Perez et al. 2013 (misc.) Hohenbergia

Micropropagation of Hohenbergia penduliflora (A. Rich.) Mez.for sustainable production of plant proteases

Author(s):—A. Pérez, T. Laudat, M. Mora, C. Carvajal, C. Aragón, J. González, M. Escalona, M. Daquinta, R. Trujillo, M. Hernández & J. Lorenzo

Publication:— (2013). — DOI

Abstract:—Hohenbergia penduliflora (A. Rich.) Mez.inhabits the protected ecological area of Cunagua High-land, Ciego de Ávila, Cuba. The availability of this plantfor experimental purposes is exceedingly limited. Tissuecultured plants of this specie, would be useful for propa-gation purposes. Experiments were carried out to optimizethe micropropagation process from the disinfection of fruitsto ex vitro hardening of regenerated plantlets. The bestresults were obtained when seeds were disinfected with2 % (v:v) sodium hypochlorite for 20 min and placedin vitro for 45 days for seed germination. Tissue culturedshoots (1 cm) with vertical wounds in the basal region(5 mm long) were placed in a medium containingMurashige and Skoog (MS) salts, 100 mg l - 1 myo-inosi-tol, 0.1 mg l - 1 thiamine-HCl, 30 g l - 1 sucrose, 8.8 l M6-benzyladenine (BA) and 1.6 l M naphthalene acetic acid(NAA). Shoots were proliferated for 45 days to obtain 8.21new shoots per explant; they were subsequently dividedand rooted on a medium containing 1.6 l M NAA for30 days. For ex vitro hardening, plastic trays containing82 cm 3 of filter-cake-sugarcane ashes were used; 100 %survival rate was recorded. After 6 months of hardening,plants were established ex vitro and ready for proteaseextraction. Comparisons between protein contents, prote-olytic activities and specific proteolytic activities of extracts from stems of macro- and micropropagated plantswere acquired. Tissue cultured stems showed statisticallylower figures which is why Ethrel was tested here toincrease proteolytic activity in micropropagated plantstems. After Ethrel applications, protein contents, proteo-lytic activities and specific proteolytic activities of extractsfrom stems were the three main indicators recorded.However, other biochemical effects of Ethrel were alsoevaluated, such as, levels of chlorophyll pigments,malondialdehyde and other aldehydes; and superoxidedismutase, and guaiacol peroxidase activities. Rising con-centrations of Ethrel (0, 1.5, 3.0, 4.5 and 6.0 mg l - 1 )decreased protein contents at 72 h but increased proteolyticand specific proteolytic activities of stem extracts. Ethrelwas effective in increasing proteolytic activity in in vitroculture-derived plant stems, at a level higher than in field-grown plant stems. Moreover, Ethrel increased superoxidedismutase and guaiacol peroxidase-specific activities inleaves; and decreased chlorophyll pigments. Ethrel did notaffect levels of malondialdehyde and other aldehydes.

Keywords:—Hohenbergia penduliflora (A. Rich.) Mez.inhabits the protected ecological area of Cunagua High-land, Ciego de A´vila, Cuba. The availability of this plantfor experimental purposes is exceedingly limited. Tissuecultured plants of this specie, would be useful for propa-gation purposes. Experiments were carried out to optimizethe micropropagation process from the disinfection of fruitsto ex vitro hardening of regenerated plantlets. The bestresults were obtained when seeds were disinfected wi