Skip to main content

Advertisement

Log in

Genome-wide SSR markers in bottle gourd: development, characterization, utilization in assessment of genetic diversity of National Genebank of India and synteny with other related cucurbits

  • Plant Genetics • Original Paper
  • Published:
Journal of Applied Genetics Aims and scope Submit manuscript

Abstract

Lagenaria siceraria (Molina) Standley is an important cultivated crop with its immense importance in pharmaceutical industry and as vegetable. Its seed, root, stem, leaves, flower, and fruit are used as an ointment for ailment of various diseases throughout Asia. Despite its worldwide importance, informative co-dominant microsatellite markers in the bottle gourd crop are very restricted, impeding genetic improvement, cultivar identification, and phylogenetic studies. Next-generation sequencing has revolutionized the approaches for discovery, assessment, and validation of molecular markers. We conducted a genome-wide analysis, for developing SSR markers by utilizing restriction site-associated DNA sequencing (RAD-Seq) data obtained from NCBI. By performing in silico mining of microsatellite repeat motifs, we developed 45,066 perfect SSR markers. Of which 207 markers were successfully validated and 120 (57.97%) polymorphic primer pairs were utilized for an in-depth genetic diversity and population structure analysis of 96 accessions from the National Genebank of India. Tetranucleotide repeats (∼34.3%) were the most prevalent followed by trinucleotide repeats (∼30.73%), further 21.03%, 9.6%, and 4.3% of di-, penta-, and hexa-nucleotide repeats in the bottle gourd genome, respectively. Synteny of SSR markers on 11 bottle gourd linkage groups was correlated with the 7 chromosomes of cucumber (93.2%), 12 chromosomes of melon (87.4%), and 11 of watermelon (90.8%). The generated SSR markers provide a valuable tool for germplasm characterization, genetic linkage map construction, studying synteny, gene discovery, and for breeding in bottle gourd and other cucurbits species.

Key Message

Development of 45,066 perfect microsatellite markers as a valuable tool for marker assisted selection (MAS) in plant breeding.

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
Fig. 5

Similar content being viewed by others

References

  • Altshuler D, Pollara VJ, Cowles CR, Van EWJ, Baldwin J, Linton L, Lander ES (2000) An SNP map of the human genome generated by reduced representation shotgun sequencing. Nature 407:513–516

    Article  CAS  PubMed  Google Scholar 

  • Andrew CC, Michael KB, Patricia AM, David LE, David P (2006) Reconstructing the origins and dispersal of the Polynesian bottle gourd (Lagenaria siceraria). Mol Biol Evol 23:893–900

    Article  Google Scholar 

  • Badmanaban R, Patel CN (2010) Studies on anthelmintic and antimicrobial activity of the leaf extracts of 359 Lagenaria siceraria. Mol J Global Pharma Technol 4:66–70

    Google Scholar 

  • Barot AM, Suneeta P, Hiral M (2014) Development of technology for manufacture of bottle gourd ice cream. J Nutr Food Sci 4:6

    Google Scholar 

  • Bhawna, Abdin MZ, Arya L, Dipnarayan S, Sureja AK, Chitra P, Verma M (2014) Population structure and genetic diversity in bottle gourd [Lagenaria siceraria (Mol) Standl] germplasm from India assessed by ISSR markers. Plant Syst Evol 300:767–773

    Article  Google Scholar 

  • Bhawna, Abdin MZ, Arya L, Verma M (2015a) Development of novel gene-based microsatellite markers for robust genotyping purposes in Lagenaria siceraria. Sci Hortic 191:15–24

    Article  CAS  Google Scholar 

  • Bhawna, Abdin MZ, Arya L, Verma M (2015b) Transferability of cucumber microsatellite markers used for phylogenetic analysis and population structure study in bottle gourd (Lagenaria siceraria (Mol) Standl). Appl Biochem Biotechnol 175:2206–2223

    Article  CAS  PubMed  Google Scholar 

  • Bowers JE, Meredith CP (1997) The parentage of a classic wine grape, Cabernet Sauvignon. Nat Genet 16:84–87

