Optimasi Sistem Regenerasi dan Transformasi Padi Varietas Elit Indonesia

Aniversari Apriana, Toto Hadiarto, Ahmad Dadang

Abstract


New rice
variety can be generated by means of transgenic approach.
Transgenic rice researches have been conducted in many
institutions worldwide using Japonica, Indica, and Javanica
varieties. The most cultivated rice in Indonesia is Indica.
Indica type is known to have low responsive tissues in
culture and transformation media when compared to
Japonica rice. This research activity is aimed to optimize
regeneration and transformation systems of the Indonesian
elite rice varieties, so that good method can be achieved to
be used in the generation of transgenic elite rice varieties of
Indica type. The research consisted of two activities:
regeneration and transformation optimizations in varieties of
Dodokan (upland rice) and Inpari 6 (irrigated rice).
Immature embryo was used as the explant in this research.
The optimization studies used 2 types of media, NBH (N6
salts and vitamins, cassamino acid 0.5 g/l, L-proline 0.5 g/l,
sucrose 20 g/l, D-glucose 10 g/l, 2.4-D 2 mg/l, NAA 1 mg/l, BA
1 mg/l, agarose Type I 5.5 g/l) and NBH-M (N6 macro salts,
B5 micro salts, and vitamins, 0.3 g/l cassamino acid, 3 g/l Lproline,
20 g/l sucrose, 3 mg/l 2.4-D, 1 mg/l NAA, 1 mg/l BAP,
5.5 agarose Type I), 2 types of regeneration media, R1 (MS
base media and vitamis, 0.3 g/l glutamine, 30 g/l sucrose, 2
mg/l kinetin, 1 mg/l NAA, 3 g/l phytagel) and R2 (MS base
media and vitamins, 2 g/l cassamino acid, 20 g/l sucrose, 30
mg/l sorbitol, 2.5 mg/l kinetin, 0.25 mg/l NAA, 3 g/l phytagel).
Optimization transformation of Indonesian elite rice varieties
used developed an empty plasmid pCAMBIA 1301 containing
hpt gene. The transformation was conducted using two
types of co-cultivation media, K1 (N6 macro salts, B5 micro
salts, and vitamins, 0.5 g/l cassamino acid, 0.5 g/l L-proline,
20 g/l sucrose, 10 g/l glucose, 2 mg/l 2.4-D, 1 mg/l NAA, 1
mg/l BAP, 0.1 mM acetosyringone) and K2 (N6 macro salts,
B5 micro salts, and vitamins, 0.5 g/l cassamino acid, 0.5 g/l Lproline,
20 g/l sucrose, 10 g/l glucose, 2 mg/l 2.4-D, 1 mg/l
NAA, 1 mg/l BAP, 0.2 mM acetosyringone). The results
showed that Inpari 6 could form embryonic calli in NBH
media and further regenerated well in R1 media (13.8%).
The co-cultivation media K1 generated more selected calli
which then generated green plant of young embryo
compared to K2. Inpari 6 showed higher regeneration rates
after transformation (3.6%) compared to Dodokan (0%).
Molecular analysis showed that all 11 transformants (Inpari

6) tested contained the hpt gene. These results are expected
to support the development of transgenic Indica rice
generation in Indonesia.


Keywords


Rice; Indica; regeneration; transformation.

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DOI: http://dx.doi.org/10.21082/jbio.v9n1.2013.p1-10

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P-ISSN : 1907-1094
E-ISSN : 2549-1547


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