Natural Introgression Research Articles

The genomes of Darwin's primroses reveal chromosome-scale adaptive introgression and differential permeability of species boundaries.

Introgression is an important source of genetic variation that can determine species adaptation to environmental conditions. Yet, definitive evidence of the genomic and adaptive implications of introgression in nature remains scarce. The widespread hybrid zones of Darwin's primroses (Primula elatior, Primula veris, and Primula vulgaris) provide a unique natural laboratory for studying introgression in flowering plants and the varying permeability of species boundaries. Through analysis of 650 genomes, we provide evidence of an introgressed genomic region likely to confer adaptive advantage in conditions of soil toxicity. We also document unequivocal evidence of chloroplast introgression, an important precursor to species-wide chloroplast capture. Finally, we provide the first evidence that the S-locus supergene, which controls heterostyly in primroses, does not introgress in this clade. Our results contribute novel insights into the adaptive role of introgression and demonstrate the importance of extensive genomic and geographical sampling for illuminating the complex nature of species boundaries.

The best of both worlds? Rattlesnake hybrid zones generate complex combinations of divergent venom phenotypes that retain high toxicity

Studying the consequences of hybridization between closely related species with divergent traits can reveal patterns of evolution that shape and maintain extreme trophic adaptations. Snake venoms are an excellent model system for examining the evolutionary and ecological patterns that underlie highly selected polymorphic traits. Here we investigate hybrid venom phenotypes that result from natural introgression between two rattlesnake species that express highly divergent venom phenotypes: Crotalus o. concolor and C. v. viridis. Though not yet documented, interbreeding between these species may lead to novel venom phenotypes with unique activities that break the typical trends of venom composition in rattlesnakes. The characteristics of these unusual phenotypes could unveil the roles of introgression in maintaining patterns of venom composition and variation, including the near ubiquitous dichotomy between neurotoxic or degradative venoms observed across rattlesnakes. We use RADseq data to infer patterns of gene flow and hybrid ancestry between these diverged lineages and link these genetic data with analyses of venom composition, biological activity, and whole animal model toxicity tests to understand the impacts of introgression on venom composition. We find that introgressed populations express admixed venom phenotypes that do not sacrifice biological activity (lethal toxicity) or overall abundance of dominant toxins compared to parental venoms. These hybridized venoms therefore do not represent a trade-off in functionality between the typical phenotypic extremes but instead represent a unique combination of characters whose expression appears limited to the hybrid zone.

A retrotransposon insertion in the Mao1 promoter results in erect pubescence and higher yield in soybean

Adaptive changes in crops contribute to the diversity of agronomic traits, which directly or indirectly affect yield. The change of pubescence form from appressed to erect is a notable feature during soybean domestication. However, the biological significance and regulatory mechanism underlying this transformation remain largely unknown. Here, we identified a major-effect locus, PUBESCENCE FORM 1 (PF1), the upstream region of Mao1, that regulates pubescence form in soybean. The insertion of a Ty3/Gypsy retrotransposon in PF1 can recruit the transcription factor GAGA-binding protein to a GA-rich region, which up-regulates Mao1 expression, underpinning soybean pubescence evolution. Interestingly, the proportion of improved cultivars with erect pubescence increases gradually with increasing latitude, and erect-pubescence cultivars have a higher yield possibly through a higher photosynthetic rate and photosynthetic stability. These findings open an avenue for molecular breeding through either natural introgression or genome editing toward yield improvement and productivity.

