Journal of Heredity 2003:94(4)
© 2003 The American Genetic Association 94:302-309
Parentage and Relatedness in Polyandrous Comb-Crested Jacanas Using ISSRs
From the USGS Forest and Rangeland Ecosystem Science Center, 3200 SW Jefferson Way, Corvallis, OR 97331 (Haig and Mullins); and the Department of Biology, University of Puget Sound, Tacoma, WA 98416 (Mace).
Address correspondence to Susan M. Haig at the address above, or e-mail: Susan_Haig{at}usgs.gov.
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In this article we present the first analysis of parentage and relatedness in a natural vertebrate population, using Intersimple Sequence Repeat (ISSR) markers. Thus, 28 ISSR markers were used in a study of a sex-role reversed, simultaneously polyandrous shorebird from northeastern Australia, the comb-crested jacana (Irediparra gallinacea). Assessment of parentage was based on comparison of field observations, novel bands, individual-specific bands found in 7/9 males and 4/6 females, and a 99% CI exclusion criteria. Integrating results from these approaches resulted in confirmation of paternity in all 36 chicks. In only one case (2.8% of chicks) was a co-mate assigned paternity. Thus, comb-crested jacanas appear to be genetically monogamous. These results showed resemblance to sequentially polyandrous birds but differed from the simultaneously polyandrous wattled jacana ( Jacana jacana; Emlen et al. 1998). A significant relationship between relatedness and ISSR similarity resulted in recognition that 14/15 adults sampled may be related to at least one other adult by 0.25 or more. Lack of dispersal may be explained by physical limitations and adequate regional habitat. ISSRs proved to be simple and helpful in resolving these issues.
In this article we use intersimple sequence repeat (ISSR) markers, also known as randomly amplified microsatellites (RAMS; Zietkiewicz et al. 1994), to examine parentage and relatedness in a sex-role reversed, simultaneously polyandrous shorebird, the comb-crested jacana (Irediparra gallinacea). With this technique, dominant markers are generated from single-primer PCR reactions where amplified regions represent the nucleotide sequence between two microsatellite priming sites oriented on opposite DNA strands (summarized in Wolfe et al. 1998a). That is, they are anonymous microsatellites generated by protocols nearly identical to RAPD techniques, except that ISSR primer sequences are designed from microsatellite regions and annealing temperatures are higher than RAPDs, reducing the amount of template-primer mismatch artifacts experienced with RAPDs. The resulting complex band patterns are similar to those produced by amplified fragment length polymorphisms (AFLPs) and minisatellite fingerprints.
Until now, use of ISSR markers has been restricted primarily to studies of cultivated species (see http://www.biosci.ohio-state.edu/
awolfe/ISSR/ISSR.bibliography.html) and have only recently been used in studies of natural plant and tree populations (Ge and Sun 1999; Wolfe et al. 1998a,b). Although species-specific amplified fragments have been found in domestic turkeys and chickens (Smith et al. 1996), the only vertebrate population study to date describes interpopulation structure in Snowy Plovers (Charadrius alexandrinus; Gorman 2000). Thus, this study represents the first use of ISSR markers to examine parentage and relatedness in a natural vertebrate population.
ISSR markers were used to examine polyandry in the comb-crested jacana (Irediparra gallinacea) from Australia. Among avian mating systems, polyandry is uncommon (Oring 1986), and the resulting parentage is not well studied (although see Dale et al. 1999; Delehanty et al. 1998; Emlen et al. 1998; Oring et al. 1992; Owens et al. 1995). Understanding paternity and a male's certainty of paternity in sex-role-reversed species is key to understanding the evolution of male uniparental care (Trivers 1972; Whittingham et al. 1992). For example, a male in a polyandrous relationship can be cuckolded by a concurrent mate of its female [termed a co-mate by Emlen et al. (1998)], by a previous mate of its female (Oring et al. 1992), or by another male not paired with its female but seeking extrapair copulations (EPCs). In this study, we investigate the extent to which cuckoldry may occur in comb-crested jacanas. We place these observations in perspective through examination of relatedness among adults in the northeastern Australian population we studied.
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Study Species
In Australia comb-crested jacana live on floating vegetation of river edges, ponds, and swamps of near-coastal regions from Sydney, north and west to the Northern Territory–Western Australia border. Like other members of the family Jacanidae, comb-crested jacana have very long toes that allow them to walk on floating vegetation. Although the sexes have similar plumage, they exhibit extreme size dimorphism; adult females average 152 g (±16.5), whereas adult males average 87 g (±6.5; Mace 2000). During breeding, comb-crested jacana females maintain simultaneous pair bonds (Garnett 1985) with one to four males and produce an average of 3.8 clutches for an average of 1.8 males (Mace 2000). Males perform all incubation and care of the precocial chicks.
