Natural Hybridization between Two Swallowtail Species Parnassius nomionand Parnassius bremeri(Lepidoptera,Papilionidae) Shown by RAPD–PCR

Genetic evidence for interspecific hybridization between Parnassius nomionand Parnassius bremeriin nature is presented. To demonstrate hybridization between these species, RAPD analysis was used. By testing 25 decamer primers, three and two diagnostic markers were revealed for P. nomionandP. bremeri, respectively. Out of 28 animals examined, 4 were shown to be interspecific hybrids. According to the distribution of diagnostic markers, the interspecific hybrids were intermediate with regard to the parental species. Ecological and biological characteristics of two butterfly species that promote their hybridization in nature are discussed.     

以绢蝶为代表的甘肃南部蝶类多样性

在甘肃南部地区的蝶类调查中,共记录到蝴蝶339种,分属于12科,157属,区系组成以广布种为主,占总种数的56.6%;古北种次之,占26.9%;东洋种最少,占16.5%;共采集到绢蝶15种,占分布于中国绢蝶总种数41种的36.6%。样线调查结果显示绢蝶在甘肃南部地区有比较明显的垂直性分布,高山亚冰雪稀疏植被带、亚高山灌丛草甸带、山地森林草原带及山地草原带均有绢蝶分布。各生境中绢蝶多样性与蝶类群落多样性进行比对得出变化趋势不一致,随着海拔升高,绢蝶物种丰富度S及Shannon-Wiener多样性指数呈逐渐上升趋势,而蝶类群落物种丰富度S及Shannon-Wiener多样性指数总体呈下降趋势。绢蝶是高原地区的特有珍稀物种,君主绢蝶(P.imperator)、依帕绢蝶(P.epaphus)、小红珠绢蝶(P.nomion)等3种绢蝶为甘肃南部地区常见种,元首绢蝶(P.cephalus)等8种绢蝶为甘肃南部地区偶见种,数量极少,其种群及生存状态需予以关注。     

三种绢蝶翅脉数字化特征的提取及初步分析

利用化学方法去除3种绢蝶翅面的鳞片和斑纹,通过扫描获取翅脉标本图片,并利用DrawWing、TPSDig和BugShape3种软件对其前翅左侧翅脉的数字特征进行了提取,并包括DrawWing提取的小红珠绢蝶右翅翅脉特征值。分析了小红珠绢蝶左右翅脉的对称性以及个体间的差异,对比了3种软件在3种绢蝶分类判别中的优劣。不仅对改进这些工具提出了建议,而且讨论了昆虫鉴定领域中数字化技术和形态测量学的应用前景。     

Hybridization and introgression between "full-fledged species". The case of Parnassius apollo and P. phoebus

Two butterfly species living in the Alps, Parnassius apollo and P. phoebus, frequently hybridize in certain localities of this region. The features of this phenomenon have been previously studied by biometry and starch gel electrophoresis, but some points remained obscure. We present them in a study combining results from cellulose acetate electrophoresis and wing pattern biometry with a determination of the mitochondrial haplotype by a PCR-RFLP analysis in a sample of butterflies from the southern French Alps. It was already known that the male hybrids are fecund and thus that interspecific gene exchange could take place via backcrosses with the parent species. In the present case, combining the identification of mtDNA with the analysis of nuclear genotypes allows us to demonstrate that hybridization can involve both sexes of both species. Moreover, it suggests that at least some female hybrids are not sterile. The impact of Haldane's rule is therefore not very strong in the present case. However, although the prerequisites for introgression between the concerned species are fulfilled, at the level of both nuclear and mitochondrial genomes, no indication of such a phenomenon could be gathered in the studied sample.     

