The Effect of Kinds PGR concentration into Culture Media to the Propagation of

PEDADA (Sonneratia Caseolaris)

Annisa Rahma Fatika

Introduction
Abrasion is a process of erosion of the coast by destructive force of ocean waves and ocean
currents. Felling trees in coastal forests or mangrove forests spur coastal abrasion faster
(Irwanto, 2010 in Naswita 2012). Moch Amron (2011) said that as much as 20 percent of the
coastline throughout the territory of Indonesia suffered damage due to various problems
including environmental changes and coastal abrasion.
Planting mangrove on the outer coastline requires a type of mangrove that can adapt to the
outer environment. One type of mangrove that is able to adapt well is the type of mangrove
pedada (Sonneratia caseolaris). Pedada mangroves are mangroves that grow in coastal areas
with high adaptation to salinity conditions. People live around mangrove forests still rely on
the results of plants that grow wild in the forest to meet the mangrove seeds pedada. To create
a vast mangrove forest, intensive propagation is needed. So that new innovations are needed
to meet the needs of mangrove pedada seedlings by means of in vitro culture.
In vitro culture is a method of plant multiplication by isolating plant parts and growing them
in media that contain nutrients and growth regulators in sterile conditions (Prabaningrum, et
al., 2016). The advantages of in vitro culture compared to other generative and vegetative
methods, which are able to produce seeds faster, more numerous, free from disease and
identical to the parent without being affected by the season (Hendaryono and Wijayani,
1994).
Suryowinoto (1996) states that the MS medium has elements and compounds that are more
complete than the other medium. Growth regulators which are widely used in tissue culture
are auxin and cytokinin. Comparison of auxin and cytokinin that is balanced in explants can
produce callus growth (Davies, 1990). Wetherell (1982) states that auxin plays a role in
stimulating cell division and enlargement at the shoots of plants. The addition of auxin in
larger amounts, or the addition of auxin which is more stable, such as 2,4-D acid tends to
cause callus growth from explants and inhibits plant shoot regeneration. Cytokines are
regulatory substances that are often used for propagation of shoots in vitro. Thidiazuron
(TDZ) is an organic compound whose activity resembles cytokinins (Pierik, 1987). The
results of the study of Azwin et al (2006) explants originating from axillary or adventitious
shoots planted on MS base media with the addition of TDZ growth regulators had a very
significant effect on the number of shoots produced.
Therefore, research needs to be carried out with the aim of obtaining the right 2.4 D and TDZ
concentrations to induce buds and mangrove shoots in vitro.

Materials and Methodds

The experiment were carried at the In Vitro Culture Laboratory, Faculty of Agriculture,
Muhammadiyah University, Yogyakarta. This research was conducted for approximately 4
months from April to JulyThe materials used consist of: explants of mangrove leaves of
sonneratia caseolaris, detergents, aquades, sunclin with active ingredients Chlorine 5.25%,
Benomil 50%, spritus, 70% alcohol, betadin, MS medium. The tools used include: Laminar
Air Flow cabinet, Bunsen lamps, autoclaves, tweezers, scalpels, scissors, plastic wrap, bunsen
lamps, aluminum foil, pH sticks, magnetic stirrer hotplates, measuring cups, measuring
pipettes, analytical scales, timers and glassware.
This research was conducted using an experimental method with a two-factor (factorial)
experiment, which is a combination experiment of giving ZPT with 2,4 D and TDZ
concentrations appropriate to induce buds and mangrove budada in vitro which was arranged
in a Completely Randomized Design (CRD) with 6 treatments. Each treatment was repeated
5 times to obtain 60 experimental units.
Sterilization of tools
Tool sterilization is done in three ways, namely: wet heat sterilization, burn sterilization, and
UV. Wet heat sterilization is carried out on glassware and tweezers by wrapping the device
that has been washed clean with umbrella paper and sterilized in an autoclave at a pressure of
1 atm at 121 ° C for 30 minutes. Fuel sterilization was carried out on dissecting kits
(tweezers, scissors and scaple) in the LAF by dipping the tool in alcohol and burning it on a
bunsen lamp before inoculation. LAF sterilization is done several hours before planting
explants by spraying 70% alcohol then turning on the UV lamp for 1 hour.
Making MS Medium
Making the medium using MS medium begins with the manufacture of macro, micro, vitamin
nutrient stock solutions and provides sucrose, growth regulators, and agar. Sterilization of the
medium was carried out using an autoclave at 121 ° C with a pressure of 1 atm for 20 minutes
and a sterile treatment medium was obtained which was ready to be used for inoculation.
Explants Sterilization
Mangrove sonneratia caseolaris leaves are washed with running water, after that it is washed
with 2g / L detergent solution for 10 minutes and rinsed 3 times. Then the leaves are soaked
with Benomil 200mg / L and streptomycin sulf 80 mg / L for 1 minute and rinsed 3 times
with sterile distilled water. Furthermore, the leaves were put into LAF and cut 10 cm then
immersed in chlorine 2.6 ml / L, 5.2 ml / L, and 7.8 ml / L for 5 minutes and 10 minutes
according to treatment. Eksplan then rinsed 3 times with sterile distilled water. After that the
explants are taken and placed in petridish containing 5 drops of betadin dissolved in 30 ml of
sterile distilled water. Eksplan cut to size 2x2 cm then drained and explants ready to be
planted in the media.

a. Pedada Leaf Explants Inoculation

After explants are sterilized, they are then planted on the media according to treatment. This
study uses a completely randomized design (CRD) of 6 treatments with 5 replications per
treatment to obtain 60 experimental units. The treatments given are:

Treatment Type 2,4 D TDZ

(Auxin) Cytokines)
A 0,5 ppm 0 ppm
B 0 ppm 0,5 ppm
C 0,5 ppm 0,5 ppm
D 0,75 ppm 0,5 ppm
E 0,5 ppm 0,75 ppm
F Without PGR (Control) Without PGR (Control)

b. Incubation of Pedada Culture

Incubation is carried out in a rack using a 40 watt TL lamp with a room temperature of 20-28
° C.
Observed parameters
The parameters observed included: Percentage of live explants (%), Percentage of explants
contaminated (%), Percentage of browning explants (%), Percentage of explants sprouting
(%), time of shoot emergence, Number of shoots, Number of leaves, number of leaves
(strands), Percentage of number of rooted explants (%)
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