Field Observation and Tissue Sampling
Two study areas were established at impoundments created by graded embankments on cattle properties near Woodstock, about 41 km south of Townsville, Queensland, Australia. The primary study site was located at Kelly's Mount View property (approximately 19°34'S, 146°47'E), and a secondary site was located at the Triangle G property, 2 km south of Kelly's. Both study areas were small during the dry (nonbreeding) season but dramatically increased in size during the wet (breeding) season. By the end of the study, Kelly's Mount View had become approximately 5 ha; Triangle G, approximately 10 ha. Both study areas were bordered by sparsely vegetated pasture land and contained areas of open water between areas of emergent vegetation consisting mostly of water lilies (Nymphaea spp.), gentians (Nymphiodes spp.), nardoo (Marsilea spp.), water hyacinth (Eichhornia spp.), and grasses.
Populations at both study areas were monitored from September 1996 to June 1997. The breeding activity of selected individuals, pairing liaisons, and subsequent clutches were recorded at each site from January to June. Adults were classified as breeding female, breeding male, co-mate, or nonbreeder. Co-mates were breeding males simultaneously pair bonded with a breeding female. Breeding females helped co-mates maintain territories, foraged within their territory without being treated as an intruder, and were occasionally involved in precopulatory presents (or at least the incipient stages), mounts, or copulations.
Adults were caught in mist nets; weighed; measured; bled; banded with an Australian Bird and Bat Banding Scheme (ABBBS) stainless steel, numbered band and a unique combination of colored plastic bands; photographed; and released. Each blood sample (160 µl) was stored in 1.5 ml Eppendorf microcentrifuge tubes containing 1X SSC and 0.01M EDTA (pH = 7) and then frozen at -20°C. These samples were later thawed and combined with a lysis buffer (50 mM TRIS-HCl, 10 mM EDTA, 200 mM NaCl, and 2% SDS in water) in 1.5 ml Eppendorf tubes and stored at 4°C.
Ten clutches were analyzed for parentage: blood was collected from three juveniles from a late clutch in 1996, and nine clutches were collected in 1997. All clutches were taken after five to seven days of incubation, allowing for substantial embryo growth. Embryos were excised, minced into small pieces, and divided between two 1.5 ml Eppendorf tubes. Tissue from the first clutch collected (four eggs) was stored at 4°C in 1.0 ml of lysis buffer. Tissue from eggs of subsequent clutches was stored at 4°C in a DMSO/NaCl buffer solution (see Seutin et al. 1991).
DNA Extraction
DNA was obtained by a standard phenol-chloroform extraction as previously reported (Haig et al. 1994b). Briefly, 10 µl of blood or 1 mm3 of tissue was digested in an Eppendorf tube with 400 µl of extraction buffer A (50 mM Tris pH 8.0, 10 mM EDTA pH 8.0, 200 mM NaCl, and 2% SDS); Proteinase K (20 mg/ml) was added to a final concentration of 600 µg/ml. Samples were vortexed and incubated overnight (18 h) at 50°C. If blood clots or other tissue was not fully dispersed, a second aliquot of Proteinase K was added and samples were incubated for an additional 2 h. Samples were extracted with equal volumes of phenol (saturated with 10 mM Tris pH 8.0) and then chloroform–isoamyl alcohol (25:1). DNA was ethanol precipitated, resuspended in TE, and quantified with a Hoefer TKO 100 fluorometer.
ISSR Amplification
Screening of primers and scoring runs were produced on a MJ Research model PTC-100 programmable thermal controller, with the following optimized conditions. A total reaction volume of 50 µl was used with the following concentrations: 50 mM KCl, 10 mM Tris-HCl pH 8.3, 3.5 mM MgCl2, 250 µM for each of the dNTPs, and 0.8uM dUTP; labeled with one of the following fluorescent rhodamine dyes (R110, R6G, or Tamra), 0.6 µM primer, acetimide at final concentration of 1%, 100 ng DNA template, and 1.75 U of Amplitaq Gold (Perkin Elmer). The following parameters were used for the amplifications: 12 min preincubation at 93°C, followed by 35 cycles of 30 s at 93°C, annealing at the appropriate temperature (49–55°C) for 30 s, and elongation at 72°C for 2 min. An additional 10 min period for elongation at 72°C followed the last cycle. Amplification products were sized on an ABI PRISM 377 DNA sequencer. Fragment information collection and analysis was performed with the software applications Genescan Analysis 2.1 and Genotyper 2.0 (Perkin Elmer). DNA PCR templates were taken from a single extraction (pool) of DNA for each sample. Replicates of samples were amplified in multiple PCR runs to verify profile reproducibility for all fragments scored.
