Regenerasi Empat Genotipe Gandum (Triticum aestivum L.) Pasca Transformasi Melalui Agrobacterium tumefaciens

Aniversari Apriana, Atmitri Sisharmini, Fitriyani Fitriyani, Sustiprijatno Sustiprijatno

Abstract


Genetic transformation of
wheat mediated by Agrobacterium tumefaciens has not
been established yet. This research aimed to obtain the
most responsive wheat genotype to be transformed, to
select the most effective combination of growth regulator for
regenerating transforman calli, and to determine the
transformation efficiency. Transformation mediated by A.
tumefaciens was performed on four genotypes of wheat,
namely Combi, Fasan, Perdix, and Naxos-Wew.
Transformed calli with green spots in selection media were
transferred to regeneration media containing 25 mg/l
hygromycin, i.e. R1H25 and R2H25. The obtained plantlets
were analyzed by PCR using specific primers for hygromycin
phosphotransferase gene. The results showed that Fasan
was the most responsive genotype in callus formation
(95.47%) and regeneration both in R1H25 (27%) and R2H25
(28.6%) media. Fourteen plantlets were succesfully
acclimatized and PCR analysis showed that there were four
positive transformants containing the hpt gene. The results
are expected to provide information on the development of
transgenic wheat in Indonesia, specifically in the success of
callus formation and regeneration of wheat after
transformation using A. tumefaciens.


Keywords


Wheat; transformation study; Agrobacterium.

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DOI: http://dx.doi.org/10.21082/jbio.v10n1.2014.p18-25

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P-ISSN : 1907-1094
E-ISSN : 2549-1547


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