Detection of hybridization between two loach species (<Emphasis Type="Italic">Paramisgurnus dabryanus</Emphasis> and <Emphasis Type="Italic">Misgurnus anguillicaudatus</Emphasis>) in wild populations

Artificial interspecific hybrids between large scale loach P. dabryanus and tetraploid pond loach M. anguillicaudatus (Cobitidae, Cypriniformes) are viable. To detect the occurrence of possible natural hybridization, genetic analyses by using microsatellite markers were performed for natural populations of large scale loach and pond loach, the reciprocal laboratory hybrids, and “supposed hybrids” with ambiguous morphology. The fertility of the artificial hybrids was also tested. At one diagnostic microsatellite (Mac50), one out of 20 “supposed hybrids” was identified to be F₁ hybrid between the two loach species because it had the same genotype as that of the laboratory hybrids. The triploid hybrids between the two species were confirmed to be female-sterile. The results show that rare hybridization has occurred between diploid large scale loach and tetraploid pond loach in nature although it may have little effect in genetic introgression. This study is helpful for fish conservation and encourages further investigation on natural hybridization and introgression of loaches.     

甘南地区小红珠绢蝶生物学特性研究初报

小红珠绢蝶Parnassius nomion Fischer von Waldheim在甘肃甘南1年发生1代。翌年5月上旬幼虫孵化,6月下旬开始化蛹,7月下旬至8月上旬,成虫开始羽化,8月中旬至9月上旬成虫开始产卵,该虫以卵越冬。     

Confirmation of cross-fertilization using molecular markers in ornamental passion flower hybrids

Several interspecific Passiflora hybrids are produced in the northern hemisphere for the ornamental plant market. In Brazil, production of passion flower hybrids is limited to the introgression of genes into the main cultivated species, yellow passion fruit, to be used as rootstocks. Confirmation of hybridization in the initial developmental stage is important for breeding perennial and sub-perennial plants, such as passion flowers, reducing time and costs in plant stock maintenance. In order to obtain F? hybrids with ornamental potential, four species of Passiflora (P. alata, P. gardneri, P. gibertii, and P. watsoniana) from the Active Germplasm Bank at UESC were hybridized. Flower buds, in pre-anthesis, of the genitors were previously protected, and the female buds were emasculated. To confirm hybridization, the genomic DNA of the genitor species and the supposed hybrids was extracted and RAPD primers were used to obtain molecular markers and select passion flower interspecific hybrids. Eight primers were used to confirm hybrids derived from P. gardneri with P. alata, P. watsoniana with P. alata, P. watsoniana with P. gardneri, and P. gardneri with P. gibertii; 75, 50, 45, and 46% of the informative bands, respectively, confirmed the hybrid nature of these plants. The RAPD technique was effective in the early identification of hybrids; this will be useful for development of hybrid Passiflora progeny.     

Erosion of interspecific reproductive barriers resulting from hatchery supplementation of rainbow trout sympatric with cutthroat trout

The frequency of hybridization between cutthroat (Onchorhynchus clarki clarki) and rainbow (O. mykiss irideus) trout from coastal habitats in British Columbia, Canada, was examined in seven populations where the two species are sympatric with no history of rainbow trout stocking and compared with areas where native rainbow trout populations have been supplemented with hatchery fish (three populations). Four nuclear markers were used to identify each species and interspecific hybrids and one mitochondrial marker showed the direction of gene exchange between species. The frequency of hybrids was significantly higher (Fisher exact test, P < 0.001) in river systems where hatchery rainbow trout have been introduced (50.6% hybrids) than in populations where the two species naturally co-occur without supplementation (9.9% hybrids).     

Population genetic structure of Picea engelmannii, P. glauca and their previously unrecognized hybrids in the central Rocky Mountains

Areas of geographic overlap between potentially hybridizing species provide the opportunity to study interspecific gene flow and reproductive barriers. Here we identified hybrids between Picea engelmannii and P. glauca by their genetic composition at 17 microsatellite markers, and determined the broad-scale geographic distribution of hybrids in the central Rocky Mountains of North America, a geographic region where hybrids and isolation between species had not previously been studied. Parameter estimates from admixture models revealed considerable variation in ancestry within and among collection sites, suggesting that within this area of geographic overlap, the interaction of the two species varies extensively. The results document a previously unrecognized patchy distribution of hybrids between P. engelmannii and P. glauca, including locations where hybrids were not known or expected to exist. Further, the ancestry of many hybrids was consistent with multiple generations of hybridization, with probable directional backcrossing to P. engelmannii, suggesting a relatively porous species boundary. The identification and characterization of hybridization between these spruce in this region raises the question of what factors maintain barriers to gene flow in these long-lived forest trees. The current research lays the groundwork for future study of the ecological and evolutionary contexts of their hybridization, as well as of differential introgression and permeability of species boundaries.     