Leishmania donovani hybridisation and introgression in nature: a comparative genomic investigation

Leishmaniasis is a neglected tropical disease transmitted by infected sandflies that results in diverse human pathologies contingent on the species of Leishmania. Leishmania donovani causes highly virulent fatal visceral leishmaniasis, whereas Leishmania major and Leishmania tropica cause less virulent, cutaneous leishmaniasis, in which the infection remains in the skin at the site of the sandfly bite. The aim of this study was to investigate the genetic basis for the emergence of L donovani strains that cause cutaneous leishmaniasis instead of visceral leishmaniasis in Sri Lanka. All available sequencing data for L donovani samples from Asia and Africa in GenBank and the Sequence Read Archive were retrieved and filtered to select for paired-end Illumina sequencing reads with no region bias and coverage of the entire reference genome. These data were used for sequence alignments against the reference L donovani genome from Sri Lanka, and sequence analysis was used to assess the presence of genomic recombination markers and the presence of foreign genetic sequences in the genomes of L donovani isolates associated with cutaneous leishmaniasis in Sri Lanka. BLAST analysis was used to compare the genetic sequences from the Sri Lankan isolates to all genomes of Leishmania species from the Old World available in TriTrypDB, including L major and L tropica. After filtering of the 1238 existing sequencing records, 684 high-quality records were used to show that 12 L donovani strains from Sri Lanka form three phylogenetic groups. In one group, the density of heterozygous variants is higher than in previously characterised Leishmania hybrid strains. BLAST analysis showed this group contains gene polymorphisms homologous with L major and L tropica genomes for 22% (2160 of 9757) to 78% (7671 of 9757) of all genes analysed. Analysis by phylogeny and BLAST showed that the L donovani-L major and L donovani-L tropica hybrid strains originated from Africa and are phylogenetically distinct from the L donovani strains in neighbouring India. Novel L donovani strains might arise in new environments through the integration of genes from another species. On the basis of the findings of this study, we hypothesise that hybridisation with genomes from L major and L tropica, followed by recombination and introgression, contributed to the emergence of L donovani offspring capable of causing cutaneous leishmaniasis in Sri Lanka. Canadian Institutes of Health Research, Fonds de recherche du Québec-Santé.

Loss of C‐genome‐specific markers during transgene introgression from Brassica napus to wild Brassica juncea

Transgene flow from engineered Brassica napus to wild weed relatives could potentially have an environmental effect. To evaluate the introgression of transgenic B. napus into wild Brassica juncea, the hybrid F1 and backcross progenies derived from B. juncea (genome constitution AABB) and transgenic B. napus (AACC) crosses were investigated. C‐genome‐specific simple sequence repeat (SSR) markers corresponding to linkage groups N11–N19 in B. napus were screened and used to estimate the marker frequency in hybrid F1 and backcross progenies. C‐genome‐specific markers could be stably detected in hybrid F1 and backcross BC1 plants, but were only rarely found in the BC2–BC5 generations. For example, a specific SSR marker for linkage group N12 segregated in BC2 generation but were completely lost in BC3–BC5, while a specific SSR marker of linkage group N15 segregated in BC1, BC2 and BC3 generations and was absent in more advanced backcrossed generations (BC4 and BC5). The results indicate that a certain gene regions in Brassica napus plants are transmitted at a relatively lower frequency to wild relatives, and more rapidly disappeared in subsequent backcross generations. We propose that a foreign gene or transgene that is integrated in the C‐chromosome of Brassica napus could reduce the risk of introgression in nature.

Pinus massoniana Introgression Hybrids Display Differential Expression of Reproductive Genes

Pinus massoniana and P. hwangshanensis are two conifer species located in southern China, which are of both economic and ornamental value. Around the middle and lower reaches of the Yangtze River, P. massoniana occurs mainly at altitudes below 700 m, while P. hwangshanensis can be found above 900 m. At altitudes where the distribution of both pines overlaps, a natural introgression hybrid exists, which we will further refer to as the Z pine. This pine has a morphological character that shares attributes of both P. massoniana and P. hwangshanensis. However, compared to the other two pines, its reproductive structure, the pinecone, has an ultra-low ripening rate with seeds that germinate poorly. In this study, we aimed to find the reason for the impaired cone maturation by comparing transcriptome libraries of P. massoniana and Z pine cones at seven successive growth stages. After sequencing and assembly, we obtained unigenes and then annotated them against NCBI’s non-redundant nucleotide and protein sequences, Swiss-Prot, Clusters of Orthologous Groups, Gene Ontology and KEGG Orthology databases. Gene expression levels were estimated and differentially expressed genes (DEGs) of the two pines were mined and analyzed. We found that several of them indeed relate to reproductive process. At every growth stage, these genes are expressed at a higher level in P. massoniana than in the Z pine. These data provide insight into understanding which molecular mechanisms are altered between P. massoniana and the Z pine that might cause changes in the reproductive process.

