Habitat Use of American Alligators in East Texas

ABSTRACT The American alligator (Alligator mississippiensis) has made a remarkable recovery throughout its range during the last half-century. In Texas, USA, current inland alligator population and harvest management strategies rely on generalized and often site-specific habitat and population data generated from coastal populations, because it is assumed that habitat and demographic similarities exist between inland and coastal populations. These assumptions have not been verified, however, and no studies have specifically examined inland alligator habitat use in Texas. We quantified alligator habitat use in East Texas during 2003–2004 to address this information gap and to facilitate development of regionally specific management strategies. Although habitat was variable among study areas, alligators used habitats with >50% open water, substantial floating vegetation, and emergent vegetation close (<12 m) to dry ground and cover. Adults used habitats further from dry ground and cover, in open water (75–85%), with less floating vegetation (6–22%) than did subadults, which used habitats that were closer to dry ground and cover, with less open water (52–68%), and more floating vegetation (8–40%). Although habitat use mirrored coastal patterns, we estimated alligator densities to be 3–5 times lower than reported in coastal Texas, likely a result of inland habitat deviations from optimal coastal alligator habitat, particularly in the preponderance of open water and floating vegetation. Our findings that 1) inland habitats varied among sites and did not exactly match assumed optimal coastal habitats, 2) alligators used these inland habitats slightly differently than coastal areas, and 3) inland alligator densities were lower than coastal populations, all highlight the need for regionally specific management approaches. Because alligator populations are influenced by habitat quality and availability, any deviations from assumed optimal habitat may magnify harvest impacts upon inland populations.     

Low mitochondrial DNA variation among American alligators and a novel non-coding region in crocodilians

We analyzed 1317-1823 base pairs (bp) of mitochondrial DNA sequence beginning in the 5' end of cytochrome b (cyt b) and ending in the central domain of the control region for 25 American alligators (Alligator mississippiensis) and compared these to a homologous sequence from a Chinese alligator (A. sinensis). Both species share a non-coding spacer between cyt b and tRNA(Thr). Chinese alligator cyt b differs from that of the American alligator by 17.5% at the nucleotide level and 13.8% for inferred amino acids, which is consistent with their presumed ancient divergence. Only two cyt b haplotypes were detected among the 25 American alligators (693-1199 bp surveyed), with one haplotype shared among 24 individuals. One alligator from Mississippi differed from all other alligators by a single silent substitution. The control region contained only slightly more variation among the 25 American alligators, with two variable positions (624 bp surveyed), yielding three haplotypes with 22, two, and one individuals in each of these groups. Previous genetic studies examining allozymes and the proportion of variable microsatellite DNA loci also found low levels of genetic diversity in American alligators. However, in contrast with allozymes, microsatellites, and morphology, the mtDNA data shows no evidence of differentiation among populations from the extremes of the species range. These results suggest that American alligators underwent a severe population bottleneck in the late Pleistocene, resulting in nearly homogenous mtDNA among all American alligators today.     

Microsatellite DNA analyses support an east-west phylogeographic split of American alligator populations

We examined the population genetic structure of American alligators (Alligator mississippiensis) sampled from 12 localities across the southeastern United States. The primary goal of this study was to determine the extent of population differentiation among alligators from four Florida lakes using eight microsatellite loci and compare the results to additional sites located at varying distances from them. Analyses of population structure revealed little differentiation (F(ST)=0.039; Rho=0.012) among the four Florida lakes, Apopka, Griffin, Orange and Woodruff, which are all located in the St. John&'s River watershed in north-central Florida. Further, there was little differentiation among these samples and samples collected from the Everglades National Park (F(ST)=0.044; Rho=0.009) and south Georgia (F(ST)=0.045; Rho=0.032). Therefore, these six samples were pooled together as a "FL/sGA group." Similarly, samples collected in the western extent of the range, Anahuac National Wildlife Refuge in Texas and Salvador Wildlife Management Area, Marsh Island Wildlife Refuge and Rockefeller Wildlife Refuge in Louisiana, also lacked population structure (F(ST)=0.024; R(ST)=0.040). These four populations were pooled into the "TX/LA group." Comparisons of these two groups with samples taken from the Santee Coastal Reserve in South Carolina and Mobile, Alabama yielded three to four times more differentiation among groups (F(ST)=0.131; Rho=0.187). These and other analyses support the hypothesis of an east-west phylogeographic split in American alligator populations and are consistent with studies of many freshwater fish and aquatic and terrestrial turtles distributed throughout this same geographic region.     

