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Knowledge of genetic identity, genetic relationships, ploidy level, and chromosome numbers can enhance the efficiency of ornamental plant breeding programs. In the present study, genome sizes, chromosome numbers, and genetic fingerprints were determined for a collection of 94 Ilex accessions, including 69 I. crenata. The genome size of the entire collection ranged from 1.50 ± 0.03 to 8.01 ± 0.18. Within the species of I. crenata, genome sizes varied (mean ± sd) between 1.98 ± 0.08 and 2.30 ± 0.04, with three outliers: 3.06 ± 0.04, 4.04 ± 0.09, and 4.19 ± 0.08. The chromosome counting results showed 2n = 40 for I. crenata accessions and confirmed the outliers as one triploid and two tetraploids. A high intra-specific genetic diversity in Ilex crenata was found, after genetic fingerprinting using genotyping-by-sequencing (GBS). The species I. crenata was separated into three clades, which coincided with intraspecific differences in genome sizes (mean ± sd) of 2.09 ± 0.006, 2.07 ± 0.05, and 2.19 ± 0.06 pg/2C per clade as mean values for the diploids. Applying a principal coordinate analysis (PCoA) to the genetic fingerprinting data of all species in the collection revealed a wide genetic variation, which has not yet been commercially exploited. These findings could form the basis for selectively breeding parents, in order to create more genetic diversity via intra- and interspecific crosses. [ABSTRACT FROM AUTHOR]

We have developed PotatoMASH (Potato Multi-Allele Scanning Haplotags), a novel low-cost, genome-scanning marker platform. We designed a panel of 339 multi-allelic regions placed at 1 Mb intervals throughout the euchromatic portion of the genome. These regions were assayed using a multiplex amplicon sequencing approach, which allows for genotyping hundreds of plants at a cost of 5 EUR/sample. We applied PotatoMASH to a population of over 700 potato lines. We obtained tetraploid dosage calls for 2012 short multi-allelic haplotypes in 334 loci, which ranged from 2 to 14 different haplotypes per locus. The system was able to diagnose the presence of targeted pest-resistance markers, to detect quantitative trait loci (QTLs) by genome-wide association studies (GWAS) in a tetraploid population, and to track variation in a diploid segregating population. PotatoMASH efficiently surveys genetic variation throughout the potato genome, and can be implemented as a single low-cost genotyping platform that will allow the routine and simultaneous application of marker-assisted selection (MAS) and other genotyping applications in commercial potato breeding programmes. [ABSTRACT FROM AUTHOR]

Columnea latent viroid (CLVd) is one of the most serious tomato diseases. In general, viroids have high mutation rates. This generates a population of variants (so-called quasi-species) that co-exist in their host and exhibit a huge level of genetic diversity. To study the population of CLVd in individual host plants, we used amplicon sequencing using specific CLVd primers linked with a sample-specific index sequence to amplify libraries. An infectious clone of a CLVd isolate Chaipayon-1 was inoculated on different solanaceous host plants. Six replicates of the amplicon sequencing results showed very high reproducibility. On average, we obtained 133,449 CLVd reads per PCR-replicate and 79 to 561 viroid sequence variants, depending on the plant species. We identified 19 major variants (>1.0% mean relative abundance) in which a total of 16 single-nucleotide polymorphisms (SNPs) and two single nucleotide insertions were observed. All major variants contained a combination of 4 to 6 SNPs. Secondary structure prediction clustered all major variants into a tomato/bolo maka group with four loops (I, II, IV and V), and a chili pepper group with four loops (I, III, IV and V) at the terminal right domain, compared to the CLVd Chaipayon-1 which consists of five loops (I, II, III, IV and V). [ABSTRACT FROM AUTHOR]