Nuclear microsatellite variation in Malagasy baobabs (Adansonia, Bombacoideae, Malvaceae) reveals past hybridization and introgression.

BACKGROUND AND AIMS: Adansonia comprises nine species, six of which are endemic to Madagascar. Genetic relationships between the Malagasy species remain unresolved due to conflicting results between nuclear and plastid DNA variation. Morphologically intermediate individuals between distinct species have been identified, indicative of interspecific hybridization. In this paper, microsatellite data are used to identify potential cases of hybridization and to provide insights into the evolutionary history of the genus on Madagascar. METHODS: Eleven microsatellites amplified with new primers developed for Adansonia rubrostipa were used to analyse 672 individuals collected at 27 sites for the six Malagasy species and morphologically intermediate individuals. Rates of individual admixture were examined using three Bayesian clustering programs, STRUCTURE, BAPS and NewHybrids, with no a priori species assignment. KEY RESULTS: Population differentiation was coherent, with recognized species boundaries. In the four Malagasy species of section Longitubae, 8·0, 9·0 and 9·5 % of individuals with mixed genotypes were identified by BAPS, NewHybrids and STRUCTURE, respectively. At sites with sympatric populations of A. rubrostipa and A. za, NewHybrids indicated these individuals to be F2 and, predominantly, backcrosses with both parental species. In northern Madagascar, two populations of trees combining A. za and A. perrieri morphology and microsatellite alleles were identified in the current absence of the parental species. CONCLUSIONS: The clear genetic differentiation observed between the six species may reflect their adaptation to different assortments of climate regimes and habitats during the colonization of the island. Microsatellite variation reveals that hybridization probably occurred in secondary contact between species of section Longitubae. This type of hybridization may also have been involved in the differentiation of a local new stabilized entity showing specific microsatellite alleles and morphological characters, suggesting a potential role of hybridization in the recent history of diversification on Madagascar.

The diversification of terpene emissions in Mediterranean oaks: lessons from a study of Quercus suber, Quercus canariensis and its hybrid Quercus afares.

Interspecific gene flow is common in oaks. In the Mediterranean, this process produced geographical differentiations and new species, which may have contributed to the diversification of the production of volatile terpenes in the oak species of this region. The endemic North African deciduous oak Quercus afares (Pomel) is considered to be a stabilized hybrid between the evergreen Quercus suber (L.) and the deciduous Quercus canariensis (Willd.), presumably being monoterpene and isoprene emitters, respectively. In a common garden experiment, we examined the terpene emission capacities, terpene synthase (TPS) activities and nuclear genetic markers in 52 trees of these three oak species. All but one of the Q. suber and Q. canariensis trees were found to be genetically pure, whereas most Q. afares trees possessed a mixed genotype with a predominance of Q. suber alleles. Analysis of the foliar terpene emissions and TPS activities revealed that all the Q. canariensis trees strongly produced isoprene while all the Q. suber trees were strong monoterpene producers. Quercus afares trees produced monoterpenes as well but at more variable and significantly lower rates, and with a monoterpene pattern different than that observed in Q. suber. Among 17 individuals tested, one Q. afares tree emitted only an insignificant amount of terpenes. No mixed isoprene/monoterpene emitter was detected. Our results suggest that the capacity and pattern of volatile terpene production in Algerian Q. afares populations have strongly diverged from those of its parental species and became quantitatively and qualitatively reduced, including the complete suppression of isoprene production.

Relevance of genetics for conservation policies: the case of Minorcan cork oaks.