    Article  CAS  PubMed  Google Scholar 

  • Bruneaux M, Johnston SE, Herczeg G, Merila J, Primmer CR, Vasemagi A (2013) Molecular evolutionary and population genomic analysis of the nine-spined stickleback using a modified restriction-site-associated DNA tag approach. Mol Ecol 22:5065–5082

    Article  Google Scholar 

  • Campomayor NB, Waminal NE, Kang BY, Nguyen TH, Lee SS, Huh JH, Kim HH (2021) Subgenome discrimination in Brassica and Raphanus allopolyploids using microsatellites. Cells 10(9):2358:103390/cells10092358

  • Cavagnaro PF, Senalik DA, Yang L et al (2010) Genome-wide characterization of simple sequence repeats in cucumber (Cucumis sativus L). BMC Genomics 11:569:101186/1471-2164-11-569

  • Danieli MN, Lauren dCD, (2020) Waterproofing of bottle gourd (Lagenaria siceraria) with castor oil polyurethane resin Belo. Horizonte 1:4

    Google Scholar 

  • Decker-Walters D, Staub J, Lopez-Sese A, Nakata E (2001) Diversity in landraces and cultivars of bottle gourd (Lagenaria siceraria; Cucurbitaceae) as assessed by random amplified polymorphic DNA. Genet Resour Crop Evol 48:369–380

    Article  Google Scholar 

  • Elshire RJ, Glaubitz JC, Sun Q, Poland JA, Kawamoto K, Buckler ES, Mitchell SE (2011) A robust, simple genotyping-by-sequencing (GBS) approach for high diversity species. PLoS ONE 6:e19379

  • Emina M, Berenji J, Ognjanov V, Mirjana L, Jelena C (2012) Genetic variability of bottle gourd Lagenaria siceraria (mol) Standley and its morphological characterization by multivariate analysis. Arch Biol Sci, Belgrade 64(2):573–583

    Article  Google Scholar 

  • Erickson DL, Smith BD, Clarke AC, Sandweiss DH, Tuross N (2005) An Asian origin for a 10,000-year-old domesticated plant in the Americas. Proc Natl Acad Sci USA 102:18315–18320

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Fard MH, Bodhankar SL, Dikshit M (2008) Cardioprotective activity of fruit of Lagenaria siceraria (Molina) Standley on Doxorubicin induced cardiotoxity in rats. Int J Pharmacol 6:466–471

    Google Scholar 

  • Garcia-Mas J, Benjak A, Sanseverino W (2012) The genome of melon (Cucumis melo L). Proc Natl Acad Sci USA 109:11872–11877

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Garza JC, Slatkin M, Freimer NB (1995) Microsatellite allele frequencies in humans and chimpanzees, with implications for constraints on allele size. Mol Biol Evol 12:594–603

    CAS  PubMed  Google Scholar 

  • Hecht BC, Campbell NR, Holecek DE, Narum SR (2013) Genome-wide association reveals genetic basis for the propensity to migrate in wild populations of rainbow and steelhead trout. Mol Ecol 22:3061–3076

    Article  CAS  PubMed  Google Scholar 

  • Hegarty M, Yadav R, Lee M, Armstead I, Sanderson R, Scollan N, Powell W, Skøt L (2013) Genotyping by RAD sequencing enables mapping of fatty acid composition traits in perennial ryegrass [Lolium perenne (L)]. Plant Biotechnology 11:572–581

    Article  CAS  Google Scholar 

  • Heiser CB (1979) The Gourd Book. University of Oklahoma Press, Norman, Oklahoma, A through and fascinating account of gourds from throughout the world

    Google Scholar 

  • Helyar SJ, Hemmer-Hansen J, Bekkevold D et al (2011) Application of SNPs for population genetics of non-model organisms: new opportunities and challenges. Mol Ecol Resour 1:123–136

