Callus Induction and Micropropagation of Lilium candidum L. Using Stem Bulbils and Confirmation of Genetic Stability via SSR-PCR

Abstract

Natural populations of Lilium candidum L. are remarkably affected by biotic and abiotic factors therefore there is a requirement to develop effective micropropagation protocol to provide mass production, multiplication and conservation of these plants. For this reason, this study was aimed to develop an efficient micropropagation method for multiple shoot production via somatic embryogenesis induced from L. candidum stem bulbils and also to determine the genetic stability of in vitro grown plants using SSR markers. The obtained results of this study are the first comprehensive reports including an investigation of genetic fidelity on somatic embryogenesis of L. candidum. After surface sterilization of bulbils, the calculated regeneration percentage of them was 89.5% and the callus induction was achieved using leaf segments of in vitro grown bulbils. The well formed somatic embryos were obtained from smooth whitish-yellow colored calli and these somatic embryos produced well formed healthy L. candidum seedlings with adventitious roots. All rooted seedlings were easily adapted to greenhouse conditions and the genetic stability of in vitro grown seedlings were determined by using SSR-PCR technique and it was calculated as 100%.

Potential utilization of filaments and ovules as explant sources in in vitro propagation of Lilium sp.

“…In other studies, Hoesen (2009) was significantly proved high survivability up to 85% of Lilium sp. plantlets in combination media of compost, sand, and soil (2:1:1), 66.7% survivability of bublets were established in combination of sand, soil, perlite and vermiculite (1:1:2:2) and fertilized with half-strength MS solution (Joshi and Dhar, 2009), plantlets were effectively acclimatized in autoclaved soil (Tang et al, 2010), 98% survivability of L. formolongo plantlets was planted in the soil substrate and covered by clear plastic for 5 weeks (Saetiew and Umamanit, 2015), 100% survival rate of plantlets was established in peat moss combined with regular watering (100 ml/plant) and pacing to a shady place (Royandazagh, 2019), rooted shoot clusters of Lilium candidum were optimally transferred ex vitro in nitrogen rich peat (Tokgoz and Altan, 2020). The lower survivability of the plantlets in the research was presumably due to no clear plastic covering them during the initial acclimatization at least for 7 days.…”

“…The method is also not suitable for scaling up the plant commercially. Therefore, producing high quality and productivity with a lower price of Lilium planting materials using tissue culture as a very useful approach for rapid propagation is importantly addressed (Tokgoz and Altan, 2020), especially in developing and establishing an in vitro mass propagation protocol (Joshi and Dhar, 2009;Naeem et al, 2013;Rafiq et al, 2021) Developing and establishing in vitro mass propagation protocols for Lilium sp. with varied results was reported previously.…”

Somaclonal Variation—Advantage or Disadvantage in Micropropagation of the Medicinal Plants

“…Moreover, SSR markers are prone to have "null alleles", i.e., no amplification of the intended PCR product, due to the mutation that can occur in the primer annealing sites, which may lead to errors in scoring. SSR markers are considered genetically stable because, when used, they did not reveal any polymorphism, or only a very low level of it, between the original plants and the regenerants [81][82][83].…”

Elicitation and Enhancement of Phenolics Synthesis with Zinc Oxide Nanoparticles and LED Light in Lilium candidum L. Cultures In Vitro

“…Little information is available in the literature on the micropropagation of Madonna lily [6][7][8][9][10][11][12][13][14][15]. Available studies mainly concern the elimination of contaminations that occur during in vitro culture initiation [9,10,12], the selection of mother plant explant [7,11,14], and growth regulators used during organogenesis [8,15]. Our previous work [6] investigated the effects of LED light on adventitious organogenesis in L. candidum.…”