De novo transcriptome assembly of pummelo and molecular marker development

PLoS One. 2015 Mar 23;10(3):e0120615. doi: 10.1371/journal.pone.0120615. eCollection 2015.

Abstract

Pummelo (Citrus grandis) is an important fruit crop worldwide because of its nutritional value. To accelerate the pummelo breeding program, it is essential to obtain extensive genetic information and develop relative molecular markers. Here, we obtained a 12-Gb transcriptome dataset of pummelo through a mixture of RNA from seven tissues using Illumina pair-end sequencing, assembled into 57,212 unigenes with an average length of 1010 bp. The annotation and classification results showed that a total of 39,584 unigenes had similar hits to the known proteins of four public databases, and 31,501 were classified into 55 Gene Ontology (GO) functional sub-categories. The search for putative molecular markers among 57,212 unigenes identified 10,276 simple sequence repeats (SSRs) and 64,720 single nucleotide polymorphisms (SNPs). High-quality primers of 1174 SSR loci were designed, of which 88.16% were localized to nine chromosomes of sweet orange. Of 100 SSR primers that were randomly selected for testing, 87 successfully amplified clear banding patterns. Of these primers, 29 with a mean PIC (polymorphic information content) value of 0.52 were effectively applied for phylogenetic analysis. Of the 20 SNP primers, 14 primers, including 54 potential SNPs, yielded target amplifications, and 46 loci were verified via Sanger sequencing. This new dataset will be a valuable resource for molecular biology studies of pummelo and provides reliable information regarding SNP and SSR marker development, thus expediting the breeding program of pummelo.

Publication types

  • Research Support, Non-U.S. Gov't

MeSH terms

  • Citrus / genetics*
  • Cluster Analysis
  • Gene Expression Profiling
  • Gene Frequency
  • Gene Library
  • Gene Ontology*
  • Genetic Markers
  • High-Throughput Nucleotide Sequencing
  • Microsatellite Repeats
  • Molecular Sequence Annotation
  • Nucleotide Motifs
  • Phenotype
  • Phylogeny
  • Physical Chromosome Mapping
  • Polymorphism, Single Nucleotide
  • Reproducibility of Results
  • Transcriptome*

Substances

  • Genetic Markers

Associated data

  • SRA/SRP051793

Grants and funding

This research was financially supported by the Ministry of Science & Technology of China (no. 2011CB100600), the National Natural Science Foundation of China (nos. 31301760, 31460507, 31330066, 31221062), the Ministry of Education Innovation Team (IRT13065), and the Fundamental Research Funds for the Central Universities (2013QC037). The funders had no role in study design, data collection and analysis, decision to publish, or preparation of the manuscript.