    Article  Google Scholar 

  • Innan H, Terauchi R, Miyashita NT (1997) Microsatellite polymorphism in natural populations of the wild plant Arabidopsis thaliana. Genetics 146:1441–1452

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Jacob M, Hussein AS, Alfred OO, Beyene AA (2016) Amelework genetic differentiation of bottle gourd [Lagenaria siceraria (Molina) Standl] landraces assessed by fruit qualitative traits and simple sequence repeat markers. Sci Hortic 216:1–11

    Google Scholar 

  • Kashi Y, King D, Soller M (1997) Simple sequence repeats as a source of quantitative genetic variation. Trends Genet 13:74–78

    Article  CAS  PubMed  Google Scholar 

  • Krzywinski M, Schein J, Birol I, Connors J, Gascoyne R, Horsman D, Jones SJ, Marra MA (2009) Circos: an information aesthetic for comparative genomics. Genome Res 19:1639–1645

    Article  CAS  PubMed  PubMed Central  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  PubMed  Google Scholar 

  • Li Q, Li H, Huang W, Xu Y, Zhou Q, Wang S, Ruan J, Huang S, Zhang Z (2019) A chromosome-scale genome assembly of cucumber (Cucumis sativus L). Gigascience 8:giz072

  • Liu K, Muse SV (2005) PowerMarker: an integrated analysis environment for genetic marker analysis. Bioinf Appl Notes 21(9):2128–2129

    Article  CAS  Google Scholar 

  • MacHugh DE, Shriver MD, Loftus RT, Cunningham P, Bradley DG (1997) Microsatellite DNA variation and the evolution, domestication and phylogeography of taurine and zebu cattle (Bos taurus and Bos indicus). Genetics 146:1071–1086

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • McCouch SR, Teytelman L, Xu YB, Lobos KB, Clare K, Walton M, Fu BY, Maghirang R, Li ZK, Xing YZ, Zhang QF, Kono I, Yano M, Fjellstrom R, DeClerck G, Schneider D, Cartinhour S, Ware D, Stein L (2002) Development and mapping of 2240 new SSR markers for rice (Oryza sativa L). DNA Res 9:199–207

    Article  CAS  PubMed  Google Scholar 

  • Mehtap Y, Hugo EC, Suat S, Ceknas E, Faheem SB (2013) Transferability of Cucurbita SSR markers for genetic diversity assessment of Turkish bottle gourd (Lagenaria siceraria) genetic resources. Biochem Syst Ecol 59:45–53

    Google Scholar 

  • Morgante M, Hanafey M, Powell W (2002) Microsatellites are preferentially associated with nonrepetitive DNA in plant genomes. Nat Genet 30:194–200

    Article  CAS  PubMed  Google Scholar 

  • Nadeau NJ, Martin SH, Kozak KM, Salazar C, Dasmahapatra KK, Davey JW, Baxter SW, Blaxter ML, Mallet J, Jiggins CD (2013) Genome-wide patterns of divergence and gene flow across a butterfly radiation. Mol Ecol 22:814–826

    Article  CAS  PubMed  Google Scholar 

  • Nidhi T, Ganesh NS, Bhirendra S (2017) Medicinal value of Lagenaria siceraria: an overview. Int J Indig Herb Drug 36–43

  • Palamthodi S, Kadam D, Lele SS (2019) Physicochemical and functional properties of ash gourd/bottle gourd beverages blended with Jamun. J Food Sci Technol 56(1):473–482

    Article  CAS  PubMed  Google Scholar 

  • Peakall R, Smouse PE (2012) GenAlEx 6.5: genetic analysis in Excel. Population genetic software for teaching and research-an update. Bioinformatics 28:2537–2539

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Pei X, Shizhong X, Xiaohua W, Ye T, Baogen W, Sha W, Dehui Q, Zhongfu L, Li G (2013) Population genomic analyses from low-coverage RAD-Seq data: a case study on the non-model cucurbit bottle gourd. Plant J 77:430–442