Marker Screening and Scoring
Initial samples were screened for variability with thirty 5'-end anchored ISSR primer (set #9) from the Nucleic Acid-Protein Unit, University of British Columbia, and primers 5'-BDB(CAA)5, 5'-DD(CCA)5, and 5'-BDN(CA)6 (Hantula et al. 1996). Each variable peak was scored as present (1) or absent (0) and was considered to represent the phenotype of a distinct locus. Peaks were scored conservatively, with our choosing only those that were distinct and highly reproducible.
ISSR markers are inherited in a dominant or codominant Mendelian fashion (Gupta et al. 1994; Tsumura et al. 1996) but are interpreted as dominant diallelic markers. Therefore, the dominant allele determines the presence of the band such that +/+ and +/- individuals have the (1) phenotype, while -/- individuals have the (0) phenotype. Absence of a band is interpreted as primer divergence, loss of a locus through deletion of the SSR site, or chromosomal rearrangement (Wolfe and Liston 1998). ISSR polymorphisms segregate independently, suggesting they represent individual loci. Thus, when scored as a dominant marker, they differ from VNTR polymorphisms that arise through slippage and unequal crossing over within a single genetic locus, producing multiple alleles that do not segregate independently (Wolfe and Liston 1998).
Statistical Analyses
Parentage was assessed in several ways. First, we assumed identification of the putative mother was correct from field observations and then compared each variable band in offspring and putative fathers. Bands found in the profile of an offspring but not in either putative parent were considered to be novel bands. Novel bands result from mutation, extrapair fertilization, incorrect field identification of the parent, or methodological aberrations. Parentage was also examined by identification of bands present in offspring and specific to one adult in the putative family group.
Exclusion criteria from Emlen et al. (1998) were used to further evaluate parentage and to provide comparisons with other parentage studies in polyandrous birds. The cutoff for acceptable similarity between putative parents and offspring was calculated, with use of the lower 99% confidence interval for similarity between putative mothers and offspring.
Many comb-crested jacanas monitored for this study were closely related (i.e., parents and offspring, half-siblings, etc.), so we were presented a unique opportunity to compare ISSR similarity within and among first-order relatives (parent-offspring, full sibs, relatedness = 0.5) and second-order relatives (half-sibs, grandparents, aunt/uncle–niece/nephews, relatedness = 0.25), and predict potential relatedness for unknown individuals (see Haig et al. 1994a, 1995). Relatedness was defined as the proportion of genes that are identical by descent between two individuals. ISSR similarity between two individuals was calculated with the similarity index of Nei and Li (1985): S = 2NAB (NA + NB), where NAB is the number of bands that individuals A and B share in common, NA is the number of bands in individual A, and NB is the number of bands in individual B.
Statistical analyses of DNA similarity data has been problematic because of lack of independence among individuals in the resulting matrices (see Lynch 1988, 1990). Thus, we used the methods of Curnow (1998) to calculate variance, SD, and SE for similarity values used in relatedness assessments. He showed that the most efficient variance estimator was obtained from an analysis of variance of all similarity values. When variance is partitioned, all similarity values are used, but their variation is separated into variation between average similarities for individuals and the residual variation. Estimates of the variance of a single similarity value can be calculated, as well as the correlation coefficient for pairs of similarities involving a common individual. This approach was used for balanced matrices (i.e., all birds, unrelated birds, males, females, all adults, full siblings, and birds with relatedness scores of 0.50). However, comparisons among parents and offspring, and half-siblings, do not result in a complete matrix with a block off–diagonal structure; thus, these methods were not appropriate, and a simple SD was calculated. Further, t tests, assuming equal variances, were used to compare mean ISSR similarities among relatedness values. Finally, relatedness among adults was categorized according to similarity values matching (mean ± 1 SD) those with known relatedness.