Microsatellite analysis of genetic diversity in the Chinese alligator (<Emphasis Type="Italic">Alligator sinensis</Emphasis>) Changxing captive population

Chinese alligator (Alligator sinensis) is a critically endangered species endemic to China. In this study, the extent of genetic variation in the captive alligators of the Changxing Reserve Center was investigated using microsatellite markers derived from American alligators. Out of 22 loci employed, 21 were successfully amplified in the Chinese alligator. Sequence analysis showed loci in American alligators had a bigger average size than that of the Chinese alligators and the longest allele of an individual locus almost always existed in the species with longer stretch of repeat units. Eight of the 22 loci were found to be polymorphic with a total of 26 alleles present among 32 animals scored, yielding an average of 3.25 alleles per polymorphic locus. The expected heterozygosity (H _E) ranged at a moderate level from 0.4385 to 0.7163 in this population. Compared to that in the American alligators, a lower level of microsatellite diversity existed in the Changxing population as revealed by about 46% fewer alleles per locus and smaller H _E at the homologous loci. The average exclusion power and the ability to detect shared genotypes and multiple paternity were evaluated for those markers. Results suggested that when the polymorphic loci were combined, they could be sensitive markers in genetic diversity study and relatedness inference within the Chinese alligator populations. The level of genetic diversity present in the current Changxing population indicated an important resource to complement reintroductions based on the individuals from the other population. In addition, the microsatellite markers and their associated diversity characterized in this population could be utilized to further investigate the genetic status of this species.     

Nest success and hatchling survival of American alligators within inland wetlands of east Texas

Because of liberalization of American alligator (Alligator mississippiensis) harvest management in Texas, estimates of nest success and hatchling survival for inland populations are essential for long-term, sustainable population and harvest management. To date, few studies have examined American alligator nest success and hatchling survival. We initiated a 3-year study from 2006 to 2008 to document alligator nest success and hatchling survival within several wetlands in east Texas. From June 2006 to August 2008, we located 30 nests from 3 wetlands within east Texas, where overall nest success was 44.2% (95% CI = 25.1–63.1%), irrespective of year. Nest circumference and day during the nesting season exerted the greatest influence on nest success. Additionally, from August 2006 to August 2008 we captured, marked, and released 271 hatchling alligators at Little Sandy National Wildlife Refuge, and recaptured an additional 192 hatchling alligators during this time. We estimated yearly apparent survival at 6.0% (95% CI = 2.0–14.6%) for hatchling alligators born in 2006 and 43.0% (95% CI = 28.4–57.8%) for those hatched in 2007. Variation in nest success and hatchling survival was likely attributed to fluctuating water levels and habitat management practices. Alligator harvest regulations need to account for variability in nest success and hatchling survival by including site-specific estimates of these metrics into harvest models. Failing to account for spatial and temporal variation in nest success and hatchling survival may result in unsustainable harvest and/or overharvest. © 2012 The Wildlife Society.     

Eleven novel microsatellite markers for the Chinese alligator (Alligator sinensis)

Chinese alligator (Alligator sinensis) is a critically endangered species endemic to China. In this study, we developed 11 novel microsatellite loci for this rare species and applied them to examine genetic variation of indigenous alligators from Changxing Nature Reserve and America-born Chinese alligators. The 11 polymorphic microsatellites presented a total of 31 alleles among 57 individuals scored, yielding an average of 2.82 alleles per locus. One allele was unique to the American population but four private alleles were detected in the Changxing population. The average expected and observed heterozygosities were 0.400 and 0.482 for the Changxing alligators and 0.520 and 0.621 for the America-born individuals, respectively. These microsatellite markers would be useful tools in the genetic examination of this endangered species.     

