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Agrobacterium-Mediated Transformation of Two Rice Varieties with Atmyb61 Gene for Lignin Content Manipulation Toward Biomass Refinery

The Atmyb61 gene is essential for lignin biosynthesis. Lignin has the promise that would be used to make renewable chemicals and biomaterials. The goal of this work was to alter the lignin content of two rice varieties: Nipponbare (Oryza sativa subsp. japonica) and Rojolele (O. sativa subsp. javanica). The AtMyb61 gene regulated by a strong constitutive CaMV 35S promoter was transformed into these rice varieties mediated by the Agrobacterium tumefaciens strain LBA 4404. Positive transgenic plants were identified using HPT leave assay and PCR analysis. A total of 192 putative transgenic plants were obtained, of which 85 and 18 putative transgenic plants for Nipponbare and Rojolele, respectively. Regeneration (57.73%) and transformation (10.20%) efficiency of Nipponbare was higher than that of regeneration (28.93%) and transformation (2.25%) efficiency of Rojolele. In planta HPT leave assay showed that 85 (68.55%) plants of Nipponbare cultivar and 18 (26.47%) plants of Rojolele cultivar expressed hygromycin phosphotransferase (hpt) indicated that the rice plants positive contained inserted genes. The results of PCR analysis using a specific primer for the hpt gene on 39 HPT-positive plants showed that all plants had transgenes. Rice genes expression involved in lignin biosynthesis, lignin content, and other physiological and agronomical characteristics will be examined in greater detail.