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Results |
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Comb-crested jacanas in the area south of Townsville, Queensland, are distributed across a mosaic of small lagoons, often with less than 100 adults per lagoon. Within a lagoon, females assist each of their mates in defending a territory. By the end of the 1996 breeding (wet) season at Kelly's Mount View lagoon, 24 breeding and nonbreeding adults had been banded, including a single polyandrous group of one female and two males that were banded and genetically sampled midway into incubation of a late clutch (Figure 1). Later, three juveniles from this clutch were banded and genetically sampled. Most birds at Kelly's dispersed as the dry season progressed and lagoon dried. In 1997 there were 17 adults present on Kelly's lagoon during the breeding season; seven were holdovers from 1996. Among the 17 adults, 14 were breeders (i.e., pair bonded and territory holders for at least some period of time) and three were nonbreeding floaters that usually stayed far from territory-holding birds and were aggressively chased if they approached territories. Two of three nonbreeders were banded but not genetically sampled. Of 14 breeding jacana, 11 were banded and genetically sampled. Seven offspring groups (families) representing 1 polyandrous female (#59) from 1996 and 4 females (two polyandrous and two monandrous) from 1997 were analyzed from Kelly's (Figure 1). Approximately 30 birds occupied the Triangle G lagoon, and three families from one polyandrous group (one female, two males) from 1997 were also included in our analyses (Figure 1). Overall, 9 males and 6 females were considered as potential parents to the 36 offspring in these 10 families.
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Parentage
Among 30 primers screened, nine variable primers produced 28 variable bands that ranged in size from 204 base pairs (bp) to 1,117 bp (Table 1). The mean number of bands scored as present per individual among 15 adults (9 males, 6 females) and 36 chicks was 9.8 (±1.8 SD).
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Assuming maternity, 35/36 chicks (97%) had a band profile that matched both putative parents (i.e., 0 novel bands). However, one novel or unattributable band (B23) was identified in one sibling in Family 10. The co-mate (#53) in Family 10 had this band.
Bands in seven of nine adult males were exclusive to a specific individual among all adults sampled (Table 2). One other band (B22) was exclusive to male 53 among all other adult males sampled. These exclusive bands identified paternity in 24/36 (67%) chicks. Further comparison of bands between putative fathers and co-mates resolved five additional paternity assignments. With these specific bands, paternity was assigned to the putative father in 28/29 (97%) chicks identified, whereas co-mate 53 was assigned paternity for chick 111 in Family 10.
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Among adult females, bird 86 had two individual-specific bands (B18, B25), which identified chicks 401, 402, 403. Bands B5, B15, and B11 were specific for females 64 (identified chick 801), 59 (identified chick 113), and 90 (identified chick 604), respectively.
The 99% paternal exclusion criteria resulting from Emlen et al. (1998) indicated 0.47 was the lower similarity cutoff point (considering a mean similarity among putative mothers and offspring of 0.72, 0.09 SD, and n = 36). Two chicks were classified as nonpaternal in this assessment: 802 (S = 0.46) and 111 (S = 0.43).
Relatedness and ISSR Similarity
Field observations and subsequent construction of a putative pedigree for the 10 families sampled suggested that two families had the same parents (Families 1 and 3) and five groups contained chicks that were half-sibs to each other: Families 1 and 5; Families 3 and 5; Families 2, 7, and 9; Families 2 and 10; and Families 6 and 9 (Figure 1).
Similarity values varied within and among relatedness categories (Table 3). There was a significant correlation between pairwise relatedness and similarity (r2 =.29; P <.001). Comparisons of mean similarities among relatedness values also resulted in significant differences in three comparisons: relatedness = 0 versus 0.25 (t = 1.96, 1,137 df, P <.001), relatedness = 0 versus 0.50 (t = 1.96, 1,176 df, P <.001), and relatedness = 0.25 versus 0.50 (t = 1.97, 231 df, P <.001). Differences in similarity among putatively unrelated adult females were not significantly different from those of putatively unrelated adult males (Table 3). Similarity also did not vary between putative mothers–offspring and putative fathers–offspring. However, similarity among siblings was significantly higher than in parent-offspring comparisons (t = 1.98, 134 df, P <.001).
