Pedigree Reconstruction and Spatial Analysis for Genetic Testing and Selection In A Larix Kaempferi (Lamb.) Carrière Plantation

Background: Larix kaempferi is one of the major timber species in Northeast Asia. Demand for the reforestation of the species is rising in South Korea due to an increase in large timber production and utilization. However, progeny trials for the species have not been explored, making it challenging to foster advanced generations of tree improvement. In the present study, genetic testing and selection for diameter growth were conducted using pedigree reconstruction and phenotypic spatial distribution analysis in a plantation of L. kaempferi. The aim of the present study was to select the superior larch individuals using the pedigree reconstruction and phenotypic spatial distribution to substitute progeny trials. The plantation of seed orchard crops was established in 1990 and one-hundred and eighty-eight trees were selected as the study material. Genetic variation was investigated first to validate its adequacy as breeding material. Genetic testing was carried out using a model considering pedigree information and spatial autoregression of the phenotypes. The major environmental effects on growth were assessed to understand the site characteristics where the breeding materials were located.Results: The expected heterozygosity of the mother trees and offspring were 0.672 and 0.681 presenting the corresponding level of genetic variation between two groups. The pedigree reconstruction using maternity analysis assigned one to six progenies to ninety-two candidate mothers. The accuracy of genetic testing was exceedingly increased with the animal model considering AR1⊗AR1 structure compared to the animal model only. The estimated genetic variance of the former was 9.086 whereas that of the latter was 4.9E-5. The predicted breeding values of the offspring were ranged from -5.937 to 5.655 and the estimated heritability of diameter growth was 0.344. Shifts in the effects of the factors were detected using geographically weighted regression analysis, which provided information on the post-sectioning of the plantation. Conclusions: The genetic testing approach based on pedigree reconstruction and phenotypic spatial distribution analysis was considered a useful analytical scheme that could replace or supplement progeny trials. Subdivision of the plantation as blocks of the progeny testing was accomplished a posteriori based on the changes of the environmental effects.

The use of shared haplotype length information for pedigree reconstruction in asexually propagated outbreeding crops, demonstrated for apple and sweet cherry

Pedigree information is of fundamental importance in breeding programs and related genetics efforts. However, many individuals have unknown pedigrees. While methods to identify and confirm direct parent–offspring relationships are routine, those for other types of close relationships have yet to be effectively and widely implemented with plants, due to complications such as asexual propagation and extensive inbreeding. The objective of this study was to develop and demonstrate methods that support complex pedigree reconstruction via the total length of identical by state haplotypes (referred to in this study as “summed potential lengths of shared haplotypes”, SPLoSH). A custom Python script, HapShared, was developed to generate SPLoSH data in apple and sweet cherry. HapShared was used to establish empirical distributions of SPLoSH data for known relationships in these crops. These distributions were then used to estimate previously unknown relationships. Case studies in each crop demonstrated various pedigree reconstruction scenarios using SPLoSH data. For cherry, a full-sib relationship was deduced for ‘Emperor Francis, and ‘Schmidt’, a half-sib relationship for ‘Van’ and ‘Windsor’, and the paternal grandparents of ‘Stella’ were confirmed. For apple, 29 cultivars were found to share an unknown parent, the pedigree of the unknown parent of ‘Cox’s Pomona’ was reconstructed, and ‘Fameuse’ was deduced to be a likely grandparent of ‘McIntosh’. Key genetic resources that enabled this empirical study were large genome-wide SNP array datasets, integrated genetic maps, and previously identified pedigree relationships. Crops with similar resources are also expected to benefit from using HapShared for empowering pedigree reconstruction.

