Introgression, Generational Expression and Salinity Tolerance Conferred by the Pea DNA Helicase 45 Transgene into Two Commercial Rice Genotypes, BR28 and BR47.

ABSTRACT

DNA helicase (PDH45) from the pea plant (Pisum sativum) is a member of the DEAD box protein family and plays a vital regulatory role in saline stress tolerance in plants. We previously reported that over-expression of PDH45 gene confers both seedling and reproductive stage salinity tolerance to a Bangladeshi rice landrace, Binnatoa (BA). In this study, transgenic BA-containing PDH45 (♂) was crossed with two different farmer-popular BRRI rice varieties (♀), BR28 and BR47, in a contained net house. F1 plants positive for the transgene and having recipient phenotype were advanced from F1 to F5. Expression of the PDH45 gene was detected in all generations. The expression level of PDH45 was 200-fold higher in the donor compared to the two recipient genotypes but without any effect on their salt stress tolerance ability in various assays. Under 120 mM NaCl stress at seedling stage, all rice genotypes showed vigorous growth, higher chlorophyll content, lower electrolyte leakage and lower LDS (Leaf Damage Score) compared to their corresponding wild types. At the reproductive stage under continuous salinity stress at 80 mM NaCl, the cross-bred lines BR28 and BR47 showed significantly better spikelet fertility and yield per plant, which were two- and 2.5-folds, respectively, than their corresponding wild types. The PDH45 transgene was observed to increase the expression of 6 salt stress-related downstream genes at 150 mM NaCl stress to similar differential degrees in the donor and recipient genotypes. However, the expression of OsLEA was significantly higher in transgenic BR28 compared to transgenic BR47, where the latter shows comparatively higher salt tolerance. The study shows stability of transgene expression across generations. It also demonstrates that there may be an effect of background genotype on transgene expression. Moreover, some downstream effects of the transgene may also be genotype-specific.