Biology subjects

Berner, T.

Publications and source records attributed to Berner, T..

4 recordsLinked to original sources

bioRxiv2023

The structure of the tetraploid sour cherry 'Schattenmorelle' (Prunus cerasus L.) genome reveals insights into its segmental allopolyploid nature

Sour cherry (Prunus cerasus L.) is an economically important allotetraploid cherry species believed to have evolved in the Caspian Sea and Black Sea regions. How, when and where exactly the evolution of this species took place is unclear. It resulted from a hybridization of the tetraploid ground cherry (Prunus fruticosa Pall.) and an unreduced (2n) pollen of the diploid ancestor sweet cherry (P. avium L.). Some indications implement that the genome of sour cherry is segmental allopolyploid, but how it is structured and to what extent is unknown. To get an insight, the genome of the sour cherry cultivar Schattenmorelle was sequenced at ~400x using Illumina NovaSeqTM short-read and Oxford Nanopore long-read technologies (ONT R9.4.1 PromethION). Additionally, the transcriptome of Schattenmorelle was sequenced using PacBio Sequel II SMRT cell sequencing at ~300x. The final assembly resulted in a ~629 Mbp long pseudomolecule reference genome, which could be separated into two subgenomes each split into eight chromosomes. Subgenome PceS_a which originates from P. avium has a length of 269 Mbp, whereas subgenome PceS_f which originates from P. fruticosa has a length of 299.5 Mbp. The length of unassembled contigs was 60 Mbp. The genome of the sour cherry shows a size-reduction compared to the genomes of its ancestral species. It also shows traces of homoeologous sequence exchanges throughout the genome. Comparative positional sequence and protein analyses provided evidence that the genome of sour cherry is segmental allotetraploid and that it has evolved in a very recent event in the past.

genomics↗

bioRxiv2022

Genome-wide identification of putative disease resistance genes (R genes) in carrot (Daucus carota subsp. sativus) by homology-based gene prediction

The cultivated carrot (Daucus carota ssp. sativus) is one of the most important root vegetable crops grown worldwide. Carrots are highly susceptible to several pests and diseases, and disease resistance is currently among the main breeding aims. The inheritance of resistance has been reported for a few carrot foliar diseases and root-knot nematodes, but no functionally characterized resistance gene (R gene) has yet been linked as a candidate gene to any resistance locus in carrot. Knowledge about the inventory of NLR genes (nucleotide-binding leucine-rich repeat receptors) and other R genes encoding transmembrane proteins such as receptor-like proteins (RLPs) and kinases (KIN) would be necessary to associate major QTLs (quantitative trait loci) identified by bi-parental QTL analyses or GWAS (genome-wide association analysis) with functional candidate R genes. In this study, we describe a combination of a genome-wide inventory of putative full-length carrot R genes based on a homology-based gene prediction approach called GeMoMa and subsequent classification by usage of the recent version of PRGdb 4.0 database (Calle-Garcia et al. 2022). A total of 320 putative carrot R genes were identified and bioinformatically characterized, including 72 newly identified gene models, that have not yet been annotated in the currently available carrot whole genome sequence. Based on the DRAGO 3 pipeline, totally 137 putative NLR genes were found, whereas 162 putative functional RLP and KIN genes were identified in the carrot genome. About one third of the R genes was found to be organized in clusters consisting only of NLR, RLP or KIN genes. To determine the evolutionary relationships of carrot R gene predictions, we generated a phylogenetic tree based on the alignment of all 320 R proteins. Three large clades (NLR, RLP and KIN) and a small clade (RLK) were identified, which reflect well the classification obtained after DRAGO 3 analysis. The presented carrot R gene inventory might be useful for resistance gene isolation, the development of (functional) molecular markers and resistance breeding in carrot.

plant biology↗

bioRxiv2021

The draft chromosome-level genome assembly of tetraploid ground cherry (Prunus fruticosa Pall.) from long reads

BackgroundCherries are stone fruits and belong to the economically important plant family of Rosaceae with worldwide cultivation of different species. The ground cherry, Prunus fruticosa Pall. is one ancestor of cultivated sour cherry, an important tetraploid cherry species. Here, we present a long read chromosome-level draft genome assembly and related plastid sequences using the Oxford Nanopore Technology PromethION platform and R10.3 pore type. FindingThe final assemblies obtained from 117.3 Gb cleaned reads representing 97x coverage of expected 1.2 Gb tetraploid (2n=4x=32) and 0.3 Gb haploid (1n=8) genome sequence of P. fruticosa were calculated. The N50 contig length ranged between 0.3 and 0.5 Mb with the longest contig being [~]6 Mb. BUSCO estimated a completeness between 98.7 % for the 4n and 96.1 % for the 1n datasets. Using a homology and reference based scaffolding method, we generated a final consensus genome sequence of 366 Mb comprising eight chromosomes. The N50 scaffold was [~]44 Mb with the longest chromosome being 66.5 Mb. The repeat content was estimated to [~]190 Mb (52 %) and 58,880 protein-coding genes were annotated. The chloroplast and mitochondrial genomes were 158,217 bp and 383,281 bp long, which is in accordance with previously published plastid sequences. ConclusionThis is the first report of the genome of ground cherry (P. fruticosa) sequenced by long read technology only. The datasets obtained from this study provide a foundation for future breeding, molecular and evolutionary analysis in Prunus studies.

genomics↗

bioRxiv2020

One-shot generation of duodecuple (12x) mutant Arabidopsis: Highly efficient routine editing in model species

Genome editing by RNA-guided nucleases, such as SpCas9, has been used in numerous different plant species. However, to what extent multiple independent loci can be targeted simultaneously by multiplexing has not been well-documented. Here, we developed a toolkit, based on a highly intron-optimized zCas9i gene, which allows assembly of nuclease constructs expressing up to 32 sgRNAs. We used this toolkit to explore the limits of multiplexing in two major model species, and report on isolation of transgene-free octuple Nicotiana benthamiana and duodecuple (12x) Arabidopsis thaliana mutant lines in a single generation (T1 and T2, respectively). We developed novel counter-selection markers for N. benthamiana, most importantly Sl-FAST2, comparable to the well-established Arabidopsis seed fluorescence marker, and FCY-UPP, based on production of toxic 5-fluorouracyl in presence of a precursor. Targeting eight genes with an array of nine different sgRNAs and relying on FCY-UPP for selection of non-transgenic T1, we identified N. benthamiana mutant lines with astonishingly high efficiencies: All analyzed plants carried mutations in all genes (~112/116 target sites edited). Furthermore, we targeted 12 genes by an array of 24 sgRNAs in A. thaliana. Efficiency was significantly lower in A. thaliana, and our results indicate Cas9 availability is the limiting factor in such higher order multiplexing applications. We identify a duodecuple mutant line by a combination of phenotypic screening and amplicon sequencing. The resources and results presented provide new perspectives for how multiplexing can be used to generate complex genotypes or to functionally interrogate groups of candidate genes.

plant biology↗