AFLP Analysis of genetic variation on captive‐bred chinese alligators: an application to select individuals for release

The Chinese alligator (Alligator sinensis) is a critically endangered species in China. Wild populations of Chinese alligator are on the edge of extinction. Through a release program, some captive‐bred alligators will be selected and released into the wild to supplement and renew natural populations. The purpose of this study was to investigate the genetic variation of captive‐bred Chinese alligators by AFLP markers and to select individuals with maximally different genetic backgrounds for release. Forty‐three captive‐bred alligators of the second filial generation from the Anhui Research Center for Chinese Alligator Reproduction (ARCCAR) were surveyed using four primer combinations, yielding 117 AFLP markers. According to AFLP fingerprints, six samples had distinctly different band patterns compared to other samples. When the six samples were removed from the analysis, there were 19 monomorphic loci and 98 polymorphic loci yielding 84% polymorphic loci. Moreover, the genetic similarity (GS) among 37 samples varied from 0.13–0.97, and the average was 0.7503±0.0064 standard error (SE). When the six samples were included, the GS value among the 43 samples declined and varied from 0.06–0.97, and the average was 0.6523±0.0079 SE. Based on a cluster analysis using UPGMA, a dendrogram of the 43 alligators was constructed. According to the cluster analysis and gender of the 43 samples, eight Chinese alligators with very different genetic backgrounds were selected and suggested for release with two groups in the future. Zoo Biol 0:1–12, 2006. © 2006 Wiley‐Liss, Inc.     

Genetic variation and population structure in native Americans

We examined genetic diversity and population structure in the American landmass using 678 autosomal microsatellite markers genotyped in 422 individuals representing 24 Native American populations sampled from North, Central, and South America. These data were analyzed jointly with similar data available in 54 other indigenous populations worldwide, including an additional five Native American groups. The Native American populations have lower genetic diversity and greater differentiation than populations from other continental regions. We observe gradients both of decreasing genetic diversity as a function of geographic distance from the Bering Strait and of decreasing genetic similarity to Siberians--signals of the southward dispersal of human populations from the northwestern tip of the Americas. We also observe evidence of: (1) a higher level of diversity and lower level of population structure in western South America compared to eastern South America, (2) a relative lack of differentiation between Mesoamerican and Andean populations, (3) a scenario in which coastal routes were easier for migrating peoples to traverse in comparison with inland routes, and (4) a partial agreement on a local scale between genetic similarity and the linguistic classification of populations. These findings offer new insights into the process of population dispersal and differentiation during the peopling of the Americas.     

Geographical differentiation inferred from flavonoid content between coastal and freshwater populations of the coastal plant Lathyrus japonicus (Fabaceae)

Lake Biwa, one of the few ancient lakes in the world, harbors many coastal species that commonly inhabit seashores. The beach pea (Lathyrus japonicus) is a typical coastal species of this freshwater lake, and morphological and genetic differentiation between inland and coastal populations of this species have been reported. Inland and coastal habitats inflict distinct environmental stresses to plants, the latter imposing salt stress and high-light intensity, which leads to physiological differentiation. These abiotic stresses affect phenolic compounds, which play an important role in the response of plants to the toxic by-products of stress metabolism. We investigated physiological differentiation of phenolic compounds of the beach pea between inland and coastal habitats using high-performance liquid chromatography (HPLC) analysis. Flavonoid composition analyses revealed that patterns of flavonoid composition of inland populations at Lake Biwa were differentiated from those of coastal populations. All Lake Biwa individuals were fixed in the same flavonols glycosylated at 3- and 7-positions. In contrast, most coastal individuals contained flavonols glycosylated at 3-position alone, and these populations exhibited higher variation in flavonoid composition compared to among/within inland populations. Variation was likely lower in inland populations because of a bottleneck during landlocked periods, which is consistent with previous phylogeographic studies. A qualitative HPLC survey of flavonoid content revealed substantial variation among individuals regardless of locality. These results suggest that changes in the habitat environment may have led to beach pea acclimation via alteration of the quantity and quality of flavonoids.     

Genetic Differentiation between Geographically Distant Populations of DROSOPHILA MELANOGASTER

We have studied allozyme variation at 26 gene loci in nine populations of Drosophila melanogaster originating on five different continents. The distant populations show significant genetic differentiation. However, only half of the loci studied have contributed to this differentiation; the other half show identical patterns in all populations. The genetic differentiation in North American, European and African populations is correlated with the major climatic differences between north and south. These differences arise mainly from seven loci that show gene-frequency patterns suggestive of latitudinal clines in allele frequencies. The clinal variation is such that subtropical populations are more heterozygous than temperate populations. These results are discussed in relation to the selectionist and neutralist hypotheses of genetic variation in natural populations.     

