Direct somatic embryogenesis and genetic homogeneity assessment of regenerated plants of Anthurium andraeanum Linden cv. Fantasia

Abstract

We present here a method for the in vitro propagation of Anthurium andraeanum cv. Fantasia through direct somatic embryogenesis. The embryos were directly formed at the cut end of most of the leaf explants when cultured on MS basal medium supplemented with N 6 -benzyladenine (BA) plus α-naphthalene acetic acid (NAA). MS basal media supplemented with BA (0.27 μM) and NAA (2.68 μM) induced maximum number of somatic embryos per leaf explant (15.33 ± 1.45) after 28 d of continuous culture. The highest numbers of embryos (12.33 ± 0.88) were matured into plantlets in MS basal medium augmented with 0.27 μM BA and 58.4 mM sucrose. Histology showed the presence of scutellum, coleoptile, welldeveloped root, and shoot initials at different stages of somatic embryo (SE) development directly on leaf explants. About 85% of the plantlets were successfully acclimatized, and of these, 80.3% plants survived after secondary hardening under greenhouse conditions. The embryo-derived plants successfully flowered. The presence of monomorphic DNA bands in random amplified polymorphic DNA (RAPD) marker analysis confirmed the genetic homogeneity of direct somatic embryo-derived plantlets in respect to their mother plant.

A regenerative protocol and SEM study for in vitro propagation of Anthurium crossed lines via indirect somatic embryogenesis

“…The stages clearly indicated the embryo maturation leading to the formation of shoots (Fig. 2f) (Bhattacharya et al, 2016).…”

“…Before inoculation, leaves were cut into 1cm squares, pricked few times with a sterilized scalpel and placed on culture media with the adaxial Bhavana, Satyan and Aswath surface down. The cultures after inoculation were incubated under cool and dark conditions at 25±2.0°C and 70% relative humidity (RH) (Bhattacharya et al, 2016).…”

Induction of somatic embryogenesis in two cultivars of anthurium analysed by scanning electron microscopy

“…In that study the author found that the NAA auxin at a concentration of 10 μM gave superior results for the establishment of embryogenic calli in nodal segments of the cultivar under study (1.23), expressed as the number of embryogenic calli formed. Bhattacharya et al (2016) achieved the best results for obtaining embryogenic calli from leaf explants in a medium supplemented with NAA (2.68 μM) and BAP -6-benzylaminopurine (0.27 μM). Other authors have been successful in inducing somatic embryos in anthurium with the 2,4-D auxin, such as in the research by Bautista et al (2008) and Bhavana et al (2018).…”

“…In anthurium, 2,4-dichlorophenoxyacetic acid (2,4-D) and naphthaleneacetic acid (NAA) are, in that order, the most commonly used auxins. The most widely used explants are the leaves, but other organs/ tissue can also be used with satisfactory results, such as nodal segments (BAUTISTA et al, 2008;XU et al, 2010;PINHEIRO et al, 2013;PINHEIRO et al, 2014;BHATTACHARYA et al, 2016). Another interesting growth regulator to be studied is the auxin Picloram (4-amino-3,5,6-trichloropyridine-2-carboxylic acid), which also promotes high callus induction (PACHECO et al, 2012).…”

“…Em antúrio, o ácido 2,4-diclorofenóxiacético (2,4-D) e o ácido naftalenoacético (ANA) são, nesta ordem, as auxinas mais utilizadas. Os explantes mais comumente empregados são as folhas, mas outros órgãos/tecidos também podem ser usados com resultados satisfatórios, como segmentos nodais (BAUTISTA et al, 2008;XU et al, 2010;PINHEIRO et al, 2013;PINHEIRO et al, 2014;BHATTACHARYA et al, 2016). Outro regulador de crescimento interessante a ser estudado é a auxina Picloram (ácido 4-amino-3,5,6tricloropiridina-2-carboxílico), que também promove alta indução de calos (PACHECO et al, 2012).…”