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Colchicine-mediated in vitro polyploidization in gerbera hybrid

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Abstract

An efficient in vitro protocol for high-frequency polyploidization for the first time in gerbera hybrid (BGC-2019-01) was developed in the present study. Two-week-old in vitro-developed shoots (tips) were treated individually with 0.1%, 0.25% and 0.5% (w/v) colchicine solutions for 4, 6, 8, and 12 h. The colchicine-treated shoot tips were then inoculated on Murashige and Skoog (MS) medium fortified with 1.5 mg/l meta-Topolin for multiple shoot proliferation and later transferred into 1.5 mg/l indole-3-acetic acid-fortified MS medium for rooting of shoots. The ploidy levels of the colchicine-treated and regenerated plantlets along with the non-treated ones were confirmed via flow cytometry analysis and metaphasic chromosome count. The highest frequency of tetraploid plantlets (50%) were obtained when shoot tips were treated with 0.1% colchicine for 4 h. Morphological observations revealed that induced tetraploid plantlets exhibited delayed fresh shoot initiation, fewer but longer shoots, as well as fewer but broader leaves. Likewise, the study of stomata revealed that in comparison to their diploid counterparts, the tetraploid plantlets exhibited less frequent yet significantly larger stomata, and higher number of chloroplasts. The tetraploids were recorded with significantly higher chlorophyll, carotenoid, and anthocyanin content during the photosynthetic pigment analyses. During ex vitro acclimatization and field growth, the tetraploid plants exhibited delayed proliferation but with higher vigor and thickened broad leaves. The genetic uniformity among the diploid and the tetraploid plants was confirmed using conserved DNA-derived polymorphism (CDDP), directed amplification of minisatellite-region DNA (DAMD), inter simple sequence repeats (ISSR), and start codon targeted (SCoT) polymorphism marker systems. The tetraploids developed in the present study would be of immense importance for the genetic improvement of gerbera as far as its ornamental values are concerned.

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Data availability

The datasets generated during and/or analyzed during the current study are available from the corresponding author on reasonable request.

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Funding

This research was funded by Department of Science & Technology and Biotechnology, Govt. of West Bengal, India [Sanction No. 1478(Sanc.)STBT-13015/15/11/2021-ST SEC].

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Authors and Affiliations

  1. Crop Research Unit (Genetics and Plant Breeding), Bidhan Chandra Krishi Viswavidyalaya, Mohanpur, Nadia, West Bengal, 741252, India

    Manisha Mahanta, Saikat Gantait, Sutanu Sarkar & Somnath Bhattacharyya

  2. All India Coordinated Research Project on Floriculture, Bidhan Chandra Krishi Viswavidyalaya, Mohanpur, Nadia, West Bengal, 741252, India

    Raghunath Sadhukhan

Authors
  1. Manisha Mahanta
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  2. Saikat Gantait
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  3. Sutanu Sarkar
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  4. Raghunath Sadhukhan
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  5. Somnath Bhattacharyya
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Contributions

SG and SB contributed to the study conception and design. MM and SG performed the material preparation, experimentations, data collection and analysis. MM and SG prepared the first draft of the manuscript. SG, SS, RS and SB commented and edited on previous versions of the manuscript. All authors read and approved the final manuscript.

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Correspondence to Saikat Gantait.

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Mahanta, M., Gantait, S., Sarkar, S. et al. Colchicine-mediated in vitro polyploidization in gerbera hybrid. 3 Biotech 13, 74 (2023). https://doi.org/10.1007/s13205-022-03457-z

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