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  1. Assessment of genetic stability on in vitro and ex vitro plants of Ficus carica var. black jack using ISSR and DAMD markers
    Parab AR, Lynn CB, Subramaniam S
    Mol Biol Rep, 2021 Nov;48(11):7223-7231.
    PMID: 34586562 DOI: 10.1007/s11033-021-06714-1
    BACKGROUND: Clonal propagation is one of the attributes of plant tissue culture. Therefore, analysis of genetic stability among the in vitro cultured plants is a crucial step. It helps to signify the clonal propagation of the micropropagated plants. Regenerated Ficus carica var. Black Jack plantlets were established using woody plant medium supplemented with 20 μM 6-Benzylaminopurine and 8 μM Indole-3-acetic acid under different light treatments such as normal fluorescent white light (60 μmol m-2 s-1), and four different LED spectra, white (400-700 nm), blue (440 nm), red (660 nm) and blue + red (440 nm + 660 nm). Genetic stability analysis was performed on the in vitro and ex vitro plants of Ficus carica var. Black Jack.

    METHODS AND RESULTS: Ten primers of each, ISSR and DAMD molecular markers, were used to assess the genetic stability of the eight samples of Ficus carica var. Black Jack. ISSR markers showed 97.87% of monomorphism whereas DAMD markers showed 100% monomorphism. Polymorphism of 2.13% was observed for the UBC840 ISSR-DNA primer which was negated under the genetic similarity index analysis for the eight samples. The findings of this study revealed that ISSR and DAMD markers are efficient in determining the polymorphism and monomorphism percentage among the in vitro and ex vitro samples of Ficus carica var. Black Jack.

    CONCLUSION: Monomorphism of 100% obtained using DAMD markers and more than 95% of monomorphism obtained using ISSR markers indicate that the regenerated plants are significantly genetically stable. These molecular markers can be used to test the genetic stability of in vitro regenerated plants. It is recommended that genetic stability analysis should be performed for long-term maintenance of such micropropagated plants.

  2. Fulltext Morphogenetic and physiological effects of LED spectra on the apical buds of Ficus carica var. Black Jack
    Parab AR, Han KY, Chew BL, Subramaniam S
    Sci Rep, 2021 12 08;11(1):23628.
    PMID: 34880352 DOI: 10.1038/s41598-021-03056-7
    The use of artificial light sources such as light-emitting diodes (LEDs) has become a prerequisite in tissue culture studies to obtain morphogenetic enhancements on in vitro plants. This technology is essential for developmental enhancements in the growing plant cultures due to its light quality and intensity greatly influencing the in vitro growing explants at a cellular level. The current study investigates the effects of different light-emitting diode (LED) spectra on the growth of apical buds of Ficus carica var. Black Jack. Ficus carica, commonly known as figs is rich in vitamins, minerals, and phytochemicals capable of treating microbial infections and gastric, inflammatory, and cardiac disorders. Apical buds of Ficus carica var. Black Jack, presented morphogenetic changes when grown under six different LED spectra. The highest multiple shoots (1.80 per growing explant) and healthy growing cultures were observed under the blue + red LED spectrum. Wound-induced callus formation was observed on apical buds grown under green LED spectrum and discolouration of the growing shoots were observed on the cultures grown under far-red LED spectrum. Multiple shoots obtained from the blue + red LED treatment were rooted using 8 µM indole-3-acetic acid (IAA), and the rooted plantlets were successfully acclimatised. Compared with the other monochromatic LEDs, blue + red proved to be significantly better for producing excellent plant morphogeny. It is apparent that blue and red LED is the most suitable spectra for the healthy development of plants. The findings have confirmed that the combination of blue + red LED can potentially be used for enhancing growth yields of medicinally and commercially important plants.
  3. Impact of culture vessels on micro-morphological features of in vitro Dendrobium Sabin Blue orchid
    Malik ANA, Uddain J, Chin CK, Chew BL, Antony JJJ, Parab AR, et al.
    BioTechnologia (Pozn), 2022;103(1):41-52.
    PMID: 36605376 DOI: 10.5114/bta.2022.113914
    Different designs of the plant tissue culture vessel, such as size, material, and shape, may alter its microenvironment atmosphere. The present study was conducted on protocorm-like bodies (PLBs) of Dendrobium Sabin Blue orchid to determine the development of PLBs on plastic and glass culture vessels of different sizes. PLBs were cultured in half-strength Murashige and Skoog (MS) medium with the same initial weight of 0.5 g in 10 replicates. The growth index of the PLBs was calculated after 11 weeks to study their growth in every vessel; additionally, biochemical analysis was performed to determine carbohydrate content, proline concentration, and photosynthesis pigments in the PLBs. Scanning electron microscopy (SEM) was performed to study stomata development on PLBs in each vessel, and histological analyses were conducted to study the cell structure. Overall, the PLBs cultured in a large 470 ml plastic vessel showed successful growth with a high growth index, high carbohydrate content, low-stress condition, and high chlorophyll content. SEM confirmed that the presence of trichome and rhizoid in PLBs cultured in the 470 ml plastic vessel. Histological analysis showed the formation of the shoot on the PLBs and the presence of starch granules. Thus, the use of plastic as a culture vessel provides a good impact for culturing PLBs and has low cost.
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