De novo assembly and characterization of transcriptome using Illumina sequencing and development of twenty five microsatellite markers for an endemic tree Juglans hopeiensis Hu in China

The Chinese walnut (Juglans hopeiensis Hu) is an endemic temperate tree species and narrowly distributed in China. However, there are still few specific molecular markers for understanding genetic diversity of this walnut. In this study, more than 44 million sequencing reads were generated using Illumina sequencing technology. De novo assembly yielded 93,822 unigenes with an average length of 731 bp. Based on sequence similarity search with known proteins, a total of 39,708 (42.3%) genes were identified. Searching against the Kyoto Encyclopedia of Genes and Genomes Pathway database (KEGG) indicated that 15,903 (17.0%) unigenes. To contribute to its conservation and management, twenty five microsatellite markers were identified in J. hopeiensis were screened for polymorphism across 27 Chinese walnut tree individuals from two locations. The number of alleles ranged from two to nine, observed heterozygosity ranged from 0.016 to 0.933 (mean 0.468), and expected heterozygosity from 0.022 to 0.823 (mean 0.462). The polymorphic information content (PIC) ranged from 0.012 to 0.831 (mean 0.437). The development of these new microsatellite markers will be useful for studying population genetic structure, evolutionary ecology, conservation, and genetic breeding of this endemic walnut tree or other Juglans species.