Genetic stability of BT cry1Ac gene in transgenic fruit and shoot borer resistant eggplant
2009
Hautea, D.M. | Narciso, J.O. | Canama, A.O. | Ripalda, R.R. | Ramil, C.P. | Canicosa, J.E.B. | Balendres, M.A.O., University of Minnesota, St. Paul, MN 55108 (USA). Dept. of Agronomy and Plant Genetics
Bioengineered eggplant varieties present a potential remedy for the chronic problem of high infestation caused by eggplant fruit and shoot borer (EFSB), Leucinodes orbonalis Green. and high pesticide use. The Maharashtra Hybrid Seed Company (Mahyco) of India has developed a highly resistant transgenic Bt eggplant event (EE-1) containing Bt cry1Ac gene. Thru a public-private partnership facilitated by the Agricultural Biotechnology Support Project II (ABSP II), Mahyco's Bt donor line containing event EE-1 was initially crossed and subsequently backcrossed to selected Philippine eggplant varieties, Mara and Dumaguete Long Purple (DLP). Molecular analyses were conducted to determine the genetic stability of event. EE-1 in three successive backcross generations and in two BC1 F2 generations using an event-specific PCR assay for EE-1 and a separate PCR assay that is specific for the selectable marker np+11 in the BC1F2 generations. In addition, the stability of Bt Cry1Ac protein expression was determined by qualitative ELISA. In the three successive BC generation (BC1F1, BC2F1 and BC3F1) to both DLP and Mara, the transgene segregated in the expected 3:1 Mendelian ratio, indicating its stable integration and inheritance as a single Mendelian unit. In the BC1F2 generations in both crosses to Mara and DLP, the transgene and its npt11 marker segregated in the expected 1:1 Mendelian ratio, further confirming the inheritance of a stably integrated single transgene. Moreover, qualitative determination of the Cry1Ac protein in the three backcross generations also show 1:1 Mendelian segregation in the BC1F2 population, of a single Bt transgene that is stably integrated and expressed.
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