Grapevine (Vitis vinifera L.) varietal assortment and evolution in the Marche region (central Italy)

Italy is one of the richest countries in terms of grapevine varietal assortment, and the Marche region in central Italy has an ancient winegrowing tradition. Increasing interest in autochthonous grapevine varieties prompted efforts to recover and identify local minor germplasm also in this region, and to search for pedigree relationships and determine the evolution of varietal assortment. In the present study this was done using nuclear and chloroplast microsatellite markers and SNP markers from the Vitis18kSNP chip. Eighteen new genotypes were found, of which seventeen belonged to well-known, less known and even unknown vines found to be specific to the Marche region. The rearranged pedigree highlighted the complex intertwining relationships between Marche varieties. Second-degree relationships were also derived. Some minor Marche varieties have increased the number of parent-offspring related members of Garganega, Sangiovese, Crepolino/Visparola and Sciaccarello, which are varieties already recognised as founders or recurrent parents of many Italian cultivars. Crepolino/Visparola was shown to be a key variety in the evolution of the Marche varietal assortment, having played the role of parent and grandparent, as explained by the presence of this genotype in the region in ancient times. Surprisingly, Semidano, a well-known Sardinian variety, played a clear role as parent or grandparent of three minor Marche varieties, testifying to its presence in central Italy in ancient times. Incrocio Bruni 54 was confirmed as an offspring of Sauvignon and Verdicchio, as declared by the breeder. The parentage of Lacrima was completed: this variety was found to be a spontaneous cross between Nera Rada and Aleatico.A dendrogram of genetic similarity clearly resembled the groups defined with the pedigree reconstruction and it gave an indication of the genetic similarity of the varieties excluded from the strictest parentage links.

SNP-based analysis reveals unexpected features of genetic diversity, parental contributions and pollen contamination in a white spruce breeding program

Accurate monitoring of genetic diversity levels of seedlots and mating patterns of parents from seed orchards are crucial to ensure that tree breeding programs are long-lasting and will deliver anticipated genetic gains. We used SNP genotyping to characterize founder trees, five bulk seed orchard seedlots, and trees from progeny trials to assess pollen contamination and the impact of severe roguing on genetic diversity and parental contributions in a first-generation open-pollinated white spruce clonal seed orchard. After severe roguing (eliminating 65% of the seed orchard trees), we found a slight reduction in the Shannon Index and a slightly negative inbreeding coefficient, but a sharp decrease in effective population size (eightfold) concomitant with sharp increase in coancestry (eightfold). Pedigree reconstruction showed unequal parental contributions across years with pollen contamination levels between 12 and 51% (average 27%) among seedlots, and 7–68% (average 30%) among individual genotypes within a seedlot. These contamination levels were not correlated with estimates obtained using pollen flight traps. Levels of pollen contamination also showed a Pearson’s correlation of 0.92 with wind direction, likely from a pollen source 1 km away from the orchard under study. The achievement of 5% genetic gain in height at rotation through eliminating two-thirds of the orchard thus generated a loss in genetic diversity as determined by the reduction in effective population size. The use of genomic profiles revealed the considerable impact of roguing on genetic diversity, and pedigree reconstruction of full-sib families showed the unanticipated impact of pollen contamination from a previously unconsidered source.

Evaluation of forest tree breeding strategies based on partial pedigree reconstruction through simulations: Pinus pinaster and Eucalyptus nitens as case studies

Despite recent developments in molecular markers, most forest tree breeding programmes do not use them routinely. One way to integrate markers would be to use them for pedigree reconstruction after a simplified mating design through polymix or open-pollinated breeding. Thanks to the latest developments in the POPSIM simulator, various breeding strategies, including some based on paternity recovery, were evaluated with specified constraints on the level of diversity over breeding cycles. These simulations were carried out in two case studies: the French Pinus pinaster (Ait.) and the New Zealand Eucalyptus nitens (H. Dean & Maiden) Maiden breeding programmes. The Pinus pinaster case study produced lower genetic gain for the polymix breeding strategy with paternity recovery compared with double-pair mating or optimal-contribution strategies. However, the polymix breeding strategy could be of interest if the mating design is faster to complete. In the Eucalyptus nitens case study, pedigree recovery was shown to be a mandatory step to controlling the erosion of diversity over breeding cycles. In both cases, the strategies based on pedigree reconstruction were applicable with a limited level of genotyping. Finally, these simulations allow some general recommendations to be drawn to help breeders when designing a strategy for forest tree breeding.