    Google Scholar 

  • Powell W, Morgante M, Andre C, Henfey M, Vogel J, Tingy S, Rafalsky A (1996) The comparison of RFLP, RAPD, AFLP and SSR (microsatellite) markers for germplasm analysis. Mol Breeding 2:225–238

    Article  CAS  Google Scholar 

  • Prashant PS, Rama CP, Birendra NR (2014) A green separation of Lagenaria siceraria seed oil. Ind Crops Prod 52:796–800

    Article  Google Scholar 

  • 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 

  • Pujolar JM, Jacobsen MW, Frydenberg J, Als TD, Larsen PF, Maes GE, Zane L, Jian JB, Cheng L, Hansen MM (2013) A resource of genome-wide single-nucleotide polymorphisms generated by RAD tag sequencing in the critically endangered European eel. Mol Ecol Resour 13:706–714

    Article  CAS  PubMed  Google Scholar 

  • Rowe HC, Renaut S, Guggisberg A (2011) RAD in the realm of next-generation sequencing technologies. Mol Ecol 20:3499–34502

    CAS  PubMed  Google Scholar 

  • Sakshi M, Akhilesh T, Sanyam G, Ajay S, Anil KG (2015) An overview on Lagenaria siceraria (bottle gourd). Journal of Biomedical and Pharmaceutical Research 4(3):04–10

    Google Scholar 

  • Santi L, Wang Y, Stile MR, Berendzen K, Wanke D, Roig C, Pozzi C, Muller K, Muller J, Rohde W, Salamini F (2003) The GA octodinucleotide repeat binding factor BBR participates in the transcriptional regulation of the homeobox gene Bkn3. Plant 34:813–826

    CAS  Google Scholar 

  • Saveliev A, Everett C, Sharpe T, Webster Z, Festenstein R (2003) DNA triplet repeats mediates heterochromatin-protein-1-sensitive variegated gene silencing. Nature 422:909–913

  • Singh P, Nath R, Venkatesh V (2021) Comparative genome-wide characterization of microsatellites in Candida albicans and Candida dubliniensis leading to the development of species-specific marker. Public Health Genomics 24(1–2):1–13

    Article  PubMed  Google Scholar 

  • Somers DJ, Isaac P, Edwards K (2004) A high-density microsatellite consensus map for bread wheat (Triticum aestivum L). Theor Appl Genet 109:1105–1114

    Article  CAS  PubMed  Google Scholar 

  • Subramanian S, Mishra RK, Singh L (2003) Genome-wide analysis of microsatellite repeats in humans: their abundance and density in specific genomic regions. Genome Biol 4:R13

    Article  PubMed  PubMed Central  Google Scholar 

  • Sunde J, Yıldırım Y, Tibblin P, Forsman A (2020) Comparing the performance of microsatellites and RADseq in population genetic studies: Analysis of data for pike (Esox lucius) and a synthesis of previous studies. Frontiers in Genetics 11:218:103389/fgene202000218

  • Tautz D, Renz M (1984) Simple sequences are ubiquitous repetitive components of eukaryotic genomes. Nucleic Acids Res 12:4127–4138

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Toth G, Gaspari Z, Jurka J (2000) Microsatellites in different eukaryotic genomes: survey and analysis. Genome Res 10:967–981

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Upaganlawar A, Balaraman R (2010) Protective effects of Lagenaria siceraria (Molina) fruit juice in isoproterenol induced myocardial infarction. Int J Pharmacol 5:645–651

    Article  Google Scholar 

  • Varshney RK, Graner A, Sorrells ME (2005) Genic microsatellite markers in plants: features and applications. Trend in Biotechonol 23:48–55

    Article  CAS  Google Scholar 

  • Varshney RK, Song C, Saxena RK (2013) Draft genome sequence of chickpea (Cicer arietinum) provides a resource for trait improvement. Nat Biotechnol 31:240–246

    Article  CAS  PubMed  Google Scholar 

  • Wang Y, Xu P, Wu X, Wu X, Wang B, Huang Y, Hu Y, Lin J, Lu Z, Li G (2018) GourdBase: a genome-centered multi-omics database for the bottle gourd (Lagenaria siceraria), an economically important cucurbit crop. Sci Rep 8:3604

