Transcriptome based high-throughput SSRs and SNPs discovery in the medicinal plant Lagenaria siceraria

Lagenaria siceraria(Molina)Standley has unique biological characteristics with high nutritional and medicinal values.It is an important pharmaceutical plant with various biologically active ingredients.Genetic improvement and deeper genomic studies require a rich resource of molecular markers.The application of next-generation sequencing technology,especially for transcriptome profiling,has greatly facilitated high throughput single nucleotide polymorphism(SNP)and simple sequence repeat(SSR)discovery.In this study,we sequenced the transcriptome of three major cultivars of L.siceraria and obtained 64.88 GB of clean data.The assembled high-quality reads were clustered into 89,347 unigenes,which were annotated by non-redundant protein database,Swiss-Port,Eukaryotic Ortholog Groups,Kyoto Encyclopedia of Genes and Genomes,and gene ontology databases.A total of 8,891 SSR and 35,873 SNP markers were predicted from unigenes by MISA and SAM tools,respectively.Characterization of the predicted markers in L.siceraria showed that the SSR and SNP densities were 60 and 243 markers per Mb of genome,respectively,and the estimated ratio of transition to transversion of SNP was 2.016.These markers will be very useful for genetic studies in L.siceraria,especially for the high-density linkage map construction and genome-wide association studies.Further genomic studies based on these results will facilitate the identification of novel genes or alleles of pharmaceutical importance.

捕虫堇叶片高效再生体系建立

以2个不同基因型的捕虫堇圆切捕虫堇(Pinguicula cyclosecta)和塞提捕虫堇(P.‘Sethos’)叶片为外植体,通过探究不定芽再生的影响因素,成功建立了捕虫堇高效再生体系。结果表明,2种捕虫堇对消毒方式和基本培养基种类要求相同,但再生能力有显著差异。圆切捕虫堇叶片在MS+1.0 mg·L^(-1)6-BA+0.2 mg·L^(-1)NAA培养基上再生能力最强,再生率为92.22%,再生系数达4.84,在MS+0.6 mg·L^(-1)6-BA+0.1 mg·L^(-1)NAA培养基上,不定芽增殖系数达4.98。塞提捕虫堇叶片在MS+2.0 mg·L^(-1)6-BA+0.2 mg·L^(-1)NAA培养基中再生率最高,为77.78%,再生系数为6.12,在增殖培养基MS+0.3 mg·L^(-1)6-BA+0.1 mg·L^(-1)NAA上,增殖系数可达4.84。2种捕虫堇在1/2MS+0.1 mg·L^(-1)IBA培养基上进行生根培养,根系生长状态最佳。该研究解决了捕虫堇繁殖系数低及工厂化育苗难等问题,为其规模化生产和育种改良提供了技术支持。