In Vitro Indirect Somatic Embryogenesis and Secondary Metabolites Production in the Saffron: Emphasis on Ultrasound and Plant Growth Regulators

Abstract

In the present study, the effects of ultrasound and plant growth regulators on in vitro callogenesis and secondary metabolites production in saffron calli were investigated. Accordingly, the saffron corms surface sterilized, sonicated and cultured on different concentrations of plant growth regulators (0.5, 1, 2 and 4 mg L-1 2,4-D or NAA in combination with 0.5 and 1 mg L-1 Kin or BAP). The percentage of callus induction, callus yield (fresh weight) and embryogenic callus formation were recorded and secondary metabolites of calli were measured by UV/VIS spectrophotometer three months after culture. The results indicated that sonication of the saffron corm explants significantly increased the in vitro callus induction and growth. So, the highest callus induction (100%) and yield (4.68 g) was achieved with sonicated explants cultured on MS medium supplemented with 2 mg L-1 1-naphthaleneacetic acid (NAA) and 0.5 mg L-1 kinetin (Kin). Somatic embryogenesis was significantly influenced by plant growth regulator regimes and the MS medium supplemented with 0.5 mg L-1 2,4-dichlorophenoxyacetic acid (2,4-D) plus 0.5 mg L-1 6-benzylaminopurine (BAP) and 0.5 mg L-1 NAA plus 0.5 mg L-1 Kin exhibited the highest percentage (75 and 72, respectively) of somatic embryogenesis. Secondary metabolite content of the callus cells was significantly different among the plant growth regulator regimes and the highest production of picrocrocin and safranal were occurred on the medium containing 0.5 mg L-1 2,4-D plus 0.5 mg L-1 BAP and 1 mg L-1 NAA plus 1 mg L-1 BAP.

Sterilization protocols and the effect of plant growth regulators on callus induction and secondary metabolites production in in vitro cultures Melia azedarach L.

“…Plants show different in vitro growth response to different types and concentrations of plant growth regulators, especially the combination of auxins and cytokinins (Guo et al 2009 ; Firoozi et al 2019 ). In the present study, all used PGRs treatment induced callogenesis from M. azedarach L. leaf explants, but with different percentage and callus growth rate.…”

Ondas Sonoras Tem Efeito Positivo Na Germinação De Sementes E Produção De Mudas De Hortaliças / Sound Waves Have a Positive Effect on Seed Germination and Seedling Production in Vegetables

“…A produção de mudas também pode ser obtida a partir da micropropagação ou cultura de tecidos e, nessas condições, as ondas sonoras também podem ser utilizadas de forma combinada a essas técnicas proporcionando aumento na organogênese. Já se sabe, por exemplo, que o ultra-som pode ser utilizado como um tipo de estímulo para se iniciar uma série de processos bioquímicos nas células das plantas (SILVA et al, 2020;FIROOZI et al, 2019).…”

“…Em uma pesquisa avaliando os efeitos de ultra-sonografia (35 kHz) e de reguladores de crescimento de plantas na produção de calogênese in vitro e de metabólitos secundários em calos de açafrão (Curcuma longa) observou-se que a sonificação dos explantes de açafrão possibilitou aumento significativo na indução e crescimento de calos in vitro, sendo a maior indução de calo (100%) e rendimento (4,68 g) obtido com explantes sonificados cultivados em meio MS suplementado com 2 mg L -1 1-naftalenoacético (ANA) e 0,5 mg L -1 de cinetina (Kin). (FIROOZI et al, 2019). Também em cultura de tecidos com segmentos de um único nó de batata (Solanum tuberosum) expostos a ultra-som de 35 kHz, 70 W, por 20 min, pesquisadores observaram que o som atuou como um estimulante de crescimento positivo.…”

Extraction, Identification And Determination of Antiviral And Anticancer Flavonoids By HPLC-DAD In Cell Suspension Culture of Melia Azedarach L.

“…Oure ndings con rm the previous studies showing that PGRs combination of the medium and explant type and source play a crucial role in plant cell suspension culture establishment and growth (Farjaminezhad et al 2013 Sgherri et al (2004) proved that the production and accumulation of secondary metabolites including phenolic compounds is associated with oxidative stress. For manipulation of production and accumulation of secondary metabolites in vitro culture, PGRs were known as one of the main factors (Firoozi et al 2019) that can activate the synthesis of phytochemicals components such as phenolic compounds and avonoids (Sharma et al 2012).…”

“…practicable way to plant regeneration, cell suspension culture and production of secondary metabolites(Ahmadpour et al 2016;Firoozi et al 2019). Different factors including plant growth regulators especially auxins and cytokinins, plant genotype and explant type in uence the callus induction, cell growth and development, and secondary metabolites production in the plant cells(Zare et al, 2009;Firoozi et al, 2019;Farjaminezhad and Garoosi, 2019) In the present study, we investigated the effect of different PGRs combinations including auxins (NAA and 2,4-D) and cytokinins (Kin and BAP) on callus induction and growth, and cell suspension culture establishment, growth and secondary metabolites production from leaf, in orescence and petiole explants of M. azedarach L. The percentage of callus induction, callus growth (biomass), color and texture of the calli were different depending on the M. azedarach L. explant type and applied PGRs combinations. Furthermore, in a given plant species different types of explants show diverse in vitro response, especially depending on PGRs combination.…”