    Article  PubMed  PubMed Central  Google Scholar 

  • WhitakerTW (1971) Endemism and pre-Columbian migration of bottle gourd, Lagenaria siceraria (Mol) Standl Man across the sea. In: Riley, C.L.K.J.C., Pennington, C.W. and Runds, R.L., Eds., Man across the Sea, University of Press, Austin, 64–69

  • Wu S, Shamimuzzaman M, Honghe S, Jerome S, Xuelian S, Alan W, Zujian W, Amnon L, Yong X, Kai-Shu L, Zhangjun F (2017) The bottle gourd genome provides insights into Cucurbitaceae evolution and facilitates mapping of a Papaya ringspot virus resistance locus. Plant J 92(5):963–975

    Article  CAS  PubMed  Google Scholar 

  • Wu S, Wang X, Reddy U, Sun H, Bao K, Gao L, Mao L, Patel T, Ortiz C, Abburi VL, Nimmakayala P, Branham S, Wechter P, Massey L, Ling KS, Kousik C, Hammar SA, Tadmor Y, Portnoy V, Gur A, Katzir N, Guner N, Davis A, Hernandez AG, Wright CL, McGregor C, Jarret R, Zhang X, Xu Y, Wehner TC, Grumet R, Levi A, Fei Z (2019) Genome of “Charleston Gray”, the principal American watermelon cultivar, and genetic characterization of 1,365 accessions in the US National Plant Germplasm System watermelon collection. Plant Biotechnol J 17(12):2246–2258

    Article  CAS  PubMed  PubMed Central  Google Scholar 

  • Xu P, Wu X, Luo J, Wang B, Liu Y, Ehlers JD, Wang S, Lu Z, Li G (2011) Partial sequencing of the bottle gourd genome reveals markers useful for phylogenetic analysis and breeding. BMC Genomics 120:467

    Article  Google Scholar 

  • Yang H, Tao Y, Zheng Z, Zhang Q, Zhou G, Sweetingham MW, Ho-wieson JG, Li C (2013) Draft genome sequence, and a sequence defined genetic linkage map of the legume crop species Lupinus angustifolius L. PLoS ONE 8: e64799

  • Yeh FC, Yang RC, Boyle T (1999) POPGENE Version 1.31. Microsoft window-based freeware for population genetic analysis

Download references

Acknowledgements

The first author Bhawna Bonthala acknowledges the award of Senior Research Fellowship from the University Grants Commission, New Delhi. Support from the National Bureau of Plant Genetic Resources, New Delhi, India, is duly acknowledged.

Author information

Authors and Affiliations

Authors

Contributions

All authors contributed to the study conception and design. Material preparation, data collection, and analysis were performed by Bhawna Bonthala, Ved Dixit, Priya Yadav, and Manjusha Verma. The first draft of the manuscript was written by Bhawna Bonthala, and all authors commented on previous versions of the manuscript. All authors read and approved the final manuscript.

Corresponding author

Correspondence to Manjusha Verma.

Ethics declarations

Conflict of interest

The authors declare no competing interests.

Additional information

Communicated by Izabela Pawłowicz.

Publisher's note

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

Supplementary Information

Below is the link to the electronic supplementary material.

Supplementary file1 (DOCX 100 KB)

Supplementary file2 (XLS 14.9 MB)

Rights and permissions

Reprints and permissions

About this article

Check for updates. Verify currency and authenticity via CrossMark

Cite this article

Bonthala, B., Abdin, M.Z., Arya, L. et al. Genome-wide SSR markers in bottle gourd: development, characterization, utilization in assessment of genetic diversity of National Genebank of India and synteny with other related cucurbits. J Appl Genetics 63, 237–263 (2022). https://doi.org/10.1007/s13353-022-00684-1

Download citation

  • Received:

  • Revised:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s13353-022-00684-1

Keywords

Navigation