Morphogenetic responses from protoplasts and tissue culture of Laminaria digitata (Linnaeus) J. V. Lamouroux (Laminariales, Phaeophyta): callus and thalloid-like structures regeneration

Abstract

The regeneration of meristematic tissues from sporophytes of Laminaria digitata was studied by protoplast and tissue culture. Sequential treatment of explants in sterile seawater with 1% Betadine for 5 min, 1% commercial bleach for 1-2 min and 2% antibiotic treatment supplemented with 1 μM GeO 2 overnight enabled viable explants as high as 55%. Different morphogenetic responses were observed from tissue culture on media supplemented with plant growth regulators alone or in combination, mainly filamentous calluses up to 50% according to the media. Dark green compact calluses were observed on two combinations: 4 μM Pi+2 μM N-(2-chloro-4-pyridyl)-N'-phenylurea (CPPU) and 0.04 μM Pi+0.44 μM 6-benzylaminopurine. Thalloid-like structures comparable to adventitious buds were regenerated on medium supplemented with 4 μM Pi+0.45 μM zeatin but at low frequency suggesting a strong genotypic effect. Friable calluses were developed from protoplasts in enriched medium with polyamines and containing 0.40 μM CPPU+0.45 μM 2,4-dichlorophenoxyacetic acid. In order to produce protoplasts, a one-step enzymatic protocol was developed and yields reached 22×10 6 protoplasts per gram of fresh weight.

Effects of ambient calcium concentration on morphological form of callus-like cells in Saccharina japonica (Phaeophyceae) sporophyte

“…On the other hand, the propagation of pigmented round callus-like cells was reported in L. digitata (Asensi et al 2001). Mussio and Rusig (2009) also reported cell masses by irregular cell division in L. digitata. The development to thallus was divided into direct development from callus-like cells in Saccharina japonica (= Laminaria japonica) and U. pinnatifida (Yan 1984) and E. cava (Notoya and Aruga 1989) and indirect development through gametophyte-like form to thallus in L. digitata, L. hyperborea (Fries 1980), and L. saccharina (Lee 1985).…”

The Influence of Abiotic Factors on the Induction of Seaweed Callus

“…PGRs are widely used in tissue cultures of higher plants [68][69][70]. Synthetic as well as natural PGRs have been utilized for seaweed callus induction, namely, 2,4dichlorophenoxyacetic acid (2,4-D); picloram (PIC); phenylacetic acid (PAA); indole-3-acetic acid (IAA); indole-3-butyric acid (IBA); α-naphthaleneacetic acid (NAA); 6-benzylaminopurine (BAP); 2-Isopentenyladenine (2iP); kinetin (KIN); uniconazole (UNI); N-(2-chloro-4-pyridyl)-N'-phenylurea (CPPU); gibberellic acid (GA); 5-aminolevulinic acid (ALA); zeatin (ZEA); and the polyamine spermine [40,44,45,60,[71][72][73][74][75][76][77][78][79][80][81]. The effect of PGRs on callus induction in seaweed is summarized in Table 4.…”

FORMULASI AUKSIN (INDOLE ACETIC ACID) DAN SITOKININ (KINETIN, ZEATIN) UNTUK MORFOGENESIS SERTA PENGARUHNYA TERHADAP PERTUMBUHAN, SINTASAN DAN LAJU REGENERASI KALUS RUMPUT LAUT, Kappaphycus alvarezii

“…Hasil yang diperoleh pada penelitian ini sesuai dengan penelitian Mussio & Rusig (2009) yang memperoleh laju regenerasi Laminaria digitata pada kisaran 25%-50% dengan penambahan zat pengatur tumbuh 2,4-dichlorophenoxyacetic acid (2,4-D). (Bradley & Cheney, 1990;Sakakibara, 2006).…”