Assessment of clonal fidelity of doubled haploid line of marigold (Tagetes erecta) using microsatellite markers

UZMA MEHRAJ; SAPNA PANWAR; KANWAR PAL SINGH; NAMITA NAMITA; RENU PANDEY; AMOLKUMAR U SOLANKE; NIHARIKA MALLICK; SURENDRA KUMAR

doi:10.56093/ijas.v89i7.91690

Authors

UZMA MEHRAJ Research Scholar, ICAR- Indian Agricultural Research Institute, New Delhi 110 012 India
SAPNA PANWAR Scientist, ICAR- Indian Agricultural Research Institute, New Delhi 110 012 India
KANWAR PAL SINGH Principal Scientist, ICAR- Indian Agricultural Research Institute, New Delhi 110 012 India
NAMITA NAMITA Scientist, ICAR- Indian Agricultural Research Institute, New Delhi 110 012 India
RENU PANDEY Principal Scientist, ICAR- Indian Agricultural Research Institute, New Delhi 110 012 India
AMOLKUMAR U SOLANKE Scientist, National Research Centre on Plant Biotechnology, New Delhi
NIHARIKA MALLICK Scientist, ICAR- Indian Agricultural Research Institute, New Delhi 110 012 India
SURENDRA KUMAR Assistant Chief Technical Officer, ICAR- Indian Agricultural Research Institute, New Delhi 110 012 India

https://doi.org/10.56093/ijas.v89i7.91690

Keywords:

Doubled haploid line, In vitro regeneration, SSR markers, Tagetes erecta

Abstract

In vitro multiplied plants of doubled haploid line of marigold (Tagetes erecta L.) derived through ovule culture, were subjected to evaluation of genetic uniformity using SSR markers in order to evaluate their genetic stability and/ or detect likely existing variations among in vitro derived plantlets. Leaf samples were taken from randomly selected 13 plants at post acclimatization stage, for checking the clonal fidelity of the in vitro derived regenerants by comparing them with the mother plant from which the leaf explants were taken. Out of 36 primers, 12 showed amplification in which clear and reproducible PCR profiles. Analysis of individual primers revealed that SSR primers produced same banding pattern which were shared by the in vitro leaf derived regenerants and the donor mother plant, which indicated that there was no variation observed within the tissue culture raised plants. All the banding profiles produced were monomorphic and were similar to those of the mother plant, and hence, showed that plants were true to type. Information presented in the study indicated that the in vitro regenerated plants were identical to the mother plant.

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References

Alizadeh M, Krishna H, Eftekhari M, Modareskia M and Modareskia M. 2015.Assessment of clonal fidelity in micropropagated horticultural plants. Journal of Chemical and Pharmaceutical Research 7(12): 977–90.

Anonymous. 2017. Indian Horticulture Database. Available Online at http://nhb.gov.in. DOI: https://doi.org/10.5194/nhess-2017-347-SC2

Bandupriya H D D, Iroshini W W M A, Perera S A C N, Vidhanaarachchi V R M, Fernando S C, Santha E S and Gunathilake T R. 2017. Genetic fidelity testing using SSR marker assay confirms trueness to type of micropropagated coconut (Cocos nucifera L.) plantlets derived from unfertilized ovaries. Open Plant Science Journal 11: 46–54. DOI: https://doi.org/10.2174/1874294701710010046

De Laia M L, Gomes E A, Esbrisse E J. and De Araujo E F. 2000. Random Amplified Polymorphic DNA (RAPD) analysis of genotypic identities in Eucalyptus clones. Silvae Genetica 49: 239–43.

Godoy-Hernandez G. and Miranda-Ham M L. 2007. Marigold biotechnology: Tissue culture and genetic transformation. Transgenic Plant Journal 1(1): 169–74.

Guha S and Maheswari S C. 1964. In vitro production of embryos from anthers of datura. Nature 204: 497–8. DOI: https://doi.org/10.1038/204497a0

Larkin P J and Scowcroft W R. 1981. Somaclonal variation a novel source of variability from cell cultures for plant improvement. Theoretical and Applied Genetics 60: 197–214. DOI: https://doi.org/10.1007/BF02342540

Mallaya N P and Ravishankar G A. 2013. In vitro propagation and genetic fidelity study of plant regenerated from inverted hypocotyl explants of eggplant (Solanum melongena L.) cv. ArkaShirish. Biotech 3(1): 45–52. DOI: https://doi.org/10.1007/s13205-012-0068-2

Murray M G and Thompson W E. 1980. Rapid isolation of high molecular weight plant DNA. Nucleic Acid Research 8: 4321–5. DOI: https://doi.org/10.1093/nar/8.19.4321

Pandey R N, Singh S P, Rastoglc J, Sharma M L, and Singh R K. 2010. One step assessment of clonal fidelity in sugarcane plantlets with RAPD and SSR marker. Australian Journal of Crop Science 6(4): 618–24.

Rahman M H and Rajora O P. 2001. Microsatellite DNA somaclonal variation in micropropagated trembling aspen (Populus tremuloides). Plant Cell Reports 20(6): 531–6. DOI: https://doi.org/10.1007/s002990100365

Rani V and Raina S N. 2000. Genetic fidelity of organized meristem-derived micropropagated plants: A critical reappraisal. In Vitro Cellular and Development Biology-Plant 36: 319–30. DOI: https://doi.org/10.1007/s11627-000-0059-6

Ravindra K, Singh K P, Raju D V S, Panwar S, Bhatia R, Kumar S. and Verma P K. 2018. Standardization of in vitro culture establishment and proliferation of micro-shoots in African and French marigold genotypes. International Journal of Current Microbiology and Applied Sciences 7(1): 2768–81. DOI: https://doi.org/10.20546/ijcmas.2018.701.332

Rawls B, Harris-Shultz K, Dhekney S, Forrester I and Sitther V. 2015.Clonal fidelity of micropropagated Psidium guajava L. plants using microsatellite markers. American Journal of Plant Sciences 6: 2385–92. DOI: https://doi.org/10.4236/ajps.2015.614241

Ray T, Dutta I, Saha P, Das S and Roy S C. 2006. Genetic stability of three economically important micropropagated banana (Musa spp.) cultivars of lower Indo-Gangetic plains as assessed by RAPD and ISSR markers. Plant Cell Tissue and Organ Culture 85: 11–21. DOI: https://doi.org/10.1007/s11240-005-9044-4

Reddy P M, Saral N. and Siddiq E A. 2002. Inter simple sequence repeat (ISSR) polymorphism and its application in plant breeding. Euphytica 128: 9–17.

Vandana, Sharma N K and Mithilesh K. 2012. Assessment of genetic fidelity of micropropagated clones of strawberry genotypes using EST-SSR markers. BioTechnology: An Indian Journal 6(4, 5): 150–3.

Whankaew S, Hasthanasombut S, Taikert R, Tappiban P, Smith D R and Triwitayakorn K. 2014. Molecular characterization and genetic relationship of marigolds (Tagetes spp.) based on simple sequence repeats markers. Plant Genetic Resources: Characterization and Utilization 12(3): 317–22. DOI: https://doi.org/10.1017/S1479262114000100