BACKGROUND AND AIMS: Marginal populations of widely distributed species can be of high conservation interest when they hold a significant or unique portion of the genetic diversity of the species. However, such genetic information is frequently lacking. Here the relevance of genetic surveys to develop efficient conservation strategies for such populations is illustrated using cork oak (Quercus suber) from Minorca (Balearic Islands, Spain) as a case study. Cork oak is highly endangered on the island, where no more than 67 individuals live in small, isolated stands in siliceous sites. As a consequence, it was recently granted protected status. METHODS: Two Bayesian clustering approaches were used to analyse the genetic structure of the Minorcan population, on the basis of nuclear microsatellite data. The different groups within the island were also compared with additional island and continental populations surrounding Minorca. KEY RESULTS: Very high genetic diversity was found, with values comparable with those observed in continental parts of the species' range. Furthermore, the Minorcan oak stands were highly differentiated from one another and were genetically related to different continental populations of France and Spain. CONCLUSIONS: The high levels of genetic diversity and inter-stands differentiation make Minorcan cork oak eligible for specific conservation efforts. The relationship of Minorcan stands to different continental populations in France and Spain probably reflects multiple colonization events. However, discrepancy between chloroplast DNA- and nuclear DNA-based groups does not support a simple scenario of recent introduction. Gene exchanges between neighbouring cork oak stands and with holm oak have created specific and exceptional genetic combinations. They also constitute a wide range of potential genetic resources for research on adaptation to new environmental conditions. Conservation guidelines that take into account these findings are provided.

Ancient and current gene flow between two distantly related Mediterranean oak species, Quercus suber and Q. ilex.

BACKGROUND AND AIMS: Quercus suber and Q. ilex are distantly related and their distributions partially overlap. They hybridize occasionally, but the complete replacement of Q. suber chloroplast DNA (cpDNA) by that of Q. ilex was identified in two specific geographical areas. The objective of this study was to determine whether the contrasting situation reflected current or recent geographical interspecies gene flow variation or was the result of ancient introgression. METHODS: cpDNA PCR-RFLPs (restriction fragment length polymorphisms) and variation at ten nuclear microsatellite loci were analysed in populations of each species, in 16 morphologically intermediate individuals and the progeny of several of them. Interspecies nuclear introgression was based on individual admixture rates using a Bayesian approach with no a priori species assignment, and on a maximum-likelihood (ML) method, using allele frequencies in the allopatric populations of each species as controls. Gene flow was compared specifically between populations located within and outside the specific areas. KEY RESULTS: High interspecies nuclear genetic differentiation was observed, with twice the number of alleles in Q. ilex than in Q. suber. According to Bayesian assignment, approx. 1 % of individuals had a high probability of being F(1) hybrids, and bidirectional nuclear introgression affected approx. 4 % of individuals in each species. Hybrid and introgressed individuals were identified predominantly in mixed stands and may have a recent origin. Higher proportions including allospecific genes recovered from past hybridization were obtained using the ML method. Similar rates of hybridization and of nuclear introgression, partially independent of cpDNA interspecies transfer suggestive of gene filtering, were obtained in the populations located within and outside the areas of complete cpDNA replacement. CONCLUSIONS: The results did not provide evidence for geographical variation in interspecies gene flow. In contrast, historical introgression is supported by palynological records and constitutes the more reliable origin of cpDNA replacement in specific regions.

Chloroplast DNA phylogeography suggests a West African centre of origin for the baobab, Adansonia digitata L. (Bombacoideae, Malvaceae).

The African baobab (Adansonia digitata L.) is an emblematic, culturally important, and physically huge tropical tree species whose natural geographical distribution comprises most of tropical Africa, but also small patches of southern Arabia, and several Atlantic and Indian Ocean islands surrounding the African continent, notably including Madagascar. We analysed the polymerase chain reaction-restriction fragment length polymorphism of five chloroplast DNA fragments obtained from 344 individuals of A. digitata collected from 74 populations covering the entire extant distribution range of the species. Our goal was to reconstruct the phylogeographical history of the species and, if possible, to identify its centre of origin, which has been a subject of controversy for many decades. We identified five haplotypes whose distribution is clearly geographically structured. Using several species of Adansonia and of closely related genera as outgroups, the haplotypes showed a clear phylogeographical pattern of three groups. Two are phylogenetically related to the outgroup taxa, and are distributed in West Africa. The third group is substantially more differentiated genetically from outgroup species, and it corresponds to southern and eastern Africa, Arabia and the Indian Ocean islands, including Madagascar. According to our results, the tetraploid A. digitata, or its diploid progenitor, probably originated in West Africa and migrated subsequently throughout the tropical parts of that continent, and beyond, by natural and human-mediated terrestrial and overseas dispersal.

Phylogeographical variation of chloroplast DNA in cork oak (Quercus suber).

