see Hurst 1992) 02:03 04 05 06 07 08 09 1 sex ratio o on Figure |. Observed (N= 41 broods, brood sizes 6 14) and expected brood sex ratios (proportion sons) under the binomial distribution. “The observed sex ratio variation Aliflers significantly from the expected; Likelihood ratio tests: G = 12889, df. = 40, p< 0.001 (all broods); G = 122.53, f= 33, p < 0.001 (broods without nestling mortality prior to blood sampling). Observed (shaded); expected (un- shaded)