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