Effects of plant growth regulators and activated charcoal on somaclonal variations of protocorm-like bodies (PLBs) of Dendrobium Sabin Blue orchid

Abstract

Protocorm-like bodies (PLBs) of Dendrobium orchids are emerging as a potential source of valuable secondary metabolites. This study examined the effect of four additives namely 1-naphthaleneacetic acid (NAA), kinetin, thidiazuron (TDZ), and activated charcoal (AC) used in culture medium on genetic variability in PLBs of Dendrobium Sabin Blue. Nine ( 9) ISSR primers and eleven (11) DAMD primers were used to assess the genetic variability of PLBs that were subcultured over a period of two years. We confirmed that the use of kinetin in culture medium for two years resulted in the highest rate of somaclonal variation in PLBs. On the other hand, TDZ and activated charcoal registered the lowest genetic variability in PLBs. The findings of this study suggest the importance of selecting additives used in the culture medium to maintain stable genetic lines of PLBs. We recommend that the assessment of somaclonal variations should be performed for long term maintenance of tissue cultures.

An Overview of Orchid Protocorm-Like Bodies: Mass Propagation, Biotechnology, Molecular Aspects, and Breeding

“…As example, the cytokinin Kinetin at 1.5 mg L −1 resulted in increases of somaclonal variations frequency of PLBs in Dendrobium Sabin Blue, detected by ISSR and DAMD molecular markers, when compared with use of TDZ at 4.0 mg L −1 added activated charcoal [158]. Although the cytokinin-like compound TDZ is appointed as one of the major causes of SV in orchid PLB induction, there were some contradictory reports.…”

Establishment of In Vitro Regeneration Protocol for Sabah’s Jewel Orchid, Macodes limii J.J. Wood & A.L. Lamb

“…Typically, plantlets regenerated via axillary buds or direct somatic embryogenesis are considered to be the most genetically uniform [25,52]. The uniformity of the in vitro cultures against genetic variations despite repeated subcultures can be linked to their respective genotype, which could be genetically resistant to alteration in their genetic materials [53]. However, artificial in vitro conditions, modified nutrients, and exposure to synthetic hormones might generate a genetic instability in cultured plants known as somaclonal variation [21,53,54].…”

“…The uniformity of the in vitro cultures against genetic variations despite repeated subcultures can be linked to their respective genotype, which could be genetically resistant to alteration in their genetic materials [53]. However, artificial in vitro conditions, modified nutrients, and exposure to synthetic hormones might generate a genetic instability in cultured plants known as somaclonal variation [21,53,54]. Furthermore, the application of TDZ and prolonged exposure to it increased the chances of heritable somaclonal variation induction within the regenerated plantlets [55][56][57].…”

ANALYSIS OF GENETIC STABILITY OF MICROPROPAGATED SUGARCANE IN DIFFERENT SUBCULTURE FREQUENCIES USING SSR MARKER / Analisis Kestabilan Genetik Tebu Hasil Mikropropagasi pada Beberapa Frekuensi Subkultur Menggunakan Marka SSR

“…Propagation of shoots in vitro PS 862 both for mass seedlings production and cryopreservation materials can be applied by lowering BAP concentrations lower than 0.3 mg/l and IBA lower than 0.5 mg/l to avoid genetic changes. It is believed that the plant growth regulator type and concentration, culture period, genotype used, and culture age can cause somaclonal variations in the culture of tissue culture results (Aydin et al 2015;Bradai, Romero and Martin 2019;Chin et al 2019).…”