Genetic biofortification: advancing crop nutrition to tackle hidden hunger.

Malnutrition, often termed "hidden hunger," represents a pervasive global issue carrying significant implications for health, development, and socioeconomic conditions. Addressing the challenge of inadequate essential nutrients, despite sufficient caloric intake, is crucial. Biofortification emerges as a promising solution by enhance the presence of vital nutrients like iron, zinc, iodine, and vitamin A in edible parts of different crop plants. Crop biofortification can be attained through either agronomic methods or genetic breeding techniques. Agronomic strategies for biofortification encompass the application of mineral fertilizers through foliar or soil methods, as well as leveraging microbe-mediated mechanisms to enhance nutrient uptake. On the other hand, genetic biofortification involves the strategic crossing of plants to achieve a desired combination of genes, promoting balanced nutrient uptake and bioavailability. Additionally, genetic biofortification encompasses innovative methods such as speed breeding, transgenic approaches, genome editing techniques, and integrated omics approaches. These diverse strategies collectively contribute to enhancing the nutritional profile of crops. This review highlights the above-said genetic biofortification strategies and it also covers the aspect of reduction in antinutritional components in food through genetic biofortification.

Phenylalanine supply alleviates the drought stress in mustard (Brassica campestris) by modulating plant growth, photosynthesis, and antioxidant defense system.

Mustard (Brassica campestris L.) is a major oilseed crop that plays a crucial role in agriculture. Nevertheless, a number of abiotic factors, drought in particular, significantly reduce its production. Phenylalanine (PA) is a significant and efficacious amino acid in alleviating the adverse impacts of abiotic stressors, such as drought. Thus, the current experiment aimed to evaluate the effects of PA application (0 and 100 mg/L) on brassica varieties i.e., Faisal (V1) and Rachna (V2) under drought stress (50% field capacity). Drought stress reduced the shoot length (18 and 17%), root length (12.1 and 12.3%), total chlorophyll contents (47 and 45%), and biological yield (21 and 26%) of both varieties (V1 and V2), respectively. Foliar application of PA helped overcome drought-induced losses and enhanced shoot length (20 and 21%), total chlorophyll contents (46 and 58%), and biological yield (19 and 22%), whereas reducing the oxidative activities of H2O2 (18 and 19%), MDA concentration (21 and 24%), and electrolyte leakage (19 and 21%) in both varieties (V1 and V2). Antioxidant activities, i.e., CAT, SOD, and POD, were further enhanced under PA treatment by 25, 11, and 14% in V1 and 31, 17, and 24% in V2. Overall findings suggest that exogenous PA treatment reduced the drought-induced oxidative damage and improved the yield, and ionic contents of mustard plants grown in pots. It should be emphasized, however, that studies examining the impacts of PA on open-field-grown brassica crops are still in their early stages, thus more work is needed in this area.

CDK9 and its repressor LARP7 modulate cardiomyocyte proliferation and response to injury in the zebrafish heart.

Cyclin dependent kinase (Cdk)9 acts through the positive transcription elongation factor-b (P-TEFb) complex to activate and expand transcription through RNA polymerase II. It has also been shown to regulate cardiomyocyte hypertrophy, with recent evidence linking it to cardiomyocyte proliferation. We hypothesised that modification of CDK9 activity could both impair and enhance the cardiac response to injury by modifying cardiomyocyte proliferation. Cdk9 expression and activity were inhibited in the zebrafish (Danio rerio) embryo. We show that dephosphorylation of residue Ser2 on the C-terminal domain of RNA polymerase II is associated with impaired cardiac structure and function, and cardiomyocyte proliferation and also results in impaired functional recovery following cardiac laser injury. In contrast, de-repression of Cdk9 activity, through knockdown of La-related protein (Larp7) increases phosphorylation of Ser2 in RNA polymerase II and increases cardiomyocyte proliferation. Larp7 knockdown rescued the structural and functional phenotype associated with knockdown of Cdk9. The balance of Cdk9 and Larp7 plays a key role in cardiomyocyte proliferation and response to injury. Larp7 represents a potentially novel therapeutic target to promote cardiomyocyte proliferation and recovery from injury.

