Pertumbuhan biak kalus dan suspensi sel tanaman kina (Cinchona ledgeriana Moens) Growth of callus and cell suspension cultures of cinchona (Cinchona ledgeriana Moens)

Abstract

Summary
In vitro technology of plants can be used to
propagate plants and to produce secondary
metabolites with a short and continuous
production cycle. Callus cultures of cinchona
(Cinchona ledgeriana Moens) on solid media and
cell cultures in liquid media have been
established. Callus could be easily initiated from
various explants of cinchona clone CB5, GA22
and QRC312. The best callus initiation and
proliferation were obtained on a Woody Plant
(WP) solid medium supplemented with 15 µM
picloram,0.5 µM BAP and 1 µM phloroglucinol.
In this medium the fresh weight of callus
increased by 12 to 14-fold within 5 to 6 weeks.
Callus that constantly grew fast was selected as a
material source for cell suspension cultures. In
WP liquid medium with the same composition,
the cells remained to grow fast where cell volume
after sedimentation (CVS) increased by almost
4-fold in two weeks. However, repeated sub-
cultures decreased cell growth rate. The cell
suspension culture was then scaled-up in a 5-L
bioreactor. The culture medium was the same as
in Erlenmeyer flasks. Cells in a bioreactor grew
very slowly, the cell biomass fresh weight and
packed cell volume (PCV) increased by 34% and
50% respectively after 21 days of culture,
although most of the cells remained viable.

Ringkasan
Teknologi in vitro tanaman dapat digunakan
untuk memperbanyak tanaman dan memproduksi
senyawa sekunder dengan siklus sangat singkat
dan berkelanjutan. Biak kalus tanaman kina
(Cinchona ledgeriana Moens) pada medium
padat dan biak sel di medium cair telah
dikembangkan. Kalus dengan mudah dapat
diinduksi dari berbagai jenis eksplan tanaman
kina klon CB5, GA22 dan QRC312. Inisiasi dan
proliferasi kalus terbaik diperoleh pada media
Woody Plant (WP) padat dengan pikloram 15
µM, BAP 0,5 µM dan floroglusinol 1 µM. Pada
medium ini bobot basah kalus meningkat 12-14
kali lipat dalam waktu 5-6 minggu. Kalus yang
tetap tumbuh cepat dipilih sebagai sumber bahan
untuk biak suspensi sel. Dalam medium cair WP
dengan komposisi yang sama, sel tetap tumbuh
dengan pesat, volume sel setelah pengendapan
(CVS) meningkat hampir empat kali lipat dalam
waktu dua minggu. Namun subkultur berulang
menurunkan laju pertumbuhan sel. Skala biak
suspensi sel kemudian diperbesar dalam bio-
reaktor kapasitas 5 L. Medium kultur yang
digunakan sama dengan medium pada labu
Erlenmeyer. Pertumbuhan sel dalam bioreaktor
sangat lambat, bobot basah sel dan packed cell
volume (PCV) hanya bertambah berturut-turut
sebesar 34% dan 50% setelah 21 hari dalam
kultur, walaupun sebagian besar sel tetap viabel.