Mervyn Shepherd

Transpecific microsatellites for hard pines

Abstract.Microsatellites are difficult to recover from large plant genomes so cross-specific utilisation is an important source of markers. Fifty microsatellites were tested for cross-specific amplification and polymorphism to two New World hard pine species, slash pine (Pinus elliottii var. elliottii) and Caribbean pine (P. caribaea var. hondurensis). Twenty-nine (58%) markers amplified in both hard pine species, and 23 of these 29 were polymorphic. Soft pine (subgenus Strobus) microsatellite markers did amplify, but none were polymorphic. Pinus elliottii var. elliottii and P. caribaea var. hondurensis showed mutational changes in the flanking regions and the repeat motif that were informative for Pinus spp. phylogenetic relationships. Most allele length variation could be attributed to variability in repeat unit number. There was no evidence for ascertainment bias.

Genetic maps for Pinus elliottii var. elliottii and P. caribaea var. hondurensis using AFLP and microsatellite markers

AbstractnGenetic maps for individual Pinus elliottii var. elliottii and P. caribaea var. hondurensis trees were generated using a pseudo-testcross mapping strategy. A total of 329 amplified fragment length polymorphic (AFLP) and 12 microsatellite markers were found to segregate in a sample of 93 interspecfic F1 progeny. The male P. caribaea var. hondurensis parent was more heterozygous than the female P. elliottii var. elliottii parent with 19% more markers segregating on the male side. Framework maps were constructed using a LOD 5 threshold for grouping and interval support threshold of LOD 2. The framework map length for the P. elliottii var. elliottii megagametophyte parent (1,170xa0cM Kosambi; 23 linkage groups) was notably smaller than the P. caribaea var. hondurensis pollen parent (1,658xa0cM Kosambi; 27 linkage groups). The difference in map lengths was assumed to be due to sex-related recombination variation, which has been previously reported for pines, as the difference in map lengths not be accounted for by the larger number of markers mapping to the P. caribaea var. hondurensis parent – 109 compared with 78 in P. elliottii var. elliottii parent. Based on estimated genome sizes for these species, the framework maps for P. elliottii var. elliottii and P. caribaea var. hondurensis covered 82% and 88% of their respective genomes. The pseudo-testcross strategy was extended to include AFLP and microsatellite markers in an intercross configuration. These comprehensive maps provided further genome coverage, 1,548 and 1,828xa0cM Kosambi for P. elliottii var. elliottii and P. caribaea var. hondurensis, respectively, and enabled homologous linkage groups to be identified in the two parental maps. Homologous linkage groups were identified for 11 out of 24 P. elliottii var. elliottii and 10 out of 25 P. caribaea var. hondurensis groups. A higher than expected level of segregation distortion was found for both AFLP and microsatellite markers. An explanation for this segregation distortion was not clear, but it may be at least in part due to genetic mechanisms for species isolation in this wide cross.

Parental and consensus linkage maps of Eucalyptus globulus using AFLP and microsatellite markers

Abstract Parental and consensus maps were constructed in an F2 inter-provenance cross of Eucalyptus globulus, using amplified fragment length polymorphism (AFLP) and microsatellite (or simple sequence repeats [SSR]) markers. The female map had 12 linkage groups and 118 markers, comprising 33 SSR and 85 AFLP loci. The male map had 14 linkage groups and 130 markers comprising 36 SSR and 94 AFLP loci. The integrated map featured 10 linkage groups and 165 markers, including 33 SSR and 132 AFLP loci, a small 11th group was identified in the male parent. Moderate segregation distortion was detected, concentrated in gender specific groups. The strongest distortion was detected in the female parent for which causal mechanisms are discussed. The inclusion of SSR markers previously mapped in several different eucalypt species within the subgenus Symphyomyrtus (E. globulus, E. camaldulensis, and predominantly E. grandis and E. urophylla), allowed comparison of linkage groups across species and demonstrated that linkage orders previously reported in E. globulus, E. grandis and E. urophylla were largely conserved.

Genetic control of propagation traits in a single Corymbia torelliana × Corymbia variegata family

Genetic control of vegetative propagation traits was described for a second-generation, outbred, intersectional hybrid family (N = 208) derived from two species, Corymbia torelliana (F. Muell.) K.D. Hill & L.A.S. Johnson and Corymbia variegata (F. Muell.) K.D. Hill & L.A.S. Johnson, which contrast for propagation characteristics and in their capacity to develop lignotubers. Large phenotypic variances were evident for rooting and most other propagation traits, with significant proportions attributable to differences between clones (broad-sense heritabilities 0.2-0.5). Bare root assessment of rooting rate and root quality parameters tended to have the highest heritabilities, whereas rooting percentage based on root emergence from pots and shoot production were intermediate. Root biomass and root initiation had the lowest heritabilities. Strong favourable genetic correlations were found between rooting percentage and root quality traits such as root biomass, volume, and length. Lignotuber development on a seedling was associated with low rooting and a tendency to poor root quality in cuttings and was in accord with the persistence of species parent types due to gametic phase disequilibrium. On average, nodal cuttings rooted more frequently and with higher quality root systems, but significant cutting type x genotype interaction indicated that for some clones, higher rooting rates were obtained from tips. Low germination, survival of seedlings, and rooting rates suggested strong hybrid breakdown in this family.

Geographical and historical determinants of microsatellite variation in Eucalyptus pilularis

Geographically distributed genetic variation is expected in species that have wide latitudinal and habitat ranges, like Eucalyptus pilularis Sm. Coastal and inland ecotypes of this tall forest tree have been distinguished in genecological studies, but patterns of regionally distributed quantitative variation are weak. At the coarsest level, variation of 12 microsatellite markers divided a rangewide sample of 424 E.xa0pilularis trees into two zones: the region to the south of Sydney forming one zone and regions to the north forming another. Genetic structuring did not correspond with ecotypes but rather with a biogeographic division, suggesting an imprint of historical isolation. Typical and uniform levels of genetic diversity (He = 0.78 ± 0.02 (mean ± SE)) were found across 10 geographic regions. Genetic structuring by regions (PhiRT = 3%), by localities within regions (PhiPT = 2%), between coastal and inland provenances (PhiPT = 2%), or due to isolation by distance was subtle. These observations, along wit...

Comparative mapping among subsection Australes (genus Pinus, family Pinaceae)

Comparative mapping in conifers has not yet been used to test for small-scale genomic disruptions such as inversions, duplications, and deletions occurring between closely related taxa. Using comparative mapping to probe this smaller scale of inquiry may provide clues about speciation in a phylogenetically problematic taxon, the diploxylon pine subsection Australes (genus Pinus, family Pinaceae). Genetic maps were constructed for two allopatric species of Australes, P. elliottii var. elliottii and P. caribaea var. hondurensis, using microsatellites and an F1 hybrid. A third map was generated directly from the meiotic products of an adult F1 hybrid, eliminating the need for an F2 generation. Numerous small-scale disruptions were detected in addition to synteny and collinearity, and these included (1) map shrinkage, (2) a paracentric inversion, (3) transmission ratio distortion, and (4) mild selection against a parental haplotype. Such cryptic signatures of genomic divergence between closely related interfertile species are useful in elucidating this problematic evolutionary history.