Triturus newts

Ben Wielstra introduces Eurasian newts of the genus Triturus.

Insights from genomes into the evolutionary importance and prevalence of hybridization in nature.

Hybridization is an evolutionary phenomenon that has fascinated biologists for centuries. Prior to the advent of whole-genome sequencing, it was clear that hybridization had played a role in the evolutionary history of many extant taxa, particularly plants. The extent to which hybridization has contributed to the evolution of Earth's biodiversity has, however, been the topic of much debate. Analyses of whole genomes are providing further insight into this evolutionary problem. Recent studies have documented ancient hybridization in a diverse array of taxa including mammals, birds, fish, fungi, and insects. Evidence for adaptive introgression is being documented in an increasing number of systems, though demonstrating the adaptive function of introgressed genomic regions remains difficult. And finally, several new homoploid hybrid speciation events have been reported. Here we review the current state of the field and specifically evaluate the additional insights gained from having access to whole-genome data and the challenges that remain with respect to understanding the evolutionary relevance and frequency of ancient hybridization, adaptive introgression, and hybrid speciation in nature.

Microsatellite analysis reveals the resilience of genetic diversity within extant populations of three Akebia species to chronic forest fragmentation in China

The species of Akebia Decne are well-known medicinal plants and have been used in Chinese herbalism for at least 2000 years. The Akebia fruits have a delicious flavor and high nutritional value and are especially rich in antineoplastic, diuretic and antiphlogistic properties, making them worthy of being exploited as a new high-value crop. However, the nature habitats of the Akebia species have been fragmented due to rapid and intensive expansion of agricultural land. In this study, population genetics based on simple sequence repeat markers were conducted on the three most economically important Akebia species in order to reveal the genetic effects of habitat fragmentation to gain useful information for conservation and management of genetic resources of Akebia. Three Akebia species maintained high within-population genetic variation and low level of genetic differentiation. Mantel’s test indicated that there was a significantly positive correlation between genetic distance and geographic distance. Our results reveal the resilience of genetic diversity within extant populations of three Akebia species to chronic forest fragmentation. For breeding purpose, germplasm collection for sampling as many individuals as possible from many populations to maximize genetic diversity of the species is recommended. In addition, natural introgression events have been revealed by Cluster and Structure analysis where admixed ancestry occurred across three marginal populations of A. trifoliata ssp. australis, suggesting these populations should be given priority for germplasm collection.

Implications of introgression for wildlife translocations: the case of North American martens

The evolutionary consequences of natural introgression provide a rare opportunity to retrospectively evaluate how the introduction of exotics or genetic rescue efforts may impact endemic faunas. Phylogeographic structure among mainland, endemic insular, and introduced North American marten (Martes americana and M. caurina) populations have been shaped by a complex history of natural, post-glacial population expansion followed by a series of anthropogenic introductions. In some cases, both natural colonization and translocations facilitated secondary contact, offering a series of replicated experiments that demonstrate how introgression, in these cases following isolation (insular and refugial), shapes genetic diversity. We test whether genetic exchange is occurring between North American marten species using mitochondrial genomes and ten nuclear loci. We present evidence of biased nuclear introgression from M. caurina into M. americana across two natural hybrid zones (insular and mainland) and found no remnant evidence of M. caurina on islands that received M. americana translocations, suggesting prior absence, potential extirpation, or genetic swamping of M. caurina from these islands. Our results highlight the importance of understanding phylogeographic variation prior to identifying source populations for wildlife translocations and caution the use of genetic rescue for North American marten populations. Although previously managed as a single species, these two species show substantial genetic divergence. When the two are placed into contact, they exhibit unidirectional, asymmetric introgression with potentially negative consequences for M. caurina, underscoring the value of mindful consideration of introgression in wildlife management.

