Bioactivity of Scytonema hofmanni (Cyanobacteria) in Lilium alexandrae in vitro propagation
María Cristina Zaccaro*
Mónica Magdalena Storni
Ana María Stella
UBACyT X64, Universidad de
Keywords: ascorbate peroxidase, bioactive substances, bulb regeneration, catalase, Cyanobacteria, glutathione reductase, Lilium alexandrae, Scytonema hofmanni.
Cyanobacteria produces bioactive compounds including plant growth regulators. Naphthalene acetic acid (NAA), a toxic substance, is a synthetic plant regulator used in micropropagation. The aim of this work was to evaluate morphogenetic and antioxidant effects produced by intra and extracellular substances from Scytonema hofmanni (Cyanobacteria) during the multiplication in vitro of Lilium alexandrae and to compare them to those produced by NAA. Intra and extracellular cyanobacterial products increased a) bulblets production reaching 83% and 78% of NAA effect, respectively; b) the bulblet diameter compared to NAA; and c) the bulblet survival due to the promotion of antioxidant activity measured as catalase, ascorbate peroxidase, and glutathione reductase activity. The cyanobacterial substances stimulated regeneration and delayed bulblet senescence. They could replace NAA, dangerous for the operator, not only during the regeneration phase but also during the storage of the viable bulblets cultivated in vitro.
regulators which accelerate the production of a number of agronomical
interesting plants are used in in vitro culture of plants.
Growth regulators are mainly obtained by chemical synthesis. Cyanobacteria
produce a variety of bioactive compounds including growth phyto-regulators
(Metting and Pyne, 1986), that could be used in the
in vitro production of vegetables, fruits, fungi and ornamental
flowers. Already in 1979, Zulpa de Caire et al. (1979)
established that Nostoc muscorum Ag. liberated into the culture
medium auxin-like substances. It has recently demonstrated that a
number of Cyanobacteria produce, accumulate, and liberate 3-indol
acetic acid (Sergeeva et al. 2002). Arthronema africanum
produces the cytokinin isopentenyl adenine (Stirk et
al. 1999). Stirk et al. (2002) found auxin and
cytokinin activity by three cyanobacterial strains. It is important
to mention that some cyanobacterial products promote regeneration
in Daucus carota (Wake et al. 1992), Santalum
album (Bapat et al. 1996), Oryza sativa
(Zaccaro et al. 2002; Storni de Cano
et al. 2003), Lilium alexandrae hort. Wallace,
(6-8 x 6-
water extracts (BWE) and extracellular products (EP).
S. hofmanni Ag. ex Born. et Flahault axenic strain Nº 58,
from the culture collection belonging to Laboratorio de Cyanobacteria,
Facultad de Ciencias Exactas y Naturales, Universidad de Buenos Aires,
Argentina, was cultivated in modified Watanabe medium (Storni
de Cano et al. 2003), photoperiod 12 hrs light, 45 µmol photon
x m-2 seg-1, 27 ±
I, 8 ml MS + 2 ml distilled water. II, 8 ml MS + 2 ml
EP. III, 8 ml MS + 2 ml distilled water + 0.1 mg/L NAA. IV,
8 ml MS + 2 ml BWE. The media contained 3% sucrose, 0.7% agar and pH
was adjusted to 6.8, before being sterilized in tubes (14 x
Enzymatic activity of catalase, ascorbate peroxidase and glutathione reductase were determined according to Beers and Sizer (1951), Nakano and Asada (1981) and Schaedler and Bassham (1977), respectively, in order to establish the bulblets oxidative stress level produced by the regeneration treatments.
Analysis of variance was performed for all data, using a completely randomized experimental design. A one-way ANOVA was performed (PC program GraphPad Prism). A Tukey HDS test (p < 0.05) was used to compare different treatments for each day, n = 14.
day 50 (Figure 1) S. hofmanni BWE increased the number
of bulblets per explant by 27% comparing with MS, representing this
increment 78% of the bulblets produced by NAA. At day 70 the increment
was 21 y 32% for EP and BWE, respectively, compared to MS. With respect
to NAA, EP produced 78% and BWE 83% of the number obtained with the
synthetic auxin (Figure 2a). No matter higher the number of
bulblets produced by NAA, cyanobacterial intracellular products increased
the number of bulblets thicker than
Intra and extracellular products from S. hofmanni increment L. alexandrae bulblets production from microscales, comparing with control without synthetic phyto-regulators as well as bulblets survival in culture because they promote a higher antioxidant activity. This effect could be due to the production and liberation of plant growth regulators such as auxin-like and cytokinin-like substances by the cyanobacterium, which produced similar results than those obtained with the synthetic auxin NAA and also because the bulblets senescence was delayed, an effect well-known for cytokinin. Besides, the cyanobacterial phyto-regulators could replace substances that result dangerous to the operator not only during the regeneration phase but also during the storage of the viable bulblets cultivated in vitro.
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