Adventitious root of T. Paniculatum Gaertn. : a‐d fresh biomass in batch,
32
2 2.2
2.4 2.6
2.8 3
3.2 3.4
3.6
7 14 -
14 + 21
28 - 28 +
35
Total su
gar
Days
4 4.2
4.4 4.6
4.8 5
5.2 5.4
5.6 5.8
6
7 14 -
14 + 21
28 - 28 +
35
pH
Days
250 260
270 280
290 300
310 320
330 340
350 360
370 380
390 400
410 420
7 14 -
14 + 21
28 - 28 +
35
Electrica l co
nd uctivity
m Scm
Days
Continuous + Buffer Batch + Buffer
Continuous + Non Buffer Batch + Non Buffer
and cells is commonly used for the production of growth associated products, typically primary metabolites and biomass. The continuous culture technique has also been
adopted for the cultivation of several plant cells such as, Coptis japonica Matsubara et al.
8 , Catharanthus roseus Park et al. , and Nicotiana tabacum ashimoto et
al. 8 .
Total sugar, conductivity and hydrogen ion concentration in the medium
Determination of hydrogen ion concentration p , total sugar and electrical conductivity EC of the médium was conducted every seven days. Basically, total sugar,
p and EC had decreased, but increased by using buffer of NaCO every two weeks figure . The total sugar value in batch system are different with continous system
which was provided new medium in days and 8 days. Meanwhile, p in batch and continuous without buffer decreased until days, except on batch and continuous with
buffer periodically in two weeks increased in days and 8 days. For electrical
conductivity in the all treatments decreased and increased when was added buffer each two weeks.
Figure 3. Adventitious root culture condition for days. Culture condition change in
A hydrogen ion p ; B Total sugar; C Electrical conductivity EC . ‐ before added buffer and + after added buffer.
Several studies described that liquid medium was examined by electrical conductivity, p and total sugar. The high conductivity of medium showed that the
adventitious root unavailable to absorb inorganic compounds. n study of Mariateresa et
32 al
. reported that sucrose availability in liquid medium bioreactor . Adelberg et
al. also reported that findings of sucrose availability in liquid medium higher
than in gelled medium. t has known that sucrose as energy source and important carbon for plant cells growth. Gertlowski and Petersen
explained that it can also affect metabolism of the cells and production of metabolites. Lee et al.
reported that sucrose is hydrolyzed into two monosaccharides, glucose and fructose, by invertase
which bounds extracellular or cell wall during the initial culture period. Meanwhile, electrical conductivity EC values of the residual media progressively increased with
increasing salt strength of the culture medium, whereas p values progressively decreased. The EC values of the residual media reflect the uptake of ions by roots and
represent an indirect method of biomass estimation Baque et al., ; Cui et al.,
. n plant tissue culture, p is an important factor affecting biomass and secondary
metabolite accumulation. For instance, in root suspension cultures of ginseng, root dry biomass and ginsenoside accumulation were strongly inhibited when p was
maintained below . or above . ahn et al., . Wu et al.,
also reported that the growth of Echinacea roots decreased when p was maintained below or above .
Lulu et al. explained that in their study after weeks of culture, the p of the
medium declined as the N
+
level increased. The EC measurements have been used as an indirect method for biomass estimation during the continuous online monitoring of
nutrient uptake in plant cell culture bioprocess engineering because of their accuracy
and efficiency.
Conclusions
n this study, we have successfully established adventitious root cultures of T. paniculatum
L capacity bioreactors containing mL medium and worked out
various treatments continuous + buffer, continuous, batch + buffer, batch . Treatment of continuous system with buffer of NaCO has shown fresh biomass .8 g and dry
biomass . g higher than the others. That treatment has produced biomass four fold of initial inoculum. The continuous system provide biomass production better than the
others and supplemented of buffer NaCO influence biomass productivity.
Acknowledgements
Thanks to ndonesia Endowment Fund for Education LPDP , Ministry of Finance, Republic of ndonesia and Plant Physiology Laboratory at Departement of
Biology, Faculty of Science and Technology, Airlangga University.
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Joint Scientific Symposium
IJJSS 2016
Chiba, 20‐24 November 2016
329
Leaf Callus Induction of Moringa oleifera with Addition Various
Concentration Napthalene Acetic Acid NAA and N
6
‐ furfuryladenine
Kinetin in MS Medium in vitro
Muhammad Evan N
a
, Erlyta Vivi P
b
a,b
Institut Teknologi Sepuluh Nopember, Kampus ITS Sukolilo, Surabaya 60111, Indonesia
b
Second affiliation, Address, City and Postcode, Country
Abstract
Moringa oleifera is the plants which have high nutritional content of minerals
and vitamins. n ndonesia, the cultivation of M. oleifera still utilizing vegetative propagation. Therefore, the resulting seedlings are limited and have a low quality of
seeds. To solve the problems, rapid M. oleifera propagation and sustainable high quality of seeds through in‐vitro culture are needed. This study aimed to determine the effect of
somatic embryogenesis induction of M. oleifera leaves due to the adding variation of plant growth regulators concentration of Napthalene Acetic Acid NAA and Kinetin in
MS medium in‐vitro. This study was conducted using a completely randomized design with four treatments, are M MS , M MS + NAA . mgL + Kinetin mgL , M MS
+ NAA mgL + Kinetin mgL , and M MS + NAA mgL + Kinetin . mgL and there were three replication of each treatments. Parameters observed that were the rate
of induction time, color and texture of callus. Data were analyzed using analysis of descriptive. The results showed that the addition of plant growth regulators NAA and
Kinetin at various concentration affected callus growth of M. oleifera leaves on MS medium in‐vitro. The medium concentration of M MS + NAA . mgL + Kinetin
mgL was the optimal concentration on callus induction on fourteenth day , the callus has white color and compact texture.
Keywords
callus induction; kinetin; Moringa oleifera; Napthalene Acetic Acid NAA