Integrating Bayesian genomic cline analyses and association mapping of morphological and ecological traits to dissect reproductive isolation and introgression in a Louisiana Iris hybrid zone.

Hybrid zones provide unique opportunities to examine reproductive isolation and introgression in nature. We utilized 45,384 single nucleotide polymorphism (SNP) loci to perform association mapping of 14 floral, vegetative and ecological traits that differ between Iris hexagona and Iris fulva, and to investigate, using a Bayesian genomic cline (BGC) framework, patterns of genomic introgression in a large and phenotypically diverse hybrid zone in southern Louisiana. Many loci of small effect size were consistently found to be associated with phenotypic variation across all traits, and several individual loci were revealed to influence phenotypic variation across multiple traits. Patterns of genomic introgression were quite heterogeneous throughout the Louisiana Iris genome, with I.hexagona alleles tending to be favoured over those of I. fulva. Loci that were found to have exceptional patterns of introgression were also found to be significantly associated with phenotypic variation in a small number of morphological traits. However, this was the exception rather than the rule, as most loci that were associated with morphological trait variation were not significantly associated with excess ancestry. These findings provide insights into the complexity of the genomic architecture of phenotypic differences and are a first step towards identifying loci that are associated with both trait variation and reproductive isolation in nature.

Pattern of natural introgression in a Nothofagus hybrid zone from South American temperate forests

Interspecific gene flow is a common phenomenon in Nothofagaceae species; however, the dynamics of introgression in hybrid zones remains largely unknown. We focused on two ecologically and morphologically different Nothofagus species from Patagonia, Nothofagus nervosa and Nothofagus obliqua. In a natural hybrid zone, we established two plots 280 m apart in altitude (ca. 1.9 °C difference in mean temperature), and two subplots which captured microsite variation (abundance and spatial distribution of species and predominance of wind direction). We used intensive sampling of individuals (2055, including adults and regeneration) and molecular genotyping of 6 highly species-specific nuclear microsatellites for the identification and classification of hybrids, based on estimates of ancestry and interclass heterozygosity. We evaluated the relative contribution of our sampling effects to variation in hybrid incidence and direction of introgression using generalized linear mixed effects models. We determined that introgressive hybridization occurs at a global rate of 7.8% and that variation was mostly explained by plots (frequency at low altitude was approximately twice that found at high altitude), while it was less influenced by subplots. The high altitude plot was dominated by late-generation backcrosses to N. obliqua (asymmetric bimodality), whereas the low altitude plot consisted of intermediate hybrids (unimodality) and showed asymmetry for introgression between subplots. Differences were not detected between adults and regeneration, suggesting early-acting reproductive isolating barriers. F1 hybrids occur at a global frequency of 3.8%, and are fertile, as the detection of first- and late-generation hybrids indicates.

Unravelling wild carrot differentiation in Europe – preliminary data on a candidate gene approach

Carrot is an outcrossing species and levels of gene flow between populations, and even between wild and domesticated relatives, are expected to be high. Cases of natural hybridization and introgression of crops and wild relatives have been reported. Have these events diluted any putative habitat-adapted genotypes? In other words, can we still find a correlation between wild carrot genotypes and regional/local environment? We have chosen to start addressing this question using a member of the alternative oxidase (AOX) gene family. AOX genes seem to be linked to all kinds of abiotic and biotic stress reactions. Wild carrots were sampled in an environmental gradient across Western Europe. This gradient included sampling points with more deviating conditions, such as Sierra de Guadarrama or the central Pyrenees and the French Massif Central. Phylogenetic reconstruction on this molecular marker is to be combined with geographic, climatic, and ecological evidence. So far, the preliminary results suggest the existence of a biogeographical barrier at the Pyrenees, and higher gene diversity than initially expected. From an applied point of view, diversity of functional traits is much more relevant than species diversity. Gene transfer from wild to cultivated plants has contributed to the evolution of crop species. Providing that deterioration of genetic resources and biodiversity loss have not been drastic, gene transfer from wild plants has the potential to further contribute to a (targeted) improvement of cultivars.

