José X. Chaparro

Genetic diversity of some wild almonds and related Prunus species revealed by SSR and EST-SSR molecular markers

Almond and its related wild species, which are widely distributed in Central and West Asia, have high genetic variation. This is an important source of genetic diversity for crop improvement. In this study a set of 32 SSR and 12 EST-SSR primer pairs were used to determine genetic diversity in 89 accessions of almond and other Prunus species. Most of the accessions (68) were collected from natural habitats of Iran. SSR primers amplified higher numbers of alleles than EST-SSR markers and discriminated genotypes more effectively. Results indicated high diversity among accessions. Observed heterozygosity (Ho) was 0.581. Nei’s index of diversity (He) and average number of alleles per locus (na) were 0.885 and 34, respectively. The mean value of polymorphism information content (PIC) was 0.874. The average Fst (F-statistics index) was 0.271 and the fixation index (Fis) was 0.151. Estimated variance among putative populations (AP) and individuals (AI) and within individuals (WI) were 5, 35, and 60%, respectively, which revealed that most of the variation was distributed among individuals rather than groups. Cultivated almonds were highly similar to P. fenzeliana, which is native to West Asia, supporting the importance of these regions in almond domestication. In the dendrogram of groups, minimum genetic distance was observed between Amygdalus and Orientalis groups from the Euamygdalus section. The Leptopus and Chameamygdalus sections were more distant from almonds than plums. The results also showed Dodecandra (Lycioides) series should be taxonomically classified closer to the section Euamygdalus.

Genetic diversity in peach [Prunus persica (L.) Batsch] at the University of Florida: past, present and future

The University of Florida (UF) stone fruit breeding and genetics program was created in 1952 to develop early ripening stone fruit cultivars with high quality, adaptation to summer rainfall, low chilling requirements, and the ability to withstand high disease pressure. Diverse germplasm sources were used to introduce desirable traits in UF breeding pool. The main objective of this research was to determine the genetic diversity and population structure of the breeding germplasm, and to search for loci under selection. A total of 195 peach genotypes were used: UF cultivars and advanced selections (n = 168), cultivars and selections from the UF-UGA-USDA joint breeding effort (n = 13), landrace cultivars (n = 4), high-chilling cultivars released by NCSU (n = 5), and related Prunus (n = 5) species. A total of 36 SSR markers distributed across the peach genome amplified 423 alleles. An average of 18 genotypes were detected per marker: A (number of observed alleles) of 11.43, Ae (effective number of alleles) of 2.58, Ho (observed heterozygosity) of 0.4, He (expected heterozygosity) of 0.52, F (Wright’s fixation index) of 0.25, and PIC (polymorphism information content) of 0.48. UPGMA cluster analysis based on Nei’s genetic distance represented best the known pedigree information for the germplasm pools. Two major groups were observed across the germplasm corresponding to melting and non-melting flesh cultivars/selections. Population structure results supported these two major groups. Several loci closely located to genome regions where different phenotypic traits have been previously mapped were detected to be under selection.

Genetic Parameters and the Impact of Off-Types for Theobroma cacao L. in a Breeding Program in Brazil

Breeding programs of cacao (Theobroma cacao L.) trees share the many challenges of breeding long-living perennial crops, and genetic progress is further constrained by both the limited understanding of the inheritance of complex traits and the prevalence of technical issues, such as mislabeled individuals (off-types). To better understand the genetic architecture of cacao, in this study, 13 years of phenotypic data collected from four progeny trials in Bahia, Brazil were analyzed jointly in a multisite analysis. Three separate analyses (multisite, single site with and without off-types) were performed to estimate genetic parameters from statistical models fitted on nine important agronomic traits (yield, seed index, pod index, % healthy pods, % pods infected with witches broom, % of pods other loss, vegetative brooms, diameter, and tree height). Genetic parameters were estimated along with variance components and heritabilities from the multisite analysis, and a trial was fingerprinted with low-density SNP markers to determine the impact of off-types on estimations. Heritabilities ranged from 0.37 to 0.64 for yield and its components and from 0.03 to 0.16 for disease resistance traits. A weighted index was used to make selections for clonal evaluation, and breeding values estimated for the parental selection and estimation of genetic gain. The impact of off-types to breeding progress in cacao was assessed for the first time. Even when present at <5% of the total population, off-types altered selections by 48%, and impacted heritability estimations for all nine of the traits analyzed, including a 41% difference in estimated heritability for yield. These results show that in a mixed model analysis, even a low level of pedigree error can significantly alter estimations of genetic parameters and selections in a breeding program.