Good species despite massive hybridization: genetic research on the contact zone between the watersnakes Nerodia sipedon and N. fasciata in the Carolinas, USA

Genomic markers generated with the amplified fragment length polymorphism method revealed extensive, panmictic-like hybridization along the narrow contact zone between the water snakes Nerodia sipedon and Nerodia fasciata in the Carolinas, USA. However, asymmetric distributions of diagnostic markers between both species and low frequencies of backcrossed hybrids with a high value of interspecific mixture infer selection against certain genotypes. This is consistent with a pronounced genetic and morphological preponderance of N. fasciata characters in the hybrid zone. Despite massive hybridization within the contact zone, the existence of nearly fixed genetic markers and the potential inferiority of certain hybrid genotypes support the species status of the two taxa and corroborate known, but nondiagnostic differences in morphology and ecology. This study stretches the applicability of species concepts to cases, where the genetic compatibility between two closely related species is very high, yet, they still evolve and persist as independent entities.     

Production and electrophoretic verification of F1 hybrids between the sibling speciesRhagoletis pomonella andR. cornivora

To investigate the potential for natural interspecific hybridization in theRhagoletis pomonella group of sibling fruit fly species, an attempt was made to hybridizeR. pomonella, the apple maggot fly, withR. cornivora, a non-pest feeding on dogwoods. Laboratory hybridization was difficult, with only five F₁ hybrids produced. Electrophoretic analysis of the hybrids confirmed their hybrid origin. Allele frequency data for three diagnostic loci in up to 5,596 individuals of the parental species, gathered from published and unpublished studies, indicate that the two sibling species hybridize in nature very infrequently, if at all.     

Interspecific hybridization in natural populations ofCyrtandra (Gesneriaceae) on the Hawaiian Islands: Evidence from RAPD markers

Interspecific hybridization among Hawaiian species ofCyrtandra (Gesneriaceae) was investigated using randomly amplified polymorphic DNA (RAPD) markers. Thirty-three different primers were used to investigate interspecific hybridization for 17 different putative hybrids based on morphological intermediacy and sympatry with putative parental species. RAPD data provided evidence for the hybrid origin of all putative hybrid taxa examined in this analysis. However, the patterns in the hybrid taxa were not found to be completely additive of the patterns found in the parental species. Markers missing in the hybrid taxa can be attributed to polymorphism in the populations of the parental species and the dominant nature of inheritance for RAPD markers. Unique markers found within hybrid taxa require further explanation but do not necessarily indicate that the taxa are not of hybrid origin. The implications suggest that these interspecific hybridization events had, and continue to have, an effect on the adaptive radiation and conservation biology ofCyrtandra.     

The interspecific hybrid Petunia parodii × P. inflata and its relevance to somatic hybridization in the genus Petunia

Summary Attempts at the reciprocal cross between Petunia parodii and P. inflata using standard emasculation and pollination techniques failed. Limited pollen tube growth down the style in reciprocal crosses led to reproductive isolation between the self-compatible P. parodii and self-incompatible P. inflata. The interspecific hybrid was successfully produced by bud-pollination of P. parodii with P. inflata as the male parent in 22 percent of attempts, but not in the opposite direction. In vitro pollination of P. parodii ovaries with P. inflata pollen also produced hybrids. The small size of the ovary made it technically impossible to use P. inflata as the female parent for in vitro pollination. The interspecific hybrids were intermediate, as compared to the two parents, for six of the seven plant and flower characters measured. Furthermore, the hybrids had high pollen fertility, set abundant seed upon self-pollination, and readily inter-crossed with the parental species. The results are consistent with a high degree of chromosomal homology in the parental species and with minor genetic divergency leading to reproductive isolation that is pre-zygotic in nature. Overcoming the barriers to cross-incompatibility by practical techniques resulted in fertile interspecific hybrids that segregated for parental characters. The potential value of employing the parental species in somatic hybridization experiments is discussed.Michigan Agricultural Experiment Station Journal Article No. 8404     

