Skip to main content
SpringerLink
Log in

Genetic homogeneity of guava plants derived from somatic embryogenesis using SSR and ISSR markers

  • Research Note
  • Published:
Plant Cell, Tissue and Organ Culture (PCTOC) Aims and scope Submit manuscript

Abstract

To evaluate genetic homogeneity of 1-year-old guava (Psidium guajava L.) plants developed from in vitro somatic embryogenesis, DNA from leaf tissues of seven randomly selected plants along with the mother plant, was isolated and subjected to molecular analysis. A total of six Simple Sequence Repeat (SSR) primer pairs, producing reproducible and clear bands ranging from 100 to 300 bp in size, resulted in amplification of single band (allele), corresponding homozygous individuals. Moreover, of 10 different inter-simple sequence repeat (ISSR) primers screened, six produced resolvable, reproducible and scorable bands. All these ISSRs produced a total of 25 bands, ranging between 300 and 1,200 bp length, and the number of scorable bands, for each primer varied from three to six with an average of 4.1 bands per primer. The amplification products were monomorphic across all the micropropagated plants produced by all SSR and ISSR primers applied. The monomorphic banding pattern in micropropagated plants and the mother plant confirms the genetic homogeneity of the in vitro raised plants and demonstrates the reliability of our in vitro propagation system for guava.

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

Access this article

Log in via an institution

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

Instant access to the full article PDF.

Institutional subscriptions

Fig. 1
Fig. 2

References

  • Aggarwal D, Kumar A, Reddy MS (2010) Shoot organogenesis in elite clones of Eucalyptus tereticornis. Plant Cell, Tissue Organ Cult 102:45–52

    Article  Google Scholar 

  • Amin MN, Jaiswal VS (1987) Rapid clonal propagation of guava through in vitro shoot proliferation on nodal explants of mature trees. Plant Cell, Tissue Organ Cult 9:235–243

    Article  Google Scholar 

  • Asthana P, Jaiswal VS, Jaiswal U (2011) Micropropagation of Sapindus trifoliatus L. and assessment of genetic fidelity of micropropagated plants using RAPD analysis. Acta Physiol Plant 33:1821–1829

    Article  CAS  Google Scholar 

  • Bairu MW, Aremu AO, Van Staden J (2011) Somaclonal variation in plants: causes and detection methods. Plant Growth Regul 63:147–173

    Article  CAS  Google Scholar 

  • Bennici A, Anzidei M, Vendramin CG (2004) Genetic stability and uniformity of Foeniculum vulgare Mill. regenerated plants through organogenesis and somatic embryogenesis. Plant Sci 166:221–227

    Article  CAS  Google Scholar 

  • Bhatia R, Singh KP, Jhang T, Sharma TR (2009) Assessment of clonal fidelity of micropropagated gerbera plants by ISSR markers. Sci Hortic 119:208–211

    Article  CAS  Google Scholar 

  • Bhatia R, Singh KP, Sharma TR, Jhang T (2011) Evaluation of the genetic fidelity of in vitro-propagated gerbera (Gerbera jamesonii Bolus) using DNA-based markers. Plant Cell, Tissue Organ Cult 104:131–135

    Article  Google Scholar 

  • Bhattacharya S, Bandopadhyay TK, Ghosh PD (2010) Somatic embryogenesis in Cymbopogon pendulus and evaluation of clonal fidelity of regenerants using ISSR marker. Sci Hortic 123:505–513

    Article  CAS  Google Scholar 

  • Biswas BK, Joshee N, Yadav A, Yadav AK (2007) Development and application of biotechnology in guava: a nutraceutical fruit. In: Desjardins Y (ed) ISHS Acta Horticulturae 744: Ist international symposium on human health effects of fruits and vegetables, pp 267–276

  • Brito G, Lopes T, Loureiro J, Rodriguez E, Santos C (2010) Assessment of genetic stability of two micropropagated wild olive species using flow cytometry and microsatellite markers. Tree Struct Funct 24:723–732

    Article  CAS  Google Scholar 

  • Carra A, Sajeva M, Abbate L, Siragusa M, Sottile F, Carimi F (2012) In vitro plant regeneration of caper (Capparis spinosa L.) from floral explants and genetic stability of regenerants. Plant Cell, Tissue Organ Cult 109:373–381