Targeted laser ablation of the zebrafish larval heart induces models of heart block, valvular regurgitation, and outflow tract obstruction.

Mammalian models of cardiac disease have provided unique and important insights into human disease but have become increasingly challenging to produce. The zebrafish could provide inexpensive high-throughput models of cardiac injury and repair. We used a highly targeted laser, synchronized to fire at specific phases of the cardiac cycle, to induce regional injury to the ventricle, atrioventricular (AV) cushion, and bulbus arteriosus (BA). We assessed the impact of laser injury on hearts of zebrafish early larvae at 72 h postfertilization, to different regions, recording the effects on ejection fraction (EF), heart rate (HR), and blood flow at 2 and 24 h postinjury (hpi). Laser injury to the apex, midzone, and outflow regions of the ventricle resulted in reductions of the ventricle EF at 2 hpi with full recovery of function by 24 hpi. Laser injury to the ventricle, close to the AV cushion, was more likely to cause bradycardia and atrial-ventricular dysfunction, suggestive of an electrical conduction block. At 2 hpi, direct injury to the AV cushion resulted in marked regurgitation of blood from the ventricle to the atrium. Laser injury to the BA caused temporary outflow tract obstruction with cessation of ventricle contraction and circulation. Despite such damage, 80% of embryos showed complete recovery of the HR and function within 24 h of laser injury. Precision laser injury to key structures in the zebrafish developing heart provides a range of potentially useful models of hemodynamic overload, injury, and repair.

Heart on a plate: histological and functional assessment of isolated adult zebrafish hearts maintained in culture.

The zebrafish is increasingly used for cardiovascular genetic and functional studies. We present a novel protocol to maintain and monitor whole isolated beating adult zebrafish hearts in culture for long-term experiments. Excised whole adult zebrafish hearts were transferred directly into culture dishes containing optimized L-15 Leibovitz growth medium and maintained for 5 days. Hearts were assessed daily using video-edge analysis of ventricle function using low power microscopy images. High-throughput histology techniques were used to assess changes in myocardial architecture and cell viability. Mean spontaneous Heart rate (HR, min(-1)) declined significantly between day 0 and day 1 in culture (96.7 ± 19.5 to 45.2 ± 8.2 min-1, mean ± SD, p = 0.001), and thereafter declined more slowly to 27.6 ± 7.2 min(-1) on day 5. Ventricle wall motion amplitude (WMA) did not change until day 4 in culture (day 0, 46.7 ± 13.0 µm vs day 4, 16.9 ± 1.9 µm, p = 0.08). Contraction velocity (CV) declined between day 0 and day 3 (35.6 ± 14.8 vs 15.2 ± 5.3 µms(-1), respectively, p = 0.012) while relaxation velocity (RV) declined quite rapidly (day 0, 72.5 ± 11.9 vs day 1, 29.5 ± 5.8 µms(-1), p = 0.03). HR and WMA responded consistently to isoproterenol from day 0 to day 5 in culture while CV and RV showed less consistent responses to beta-agonist. Cellular architecture and cross-striation pattern of cardiomyocytes remained unchanged up to day 3 in culture and thereafter showed significant deterioration with loss of striation pattern, pyknotic nuclei and cell swelling. Apoptotic markers within the myocardium became increasingly frequent by day 3 in culture. Whole adult zebrafish hearts can be maintained in culture-medium for up to 3 days. However, after day-3 there is significant deterioration in ventricle function and heart rate accompanied by significant histological changes consistent with cell death and loss of cardiomyocyte cell integrity. Further studies are needed to assess whether this preparation can be optimised for longer term survival.