oa Isolation of promoters for specific temporal and spatial genetic engineering against nematodes (Heterodera glycines) in soybean (glycine max)

Nematodes cause billions of dollars of damage each year to agricultural crops worldwide. Nematicides are costly and environmentally detrimental. Crop rotation does not eliminate nematodes, but merely decreases crop damage. For successful engineering of transgenic plants for nematodes resistance, it is important to regulate plant resistance through promoters that over-express resistance genes or produce RNAi in the correct cells at the proper time. Data from our previous microarray experiments showed a number of genes that are responsive and activated at feeding site upon nematode infection after 3, 6 and 9 days. In this investigation, we identified and isolated seven different promoter regions, supposedly harboring different regulatory elements upstream to seven responsive genes that are highly expressed in the nematode feeding site. The upstream promoter regions were cloned, sequenced and tested in our newly developed expression vectors series, pJan25S (NCBI: KC416200), pJan25T (NCBI: KC416201) and pJan25X (NCBI: KC416202), that are an enhancements of pGWB533 for pro- moter testing.. Accumulation of the products of reporter genes encoding green fluorescent protein and b-glucuronidase (GUS) was monitored after transformation of onion cells using biolistics gene transfer. These promoter constructs were also prepared for transformation in Arabidopsis and soybean. These promoters will provide scientists with useful regulators for expressing genes and gene products that may inhibit the life cycle of the nematode at nematode feeding site. The same approach can be used for enhancing resistance of Qatari important plants against different pathogen.