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Significant differences in ISSR similarity among putative relatedness categories provided a means to better assess relationships among individuals whose relatedness was unknown. Thus, birds with ISSR similarity values as high as levels observed for birds where relatedness
0.25 (i.e., mean + 1 SD) were identified in 13.7% (143/1,042) of comparisons. Previously these birds were classified (by field observations) as unrelated. All but one (#85) of 15 adults sampled had a similarity value (with at least one other adult) that fell within the relatedness category of 0.25. Among these, six adults (58 and 93, 59 and 84, and 63 and 83) fell within the relatedness category of 0.50. Recognizing the high similarity among breeding adults provided useful information regarding relatedness and parentage. For example, two pairs of potentially closely related adults bred (Family 5: 83 and 84; Family 9: 53 and 58). This is reflected by high similarity between parents and offspring and among siblings. The highest similarity (0.87) among all pairs of putatively unrelated birds was between co-mate 53 and chick 111 from Family 10.
Defining potential relatedness among putatively unrelated birds was most helpful in confirming maternity. In a number of cases, similarity between putative mothers and offspring was the same or less than with other females sampled. For example, in Family 5, chick 502 had higher similarity with female 59 (S = 0.75) than with its putative mother 83 (S = 0.67); however, this may be explained by high similarity between 59 and the father, male 84 (S = 0.70). Similarly, in Family 6, lack of resolution between the putative mother 90 and female 58 could be due to their similarity (S = 0.70). This relationship was further complicated by high similarity between female 58 and putative father 53 (S = 0.67). In Family 7, male 93 had high similarity with putative mother 58 (S = 0.75), which may help explain high similarity between it and chick 704 (S = 0.82). Thus, without consideration of adult relationships, 93 could be assigned paternity when it most likely belonged to 63. This also occurred in Family 8 where chick 802 had higher similarity with male 91 (S = 0.67) than with its putative father, but male 91 also had a high similarity with 802's mother (S = 0.67).
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Discussion |
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ISSR Markers in Studies of Vertebrate Populations
Results of this study provide evidence for the usefulness of ISSR markers in defining parentage and relatedness, and hence insight into the structure of vertebrate populations. The power of these markers will be particularly useful in avian studies in which development of microsatellites can be problematic (Longmire et al. 1999; Primmer et al. 1997). Further, ISSRs are more cost effective than AFLPs and more reliable than RAPDs, so they present a simple alternative to these nuclear methods (see review in Zhang and Hewitt 2003).
One potential drawback to evaluating parentage by using dominant markers such as ISSRs concerns the number of loci used and manner in which data are analyzed (e.g., Epifanio and Phillip 1997; Gerber et al. 2000; Lewis and Snow 1992; Nason and Ellstrand 1993; Neff et al. 2000a,b). However, given the ease and confidence with which variable ISSR markers can be generated, meeting or exceeding any criterion currently proposed may be a viable alternative to developing the appropriate number of variable microsatellite markers.
One unique aspect of our study was identification of individual-specific ISSR loci (also see Scott and Williams 1993). These bands were diagnostic for paternity in 90% of families examined and diagnostic for maternity in several families. The large number of individual-specific loci in this study may be an artifact of limited sample size. However, in a larger study, identification of specific bands for even a few individuals in a population will strengthen assurance in some parentage assignments; hence, it is worthwhile to examine data for this possibility.
Parentage and Polyandry in Comb-Crested Jacanas
Polyandry in comb-crested jacanas may have evolved in response to high rates of clutch loss, the possibility of uniparental care, and high assurance of paternity by incubating males (Mace 2000). The present study provided genetic evidence for paternity assurance in a population where there were always additional males available for copulation: in 8/10 families sampled, breeding females had 1–3 co-mates present. Thus, a female may assure paternity by spending much of her time during laying in the territory of the male for whom she is laying eggs and more or less restricting copulations to that male. Without this implied confidence in paternity, there could be an increased probability of a male's deserting a nest and loss of eggs for the female.
Extrapair paternity (2.8% of chicks; 10% of nests) was attributed only to one male and chick and occurred in a late nest. The mother, female 59, was paired to males 69 and 53 late in the breeding season, when there were no other birds breeding. She produced a clutch for male 69 (Family 10), and co-mate 53 apparently sired one of the chicks.
This single example of extrapair paternity may be attributed to sperm storage from a previous mating, fertilization from an extrapair copulation, or misidentification of a chick or parent in the field or lab. Although sperm storage is possible (male 53 was pair bonded to female 59), it is unlikely, given the low frequency with which extrapair paternity was observed in the population. That is, if sperm storage were a factor in this population, the individual-specific ISSR markers would have detected it. Thus, we conclude that chick 111 resulted from an extrapair copulation between co-mate 53 and female 59 at the time clutch 10 was being laid.
