De novo assembly, functional annotation, and marker development of Asian pear (Pyrus pyrifolia) fruit transcriptome through massively parallel sequencing.

This study investigated the Asian pear transcriptome using the RNA-Seq normalized fruit cDNA library to create a transcriptomic resource for unigene and marker discovery. Following the removal of lowquality reads, 127,085,054 trimmed reads were assembled de novo to yield 37,649 non-redundant unigenes with an average length of 599 bp. Alternative splicing events were detected in 4121 contigs. A total of 30,560 single nucleotide polymorphisms (SNPs) and 7443 simple sequence repeat (SSR) makers were obtained. Approximately 21,449 (56.9%) unigenes were categorized into three gene ontology groups; 3682 (9.8%) were classified into 25 cluster of orthologous groups; and 10,451 (27.8%) were assigned to six Kyoto Encyclopedia of Genes and Genomes pathways. Differentially expressed genes were investigated using the reads per kilobase of the exon model per million reads methodology. A total of 546 unigenes showed significant differences in expression levels at different fruit developmental stages. Gene ontology categories associated with various aspects, including carbohydrate metabolic processes, transmembrane transport, and signal transduction, were enriched with genes with divergent expressions. These Pyrus pyrifolia transcriptome data provide a rich resource for the discovery and identification of new genes. Furthermore, the numerous putative SSRs and SNPs detected in this study will be important resources for the future development of a linkage map or of marker-assisted breeding programs for the Asian pear.

[Identification of mutations associated with coronary artery lesion susceptibility in Kawasaki disease by targeted enrichment of genomic region sequencing technique].

Objective: To screen and identify the mutations in Kawasaki disease by targeted enrichment of genomic region sequencing technique and investigate susceptibility genes associated with coronary artery lesion. Method: This was a case-control study.A total of 114 patients diagnosed as Kawasaki disease treated in Shanghai Children's Hospital between December 2015 and November 2016 were studied and another 45 healthy children who were physically examined in outpatient department were enrolled as control group. Patients were divided into two groups based on the results of echocardiogram. Peripheral venous blood was obtained from patients and controls. Genomic DNA was extracted. SeqCap EZ Choice libraries were prepared by targeted enrichment of genomic region technology. Then the libraries were sequenced to identify susceptibility genes associated with coronary artery lesion in patients diagnosed as Kawasaki disease.Susceptible genes were identified by Burden test, Pearson chi-square test or Fisher's exact probability test. Result: There was statistically significant difference in TNFRSF11B(rs2073618)G>C(p.N3K)mutation and GG/GC/CC genotype between Kawasaki disease group and control group(χ(2)=15.52, P=0.00). There was statistically significant difference in TNFRSF13B(rs34562254)C>T(p.P251L)mutation(χ(2)=10.40, P=0.01)and LEFTY1(rs360057)T>G(p.D322A)mutation(χ(2)=8.505, P=0.01)between patients with coronary artery lesions and those without. Conclusion: Targeted enrichment of genomic region sequencing technology can be used to do primary screening for the susceptible genes associated with coronary artery lesions in Chinese Kawasaki patients and may provide theoretical basis for larger sample investigation of risk prediction score standard in Kawasaki disease.