Expression of TGMV antisense RNA in transgenic tobacco inhibits replication of BCTV but not ACMV geminiviruses.

Transgenic tobacco plants expressing an antisense RNA targeted against tomato golden mosaic virus (TGMV) show reduced/no symptoms and viral DNA accumulation upon TGMV infection [5]. The targeted region includes the AL1 gene, encoding an essential viral replication protein. This DNA sequence is conserved in various other geminiviruses, suggesting they too might show inhibition of replication in these plants. We infected leaf material with African cassava mosaic virus (ACMV) and beet curly top virus (BTCV) and saw a 4-fold reduction of BCTV, but not ACMV, DNA accumulation, compared to controls. The equivalent regions of BCTV and ACMV show similar overall homology to the TGMV target (63% and 64% respectively), but within this, BCTV displays a 280 nucleotide region of high homology (82%). In contrast, for ACMV, the homology is more dispersed. This indicates that a critical stretch of good complementarity is needed to block expression of the target mRNA, that is effective even within along antisense transcript. These studies indicate the potential for developing a multifunctional antisense cassette.