Genic homozygosity in an ancient reptile (Alligator mississippiensis)

Proteins assayed electrophoretically showed variation at only three of 49 presumed genetic loci in alligators from southwestern Louisiana. Average heterozygosity per individual was 0.021±0.012; proportion of polymorphic loci was 0.06. Data on the history, structure, and ecology of this alligator population are consistent with natural selection as the primary factor accounting for this low genetic variability. However, neither a historic population bottleneck nor some genetic mechanism limiting variability can be dismissed as a possible factor.The study was supported by NSF Grant BMS 73-0125 to H.C.D.     

使用SSR和mtVNTR分子标记识别扬子鳄个体

扬子鳄是中国特有的濒危物种,为有效避免种群衰退,最大限度地保持该物种现有的遗传多样性,有必要对种群的个体进行个体识别研究,以便重建遗传谱系,指导现有繁育工作。应用SSR(Simple sequence repeats)与mtVNTR(Variable number tandem repeats on mitochondrial DNA)两种分子标记对扬子鳄39个个体进行了个体识别分析,结果显示：8个SSR座位的累计个体识别率与累计父权排除率分别达0．9968、0．7697,mtVNTR的个体识别率为0．9146,联合SSR与mtVNTR两种分子标记的累计个体识别率理论值达0．9997,并在实际分析中将39个扬子鳄个体完全区分开,其区分能力较RAPD(Random amplified polymorphic DNA)、AFLP(Amplified fragment length polymorphism)及mtDNA控制区5’端序列分析等分子标记要高。此外,SSR和mtvNTR还可对某些低频等位基因及其携带个体做有效的筛查,这对今后进行大量扬子鳄个体的分子标记识别和群体的遗传谱系建立等工作将具有一定实际意义[动物学报51(3)：501—506,2005]。     

Microsatellite markers reveal a spectrum of population structures in the malaria parasite Plasmodium falciparum

Multilocus genotyping of microbial pathogens has revealed a range of population structures, with some bacteria showing extensive recombination and others showing almost complete clonality. The population structure of the protozoan parasite Plasmodium falciparum has been harder to evaluate, since most studies have used a limited number of antigen-encoding loci that are known to be under strong selection. We describe length variation at 12 microsatellite loci in 465 infections collected from 9 locations worldwide. These data reveal dramatic differences in parasite population structure in different locations. Strong linkage disequilibrium (LD) was observed in six of nine populations. Significant LD occurred in all locations with prevalence <1% and in only two of five of the populations from regions with higher transmission intensities. Where present, LD results largely from the presence of identical multilocus genotypes within populations, suggesting high levels of self-fertilization in populations with low levels of transmission. We also observed dramatic variation in diversity and geographical differentiation in different regions. Mean heterozygosities in South American countries (0.3-0.4) were less than half those observed in African locations (0. 76-0.8), with intermediate heterozygosities in the Southeast Asia/Pacific samples (0.51-0.65). Furthermore, variation was distributed among locations in South America (F:(ST) = 0.364) and within locations in Africa (F:(ST) = 0.007). The intraspecific patterns of diversity and genetic differentiation observed in P. falciparum are strikingly similar to those seen in interspecific comparisons of plants and animals with differing levels of outcrossing, suggesting that similar processes may be involved. The differences observed may also reflect the recent colonization of non-African populations from an African source, and the relative influences of epidemiology and population history are difficult to disentangle. These data reveal a range of population structures within a single pathogen species and suggest intimate links between patterns of epidemiology and genetic structure in this organism.     

THE RELATIVE IMPORTANCE OF HISTORICAL EVENTS AND GENE FLOW ON THE POPULATION STRUCTURE OF A MEDITERRANEAN RAGWORT,SENECIO GALLICUS (ASTERACEAE)