Species-specific markers provide molecular genetic evidence for natural introgression of bullhead catfishes in Hungary.

Since three bullhead catfish species were introduced to Europe in the late 19th century, they have spread to most European countries. In Hungary, the brown bullhead (Ameiurus nebulosus) was more widespread in the 1970s–1980s, but the black bullhead (Ameiurus melas) has gradually supplanted since their second introduction in 1980. The introgressive hybridization of the two species has been presumed based on morphological examinations, but it has not previously been supported by genetic evidence. In this study, 11 different Hungarian habitats were screened with a new species-specific nuclear genetic, duplex PCR based, marker system to distinguish the introduced catfish species, Ameiurus nebulosus, Ameiurus melas, and Ameiurus natalis, as well as the hybrids of the first two. More than 460 specimens were analyzed using the above markers and additional mitochondrial sequence analyses were also conducted on >25% of the individuals from each habitat sampled. The results showed that only 7.9% of the specimens from two habitats belonged to Ameiurus nebulosus, and 92.1% were classified as Ameiurus melas of all habitats, whereas the presence of Ameiurus natalis was not detected. Two specimens (>0.4%) showed the presence of both nuclear genomes and they were identified as hybrids of Ameiurus melas and Ameiurus nebulosus. An additional two individuals showed contradicting results from the nuclear and mitochondrial assays as a sign of a possible footprint of introgressive hybridization that might have happened two or more generations before. Surprisingly, the level of hybridization was much smaller than expected based on the analyses of the North American continent’s indigenous stock from the hybrid zones. This phenomenon has been observed in several invasive fish species and it is regarded as an added level of complexity in the management of their rapid adaptation.

Introgression Threatens the Genetic Diversity of Quercus austrocochinchinensis (Fagaceae), an Endangered Oak: A Case Inferred by Molecular Markers.

Natural introgression can cause negative effects where rare species experience genetic assimilation and invade by their abundant congeners. Quercus austrocochinchinensis and Q. kerrii (subgenus Cyclobalanopsis) are a pair of closely related species in the Indo-China area. Morphological intermediates of the two species have been reported in this region. In this study, we used AFLP, SSR and two key leaf morphological diagnostic traits to study the two Q. austrocochinchinensis populations, two pure Q. kerrii and two putative hybrid populations in China. Rates of individual admixture were examined using the Bayesian clustering programs STRUCTURE and NewHybrids, with no a priori species assignment. In total, we obtained 151 SSR alleles and 781 polymorphic loci of AFLP markers. Population differentiation inferred by SSR and AFLP was incoherent with recognized species boundaries. Bayesian admixture analyses and principal coordinate analysis identified more hybrids and backcrossed individuals than morphological intermediates in the populations. SSR inferred a wide genetic assimilation in Q. austrocochinchinensis, except for subpopulation D2 in the core area of Xi-Shuang-Ban-Na Nature Reserve (XSBN). However, AFLP recognized more Q. austrocochinchinensis purebreds than SSR. Analysis using NewHybrids on AFLP data indicated that these hybridized individuals were few F2 and predominantly backcrosses with both parental species. All these evidences indicate the formation of a hybrid swarm at XSBN where the two species co-exist. Both AFLP and SSR recognized that the core protected area of XSBN (D2) has a high percentage of Q. austrocochinchinensis purebreds and a unique germplasm. The Hainan population and the other subpopulations of XSBN of the species might have lost their genetic integrity. Our results revealed a clear genetic differentiation in the populations and subpopulations of Q. austrocochinchinensis and ongoing introgression between Q. austrocochinchinensis and Q. kerrii at the disturbed contact areas. Combining the results from genetic and morphological analyses, the conservation of subpopulation D2 should be prioritized. Conservation and restoration of the integrity of tropical ravine rainforest is an important long-term goal for the successful conservation of Q. austrocochinchinensis. The fine-scale landscape might play an essential role in shaping the spatial patterns of hybridization. Further studies are needed to evaluate these patterns and dynamics.

