Nitric oxide mitigates the effect of water deficit in Crambe abyssinica.

Crambe abyssinica is widely cultivated in the off-season in the Midwest region of Brazil with great potential for biodeisel production. Low precipitation is characteristic of this region, which can drastically affect the productivity of C. abyssinica. Signaling molecules, such as nitric oxide (NO), can potentially alleviate the effects of water stress on plants. Here we test whether nitric oxide, applied by donor sodium nitroprusside (SNP), can alleviate the occurrence of water deficit damages in Crambe plants and maintain physiological and biochemical processes. Crambe plants were sprayed with three doses of SNP (0, 75, and 150 µM) and were submitted to two water levels (100% and 50% of the maximum water holding capacity). After 32 and 136 h, leaves were analyzed to evaluate the concentration of NO, water relations, gas exchange, chlorophyll a fluorescence, chloroplastidic pigments, proline, malondialdehyde, hydrogen peroxide, superoxide anions, and the antioxidant enzymes activity. Application of SNP allowed the maintenance of gas exchange, chlorophyll fluorescence parameters, and activities of antioxidant enzymes in plants exposed to water deficit, as well as increased the concentration of NO, proline, chloroplastidic pigments and osmotic potential. The application of SNP also decreased the concentration of malondialdehyde and reactive oxygen species in plants submitted to water deficit. Thus, the application of SNP prevented the occurrence of symptoms of water deficit in Crambe plants, maintaining the physiological and biochemical responses at reference levels, even under stress conditions.

Genetic stability of micropropagated plants of Crambe abyssinica Hochst using ISSR markers.

Crambe (Crambe abyssinica) is a non-edible annual herb, which was first cultivated to extract oil for industry, and now has great potential for biodiesel production. The objective of this investigation was to evaluate the genetic stability of micropropagated plants of the C. abyssinica Hochst cultivar 'FMS brilhante' using polymerase chain reaction techniques based on inter-simple sequence repeat (ISSR) molecular markers. The aim was to develop a protocol for the in vitro regeneration of these plants with low genetic variation as compared to the donor plant. For micropropagation, shoot tips from in vitro germinated seedlings were used as explants and were initially cultivated for 90 days on MS medium with 5.0 µM 6-benzylaminopurine (BAP), which at 90 days, led to the highest number of shoots per explant (NSE) (12.20 shoots) being detected. After 120 days, the interaction between BAP concentration and naphthalene acetic acid (NAA) was tested, and the highest NSE was observed following exposure to 0.0/0.5 µM BAP/NAA (11.40 shoots) and 1.0/0.0 µM BAP/NAA (11.00 shoots). The highest proportion of rooting phase were observed following exposure to 0.5 µM NAA (30%). The 13 ISSR primers used to analyze genetic stability produced 91 amplification products, of which only eight bands were polymorphic and 83 were monomorphic for all 10 regenerated crambe plants, compared to the donor plant explant. These results indicate that crambe shoot tips are a highly reliable explant that can be used to micropropagate genetically true-to-type plants or to maintain genetic stability, as verified using ISSR markers.

Reduced Silver Nanoparticle Phytotoxicity in Crambe abyssinica with Enhanced Glutathione Production by Overexpressing Bacterial γ-Glutamylcysteine Synthase.

Silver nanoparticles (Ag NPs) are widely used in consumer products, and their release has raised serious concerns about the risk of their exposure to the environment and to human health. However, biochemical mechanisms by which plants counteract NP toxicity are largely unknown. We have previously engineered Crambe abyssinica plants expressing the bacterial γ-glutamylecysteine synthase (γ-ECS) for enhancing glutathione (GSH) levels. In this study, we investigated if enhanced levels of GSH and its derivatives can protect plants from Ag NPs and AgNO3 (Ag(+) ions). Our results showed that transgenic lines, when exposed to Ag NPs and Ag(+) ions, were significantly more tolerant, attaining a 28%-46% higher biomass and 34-49% more chlorophyll content, as well as maintaining 35-46% higher transpiration rates as compared to those of wild type (WT) plants. Transgenic γ-ECS lines showed 2-6-fold Ag accumulation in shoot tissue and slightly lower or no difference in root tissue relative to levels in WT plants. The levels of malondialdehyde (MDA) in γ-ECS lines were also 27.3-32.5% lower than those in WT Crambe. These results indicate that GSH and related peptides protect plants from Ag nanotoxicity. To our knowledge, this is the first direct report of Ag NP detoxification by GSH in transgenic plants, and these results will be highly useful in developing strategies to counteract the phytotoxicty of metal-based nanoparticles in crop plants.