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Effect of Silver Nitrate and Thidiazuron on Shoot Proliferation, Hyperhydricity and Assessment of Genetic Fidelity of Microplants in Carnation (Dianthus caryophyllus L.)

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Abstract

In the present investigation, optimization of the best concentration combinations of BAP (Benzyl Amino Purine), TDZ (Thidiazuron) and Silver Nitrate (AgNO3) for micropropagation of carnation (Dianthus caryophyllus L.) cv. Irene was performed. Genetic fidelity in the microplants of different generation was determined with the help of ISSR markers. Results of the present investigation showed that the MS gelled media supplemented with BAP (0.5 mg L–1) and AgNO3 (1.5 mg L–1) showed maximum shoot proliferation, good shoot length and reduced hyperhydricity significantly. Our study also indicates that hyperhydricity decreased by substantially by increasing the concentration of AgNO3 compared to control. High level of AgNO3 in MS medium did not helped to reduce hyperhydricity. Another chemical combination of TDZ (1.0 mg L–1) and AgNO3 (0.5 mg L–1) in MS gelled media gave the maximum shoot proliferation, shoot length with minimum hyperhydricity which showed normal growth and development compared to control and other treatments. This concentration combination proved effective to reduce hyperhydricity and to enhance shoot proliferation. Because TDZ with AgNO3 had pronounced effect to reduce hyperhydricity, therefore, this concentration combination was used to advance the progeny up to subculture (SC10) using donor mother microplants (SC1) selected from the best concentration response to know the genetic fidelity among the different generations. Microplants which were non hyperhydrified were used to assess the genetic fidelity using ISSR markers. Fourteen ISSR primers were used that gave good amplification and produced 616 bands with an average of 44 amplicon per primer from in vitro raised microplants including mother plant where the banding patterns exhibited uniform and homogeneous banding patterns. This study may help us to know applicability of ISSR markers to establish genetic fidelity among in vitro—raised microplants of carnation.

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ACKNOWLEDGMENTS

Author of the present work is thankful to Dr. Manoj Kumar Yadav for providing the support to complete the work in his lab with proper guidance. Research support provided by SVPUA&T Meerut is acknowledged by the authors. All authors had to have the equal contribution in the research work done.

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Authors and Affiliations

  1. Department of Horticulture, College of Agriculture, SVP University of Agriculture and Technology, 250110, Meerut, Uttar Pradesh, India

    Ram Lakhan Maurya & Mukesh Kumar

  2. Department of Biotechnology, SVP University of Agriculture and Technology, Meerut, 250110, Uttar Pradesh, India

    Ujjwal Sirohi & Manoj Kumar Yadav

  3. National Institute of Plant Genome Research (NIPGR), 110067, New Delhi, India

    Ujjwal Sirohi

  4. College of Biotechnology, SVP University of Agriculture and Technology, 250001, Meerut, Uttar Pradesh, India

    Priya

  5. Department of Chemical Technology, Institute of Chemical Technology, 400019, Matunga, Mumbai, India

    Priya

  6. Department of Chemistry, Meerut College Meerut, 250001, Uttar Pradesh, India

    Veena Chaudhary

  7. Plant Genetic Resources and Improvement, CSIR-National Botanical Research Institute, Rana Pratap Marg, Lucknow, 226001, Uttar Pradesh, India

    Venukonda Rakesh Sharma

  8. Department of Biotechnology, DDU University Gorakhpur, Uttar Pradesh, India

    Dinesh Yadav

Authors
  1. Ram Lakhan Maurya
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  2. Mukesh Kumar
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  3. Ujjwal Sirohi
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  4. Priya
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  5. Veena Chaudhary
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  6. Venukonda Rakesh Sharma
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  7. Dinesh Yadav
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  8. Manoj Kumar Yadav
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Contributions

All the authors contributed equally in the present research. RLM, US and VRS did the wet experiments related to in vitro culture and molecular analysis using ISSR markers to check the genetic fidelity. VC, DY and P contributed to process the data, manuscript compilation andfinalization. MK, DY and MKY conceived the research idea and helped to design the experiments and revised the manuscript in the present form.

Corresponding author

Correspondence to Manoj Kumar Yadav.

Ethics declarations

The authors declare that they have no conflicts of interest. There is no ethical issues arise or involved in the present research work. No animal were involved or killed during the experiment. No human sentiments were involved or broken during the research work. The manuscript is not submitted anywhere else.

Additional information

Abbreviations-TDZ (Thidiazuron), AgNO3 (Silver Nitrate), RAPD (Randomly Amplified Polymorphic DNA), SSR (Simple Sequence Repeats), ISSR (Inter Simple Sequence Repeats), Cetyl Triethyl Ammonium Bromide (CTAB), Murashige and Skoog (MS).

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Maurya, R.L., Kumar, M., Sirohi, U. et al. Effect of Silver Nitrate and Thidiazuron on Shoot Proliferation, Hyperhydricity and Assessment of Genetic Fidelity of Microplants in Carnation (Dianthus caryophyllus L.). Cytol. Genet. 57, 87–94 (2023). https://doi.org/10.3103/S0095452723010061

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