Skip to main content
Log in

Genome-wide distribution of simple sequence repeats in pomegranate and their application to the analysis of genetic diversity

  • Original Article
  • Published:
Tree Genetics & Genomes Aims and scope Submit manuscript

Abstract

Pomegranate (Punica granatum L.) germplasm collections are increasingly appreciated as a repository for the genetic improvement of species, and their genetic analysis is an essential prerequisite for their utilization in pomegranate breeding. Sufficient markers are a prerequisite for genetic analysis. In this study, we identified 36,792 simple sequence repeat (SSR) markers in the pomegranate genome. Dinucleotide repeats are the most abundant SSRs, and of the mono-, di-, tri-, and tetranucleotide repeats, A/T, AT/AT, AAT/ATT, and AAAT/ATTT were the most dominant motif type. There were only 1.80% SSR motifs located in the coding regions, while most (98.20%) SSR motifs were located in noncoding regions. We confirmed the reliability of 50 SSR markers by PCR amplification, from which 11 highly polymorphic SSR markers were selected to study the genetic diversity of 218 pomegranate accessions. A total of 63 alleles was detected for the 11 SSR primers, ranging from 2 (PG152) to 9 (PG093, PG080) across the pomegranate accessions. The polymorphism information content values of each SSR primer pair ranged from 0.22 to 0.70, with a mean value of 0.45. Based on population structure analysis, these accessions were clustered into three main populations and 42 accessions were chosen as the core set. The availability of these SSR markers and the clustering result for the pomegranate germplasm provide new important information for pomegranate genetics and breeding programs.

This is a preview of subscription content, log in via an institution to check access.

Access this article

Price excludes VAT (USA)
Tax calculation will be finalised during checkout.

Instant access to the full article PDF.

Fig. 1
Fig. 2
Fig. 3
Fig. 4

Similar content being viewed by others

References

  • Botstein D, White RL, Skolnick M, Davis RW (1980) Construction of a genetic linkage map in man using restriction fragment length polymorphisms. Am J Hum Genet 32:314–331

    CAS  PubMed  Google Scholar 

  • Cui J, Cheng J, Nong D, Peng J, Hu Y, He W, Zhou Q, Dhillon NPS, Hu K (2017) Genome-wide analysis of simple sequence repeats in bitter gourd (Momordica charantia). Front Plant Sci 8:1103

    Article  Google Scholar 

  • Curro S, Caruso M, Distefano G, Gentile A, La Malfa S (2010) New microsatellite loci for pomegranate, Punica granatum (Lythraceae). Am J Bot 97:e58–e60

    Article  Google Scholar 

  • Da Silva JA, Rana TS, Narzary D, Verma N, Meshram DT, Ranade SA (2013) Pomegranate biology and biotechnology: a review. Sci Hortic 160:85–107

    Article  Google Scholar 

  • Doyle J, Doyle J (1990) Isolation of DNA from small amounts of plant tissues. BRL Focus 12:13–15

    Google Scholar 

  • Evanno G, Regnaut S, Goudet J (2005) Detecting the number of clusters of individuals using the software STRUCTURE: a simulation study. Mol Ecol 14:2611–2620

    Article  CAS  Google Scholar 

  • Ferrara G, Giancaspro A, Mazzeo A, Giove SL, Matarrese AMS, Pacucci C, Punzi R, Trani A, Gambacorta G, Blanco A (2014) Characterization of pomegranate (Punica granatum L.) genotypes collected in Puglia region, Southeastern Italy. Sci Hortic 178:70–78

    Article  Google Scholar 

  • Hancock JM (1996) Simple sequences and the expanding genome. Bioessays 18:421–425

    Article  CAS  Google Scholar 

  • Hasnaoui N, Buonamici A, Sebastiani F, Mars M, Trifi M, Vendramin G (2010) Development and characterization of SSR markers for pomegranate (Punica granatum L.) using an enriched library. Conserv Genet Resour 2:283–285

    Article  Google Scholar 

  • Jbir R, Melgarejo P, Hernández F, Haddioui A, Hannachi AS (2014) Efficiency of Inter Simple Sequence Repeat (ISSR) markers for the assessment of genetic diversity of Moroccan pomegranate (Punica granatum L.) cultivars. Biochem Syst Ecol 56:24–31

    Article  Google Scholar 

  • Kantety RV, La Rota M, Matthews DE, Sorrells ME (2002) Data mining for simple sequence repeats in expressed sequence tags from barley, maize, rice, sorghum and wheat. Plant Mol Biol 48:501–510

    Article  CAS  Google Scholar 

  • Kim KW, Chung HK, Cho GT, Ma KH, Chandrabalan D, Gwag JG, Kim TS, Cho EG, Park YJ (2007) PowerCore: a program applying the advanced M strategy with a heuristic search for establishing core sets. Bioinformatics 23:2155–2162