The extrapair fertilization (EPF) between male 53 and female 59 came at the end of the breeding season, when there were no apparent females available to breed with male 53. Valle (1994) predicted that males in polyandrous mating systems who are being deserted by a mate or are at the end of the breeding season might benefit from copulation-fertilization with the deserting female. This prediction was borne out in our study and in other studies of avian polyandry. For example, 6.5% of red phalarope (Phalaropus fulicarious; Dale et al. 1999) chicks and 4.6% of Eurasian dotterel (Charadrius morinellus; Owens et al. 2000) chicks resulted from EPFs that occurred late in the breeding season. Results from these studies suggest that cuckoldry is not a significant factor for males. Further, these rates of EPF are quite low relative to those reported for many socially monogamous or polygynous species (see Möller and Birkhead 1993; Petrie and Kempenaers 1998; Westneat et al. 1990).
A different situation arose in the other simultaneously polyandrous avian species for which there are genetic parentage data. Emlen et al. (1998) reported that 18/74 (24%; corrected to 17.9% because of larger groups in the sample) nests had one or more chicks by EPF, and, overall, 24/235 (10%) of chicks were sired by EPF in the wattled jacana (Jacana jacana). This resulted in a 7.5% (n = 235 chicks) population-wide frequency of extrapair paternity. EPFs took place throughout the breeding season, not at the end as with the other species described, and resulted in a serious cost of cuckoldry to males. They concluded that a male wattled jacana's chance of cuckoldry was 0% in monandrous matings, 41% in broods when another male was "available," and 74% when a female was observed copulating with multiple mates.
Female wattled jacanas copulating with multiple males did so within a short period of time (median time between copulations was 19 min), so sperm-mixing would be expected. Not surprisingly, the rate of copulation between a male wattled jacana and his mate was much higher than among comb-crested jacanas. Emlen et al. reported an average of 1.3 copulations per h during egg-laying, or approximately 65 copulations per clutch. Although obvious when they occur during daylight hours, copulations were rarely observed in comb-crested jacanas (Mace TR, unpublished data). There were 0.1 copulations observed per h for the 4 days before laying and the laying period, 0.24 sexual presents or mounts during prelaying, and 0.14 sexual presents or mounts during laying (n = 17 nests produced by five females and eight males). Assuming 4 days prelaying and 12 h of observation per day, we observed approximately five copulations per clutch during prelaying and laying. Emlen et al. suggest that, though a high certainty of paternity may be necessary for the evolution of male uniparental care, once established, such certainty may not be necessary for its maintenance. Thus, perhaps comb-crested jacanas and the other polyandrous species exhibiting low frequency of EPFs are at an earlier evolutionary stage on the path to classical polyandry than wattled jacanas.
Population Structure and Relatedness
High levels of ISSR similarity among seemingly unrelated adults may reflect the poor dispersing capabilities of comb-crested jacanas, variable habitat viability, their mating system, or some combination. Inferring a previous bottleneck is not appropriate here, because there was not high similarity among all adults, but just between specific individuals. This suggests a population with a high level of coancestry and low level of breeding or natal dispersal (or both). The result of this population structure is a further reduction in effective population size from what would be expected in an outbred polyandrous population where already there is unequal breeding among males and females.
Field observations provide support for limited dispersal leading to an increase in relatedness among individuals. Comb-crested jacanas are weak fliers, are nonmigratory, and have numerous breeding sites available to them in the local area. Thus, long-distance dispersal may not occur, but local movement may be common. There is seasonality to the area, and birds disperse from evaporating wetlands to more permanent wetlands in the peak of the dry season. In 1996, 24 birds were banded before the dry season at the Kelly's Mount View site, but, by the time the wet season began, all but seven had dispersed and new birds were moving in.
The rate of inbreeding in this population will be slowed by matings among nonrelatives or more distant relatives. This appears to be the case in all but two pairs of comb-crested jacanas (22%). These two pairs have a potential relatedness value of 0.25 or more. Given our small sample size, it is difficult to evaluate whether this rate would change with observations of additional pairs. If it remained the same, with almost a quarter of matings among close relatives, the negative aspects of inbreeding would come to the forefront rather quickly in the absence of immigration into the population. However, comb-crested jacanas may have adequate immigration. Conversely, they may have some as yet unidentified mechanisms for inbreeding avoidance in a population with a high percentage of close relatives.
