The vacuolar proton pyrophosphatase gene (SbVPPase) from the Sorghum bicolor confers salt tolerance in transgenic Brahmi [Bacopa monnieri (L.) Pennell].

Plants overcome the effect of Na+ toxicity either by excluding Na+ at the plasma membrane or by sequestering them into the vacuoles. Influx of Na+ ions into the plant vacuoles is usually driven by H+ generated by vacuolar-type H+-ATPase as well as vacuolar proton pyrophosphatse (VPPase). In the present study, we have developed Bacopa monnieri transgenics via Agrobacterium tumefaciens containing the recombinant vector pCAMBIA2300-SbVPPase gene. Transformants were produced using nodal explants. Transformants were confirmed by PCR and DNA blot analysis. qPCR analysis showed higher transcript levels of SbVPPase compared to untransformed control (UC). Higher VPPase activity was recorded in transgenics compared to UC. Under 150 mM salt stress, transgenic shoots showed enhanced Na+ accumulation with better biomass production, increased glycine betaine content, and total soluble sugar levels than UC. Transgenic shoots showed 2.9-3.8-folds lower levels of malondialdehyde content indicating lesser membrane damage. Increase in antioxidant enzyme activities (1.4-3.2-folds) was observed in transgenics compared to UC. Transgenics also displayed 7.3-9.0-folds enhanced accumulation of the medicinally important compound bacoside A. Increased biomass production, accumulation of Na+, osmolytes (glycine betaine, sugars etc.), and elevated antioxidant enzyme activities indicate better osmotic adjustment in transgenics by compartmentalization of Na+ into the vacuoles under salt stress conditions. Thus, overexpression of SbVPPase in Bacopa alleviated salt stress by sequestering Na+.

Overexpression of SbAP37 in rice alleviates concurrent imposition of combination stresses and modulates different sets of leaf protein profiles.

KEY MESSAGE: SbAP37 transcription factor contributes to a combination of abiotic stresses when applied simultaneously in rice. It modulates a plethora of proteins that might regulate the downstream pathways to impart salt stress tolerance. APETALA type of transcription factor was isolated from Sorghum bicolor (SbAP37), overexpressed in rice using a salt inducible abscisic acid 2 (ABA2) promoter through Agrobacterium tumefaciens following in planta method. Transgenics were confirmed by PCR amplification of SbAP37, hygromycin phosphotransferase (hptII) marker and ABA2 promoter and DNA blot analysis. Plants were exposed to 150 mM NaCl coupled with high day/night 36 ± 2/25 ± 2 °C temperatures and also drought stress by withholding water for 1-week separately at the booting stage. SbAP37 expression was 2.8- to 4.7-folds higher in transgenic leaf under salt, but 1.8- to 3.3-folds higher in roots under drought stress. Native gene expression analysis showed that it is expressed more in stem than in roots and leaves under 150 mM NaCl and 6% PEG stress. In the present study, proteomic analysis of transgenics exposed to 150 mM NaCl coupled with elevated temperatures was taken up using quadrupole time-of-flight (Q-TOF) mass spectrometry (MS). The leaf proteome revealed 11 down regulated, 26 upregulated, 101 common (shared), 193 newly synthesized proteins in transgenics besides 368 proteins in untransformed plants. Some of these protein sets appeared different and unique to combined stresses. Our results suggest that the SbAP37 has the potential to improve combined stress tolerance without causing undesirable phenotypic characters when used under the influence of ABA2 promoter.