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Characterizing the transcriptome and microsatellite markers for almond (Amygdalus communis L.) using the Illumina sequencing platform.

ABSTRACT: The almond tree (Prunus amygdalus Batsch) is an important nut tree grown in subtropical regions that produces nutrient-rich nuts. However, a paucity of genomic information and DNA markers has restricted the development of modern breeding technologies for almond trees.In this study, almonds were sequenced with Illumina paired-end sequencing technology to obtain transcriptome data and develop simple sequence repeats (SSR) markers. We generated approximately 64 million clean reads from the various tissues of mixed almonds, and a total of 42,135 unigenes with an average length of 988 bp were obtained in the present study. A total of 27,586 unigenes (57.7% of all unigenes generated) were annotated using several databases. A total of 112,812 unigenes were annotated with the Gene Ontology (GO) database and assigned to 82 functional sub-groups, and 29,075 unigenes were assigned to the KOG database and classified into 25 function classifications. There were 9470 unigenes assigned to 129 Kyoto Encyclopaedia of Genes and Genomes (KEGG) pathways from five categories in the KEGG pathway database. We further identified 8641 SSR markers from 48,012 unigenes. A total of 100 SSR markers were randomly selected to validate quality, and 82 markers could amplify the specific products of A. communis L., whereas 70 markers were successfully transferable to five species (A. ledebouriana, A. mongolica, A. pedunculata, A. tangutica, and A. triloba).Our study was the first to produce public transcriptome data from almonds. The development of SSR markers will promote genetics research and breeding programmes for almonds.

SUBMITTER: Zhang L

PROVIDER: S-EPMC5649074 | biostudies-literature | 2018

REPOSITORIES: biostudies-literature

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Characterizing the transcriptome and microsatellite markers for almond (<i>Amygdalus communis</i> L.) using the Illumina sequencing platform.

Zhang Linsen L Yang Xiaoni X Qi Xiangning X Guo Chunhui C Jing Zhaobin Z

Hereditas 20171019

<h4>Background</h4>The almond tree (<i>Prunus amygdalus</i> Batsch) is an important nut tree grown in subtropical regions that produces nutrient-rich nuts. However, a paucity of genomic information and DNA markers has restricted the development of modern breeding technologies for almond trees.<h4>Results</h4>In this study, almonds were sequenced with Illumina paired-end sequencing technology to obtain transcriptome data and develop simple sequence repeats (SSR) markers. We generated approximatel ...[more]

PMID: 29075165

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Project description:BACKGROUND: Amorphophallus is a genus of perennial plants widely distributed in the tropics or subtropics of West Africa and South Asia. Its corms contain a high level of water-soluble glucomannan; therefore, it has long been used as a medicinal herb and food source. Genetic studies of Amorphophallus have been hindered by a lack of genetic markers. A large number of molecular markers are required for genetic diversity study and improving disease resistance in Amorphophallus. Here, we report large scale of transcriptome sequencing of two species: Amorphophallus konjac and Amorphophallus bulbifer using deep sequencing technology, and microsatellite (SSR) markers were identified based on these transcriptome sequences. RESULTS: cDNAs of A. konjac and A. bulbifer were sequenced using Illumina HiSeq™ 2000 sequencing technology. A total of 135,822 non-redundant unigenes were assembled from about 9.66 gigabases, and 19,596 SSRs were identified in 16,027 non-redundant unigenes. Di-nucleotide SSRs were the most abundant motif (61.6%), followed by tri- (30.3%), tetra- (5.6%), penta- (1.5%), and hexa-nucleotides (1%) repeats. The top di- and tri-nucleotide repeat motifs included AG/CT (45.2%) and AGG/CCT (7.1%), respectively. A total of 10,754 primer pairs were designed for marker development. Of these, 320 primers were synthesized and used for validation of amplification and assessment of polymorphisms in 25 individual plants. The total of 275 primer pairs yielded PCR amplification products, of which 205 were polymorphic. The number of alleles ranged from 2 to 14 and the polymorphism information content valued ranged from 0.10 to 0.90. Genetic diversity analysis was done using 177 highly polymorphic SSR markers. A phenogram based on Jaccard's similarity coefficients was constructed, which showed a distinct cluster of 25 Amorphophallus individuals. CONCLUSION: A total of 10,754 SSR markers have been identified in Amorphophallus using transcriptome sequencing. One hundred and seventy-seven polymorphic markers were successfully validated in 25 individuals. The large number of genetic markers developed in the present study should contribute greatly to research into genetic diversity and germplasm characterization in Amorphophallus.

Dataset Information

Characterizing the transcriptome and microsatellite markers for almond (Amygdalus communis L.) using the Illumina sequencing platform.

Publications

Characterizing the transcriptome and microsatellite markers for almond (<i>Amygdalus communis</i> L.) using the Illumina sequencing platform.

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