BACKGROUND AND AIMS: In the last decades, the geographical location of the centre of origin of Quercus suber (cork oak), a strictly western Mediterranean oak species, has been the subject of controversy. METHODS: RFLP variation over the whole chloroplast DNA molecule and PCR-RFLPs over seven specific cpDNA fragments were analysed phylogeographically to reconstruct the evolutionary history of cork oak. KEY RESULTS: Nine chlorotypes of the 'suber' cpDNA lineage were identified throughout the species range. Using closely related Mediterranean oak species as outgroup, the chlorotypes showed a clear phylogeographical pattern of three groups corresponding to potential glacial refuges in Italy, North Africa and Iberia. The most ancestral and recent groups were observed in populations located in the eastern and western parts of the species range, respectively. Several unrelated chlorotypes of the 'ilex' cpDNA lineage were also identified in specific western areas. CONCLUSIONS: The results support a Middle-Eastern or a central Mediterranean origin for cork oak with subsequent westward colonization during the Tertiary Period, and suggest that the 'ilex' chlorotype variation does not reflect entirely cytoplasmic introgression by Q. ilex but originated partly in Q. suber.

Isoprenoid emissions of Quercus spp. (Q. suber and Q. ilex) in mixed stands contrasting in interspecific genetic introgression.

• Among oak species, Quercus ilex is classified as a monoterpene emitter and Q. suber is mainly known as a nonisoprenoid emitter. The extent and origin of this diversification is unknown. • We examined intra- and interspecific emission variability in two mixed stands which differed in their level of hybridization and reciprocal genetic introgression based on variations in cytoplasmic (chloroplast DNA) and nuclear (allozyme) markers. • At both sites all trees identified as Q. ilex, or as recent descendants from Q. ilex × Q. suber hybrids, emitted monoterpenes. Of Q. suber trees (genetically introgressed or not by Q. ilex), 91% were also monoterpene emitters, and the remainder nonemitters. One tree identified as a Q. canariensis × Q. ilex hybrid emitted both isoprene and monoterpenes. Compared with Q. ilex, the standard emission rate of Q. suber was higher in summer and lower in autumn. Both species emitted the same monoterpenes, proportions of which showed significant intra- and interspecific variability. • The results suggest that Q. suber populations in the French Mediterranean intrinsically emit monoterpenes, and that gene flow between oak species contributes to diversification of emission signatures.

ASSOCIATION BETWEEN QUANTITATIVE TRAITS AND ALLOZYME HETEROZYGOSITY IN A TETRASOMIC SPECIES: DACTYLIS GLOMERATA.

Tetraploid individuals of orchardgrass (Dactylis glomerata L. subsp. hispanica Roth.) sampled from a natural population were used to evaluate the correlation between both single and multilocus heterozygosity at 7 enzyme loci, and several quantitative traits (plant size at time of collection, leaf weight, and panicle number in experimental trials). Four hundred and forty-eight plants were studied at the 7 loci and 288 of these individuals were scored for an additional eighth locus. Five genotype classes (monogenic, simplex, and duplex digenic, trigenic, and tetragenic) were distinguished according to their heterozygosity level. Multilocus heterozygosity showed a significant positive correlation with both leaf and panicle yield in experimental conditions, but not with original plant size, which was found to be markedly influenced by environmental microheterogeneity. Multilocus heterozygosity, estimated from both the number of heterozygous loci and the number of distinct alleles per locus, had a significant influence on plant performance. Individual locus effects were positive and significant at two loci (GOT1 and PX1). Panicle number increased regularly with heterozygosity level (from monogenics to tetragenics) at the GOT1 locus, as did leaf weight and panicle number at the PX1 locus. Such variation would be predicted by overdominance at these loci or at linked loci. Significant relationships between leaf yield and heterozygosity level at the GOT1 locus distinguished the homozygotes from the heterozygotes (of any class) and was thus more consistent with inbreeding effects. No significant differences were observed among the five genotype classes for any quantitative trait at the six remaining loci. At both the GOT1 and PX1 loci, heterozygosity had a significant independent effect on leaf weight and panicle number even when the correlation between these traits was removed by analysis of covariance.