Skip to main content
Log in

Development of EST-SSR markers based on transcriptome sequencing for germplasm evaluation of 65 lilies (Lilium)

  • Original Article
  • Published:
Molecular Biology Reports Aims and scope Submit manuscript

Abstract

Background

Lilium genus consists of approximately 100 species and numerous varieties, many of which are interspecific hybrids, which result in a complicated genetic background. The germplasm identification, genetic relationship analysis, and breeding of Lilium rely on exploiting genetic information among different accessions. Hence, an attempt was made to develop new EST-SSR markers and study the molecular divergence among 65 genotypes of Lilium.

Methods and results

A total of 5509 EST-SSRs were identified from the high-throughput sequencing database of L. ‘Elodie’. After primer screening, six primer pairs with the most abundant polymorphic bands were selected from 100 primer pairs. Combined with the other 10 reported SSR primers, a total of 16 pairs detected genetic information with an average PIC value of 0.7583. The number of alleles per locus varied from four to 33, the expected heterozygosity varied from 0.3289 to 0.9231, and the observed heterozygosity varied from 0.2857 to 0.5122. Based on the phylogenic results, 22 Asiatic hybrids (A), seven Longiflorum × Asiatic hybrids (LA), as well as two native species were grouped. Eighteen Oriental hybrids (O) and nine Oriental × Trumpet (OT) hybrids, four native species, one LO, and one LL (L. pardalinum × L. longiflorum) variety were grouped.

Conclusions

Two major clusters were reported and a large number of genotypes were grouped based on UPGMA and STRUCTURE analysis methods. The PIC value as well as other parameters revealed that the EST-SSR markers selected were informative. In addition, the clustering pattern displayed better agreement with the cultivar’s pedigree. The newly identified SSRs in this study provides molecular markers for germplasm characterization and genetic diversity for Lilium.

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

Similar content being viewed by others

References

  1. Long YY, Zhang JZ, Zhang LN (1999) Lily-the king of corm flowers. Jindun Press, Beijing, pp 1–2

    Google Scholar 

  2. Van Tuyl JM, Arens P, Shahin A, Marasek-Ciołakowska A, Barba-Gonzalez R, Kim HT, Lim KB (2018) Lilium. Ornamental crops. Handbook Plant Breeding 11:481–512

    Google Scholar 

  3. Comber HF (1949) A new classification of the genus Lilium. Lily Yearbook. R Hort Soc 13:86–105

    Google Scholar 

  4. Shahin A, Smulders MJM, van Tuyl JM, Arens P, Bakker FT (2014) Using multi-locus allelic sequence data to estimate genetic divergence among four Lilium (Liliaceae) cultivars. Front Plant Sci 5:567

    Article  PubMed  PubMed Central  Google Scholar 

  5. Kong Y, Bai J, Lang L, Bao F, Dou X, Wang H, Shang H (2017) Floral scents produced by Lilium and Cardiocrinum species native to China. Biochem Syst Ecol 70:222–229

    Article  CAS  Google Scholar 

  6. Lee CS, Kim SC, Yeau SH, Lee NS (2011) Major lineages of the genus Lilium (Liliaceae) based on nrDNA ITS sequences, with special emphasis on the Korean species. J Plant Biol 54:159–171

    Article  Google Scholar 

  7. Kong Y, Wang H, Lang L, Dou X, Bai J (2021) Metabolome-based discrimination analysis of five Lilium bulbs associated with differences in secondary metabolites. Molecules 26:1340

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  8. Munafo JP, Gianfagna TJ (2011) Quantitative analysis of steroidal glycosides in different organs of Easter lily (Lilium longiflorum Thunb.) by LC-MS/MS. J Agric Food Chem 59:995–1004

    Article  CAS  PubMed  Google Scholar 

  9. Francis JA, Rumbeiha W, Nair MG (2004) Constituents in Easter lily flowers with medicinal activity. Life Sci 76(6):671–683

    Article  CAS  PubMed  Google Scholar 

  10. Kumar SPJ, Susmita C, Sripathy KV, Agarwal DK, Pal G, Singh AN, Kumar S, Rai AK, Simal–Gandara J (2022) Molecular characterization and genetic diversity studies of Indian soybean (Glycine max (L.) Merr.) cultivars using SSR markers. Mol Biol Rep 49:2129–2140

    Article  CAS  PubMed  Google Scholar 

  11. Ma L, Wang X, Yan M, Liu F, Zhang S, Wang X (2022) Genome survey sequencing of common vetch (Vicia sativa L.) and genetic diversity analysis of Chinese germplasm with genomic SSR markers. Mol Biol Rep 49:313–320