    Article  Google Scholar 

  • Chandra R, Bajpai A, Gupta S, Tiwari RK (2004) Embryogenesis and plant regeneration from mesocarp of Psidium guajava L. (guava). Indian J Biotech 3:246–248

    Google Scholar 

  • Doyle JJ, Doyle JL (1990) Isolation of plant DNA from fresh tissue. Focus 12:13–15

    Google Scholar 

  • George EF, Debergh PC (2008) Micropropagation: uses and methods. In: George EF, Hall MA, De Klerk GJ (eds) Plant propagation by tissue culture, 3rd edn. Springer, Dordrecht, The Netherlands, pp 29–64

  • Gupta AK, Harish, Rai MK, Phulwaria M, Shekhawat NS (2011) Isolation of genomic DNA suitable for community analysis from mature trees adapted to arid environment. Gene 487:156–159

    Article  PubMed  CAS  Google Scholar 

  • Jain SM (2001) Tissue culture-derived variation in crop improvement. Euphytica 118:153–166

    Article  CAS  Google Scholar 

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

    Article  CAS  Google Scholar 

  • Kumar S, Mangal M, Dhawan AK, Singh N (2011) Assessment of genetic fidelity of micropropagated plants of Simmondsia chinensis (Link) Schneider using RAPD and ISSR markers. Acta Physiol Plant 33:2541–2545

    Article  CAS  Google Scholar 

  • Loh CS, Rao AN (1989) Clonal propagation of guava (Psidium guajava L.) from seedlings and grafted plants and adventitious shoot formation in vitro. Sci Hortic 39:31–39

    Article  Google Scholar 

  • Lopes T, Pinto G, Loureiro J, Costa A, Santos C (2006) Determination of genetic stability in long-term somatic embryogenic cultures and derived plantlets of cork oak using microsatellite markers. Tree Physiol 26:1145–1152

    Article  PubMed  CAS  Google Scholar 

  • Marum L, Rocheta M, Maroco J, Oliveira MM, Miguel C (2009) Analysis of genetic stability at SSR loci during somatic embryogenesis in maritime pine (Pinus pinaster). Plant Cell Rep 28:673–682

    Article  PubMed  CAS  Google Scholar 

  • Murashige T, Skoog FA (1962) Revised medium for rapid growth and bioassays with tobacco tissue cultures. Physiol Plant 15:473–497

    Article  CAS  Google Scholar 

  • Nookaraju A, Agrawal DC (2012) Genetic homogeneity of in vitro raised plants of grapevine cv. Crimson Seedless revealed by ISSR and microsatellite markers. S Afr J Bot 78:302–306

    Article  CAS  Google Scholar 

  • Papadatau P, Pontikis CA, Ephtimiadou E, Lydaki M (1990) Rapid multiplication of guava seedlings by in vitro shoot tip culture. Sci Hortic 45:99–108

    Article  Google Scholar 

  • Rahman MH, Rajora OP (2001) Microsatellite DNA somaclonal variation in micropropagated trembling aspen (Populus tremuloides). Plant Cell Rep 20:531–536

    Article  CAS  Google Scholar 

  • Rai MK, Akhtar N, Jaiswal VS (2007) Somatic embryogenesis and plant regeneration in Psidium guajava L. cv. Banarasi local. Sci Hortic 113:129–133

    Article  CAS  Google Scholar 

  • Rai MK, Jaiswal VS, Jaiswal U (2008) Effect of ABA and sucrose on germination of encapsulated somatic embryos of guava (Psidium guajava L.). Sci Hortic 117:302–305

    Article  CAS  Google Scholar 

  • Rai MK, Jaiswal VS, Jaiswal U (2009) Effect of selected amino acids and polyethylene glycol on maturation and germination of somatic embryos of guava (Psidium guajava L.). Sci Hortic 121:233–236

    Article  CAS  Google Scholar 

  • Rai MK, Asthana P, Jaiswal VS, Jaiswal U (2010) Biotechnological advances in guava (Psidium guajava L.): recent developments and prospects for further research. Tree Struct Funct 24:1–12

    Article  CAS  Google Scholar 

  • Rai MK, Shekhawat NS, Harish, Gupta AK, Phulwaria M, Ram K, Jaiswal U (2011) The role of abscisic acid in plant tissue culture—a review of recent progress. Plant Cell, Tissue Organ Cult 106:179–190

