Protocols for ferns

Hello, could you recommend plant tissue culture protocol for propagation of Polypodium subauriculatum ‘Knightieae and Drynaria Rigidula Whitei ferns?

Thanks in advance.


Hello Rita,

some worked on micropropagation possibilities of Polypodium vulgare L. You may try to adapt their protocols for P. subauriculatum. Here are exemplary protocols for two different inoculum sources:


Primary culture of Polypodium vulgare has been initiated by using a homogenate of gametophyte fragments, grinded under aseptic conditions (Aldea et al. 2016). Sterilization has been done using 70 % ethanol and 6 % calcium hypochlorite solution. The homogenate has been inoculated on liquid MS with 1/2 minerals concentration, without hormones, pH 5.8 (usage of sucrose has not been mentioned). Cultivation has been proceeded while stirring at 75 rpm, 22-24 ± 2 °C, 16 h photoperiod at 3000 lux. The gametophytes differentiated into sporophytes after 3 weeks. Faster rooting, in comparison with liquid medium, has been reported by usage of agar-solidified 1/2 MS with 2 mg/l IBA.


After surface disinfection of Polypodium vulgare spores (10 min in 5 % NaOCl + some drops Polysorbate 20, 3 times washing with distilled sterile water), cultures have been initiated in 1/2 MS + 30 g/l sucrose, solidified with 8 g/l Agar, pH 5.7, 25 ± 2 °C, 16 h photoperiod at 37µM/sm². The developing prothallus clusters have been used for subcultivation after dividing them into 4 sections.


Aldea, Florentina, et al. “In vitro micropropagation of fern species (Pteridophyta) of biotechnological interest, for ex situ conservation.” Oltenia. Studii și comunicări. Ştiinţele Naturii. Muzeul Olteniei Craiova 32.2 (2016): 27-35. (PDF, 1.6 MB)

Messeguer, J., et al. “Polypodium vulgare L.(Wood Fern): In vitro cultures and the production of phytoecdysteroids.” Medicinal and Aromatic Plants X. Springer, Berlin, Heidelberg (1998): 333-348.

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Thank you Orexis, I’ll try these.