Genetic and morphological differentiation patterns between sister species: the case of Onthophagus taurus and Onthophagus illyricus (Coleoptera, Scarabaeidae)

The present study investigated the morphological and genetic differentiation pattern between two sympatric dung beetle sister species, Onthophagus taurus and Onthophagus illyricus . The geometric morphometric approach coupled with the use of molecular markers allowed examination of the nature of interspecific relationships and an outline of the evolutionary and geographical processes that might have led to interspecific differentiation and the present-day partial sympatric and syntopic pattern of distribution. Geometric morphometrics failed to discrimininate the two species on the ground of external morphological traits, but revealed interspecific differences when the shape of the primary sexual traits was taken into account. The use of two different molecular markers (cytochrome oxidase subunit I gene and amplified fragment-length polymorphism) demonstrates that the two species are genetically well differentiated, forming two distinguishable lineages probably diverged during the Pliocene by allopatric speciation. No evidence of past or recent introgression and no support for hybridization were found, suggesting that sympatry was established subsequently, when speciation was accomplished. Both molecular markers and genital shape indicate that phenotypically intermediate individuals, despite their ambiguous appearance, are members of O. illyricus species rather than hybrids.  © 2006 The Linnean Society of London, Biological Journal of the Linnean Society , 2006, 89 , 197–211.     

Hybridization and Introgression among Species of Sunfish (Lepomis): Analysis by Mitochondrial DNA and Allozyme Markers

We explore the potential of mitochondrial DNA (mtDNA) analysis, alone and in conjunction with allozymes, to study low-frequency hybridization and introgression phenomena in natural populations. MtDNAs from small samples of nine species of sunfish (Lepomis, Centrarchidae) were purified and digested with each of 13 informative restriction enzymes. Digestion profiles for all species were highly distinct: estimates of overall fragment homology between pairs of species ranged from 0-36%. Allozymes encoded by nine nuclear genes also showed large freqency differences among species and together with mtDNA provided many genetic markers for hybrid identification. A genetic analysis of 277 sunfish from two locations in north Georgia revealed the following: (1) a low frequency of interspecific hybrids, all of which appeared to be F1's; (2) the involvement of five sympatric Lepomis species in the production of these hybrids; (3) no evidence for introgression between species in our study locales (although for rare hybridization, most later-generation backcrosses would not be reliably distinguished from parentals); (4) a tendency for hybridizations to take place preferentially between parental species differing greatly in abundance; (5) a tendency for the rare species in a hybrid cross to provide the female parent. Our data suggest that absence of conspecific pairing partners and mating stimuli for females of rarer species may be important factors in increasing the likelihood of interspecific hybridization. The maternal inheritance of mtDNA offers at least two novel advantages for hybridization analysis: (1) an opportunity to determine direction in hybrid crosses; and (2) due to the linkage among mtDNA markers, an increased potential to distinguish effects of introgression from symplesiomorphy or character convergence.     