In summary, results of this study not only shed light on the complex social system of comb-crested jacanas, but demonstrate the heritability of ISSR markers on a fine scale. Future studies may benefit from the efficiency of these markers.
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Acknowledgments |
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We are most grateful to M. Huso for advice on statistical analyses of relatedness matrices and to L. Oring, D. Queller, and M. Webster for comments related to this manuscript. We thank D. Jenni for finding our study areas; J. Kelly and B. Gravener for access to lagoons on their properties; M. Hall, Australian Institute of Marine Science, for use of his lab during tissue preparation; and the staffs of the ABBBS and the Queensland Wildlife Management Office for their help with permits. Support for this project was provided by the John Lantz Fellowship at the University of Puget Sound and the USGS Forest and Rangeland Ecosystem Science Center.
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Footnotes |
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Corresponding Editor: Stephen J. O'Brien
Received November 21, 2002
Accepted March 28, 2003
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References |
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Curnow RN, 1998. Estimating genetic similarities within and between populations. J Agric Biol Environ Stat. 3:347-358.
Dale J, Montgomerie R, Michaud D, Boag P, 1999. Frequency and timing of extrapair fertilizations in polyandrous red phalarope. Behav Ecol Sociobiol. 46:50-56.
Delehanty DJ, Fleischer RC, Colwell MA, Oring LW, 1998. Sex-role reversal and the absence of extra-pair fertilization in Wilson's Phalarope. Anim Behav. 55:995-1002.[CrossRef][Web of Science][Medline]
Emlen ST, Wrege PH, Webster MS, 1998. Cuckoldry as a cost of polyandry in the sex-role-reversed Wattled Jacana, Jacana jacana. Proc Royal Soc Lond B. 265:2359-2364.
Epifanio JM, Phillipp DP, 1997. Sources for misclassifying genealogical origins in mixed hybrid populations. J Hered. 88:62-65.
Garnett S, 1985. Evidence for polyandry in the comb-crested jacana. Stilt. 7:24.
Ge XJ, Sun M, 1999. Reproductive biology and genetic diversity of a cryptoviviparous mangrove Aegiceras corniculatum (Myrsinaceae) using allozyme and intersimple sequence repeat (ISSR) analysis. Mol Ecol. 8:2061-2069.[CrossRef][Medline]
Gerber S, Mariette S, Streiff R, Bodenes C, Kremer A, 2000. Comparison of microsatellites and amplified fragment length polymorphism markers for parentage analysis. Mol Ecol. 9:1037-1048.[CrossRef][Medline]
Gorman LR, 2000. Population differentiation among snowy plovers (Charadrius alexandrinus) in North America (MS thesis). Corvallis, OR: Oregon State University.
Gupta M, Chyi Y-S, Romero-Severson J, Owen JL, 1994. Amplification of DNA markers from evolutionary diverse genomes using single primers of simple-sequence repeats. Theor Appl Genet. 89:998-1006.[Web of Science]
Haig SM, Ballou JD, Casna NJ, 1994a. Identification of kin structure among Guam rail founders: a comparison of pedigrees and DNA profiles. Mol Ecol. 3:109-119.
Haig SM, Ballou JD, Casna NJ, 1995. Genetic identification of kin in Micronesian kingfishers. J Hered. 86:423-431.
Haig SM, Rhymer JM, Heckel DG, 1994b. Population differentiation in randomly-amplified polymorphic DNA of red-cockaded woodpeckers. Mol Ecol. 3:581-595.[Medline]
Hantula J, Dusabenyagasani M, Hamelin RC, 1996. Random amplified microsatellites (RAMS)—a novel method for characterizing genetic variation within fungi. Eur J For Path. 26:159-166.
Lewis PO, Snow AA, 1992. Deterministic paternity exclusion using RAPD markers. Mol Ecol. 1:155-160.[Medline]
Longmire JL, Hahn DC, Roach JL, 1999. Low abundance of microsatellite repeats in the genome of the brown-headed cowbird (Molothrus alter). J Hered. 90:574-578.
Lynch M, 1988. Estimation of relatedness by DNA fingerprinting. Mol Biol Evol. 5:584-599.[Abstract]
Lynch M, 1990. The similarity index and DNA fingerprinting. Mol Biol Evol. 7:478-484.[Abstract]
Mace TR, 2000. Time budget and pair-bond dynamics of the comb-crested jacana Irediparra gallinacea: a test of hypotheses. Emu. 100:31-42.[CrossRef]
Möller AP, Birkhead TR, 1993. Cuckoldry and sociality: a comparative study of birds. Am Nat. 142:118-140.[CrossRef][Web of Science]
Nason JD, Ellstrand NC, 1993. Estimating the frequencies of genetically distinct classes of individuals in hybridized populations. J Hered. 84:1-12.