Seed germination and seedling establishment of some wild almond species

Wild almond species are important genetic resources for resistance to unsuitable condition, especially drought stress. They have been used traditionally as rootstocks in some areas of Iran. So far, 21 wild almond species and 7 inter species hybrids have been identified in Iran. To study seed germination and seedling establishment of some of these species, three separate experiments were designed. In the first experiment, the application of gibberellic acid (GA 3 ) (0, 250, 500 and 750 ppm) for 24 h was studied on germination characteristics of four wild almond accessions after stratification at 5 ± 0.5°C in Perlite media. Germination percentage, index vigor and root initiation factors were different in almond accessions, but were not affected by hormonal treatments. In the second experiment, seeds of another six wild almond accessions were stratified to compare their germination ability. Germination percentage, index vigor and root initiation were different among accessions significantly. In the last experiment, the establishment and vigor of 14 accessions from eight almond species have been evaluated in plastic bags in outdoor conditions. Two ecotypes of Prunus spp. had the highest stem diameter and length at all growing stages. Keywords: Amygdalus, germination percentage, index vigor, root initiation, stem length, stem diameter.

Phylogenetic relationships among the first and second introns of selected Prunus S-RNase genes

Rahemi, A., Gradziel T.M., Chaparro J.X., Folta, K.M., Taghavi, T., Fatahi, R., Ebadi, A. and Hassani, D. 2015. Phylogenetic relationships among the first and second introns of selected Prunus S-RNase genes. Can. J. Plant Sci. 95: 1145-1154. To identify and evaluate self-incompatible alleles in almonds and related germplasm, DNA from 15 Prunus species was amplified using two degenerate consensus primer pairs flanking first and second S-locus introns (PaConsI-FD+EM-Pc1ConsRD and EM-Pc2ConsFD+EM-Pc3ConsRD). Twenty-eight amplified PCR products were analyzed by automated sequencer capillary electrophoresis. Sequenced fragments were aligned against available Prunus S-locus sequences in the National Center for Biotechnology Information and S-alleles identities were determined. The phylogenetic relationships between S-alleles in the germplasm studied were determined by the homology between their sequences and dendrograms were obtained for each primer pair. The Maximum Likelihood (homology) ranged from 84 to 100%. Most sequences were similar to cultivated almond (Prunus dulcis) or to the European wild almond (P. webbii). Twenty-six alleles for the first and the second introns were registered in the database in the GenBank. Two sequences of the first and second introns, which were taken from Prunus nairica and had similarity in GenBank, were registered in the database under a common sequence of the first and second intron. Analysis of phylogenetic relationships (dendrograms) among S-alleles from wild almond species as well as S-alleles cluster relations showed most pairs of alleles well supported by bootstrap.

A major locus for resistance to Botryosphaeria dothidea in Prunus

Species in the fungal family Botryosphaeriaceae are significant pathogens of peach. The climatic conditions in the Southeastern USA are conducive to the development of peach fungal gummosis (PFG) with an estimated yield reduction of up to 40% in severe cases. Genotypes with resistance to this PFG were identified in interspecific crosses and segregating backcross populations generated using Kansu peach (Prunus kansuensis Rehder), almond [Prunus dulcis (Mill.) D.A. Webb], and peach [Prunus persica (L.) Batsch]. Hybrids were evaluated for four consecutive years in field conditions. Data generated was validated in different environments using clonal replicates of the hybrids. The F1 and BC1F1 segregation population data suggest a dominant allele for PFG resistance originating from almond. Segregation and mapping analysis located the PFG resistance locus on a chimeric linkage groups 6–8 near the leaf color locus. The molecular markers identified will facilitate marker-assisted selection (MAS) and introgression of this resistance trait into commercial peach germplasm.