DNA fingerprinting of individual species and intergeneric and interspecific hybrids of genera Bos and Bison,subfamily Bovinae

Genome fingerprinting with a hypervariable minisatellite sequence of phage M13 DNA was used to study the genetic variation in individual species of the genera Bos and Bison (subfamily Bovinae) and in their interspecific and intergeneric hybrids. DNA fingerprints were obtained for domestic cow Bos taurus primigenius, vatussy Bos taurus macroceros, banteng Bos javanicus, gaur Bos gaurus, wisent Bison bonasus, bison Bison bison, and for the interspecific and intergeneric hybrids. Compared with the original species, most hybrids showed a greater variation in number and size of hybridization fragments. An association was revealed between the number of hybridization fragments and blood composition of interspecific hybrids resulting from unique crossing of domestic cow and banteng. Pairwise similarity coefficients were calculated to construct a dendrogram of genetic similarity, which reflected the relationships between the parental species and hybrids varying in blood composition. The applicability of the method for identifying interspecific and intergeneric hybrids and for studying the consequences of distant hybridization in the subfamily Bovinae is discussed.     

Enhanced hybridization rates in a <Emphasis Type="Italic">Larix gmelinii</Emphasis> var. <Emphasis Type="Italic">japonica</Emphasis> × <Emphasis Type="Italic">L. kaempferi</Emphasis> interspecific seed orchard with a single maternal clone revealed by cytoplasmic DNA markers

We identified diagnostic chloroplast DNA and mitochondrial DNA markers that can (a) discriminate between Larix gmelinii var. japonica and L. kaempferi, and (b) determine the maternal and paternal species of hybrids between them by exploiting the difference in inheritance mode between the two genomes. We also investigated the hybridization rates at a site with two types of interspecific seed orchard—a new type with rows of a single maternal clone of L. gmelinii var japonica among rows of L. kaemferi and a “traditional” type with multiple, intimately mixed clones—in 2 years using chloroplast diagnostic DNA markers. The average hybridization rates in the single maternal clone (SMC) interspecific seed orchards [84.2% (±9.4%) in 2004 and 94.1% (±3.9%) in 2005) were higher than that in the traditional interspecific seed orchards [15.9% (±13.4%) in 2004 and 30.0% (±25.5%) in 2005] because of the self-incompatibility of the L. gmelinii var. japonica clone. We detected significant differences in hybridization rates between the orchard types in both investigated years (P < 0.001, analysis of variance, ANOVA). This finding suggests that SMC interspecific seed orchards can reliably provide seeds with high proportions of hybrids. In the traditional interspecific seed orchard, there were significant differences in the proportions of hybrids among L. gmelinii var. japonica seeds between the two years (P < 0.005, ANOVA), which may have been partly due to differences in the relative amounts of pollen cones produced by L. gmelinii var. japonica and L. kaempferi.     

An examination of hybridization between the cattail species typha latifolia and typha angustifolia using random amplified polymorphic DNA and chloroplast DNA markers

Typha glauca represents a significant portion of the biomass of the wetlands surrounding the Great Lakes, USA. It is generally accepted to be a form of hybrid between T. latifolia and T. angustifolia, which itself appears to be an exotic introduction from Europe. Based on morphological and isozyme data, conflicting theories have been proposed for the hybrid nature of T. glauca: it has been described as a hybrid swarm, a distinct hybrid species and an F1 hybrid. Therefore, we developed random amplified polymorphic DNA (RAPD) and chloroplast DNA markers, specific to the parental species, to assess hybrids. Ten RAPD primers gave 17 fragments specific to T. angustifolia and 13 fragments specific to T. latifolia. All of the interspecific hybrids contained each of the species-specific markers, indicating an F1 hybrid status. Furthermore, all hybrids tested contained the T. angustifolia chloroplast haplotype, which is consistent with differential interspecific crossing success found previously. Additional confirmation of an F1 hybrid status was gained by examining seedlings from T. glauca. These progeny were expected to be advanced-generation hybrids, as opposed to the F1 hybrid parent. Analysis of the seedlings revealed segregating marker patterns consistent with patterns observed in experimental advanced-generation hybrids, although these advanced hybrids do not appear to be a significant part of mature stands. Our data do not provide support for extensive gene flow between T. latifolia and T. angustifolia. However, our results suggest that hybridization between the native and introduced Typha species has impacted the native population through the spread of the F1 hybrid, T. glauca.