PROMOTER MeEF1A6 ACTIVITY ON Nicotiana tabacum L. PUTATIVE TRANSGENIC

• Main Author: Oktaviani, Indah
• Format: Theses
• Language: Indonesia
• Online Access: https://digilib.itb.ac.id/gdl/view/36241
• Institution: Institut Teknologi Bandung

Promoter MeEF1A6 constitute as one of the promoters of which was isolated from cassava eEF1A gene family (Manihot esculenta Crantz cv. Adira). Previous research suggests that transient transformation of the promoter MeEF1A6 that mediated by Agrobacterium tumefaciens strain GV3101, could control the expression of the reporter gene uidA which encodes enzim ?-glucuronidase (GUS) in all parts of tobacco sprouts, leaves organs, stems, and roots of corn plants, as well as tomatoes. However, MeEF1A6 promoter activity test in the regulation of gene reporter uidA expression on putative transgenic plants has not been performed. This study aimed to test the promoter activity MeEF1A6 in the regulation of the reporter gene uidA expression on transformed putative transgenic plants. Transformation technique used for Nicotiana tabacum L. by leaf discs transformation method with modification using A. tumefaciens strain GV3101 containing the binary vector pBI121-MeEF1A6, pBI121-CaMV35S (positive control), and A. tumefaciens without binary vector (negative control) as intermediary. Transformation efficiency result for promoter MeEF1A6 was 8,78% and 5,6% for positive control (CaMV35S). Promoter activity test was conducted qualitatively using histochemical GUS assays. Localization of uidA gene expression in the putative transgenic plants tissues could be observed directly using a light microscope and sectioning method. Observations on the root organs showed that promoter MeEF1A6 activity were not limited only in the cortex, epidermis and hair roots, but it also active in the cortex, xylem, and pith in the stem, and whole leaf tissue. GUS expression that regulated by promoter MeEF1A6 on the leaf surface, consistently seen more strongly in the area of stomata and the leaf veins. Based on these results we can conclude promoter MeEF1A6 took an active role in regulating expression of gene reporter uidA in the roots, stems, and leaves of putative transgenic tobacco plants.