    Article  CAS  Google Scholar 

  • Lassois L, Denancé C, Ravon E, Guyader A, Guisnel R, Hibrand-Saint-Oyant L, Poncet C, Lasserre-Zuber P, Feugey L (2016) Genetic diversity, population structure, parentage analysis, and construction of core collections in the French apple germplasm based on SSR markers. Plant Mol Biol Report 34(4):827–844

    Article  CAS  Google Scholar 

  • Li YC, Korol AB, Fahima T, Beiles A, Nevo E (2002) Microsatellites: genomic distribution, putative functions and mutational mechanisms: a review. Mol Ecol 11:2453–2465

    Article  CAS  Google Scholar 

  • Liang W, Dondini L, De Franceschi P, Paris R, Sansaviniet S, Tartarini S (2015) Genetic diversity, population structure and construction of a core collection of apple cultivars from Italian germplasm. Plant Mol Biol Report 33(3):458–473

    Article  CAS  Google Scholar 

  • Liu CY, Zhang Q, Yao XH, Zhong CH, Yan CL, Huang HW (2016) Characterization of genome-wide simple sequence repeats and application in interspecific genetic map integration in kiwifruit. Tree Genet Genomes 12:21

    Article  Google Scholar 

  • McColl R (1991) China’s silk roads: a modern journey to China’s Western regions. Focus Geogr 41:1

    Google Scholar 

  • Melgarejo P, Martínez J, Hernández F, Martínez R, Legua P, Oncina R, Martínez-Murcia A (2009) Cultivar identification using 18S–28S rDNA intergenic spacer-RFLP in pomegranate (Punica granatum L.). Sci Hortic 120:500–503

    Article  CAS  Google Scholar 

  • Metzgar D, Bytof J, Wills C (2000) Selection against frameshift mutations limits microsatellite expansion in coding DNA. Genome Res 10:72–80

    CAS  PubMed  PubMed Central  Google Scholar 

  • Morgante M, Hanafey M, Powell W (2002) Microsatellites are preferentially associated with nonrepetitive DNA in plant genomes. Nat Genet 30:194

    Article  CAS  Google Scholar 

  • Narzary D, Mahar KS, Rana TS, Ranade SA (2009) Analysis of genetic diversity among wild pomegranates in Western Himalayas, using PCR methods. Sci Hortic 121:237–242

    Article  CAS  Google Scholar 

  • Ophir R, Sherman A, Rubinstein M, Eshed R, Schwager MS, Harel-Beja R, Bar-Ya’akov I, Holland D (2014) Single-nucleotide polymorphism markers from de-novo assembly of the pomegranate transcriptome reveal germplasm genetic diversity. PLoS One 9(2):e88998

    Article  Google Scholar 

  • Pan Y-B, Cordeiro GM, Richard E, Henry R (2003) Molecular genotyping of sugarcane clones with microsatellite DNA markers. Maydica 48:319–329

    Google Scholar 

  • Parvaresh M, Talebi M, Sayed-Tabatabaei B-E (2012) Molecular diversity and genetic relationship of pomegranate (Punica granatum L.) genotypes using microsatellite markers. Sci Hortic 138:244–252

    Article  CAS  Google Scholar 

  • Pirseyedi SM, Valizadehghan S, Mardi M, Ghaffari MR, Mahmoodi P, Zahravi M, Zeinalabedini M, Nekoui SMK (2010) Isolation and characterization of novel microsatellite markers in pomegranate (Punica granatum L.). International Journal of Molecular Sciences 11

  • Pritchard JK, Stephens M, Donnelly P (2000) Inference of population structure using multilocus genotype data. Genetics 155:945–959

    Article  CAS  Google Scholar 

  • Qin G, Xu C, Ming R, Tang H, Guyot R, Kramer EM, Hu Y, Yi X, Qi Y, Xu X (2017) The pomegranate (Punica granatum L.) genome and the genomics of punicalagin biosynthesis. Plant J 91:1108–1128

    Article  CAS  Google Scholar 

  • Roy A, Bandyopadhyay A, Mahapatra AK, Ghosh SK, Singh NK, Bansal KC, Koundal KR, Mohapatra T (2006) Evaluation of genetic diversity in jute (Corchorus species) using STMS, ISSR, and RAPD markers [J]. Plant breed 125(3):292–297

  • Sonah H, Deshmukh RK, Sharma A, Singh VP, Gupta DK, Gacche RN, Rana JC, Singh NK, Sharma TR (2011) Genome-wide distribution and organization of microsatellites in plants: an insight into marker development in Brachypodium. PLoS One 6:e21298

