Over-ekspresi Gen OsWRKY76 untuk ketahanan terhadap cendawan blas (Pyricularia grisea Sacc.) pada padi

Abstract

Rice plant is estimated to have 109 transcription factor genes of OsWRKY family. Though few of these genes has been known involved in the defense mechanism, the function of the most of them has not been uncovered yet. This study was intended to show that over-expression of OsWRKY76 gene in transgenic rice increase the resistance to blast infection. 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 higher efficiency than EHA 105. PCR analysis of randomly selected 25 independent lines showed that all of them positively contained the over-expression construct of the OsWRKY76 gene. DNA blot analysis showed that a range of 1-4 copies of the over-expression construct of the OsWRKY76 gene was integrated in the genoms of randomly selected 26 independent lines. Out of 6 independent lines containing single copy of the construct, 3 independent lines showed better resistance than non-transgenic plants upon the inoculation using dominant Pyricularia grisea isolat 173.