Distorted segregation in plant hybrids and its implication for evolution.

During natural hybridization and introgression, offspring may obtain alleles from both parents unevenly, resulting in allelic segregation that significantly deviated from the Mendelian ratio, referred to as segregation distortion (SD). Segregation distortion is a common phenomenon and a number of factors can influence the ratio and pattern of segregation distortion in hybrid descendants. However, knowledge on the evolutionary implication of SD is still very limited. In this review, we summarize the current knowledge regarding mechanisms of SD such as genetic interaction, genetic divergence, cytoplasmic background, and environmental effect, and analyze its implication for evolution. In short, SD affects evolutionary potential, sex ratio, genetic diversity and maintenance of stable genetic divergence. Therefore, the effect of transgene on the evolutionary potential of wild relative populations through SD and the changes of the pattern of SD in continuous generations should be paid attention to in future studies.

Hybrid Incompatibilities, Local Adaptation, and the Genomic Distribution of Natural Introgression between Species.

Under allopatric speciation, geographic barriers eliminate gene flow between eventual species at all loci in the genome simultaneously. There is increasing evidence, however, that speciation can be complex, with some loci experiencing gene flow during speciation or during bouts of secondary contact. In taxa with heteromorphic sex chromosomes-birds, butterflies, mammals, and Drosophila-the X (or Z) chromosome generally shows reduced levels of gene flow compared to autosomes. To investigate why, we develop population genetic models of secondary contact and gene flow at a neutral locus that is genetically linked to selected loci involved in hybrid incompatibilities and/or local adaptation. Using models that assume weak migration and strong selection, we compare gene flow at X-linked versus autosomal neutral loci as a function of linkage, dominance, sex-specific selection, and sex-specific recombination. For most cases, gene flow at neutral loci on the X is reduced relative to autosomes, as the greater efficacy of hemizygous selection in XY hybrids reduces the opportunity for neutral migrant alleles to escape their genetically linked, locally disfavored alleles via recombination. There are some circumstances, however, involving sex-limited selection and sex-limited recombination that allow neutral loci on the X to introgress more readily than those on autosomes.

Interspecific introgressive origin of genomic diversity in the house mouse.

We report on a genome-wide scan for introgression between the house mouse (Mus musculus domesticus) and the Algerian mouse (Mus spretus), using samples from the ranges of sympatry and allopatry in Africa and Europe. Our analysis reveals wide variability in introgression signatures along the genomes, as well as across the samples. We find that fewer than half of the autosomes in each genome harbor all detectable introgression, whereas the X chromosome has none. Further, European mice carry more M. spretus alleles than the sympatric African ones. Using the length distribution and sharing patterns of introgressed genomic tracts across the samples, we infer, first, that at least three distinct hybridization events involving M. spretus have occurred, one of which is ancient, and the other two are recent (one presumably due to warfarin rodenticide selection). Second, several of the inferred introgressed tracts contain genes that are likely to confer adaptive advantage. Third, introgressed tracts might contain driver genes that determine the evolutionary fate of those tracts. Further, functional analysis revealed introgressed genes that are essential to fitness, including the Vkorc1 gene, which is implicated in rodenticide resistance, and olfactory receptor genes. Our findings highlight the extent and role of introgression in nature and call for careful analysis and interpretation of house mouse data in evolutionary and genetic studies.

