Characterization and Application of EST-SSR Markers Developed from Transcriptome Sequences in Elymus breviaristatus (Poaceae: Triticeae)

Author:

Li Jin1ORCID,Zhang Changbing2,Chen Shiyong13,Jiang Keke1,Guan Hao1,Liu Wenhui4

Affiliation:

1. College of Animal and Veterinary Sciences, Southwest Minzu University, Chengdu 610041, China

2. Sichuan Academy of Grassland Science, Chengdu 611731, China

3. Key Laboratory of Animal Science of State Ethnic Affairs Commission, Southwest Minzu University, Chengdu 610041, China

4. Key Laboratory of Superior Forage Germplasm in the Qinghai-Tibetan Plateau, Qinghai Academy of Animal Science and Veterinary Medicine, Qinghai University, Xining 810016, China

Abstract

Background: Elymus L. is the largest genus in the Triticeae tribe. Most species in this genus are highly stress resistant, with excellent forage value. Elymus breviaristatus, a rare species endemic to the Qinghai-Tibet Plateau (QTP), is declining due to habitat fragmentation. However, genetic data for E. breviaristatus are limited, with expressed sequence tag (EST) markers being particularly rare, hampering genetic studies and protection measures. Results: We obtained 9.06 Gb clean sequences from the transcriptome of E. breviaristatus, generating 171,522 unigenes, which were assembled and functionally annotated against five public databases. We identified 30,668 SSRs in the E. breviaristatus transcriptome, from which 103 EST-SSR primer pairs were randomly selected. Of these, 58 pairs of amplified products of the expected size, and 18 of the amplified products were polymorphic. Model-based Bayesian clustering, the unweighted pair group method with arithmetic average (UPGMA), and principal coordinate analysis (PCoA) of 179 wild E. breviaristatus in 12 populations using these EST-SSRs were generally consistent, grouping the 12 populations into two major clades. Analysis of molecular variance (AMOVA) found 70% of the genetic variation among the 12 populations and 30% within the populations, indicating a high level of genetic differentiation (or low gene exchange) among the 12 populations. The transferability of the 58 successful EST-SSR primers to 22 related hexaploid species was 86.2–98.3%. UPGMA analysis generally grouped species with similar genome types together. Conclusions: Here, we developed EST-SSR markers from the transcriptome of E. breviaristatus. The transferability of these markers was evaluated, and the genetic structure and diversity of E. breviaristatus were explored. Our results provide a basis for the conservation and management of this endangered species, and the obtained molecular markers represent valuable resources for the exploration of genetic relationships among species in the Elymus genus.

Funder

Double First-Class program of Southwest Minzu University

Key Laboratory of Superior Forage Germplasm in the Qinghai-Tibetan plateau

Publisher

MDPI AG

Subject

Genetics (clinical),Genetics

Reference59 articles.

1. Gustafson, J.P. (1984). The Genomic System of Classification as a Guide to Intergeneric Hybridization with the Perennial Triticeae, Columbia University Press.

2. Conspectus of the Triticeae;Feddes Repert.,1984

3. Chromosome engineering, mapping, and transferring of resistance to Fusarium head blight disease from Elymus tsukushiensis into wheat;Cainong;Theor. Appl. Genet.,2015

4. Keng, Y.L. (1959). Flora Illustralis Plantarum Primarum Sinicarum, Gramineae, Science Press.

5. Wu, Z.Y., Raven, P.H., and Hong, D.Y. (2006). Tribus Triticeae, Poaceae, Science Press.

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