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Clonal In Vitro Propagation of Peat Mosses

HORT-501 In Vitro Propagation 2(1-2)
(Sphagnum L)

Submitted by
Aleena Khalid (17-arid-4787)

B.Sc (Hons) 5th Semester

Submitted to
Dr. Touqeer Ahmad

Department of horticulture
PMAS-Arid Agriculture University Rawalpindi
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Content: Introduction

Why we need in vitro?

Objectives

Materials and Methods

Results

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 INTRODUCTION
Peat mosses are a class of mosses Peatlands are an important sink of
with unique morphological, carbon sequestrated in peat. carbon
developmental and physiological uptake by peat moss is about 1.5
characteristics. million km

Sphagnopsida are real ecosystem The cultivation of peat mosses on

engineers as they acidify their degraded peatlands to produce
surrounding habitat, thus creating Sphagnum biomass, called
conditions unsuitable for many Sphagnum farming
competitive plants

Sphagnum species are highly A common technique for active

suitable as biomonitors for the biomonitoring with these plants is
assessment of air quality. called ‘‘moss bag technique’’

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 Why we need in vitro?

The availability of Sphagnum Sphagnopsida in a scalable,

species is limited as most species standardized in vitro culture like in
are rare in Western and Central photobioreactors can enhance their
Europe and protected scientific potential.

Culture techniques with high multiplication rate of

Sphagnum material is also of economic interest as
it depends on the availability of sufficient peat
mosses as ‘seeding’ material. In vitro make it
possible to produce disease free seeding materia

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OBJECTIVES To standardized its
cultivation and
application

To improve the
‘‘moss bag
technique’’
towards a highly
reproducible

To produce cost-
effective
alternative to the
use of automatic
measuring de
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 MATERIALS AND METHODS

Explant Sterilization Incubation

Sporangia from five For spore sterilization, 0.1 % sodium
Sphagnum species hypochlorite /600 L is used .
were collected in the Sodium hypochlorite solution was
field. prepared freshly and 1 drop of Tween20
was added per 10 mL of the solution
After incubation series of 45 s, 1, 1.5,
2, 2.5, 3, 3.5 and 4 min each 75/L of
the mixture were transferred to 4 mL
autoclaved water.
From this dilution 1 mL was
transferred to a sterile Petri dish
containing solid Knop medium

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 RESULTS

1: Establishment of Axenic In vitro culture

2: Sucrose supplementation enhanes biomass yields
3: Optimized in vitro cultivation of Sphagnum palustre in bioreactors
4: Morphological characterization of in vitro-cultivated Sphagnum
palustre

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1: Establishment of axenic in vitro cultures of Sphagnum palustre

Sphagnum palustre After sterilization, spores From thalloid protonema

sporangia were collected germinated within gametophores developed
in the field and the spores approximately 1–2 weeks
were surface sterilized

Gametophores were cultivated as Filaments as well as

independent clones and can be thalloid protonema
cultivated on solid Knop medium developed,
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 2: Cultivation Techniques of Sphagnum Palustre

 Photo-bioreactor containing
Solid Knop medium 5 L aerated flasks Erlenmeyer flasks
liquid Knop medium with
on Petri dishes
microelements 0.3 % sucrose
& 1.25 mM ammonium nitrate

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3: Sucrose Supplementation Enhances Biomass
Yields

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4: Morphological Characterization of In Vitro
Cultivated Sphagnum Palustre

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 Conclusion
For advanced production of S. palustre they tested different cultivation
techniques, growth media and inocula, and analyzed the effects of tissue
disruption.
Cultivation on solid medium is suitable for long term storage, submerse
cultivation in liquid medium yielded highest amounts of biomass.
By addition of sucrose and ammonium nitrate there is an increase in the biomass
by around 10- to 30-fold within 4 weeks. The morphology of in vitro-cultivated
gametophores showed similar phenotypic characteristics compared Electronic
supplementary material

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References

Beike, A. K., Spagnuolo, V., Lüth, V., Steinhart, F., Ramos-Gómez, J., Krebs, M., ... &
Reski, R. (2015). Clonal in vitro propagation of peat mosses (Sphagnum L.) as novel
green resources for basic and applied research. Plant Cell, Tissue and Organ Culture
(PCTOC), 120(3), 1037-1049.

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