Evidence to support natural hybridization between Anopheles sinensis and Anopheles kleini (Diptera: Culicidae): possibly a significant mechanism for gene introgression in sympatric populations

BackgroundMalaria caused by Plasmodium vivax is still a public health problem in the Republic of Korea (ROK), particularly regarding the recent re-emergence of this malarial species near the demilitarized zone in northwestern Paju City, Gyeonggi-do Province. Currently, at least 4 species (An. kleini, An. pullus, An. belenrae and An. lesteri) of the Hyrcanus Group are reported as possible natural vectors of vivax malaria in the ROK, and An. sinensis, which is the most dominant species, has long been incriminated as an important natural vector of this P. vivax. However, An. sinensis was ranked recently as a low potential vector. According to the discovery of natural hybrids between An. sinensis (a low potential vector for P. vivax) and An. kleini (a high potential vector for P. vivax) in Paju City, intensive investigation of this phenomenon is warranted under laboratory conditions.MethodsMosquitoes were collected during 2010-2012 from Paju City, ROK. Hybridization experiments used iso-female line colonies of these anophelines together with DNA analysis of ribosomal DNA [second internal transcribed spacer (ITS2)] and mitochondrial DNA [cytochrome c oxidase subunit I (COI)] of the parental colonies, F1-hybrids and repeated backcross progenies were performed intensively by using a PCR-based assay and pyrosequencing technology.ResultsThe results from hybridization experiments and molecular investigations revealed that the mitochondrial COI gene was introgressed from An. sinensis into An. kleini. The An. sinensis progenies obtained from consecutive repeated backcrosses in both directions, i.e., F2-11 progeny [(An. sinensis x An. kleini) x An. sinensis] and F3-5 progeny [(An. kleini x An. sinensis) x An. kleini] provided good supportive evidence.ConclusionsThis study revealed introgression of the mitochondrial COI gene between An. sinensis and An. kleini through consecutive repeated backcrosses under laboratory conditions. This new body of knowledge will be emphasized in reliable promising strategies in order to replace the population of An. kleini as a high potential vector for P. vivax, with that of a low potential vector, An. sinensis, through the mechanism of gene introgression in nature.

Evidence of low-levels of natural introgression between Pinus jaliscana and Pinus oocarpa in an open-pollinated progeny trial using near-infrared spectroscopy

There have been no reports of natural introgression between Pinus jaliscana and Pinusoocarpa where they occur sympatrically in the highlands of western Mexico. However, a small number of putative hybrid trees were observed in a progeny trial of 57 open-pollinated families of P. jaliscana at De Rust, South Africa that represented all nine known natural populations of the species in its native geographic range. The objectives of the study were to confirm hybridity using traditional botanical analysis of needle traits (no cones were available) and once admixtures were confirmed to develop near infrared spectroscopy (NIRS) models to define the spectral signature of “pure P. jaliscana” and the hybrid. The calibration models were then run on a large subsample of 260 trees that served as a validation data set to predict the range-wide occurrence of introgression in P. jaliscana. The calibration data set for the NIRS included 57 pure P. jaliscana and 31 hybrid trees identified in the trial that were morphologically separated using canonical analysis based on four external needle traits: number of needles per fascicle, needle length, needle width, fascicle sheath length and three needle color traits: hue, value and chroma. NIRS models were developed using 1g of ground, oven-dried needles/tree that were scanned at visible and NIR wave lengths between 400 and 2500nm. The raw spectral data, log(1/reflectance) were processed using standard normal variate and de-trending transformations. The NIRS calibration model identified six hybrid trees in the validation population for an overall species admixture rate of 2.3% versus 7.7% based on subjective scoring in the field. The implications of P. oocarpa introgression in P. jaliscana in natural stands in western Mexico are discussed. NIRS provides a quick way to obtain reasonable estimates of species admixtures in progeny trials and is an inexpensive surrogate to using molecular markers. NIRS seems most practical when species are very closely related and/or there is unbalanced representation of parental species in progeny tests to make comparisons with putative hybrids difficult.