Comparisons of cytoplasmic and nuclear diversity within and among natural plant populations have the potential to distinguish the relative influences of seed and pollen dispersal on contemporary gene flow, or alternatively, may permit inferences of the colonization history of a species via seed. We examined patterns of cpDNA and allozyme variation in Senecio gallicus, a diploid, annual plant that occurs in both coastal and ruderal inland areas of the Iberian Peninsula and southern France. The species appears to have a strong propensity for long-distance seed dispersal. Five cpDNA haplotypes were found by RFLP analysis among a sample of 111 individuals derived from 11 populations. Differences in haplotype frequencies across populations were most evident with respect to a dramatic increase in the frequency of a derived haplotype from coastal to inland localities. The level of cpDNA differentiation among populations within the inland group (θ₀ = 0.07) was significantly less than that seen within the coastal group (θ₀ = 0.41). In contrast, for allozymes, no significant difference in population structure was evident between collections from coastal and inland habitats. At the rangewide geographic scale, there was only a very weak association between inferred levels of gene flow and geographic distance for cpDNA, and no such association was found for allozymes. It appears that while seed movement in the species might be sufficiently great to disturb the pattern of isolation by distance for cpDNA, it cannot fully account for the nearly randomized spatial structure at polymorphic allozyme loci. It is suggested that isolation of populations in Atlantic-Mediterranean coastal refugia during previous glacial maxima, and the effects of subsequent colonization events in inland areas, have had an important effect on molding the present genetic structure of the species.     

Genetic diversity and differentiation in a wide ranging anadromous fish,American shad (Alosa sapidissima), is correlated with latitude

Studies that span entire species ranges can provide insight into the relative roles of historical contingency and contemporary factors that influence population structure and can reveal patterns of genetic variation that might otherwise go undetected. American shad is a wide ranging anadromous clupeid fish that exhibits variation in demographic histories and reproductive strategies (both semelparity and iteroparity) and provides a unique perspective on the evolutionary processes that govern the genetic architecture of anadromous fishes. Using 13 microsatellite loci, we examined the magnitude and spatial distribution of genetic variation among 33 populations across the species' range to (i) determine whether signals of historical demography persist among contemporary populations and (ii) assess the effect of different reproductive strategies on population structure. Patterns of genetic diversity and differentiation among populations varied widely and reflect the differential influences of historical demography, microevolutionary processes and anthropogenic factors across the species' range. Sequential reductions of diversity with latitude among formerly glaciated rivers are consistent with stepwise postglacial colonization and successive population founder events. Weak differentiation among U.S. iteroparous populations may be a consequence of human‐mediated gene flow, while weak differentiation among semelparous populations probably reflects natural gene flow. Evidence for an effect of reproductive strategy on population structure suggests an important role for environmental variation and suggests that the factors that are responsible for shaping American shad life history patterns may also influence population genetic structure.     

A comparison of variability and population structure for major histocompatibility complex and microsatellite loci in California coastal steelhead (Oncorhynchus mykiss Walbaum)

The major histocompatibility complex (MHC) contains genes integral to immune response in vertebrates. MHC genes have been shown to be under selection in a number of vertebrate taxa, making them intriguing for population genetic studies. We have conducted a survey of genetic variation in an MHC class II gene for steelhead trout from 24 sites in coastal California and compared this variation to that observed at 16 presumably neutral microsatellite loci. A high amount of allelic variation was observed at the MHC when compared to previously published studies on other Pacific salmonids. Elevated nonsynonymous substitutions, relative to synonymous substitutions, were detected at the MHC gene, indicating the signature of historical balancing selection. The MHC data were tested for correlations to and deviations from the patterns found with the microsatellite data. Estimates of allelic richness for the MHC gene and for the microsatellites were positively correlated, as were estimates of population differentiation (F(ST)). An analysis for F(ST) outliers indicates that the MHC locus has an elevated F(ST) relative to the neutral expectation, although a significant result was found for only one particular geographical subgroup. Relatively uniform allele frequency distributions were detected in four populations, although this finding may be partially due to recent population bottlenecks. These results indicate that, at the scale studied here, drift and migration play a major role in the observed geographical variability of MHC genes in steelhead, and that contemporary selection is relatively weak and difficult to detect.     

