Introduksi Konstruk Over-Ekspresi Kandidat Gen OsWRKY76 melalui Agrobacterium tumefaciens pada Tanaman Padi Nipponbare

Aniversari Apriana, Atmitri Sisharmini, Wening Enggarini, Sudarsono Sudarsono, Nurul Khumaida, Kurniawan Rudi Trijatmiko

Abstract


Delivering of Over-Expression Construct OsWRKY76
Candidate Gene in Rice cv. Nipponbare through
Agrobacterium tumefaciens. Aniversari Apriana, Atmitri
Sisharmini, Wening Enggarini, Sudarsono, Nurul.
Khumaida, and Kurniawan R. Trijatmiko. Plant genetic
improvement can be done through classical breeding or
genetic engineering. WRKY is a transcription factor involved
in regulating plant defense responses. OsWRKY76 gene is
located in a narrow segment of chromosome 9 which is
identified previously to be related to wide spectrum
resistance in rice. A sequence of OsWRKY76 (+1.200 bp)
has available in the gene bank and it makes possible to
isolate, clone, and construct the gene into over-expression
vector. The aim of this research was to assemble an overexpression
construct of OsWRKY76 candidate gene and
introduce it into rice through Agrobacterium-mediated
transformation. A construct of pCAMBIA-
1301::35S::OsWRKY76 has been successfully assembled and
transformed into embryogenic calli of rice cv. Nipponbare
using A. tumefaciens strain Agl-1 and EHA 105. A number of
126 independent lines has been produced, in which Agl-1
showed 3.8 times more efficient than EHA 105. PCR analysis
of randomly selected 25 independent lines showed that all
of them positively contained hptII gene, a selectable marker
used in the over-expression construct of the OsWRKY76
candidate gene. Based on the result, it could be concluded
that the over-expression construct of OsWRKY76 candidate
gene have been successfully introduced into the tissue of
Nipponbare.


Keywords


Transcription factor; OsWRKY gene; blast resistance; rice; Nipponbare

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DOI: http://dx.doi.org/10.21082/jbio.v7n1.2011.p19-27

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