    Article  CAS  PubMed  Google Scholar 

  12. Nyabera LA, Nzuki IW, Runo SM, Amwayi PW (2021) Assessment of genetic diversity of pumpkins (Cucurbita spp.) from western Kenya using SSR molecular markers. Mol Biol Rep 48:2253–2260

    Article  CAS  PubMed  Google Scholar 

  13. Liu C, Dou Y, Guan X, Fu Q, Zhang Z, Hu Z, Zheng J, Lu Y, Li W (2017) De novo transcriptomic analysis and development of EST-SSRs for Sorbus pohuashanensis (Hance) Hedl. PLoS ONE 12(6):e0179219

    Article  PubMed  PubMed Central  Google Scholar 

  14. Zietkiewicz E, Rafalski A, Labuda D (1994) Genome fingerprinting by simple sequence repeat (SSR) anchored polymerase chain reaction amplification. Genomics 20(2):176–183

    Article  CAS  PubMed  Google Scholar 

  15. Ukoskit K, Posudsavang G, Pongsiripat N, Chatwachirawong P, Klomsaard P, Poomipant P, Tragoonrung S (2018) Detection and validation of EST-SSR markers associated with sugar-related traits in sugarcane using linkage and association mapping. Genomics 111:1–9

    Article  PubMed  Google Scholar 

  16. Du F, Jiang J, Jia H, Zhao X, Wang W, Gao Q, Mao W, Wu Y, Zhang L, Grierson D, Xia Y, Gao Z (2015) Selection of generally applicable SSR markers for evaluation of genetic diversity and identity in Lilium. Biochem Syst Ecol 61:278–285

    Article  CAS  Google Scholar 

  17. Lee SI, Park KC, Song YS, Son JH, Kwon SJ, Na JK, Kim JH, Kim NS (2011) Development of expressed sequence tag derived-simple sequence repeats in the genus Lilium. Genes Genom 33:727–733

    Article  CAS  Google Scholar 

  18. Yuan S, Ge L, Liu C, Ming J (2013) The development of EST-SSR markers in Lilium regale and their cross-amplification in related species. Euphytica 189:393–419

    Article  CAS  Google Scholar 

  19. Shahin A, van Kaauwen M, Esselink D, Bargsten JW, van Tuyl JM, Visser RG, Arens P (2012) Generation and analysis of expressed sequence tags in the extreme large genomes Lilium and Tulipa. BMC Genomics 13:640

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  20. Chen M, Nie G, Yang L, Zhang Y, Cai Y (2021) Homeotic transformation from stamen to petal in Lilium is associated with MADS-box genes and hormone signal transduction. Plant Growth Regul 95:49–64

    Article  CAS  Google Scholar 

  21. Thiel T, Michalek W, Varshney RK, 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  PubMed  Google Scholar 

  22. Rozen S, Skaletsky H (2000) Primer3 on the WWW for general users and for biologist programmers. Methods Mol Biol 132:365–386

    CAS  PubMed  Google Scholar 

  23. Du F, Wu Y, Zhang L, Li XW, Zhao XY, Wang WH, Gao ZS, Xia YP (2015) De novo assembled transcriptome analysis and SSR marker development of a mixture of six tissues from Lilium Oriental hybrid ‘Sorbonne’. Plant Mol Biol Rep 33:281–293

    Article  CAS  Google Scholar 

  24. Glaubitz JC (2004) CONVERT: a user friendly program to reformat diploid genotypic data for commonly used population genetic software packages. Mol Ecol Notes 4:309–310

    Article  CAS  Google Scholar 

  25. Yeh FC, Boyle TJB (1997) Population genetic analysis of co-dominant and dominant markers and quantitative traits. Belg J Bot 129:157–163

    Google Scholar 

  26. Langella O (2007) Populations 1.2.30: population genetic software (individuals or populations distances, phylogenetic trees). France. http://bioinformatics.org/~tryphon/populations/. Accessed 14 Sept 2012

  27. Letunic I, Bork P (2019) Interactive tree of life (iTOL) v4: recent updates and new developments. Nucleic Acid Res 47:W256–W259

    Article  CAS  PubMed  PubMed Central  Google Scholar 

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

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  29. 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  PubMed  Google Scholar 

  30. Earl DA, vonHoldt BM (2012) STRUCTURE HARVESTER: a website and program for visualizing STRUCTURE output and implementing the Evanno method. Conserv Genet Resour 4:359–361

    Article  Google Scholar 

  31. Kalia RK, Rai MK, Kalia S, Singh R, Dhawan AK (2011) Microsatellite markers: an overview of the recent progress in plants. Euphytica 177:309–334