    Article  CAS  Google Scholar 

  • Soriano JM, Zuriaga E, Rubio P, Llácer G, Infante R, Badenes ML (2011) Development and characterization of microsatellite markers in pomegranate (Punica granatum L.). Mol Breed 27:119–128

    Article  Google Scholar 

  • Tamura K, Peterson D, Peterson N, Stecher G, Nei M, Kumar S (2011) MEGA5: molecular evolutionary genetics analysis using maximum likelihood, evolutionary distance, and maximum parsimony methods. Mol Biol Evol 28:2731–2739

    Article  CAS  Google Scholar 

  • Thiel T, Michalek W, Varshney R, Graner A (2003) Exploiting EST databases for the development and characterization of gene-derived SSR-markers in barley (Hordeum vulgare L.). Theor Appl Genet 106:411–422

    Article  CAS  Google Scholar 

  • Van Oosterhout C, Weetman D, Hutchinson WF (2006) Estimation and adjustment of microsatellite null alleles in nonequilibrium populations. Mol Ecol Notes 6(1):255–256

    Article  Google Scholar 

  • Wu H-B, Gong H, Liu P, He X-L, Luo S-B, Zheng X-M, Zhang C-Y, He X-M, Luo J (2014) Large-scale development of EST-SSR markers in sponge gourd via transcriptome sequencing. Mol Breed 34:1903–1915

    Article  CAS  Google Scholar 

  • Yeh FC, Yang R, Boyle TB, Ye Z, Mao JX (1997) POPGENE, the user-friendly shareware for population genetic analysis. Molecular Biology and Biotechnology Centre, University of Alberta, Canada 10:295–301

  • Yuan Z, Yin Y, Qu J, Zhu L, Li Y (2007) Population genetic diversity in Chinese pomegranate (Punica granatum L.) cultivars revealed by fluorescent-AFLP markers. Journal of Genetics and Genomics 34:1061–1071

    Article  CAS  Google Scholar 

  • Yuan ZH, Fang YM, Zhang TK, Fei Z, Han F, Liu C, Liu M, Xiao W, Zhang W, Wu S, Zhang M, Ju Y, Xu H, Dai H, Liu Y, Chen Y, Wang L, Zhou J, Guan D, Yan M, Xia Y, Huang X, Liu D, Wei H, Zheng H (2018) The pomegranate (Punica granatum L.) genome provides insights into fruit quality and ovule developmental biology. Plant Biotechnol J 16(7):1363–1374

    Article  CAS  Google Scholar 

  • Zhang Q, Ma B, Li H, Chang Y, Han Y, Li J, Wei G, Zhao S, Khan MA, Zhou Y (2012) Identification, characterization, and utilization of genome-wide simple sequence repeats to identify a QTL for acidity in apple. BMC Genomics 13:537

    Article  Google Scholar 

  • Zhao L, Li M, Cai G, Pan T, Shan C (2013) Assessment of the genetic diversity and genetic relationships of pomegranate (Punica granatum L.) in China using RAMP markers. Sci Hortic 151:63–67

    Article  CAS  Google Scholar 

  • Zhou GJ, Yuan YY, Zeng FZ, Xian KZ, Chen ZG (1995) Status and prospect of pomegranate production in China. Southwest China Journal of Agricultural Sciences 8:111–116 (in Chinese with an English abstract)

    Google Scholar 

Download references

Acknowledgments

We are thankful for Professor Pan Yongbao (Agricultural Research Services, United States Department of Agriculture) for the help in polishing the language.

Funding

This research was funded by the Project of the Key Laboratory of Anhui Academy of Agricultural Sciences (2019YL013) and the Natural Science Foundation of Anhui Province (1708085MC85).

Data archiving statement

This research contains no data that requires submission to a public database. The 36,792 SSR motifs found in this study are listed in Supplementary Table S3.

Author information

Authors and Affiliations

Authors

Corresponding author

Correspondence to Gaihua Qin.

Additional information

Communicated by G. G. Vendramin

Publisher’s note

Springer Nature remains neutral with regard to jurisdictional claims in published maps and institutional affiliations.

Electronic supplementary material

ESM 1

(XLSX 15 kb)

ESM 2

(XLSX 10 kb)

ESM 3

(XLSX 1899 kb)

ESM 4

(XLSX 12 kb)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Liu, C., Li, J. & Qin, G. Genome-wide distribution of simple sequence repeats in pomegranate and their application to the analysis of genetic diversity. Tree Genetics & Genomes 16, 36 (2020). https://doi.org/10.1007/s11295-020-1428-4

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • DOI: https://doi.org/10.1007/s11295-020-1428-4

Keywords

Navigation