基于ISSR数据探讨卷叶凤尾藓(Fissidens dubius P.Beauv.)遗传多样性

卷叶凤尾藓(Fissidens dubius P.Beauv.)是凤尾藓科凤尾藓属植物,该种分布广泛,形态变异强烈。为了解其遗传多样性及种群遗传结构特点,本研究利用ISSR分子标记对采集于浙江、福建、广西、四川、辽宁5个省区的卷叶凤尾藓14个自然种群的遗传多样性进行了评价。结果显示:筛选出的12对引物共扩增出259条清晰、重复性高的条带,其中多态性位点有248个,多态位点百分率为95.75%;种群总的Nei's基因多样性指数为0.2327,Shannon's信息指数为0.3701,说明卷叶凤尾藓遗传多样性水平较高;14个种群的遗传分化系数(Gst)为0.7078,种群间的基因流(Nm)为0.1864,表明大部分遗传变异(72.01%)存在于种群间,27.99%的遗传变异存在于种群内,即卷叶凤尾藓种群间遗传分化明显。基于ISSR数据的聚类分析表明,在遗传距离50为分组阈值处,14个种群可被分为6组(G1~G6):G1为来自于浙江省不同采集地点的8个种群(JHBS除外);G2由浙江金华北山种群(JHBS)组成;G3包括福建武夷山(WYS)和天宝岩种群(TBY);G4、G5、G6分别由广西龙胜县花坪种群(GX)、四川龙池种群(LC)、辽宁白石砬子保护区种群(BSLZ)组成,表明卷叶凤尾藓种群间的遗传分化主要由地理距离造成,种群内的遗传分化可能与其生境的异质性有关。     

我国扬子鳄种群及栖息地保护现状

扬子鳄(Alligator sinensis)是我国特有的古老而珍稀的爬行动物,现阶段野生扬子鳄的分布区域进一步萎缩,成为彼此孤立的点状,残存栖息地的生态环境趋于恶化,野生鳄数量估计为120～150条,老年化程度高。扬子鳄的保护管理依据现实状况及时加以调整,加大了野外保护力度,逐步改善野生鳄的栖息生境,实施野外放归工程,初步遏制了野生鳄数量迅速下滑的局面。人工饲养种群数量已逾10 000条,当前人工饲养繁殖的重点是管理好有限的遗传多样性资源。扬子鳄的研究主要涉及形态学、解剖学、组织胚胎学、生态学、生理生化、细胞及分子生物学,人工饲养繁殖技术等方面。营养生理和保护遗传学的应用研究有待加强。     

Population genetic structure of savannah elephants in Kenya: conservation and management implications

We investigated population genetic structure and regional differentiation among African savannah elephants in Kenya using mitochondrial and microsatellite markers. We observed mitochondrial DNA (mtDNA) nucleotide diversity of 1.68% and microsatellite variation in terms of average number of alleles, expected and observed heterozygosities in the total study population of 10.20, 0.75, and 0.69, respectively. Hierarchical analysis of molecular variance of mtDNA variation revealed significant differentiation among the 3 geographical regions studied (F(CT) = 0.264; P < 0.05) and a relatively lower differentiation among populations within regions (F(SC) = 0.218; P < 0.0001). Microsatellite variation significantly differentiated among populations within regions (F(SC) = 0.019; P < 0.0001) but not at the regional levels (F(CT) = 0.000; P > 0.500). We attribute the high differentiation at the mitochondrial genome to the matrilineal social structure of elephant populations, female natal philopatry, and probably ancient vicariance. Lack of significant regional differentiation at the nuclear loci vis-a-vis strong differences at mtDNA loci between regions is likely the effect of subsequent homogenization through male-mediated gene flow. Our results depicting 3 broad regional mtDNA groups and the observed population genetic differentiation as well as connectivity patterns should be incorporated in the planning of future management activities such as translocations.     

Genetic diversity of Chinese alligator (Alligator sinensis) revealed by AFLP analysis: an implication on the management of captive conservation

Chinese alligator (Alligator sinensis) is one of the most critically endangered species among 23 extant crocodiles in the world. To prevent the extinction of the species, a captive propagation started at early 1980s, and the total number of alligator was brought up to 10 thousands from dozens of founder in 2000. But several genetic investigations showed those alligators were under an extremely low genetic diversity status with few detectible polymorphic loci. To get more insight into its genetic diversity for the management of captive Chinese alligator, AFLP was adopted to characterize variations in the population. Total of 347 bands were generated from 47 individuals using 3 primer combinations, of which 203 (58.50%) were polymorphic, and 35 AFLP phenotypes were revealed from those individuals. Comparing the results between RAPD and AFLP analysis on almost same sample set clearly indicated that AFLP is more efficient in revealing polymorphic loci, especially in those populations with extremely low genetic diversity. In present three assays, electrophoresis profile also displayed 3 individuals possessing very highly polymorphic AFLP phenotypes that were never been found by RAPD and mtDNA D-loop sequencing, implicating that we should offer these individuals more breeding opportunities to maintain the genetic diversity in the population and restrict those carrying few polymorphic loci from reproduction.