Neff BD, Repka J, Gross MR, 2000a. Parentage analysis with incomplete sampling of candidate parents and offspring. Mol Ecol. 9:515-528.[CrossRef][Medline]
Neff BD, Repka J, Gross MR, 2000b. Statistical confidence in parentage analysis with incomplete sampling: how many loci and offspring are needed? Mol Ecol. 9:529-539.[CrossRef][Medline]
Nei M, Li WH., 1985. Mathematical model for studying genetic variation in terms of restricted nucleases. PNAS USA. 76:5269-5273.
Oring LW, 1986. Avian polyandry. In: Current ornithology ( Johnson RF, ed). New York: Plenum; 309–351.
Oring LW, Fleischer RC, Reed JM, Marsden KE, 1992. Cuckoldry through stored sperm in the sequentially polyandrous spotted sandpiper. Nature. 359:631-633.[CrossRef]
Owens IPF, Dixon A, Burke T, Thompson DBA., 1995. Strategic paternity assurance in the sex-role reversed Eurasian dotterel (Charadrius morinellus): behavioral and genetic evidence. Behav Ecol. 6:14-21.
Petrie M, Kempenaers B., 1998. Extra-pair paternity in birds: explaining variation between species and populations. TREE. 13:52-58.
Primmer CR, Raudsepp T, Chowdhary BP, Möller AP, Ellegren H, 1997. Low frequency of microsatellites in the avian genome. Gen Res. 7:471-482.
Scott MP, Williams SM, 1993. Comparative reproductive success of communally breeding burying beetles as assessed by PCR with randomly amplified polymorphic DNA. PNAS USA. 90:2242-2245.
Seutin G, White BN, Boag PT, 1991. Preservation of avian blood and tissue samples for DNA analysis. Can J Zool. 69:82-90.[CrossRef][Web of Science]
Smith, EJ, Ray SA, Bakst MR, Teuscher C, Savage TF, 1996. Simple sequence repeat-based single primer amplifications of genomic DNA in random bred populations of turkeys and chickens. Anim Biotech. 7:47-58.
Trivers RL, 1972. Parental investment and sexual selection. In: Sexual selection and the descent of man (Campbell B, ed). Chicago: Aldine-Atherton; 136–179.
Tsumura Y, Ohba K, Strauss SH, 1996. Diversity and inheritance of inter-simple sequence repeat polymorphisms in Douglas fir (Pseudotsuga menziesii) and sugi (Cryptomeria japonica). Theor Appl Genet. 92:40-45.[CrossRef][Web of Science]
Valle CA, 1994. Parental role-reversed polyandry and paternity. Auk. 111:476-478.[Web of Science]
Westneat DF, Sherman PW, Morton ML, 1990. The ecology and evolution of extra-pair copulations in birds. Curr Ornithol. 7:331-369.
Whittingham LA, Taylor PD, Robertson RJ, 1992. Confidence of paternity and male parental care. Am Nat. 139:1115-1125.[CrossRef][Web of Science]
Wolfe AD, Liston A, 1998. Contributions of PCR-based methods to plant systematics and evolutionary biology. In: Plant molecular systematics II (Soltis DE, Soltis PS, and Doyle JJ, eds). New York, Chapman and Hall; 43–86.
Wolfe AD, Xiang Q-Y, Kephart SR, 1998a. Assessing hybridization in natural populations of Penstemon (Scrophulariaceae) using hyperviariable inter-simple sequence repeat (ISSR) bands. Mol Ecol. 7:1107-1125.[CrossRef][Medline]
Wolfe AD, Xiang Q-Y, Kephart SR, 1998b. Diploid hybrid speciation in Penstemon (Scrophulariaceae). PNAS USA. 95:5112-5115.
Zhang DX, Hewitt GM, 2003. Nuclear DNA analyses in genetic studies of populations: practice, problems, and prospects. Mol Ecol. 12:563-584.[CrossRef][Medline]
Zietkiewicz E, Rafalski A, Labuda D, 1994. Genome fingerprinting by Simple Sequence Repeat (SSR)–anchored polymerase chain reaction amplification. Genomics. 20:176-183.[CrossRef][Web of Science][Medline]
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