  • Rani V, Raina SN (2000) Genetic fidelity of organized meristem derived micropropagated plants: a critical reappraisal. In vitro Cell Dev Biol Plant 36:319–330

    Article  CAS  Google Scholar 

  • Rani V, Parida A, Raina SN (1995) Random amplified polymorphic DNA (RAPD) markers for genetic analysis in micropropagated plants of Populus deltoides Marsh. Plant Cell Rep 14:459–462

    Article  CAS  Google Scholar 

  • Risterucci AM, Duval MF, Rohde W, Billotte N (2005) Isolation and characterization of microsatellite loci from Psidium guajava L. Mol Ecol Notes 5:745–748

    Article  CAS  Google Scholar 

  • Singh SK, Meghwal PR, Sharma HC, Singh SP (2002) Direct shoot organogenesis on explants from germinated seedlings of Psidium guajava L. cv. Allahabad Safeda. Sci Hortic 95:213–221

    Article  CAS  Google Scholar 

  • Singh SK, Rai MK, Sahoo L (2012) An improved and efficient micropropagation of Eclipta alba through transverse thin cell layer culture and assessment of clonal fidelity using RAPD analysis. Ind Crop Prod 37:328–333

    Article  CAS  Google Scholar 

  • Varshney A, Lakshmikumaran M, Srivastava PS, Dhawan V (2001) Establishment of genetic fidelity of in vitro-raised Lilium bulblets through RAPD markers. In Vitro Cell Dev Biol Plant 37:227–231

    Article  CAS  Google Scholar 

  • Wang QM, Gao FZ, Gao X, Zou FY, Sui X, Wang M, Hui YJ, Wang L (2012) Regeneration of Clivia miniata and assessment of clonal fidelity of plantlets. Plant Cell, Tissue Organ Cult 109:191–200

    Google Scholar 

  • Yasseen M, Barringer SA, Schnell RJ, Splittoesser WE (1995) In vitro shoot proliferation of guava (Psidium guajava L.) from germinated seedlings. Plant Cell Rep 14:525–528

    Google Scholar 

Download references

Acknowledgments

The authors (M. K. Rai, Harish, Mahendra Phulwaria) wish to acknowledge to University Grants Commission (UGC), New Delhi for the award of Dr. D.S. Kothari Post Doctoral Fellowship and UGC Post Doctoral Fellowship.

Author information

Author notes

  1. Harish

    Present address: Department of Botany, Mohanlal Sukhadia University, Udaipur, Rajasthan, 313001, India

Authors and Affiliations

  1. Biotechnology Centre, Department of Botany, Jai Narain Vyas University, Jodhpur, Rajasthan, 342033, India

    Manoj K. Rai, Mahendra Phulwaria, Amit K. Gupta & N. S. Shekhawat

  2. Department of Botany, Mahila Mahavidyalaya, Banaras Hindu University, Varanasi, Uttar Pradesh, 221005, India

    U. Jaiswal

Authors
  1. Manoj K. Rai
    View author publications

    You can also search for this author in PubMed Google Scholar

  2. Mahendra Phulwaria
    View author publications

    You can also search for this author in PubMed Google Scholar

  3. Harish
    View author publications

    You can also search for this author in PubMed Google Scholar

  4. Amit K. Gupta
    View author publications

    You can also search for this author in PubMed Google Scholar

  5. N. S. Shekhawat
    View author publications

    You can also search for this author in PubMed Google Scholar

  6. U. Jaiswal
    View author publications

    You can also search for this author in PubMed Google Scholar

Corresponding author

Correspondence to Manoj K. Rai.

Rights and permissions

Reprints and permissions

About this article

Cite this article

Rai, M.K., Phulwaria, M., Harish et al. Genetic homogeneity of guava plants derived from somatic embryogenesis using SSR and ISSR markers. Plant Cell Tiss Organ Cult 111, 259–264 (2012). https://doi.org/10.1007/s11240-012-0190-1

Download citation

  • Received:

  • Accepted:

  • Published:

  • Issue Date:

  • DOI: https://doi.org/10.1007/s11240-012-0190-1

Keywords

Search

Navigation