    Article  CAS  Google Scholar 

  32. Ma JQ, Ma CL, Yao MZ, Jin JQ, Wang ZL, Wang XC, Chen L (2012) Microsatellite markers from tea plant expressed sequence tags (ESTs) and their applicability for cross-species/genera amplification and genetic mapping. Sci Hortic 134:167–175

    Article  CAS  Google Scholar 

  33. Qiu L, Yang C, Tian B, Yang JB, Liu A (2010) Exploiting EST databases for the development and characterization of EST-SSR markers in castor bean (Ricinus communis L.). BMC Plant Biol 10:278–287

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  34. Aggarwal RK, Hendre PS, Varshney RK, Bhat PR, Krishnakumar V, Singh L (2007) Identification, characterization and utilization of EST-derived genic microsatellite markers for genome analyses of coffee and related species. Theor Appl Genet 114:359–372

    Article  CAS  PubMed  Google Scholar 

  35. Baisakh N, Subudhi PK, Arumuganathan K, Parco AP, Harrison SA, Knott CA, Materne MD (2009) Development and interspecific transferability of genic microsatellite markers in Spartina spp. with different genome size. Aquat Bot 91:262–266

    Article  CAS  Google Scholar 

  36. Varshney RK, Thiel T, Stein N, Langridge P, Graner A (2002) In silico analysis on frequency and distribution of microsatellites in ESTs of some cereal species. Cell Mol Biol Lett 7:537–546

    CAS  PubMed  Google Scholar 

  37. Wang YW, Samuels TD, Wu YQ (2011) Development of 1030 genomic SSR markers in switchgrass. Theor Appl Genet 122:677–686

    Article  CAS  PubMed  Google Scholar 

  38. Wang Z, Yu G, Shi B, Wang X, Qiang H, Gao H (2014) Development and characterization of simple sequence repeat (SSR) markers based on RNA-sequencing of Medicago sativa and in silico mapping onto the M. truncatula genome. PLoS ONE 9(3):e92029

    Article  PubMed  PubMed Central  Google Scholar 

  39. Tantasawat P, Trongchuen J, Prajongjai T, Seehalak W, Jittayasothorn Y (2011) Variety identification and comparative analysis of genetic diversity in yardlong bean (Vigna unguiculata spp. sesquipedalis) using morphological characters, SSR and ISSR analysis. Sci Hort 124:204–216

    Article  Google Scholar 

  40. Lim KB, Barba-Gonzalez R, Zhou S, Ramanna MS, van Tuyl JM (2008) Interspecific hybridization in lily (Lilium): taxonomy and commercial aspects of using species hybrids in breeding. Floriculture, ornamental and plant biotechnology, vol 5. Global Science Books, Japan, pp 146–151

    Google Scholar 

  41. McRae EA (1998) Lilies: a guide for growers and collectors. Timber Press, Portland

    Google Scholar 

  42. Lim KB, Van Tuyl JM (2006) Lily, Lilium hybrids. Flower Breed Genet 19:517–537

    Google Scholar 

  43. Asano Y (1989) Lilium L. In: Tsukumoto Y (ed) The great dictionary of horticulture, vol 5. Sykakukum, Tokyo, pp 198–209

    Google Scholar 

Download references

Funding

This work was supported by the Shanghai Agriculture Applied Technology Development Program, China (Grant No. X2020-02-08-00-12-F01463).

Author information

Authors and Affiliations

Authors

Contributions

MC and YZ conceived the theme of the study and designed the experiment; MC, GN and XL performed the experiment; MC, LY and YC analyzed the SSR data; MC wrote the manuscript. All co-authors read and approved the final manuscript.

Corresponding author

Correspondence to Yongchun Zhang.

Ethics declarations

Conflict of interest

The authors have no relevant financial or non-financial interests to disclose.

Ethics approval

The article does not contain any studies with human participants or animals performed by any of the authors.

Consent for publication

All the authors have read and consented to submit the manuscript.

Additional information

Publisher’s Note

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

Supplementary Information

Rights and permissions

Springer Nature or its licensor (e.g. a society or other partner) holds exclusive rights to this article under a publishing agreement with the author(s) or other rightsholder(s); author self-archiving of the accepted manuscript version of this article is solely governed by the terms of such publishing agreement and applicable law.

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Chen, M., Nie, G., Li, X. et al. Development of EST-SSR markers based on transcriptome sequencing for germplasm evaluation of 65 lilies (Lilium). Mol Biol Rep 50, 3259–3269 (2023). https://doi.org/10.1007/s11033-022-08083-9

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11033-022-08083-9

Keywords

Navigation