The revolutionary technology of CRISPR/Cas systems and their extraordinary potential to address fundamental questions in every field of biological sciences has led to their developers being awarded the 2020 Nobel Prize for Chemistry. In agriculture, CRISPR/Cas systems have accelerated the development of new crop varieties with improved traits—without the need for transgenes. However, the future of this technology depends on a clear and truly global regulatory framework being developed for these crops. Some CRISPR-edited crops are already on the market, and yet countries and regions are still divided over their legal status. CRISPR editing does not require transgenes, making CRISPR crops more socially acceptable than genetically modified crops, but there is vigorous debate over how to regulate these crops and what precautionary measures are required before they appear on the market. This article reviews intended outcomes and risks arising from the site-directed nuclease CRISPR systems used to improve agricultural crop plant genomes. It examines how various CRISPR system components, and potential concerns associated with CRISPR/Cas, may trigger regulatory oversight of CRISPR-edited crops. The article highlights differences and similarities between GMOs and CRISPR-edited crops, and discusses social and ethical concerns. It outlines the regulatory framework for GMO crops, which many countries also apply to CRISPR-edited crops, and the global regulatory landscape for CRISPR-edited crops. The article concludes with future prospects for CRISPR-edited crops and their products.

Barley (Hordeum vulgare, L.) is the fourth most important cereal crop in the world. Salinity decreases the productivity of plants grown under salinity conditions. It leads to deficiency and limited absorption of water and nutrients, ionic stress, oxidative stress, and osmotic imbalance. In saline soil, a field experiment was conducted to verify the effects of nine combinations among three levels of bio-fertilizers, i.e., control (without), arbuscular mycorrhizal fungi (AMF), and phosphate solubilizing bacteria (PSB), as well as three levels of phosphorus fertilizer recommended dose (RDP) on barley yield, its components and nutrients uptake, to evaluate the useful influences of these combinations to improve P management under salinity stress related to yield and its components as well as N, P, and K uptake in barley. Findings revealed that the combination AMF + 100% RDP improved plant height, length of spike, spikes weight, number of spikes plant⁻¹, weight of 1000-grain, straw yield, grain yield, uptake of nitrogen (N), phosphorus (P), potassium (K) in grain and uptake of nitrogen (N), phosphorus (P), potassium (K) in straw by 19.76, 33.21, 40.08, 33.76, 14.82, 24.95, 47.52, 104.54, 213.47, 168.24, 124.30, 183.59, and 160.84% in the first season, respectively. Meanwhile, the increase was 19.86, 29.73, 40.47, 39.94, 14.92, 24.95, 47.94, 104.73, 213.33, 168.64, 124.47, 183.86, and 161.09% in the second season, respectively. AMF showed greater efficiency and effectiveness compared to PSB in improving yield and its components for all studied traits. The results of principle component analysis indicated that all combinations except AMF + zero% RDP, PSB + zero% RDP, control + zero% RDP, and control + 66% RDP showed high scores on positive PC1, where all studied traits were high. Therefore, it is recommended to inoculate the soil with AMF or PSB with the addition of phosphate fertilizer at the recommended dose under salinity conditions, i.e., AMF + 100% RDP (T1) or AMF + 66% RDP (T2) or PSB + 100% RDP (T4). The use of bio-fertilizers has increased plant tolerance to salt stress, and this was evident from the increase in different traits with the use of treatments that include bio-fertilizers.

Biofilm control by essential oil (EO) application has recently increased to preclude biofilm production on foods and environmental surfaces. In this work, the anti-biofilm effects of garlic and thyme essential oils using the minimum inhibitory concentration (MIC) method against Salmonella typhimurium recovered from different abattoir samples were investigated along with the virulence genes (InvA, SdiA and Stn genes), and the antimicrobial susceptibility profile of S. typhimurium as well. The obtained results revealed that S. typhimurium contaminated abattoir samples to varying degrees. The InvA gene was investigated in all isolates, whereas the SdiA and Stn genes were observed in four and three isolates, respectively. Utilizing the disc diffusion method, S. typhimurium isolates demonstrated substantial resistance to most of the examined antibiotics with a high multiple antibiotic resistance index. S. typhimurium isolates demonstrated biofilm formation abilities to various degrees at varied temperatures levels (4 °C and 37 °C). In conclusion, the obtained samples from the research area are regarded as a potential S. typhimurium contamination source. Furthermore, garlic essential oil (GEO) has more potential to inhibit S. typhimurium biofilm at different sub-minimum inhibitory concentrations as compared to thyme essential oil (TEO). Therefore, these EOs are considered as potential natural antibacterial options that could be applied in food industry.

In recent decades, Pakistan has suffered a decline in cotton production due to several factors, including insect pests, cotton leaf curl disease (CLCuD), and multiple abiotic stresses. CLCuD is a highly damaging plant disease that seriously limits cotton production in Pakistan. Recently, genome editing through CRISPR/Cas9 has revolutionized plant biology, especially to develop immunity in plants against viral diseases. Here we demonstrate multiplex CRISPR/Cas-mediated genome editing against CLCuD using transient transformation in N. benthamiana plants and cotton seedlings. The genomic sequences of cotton leaf curl viruses (CLCuVs) were obtained from NCBI and the guide RNA (gRNA) were designed to target three regions in the viral genome using CRISPR MultiTargeter. The gRNAs were cloned in pHSE401/pKSE401 containing Cas9 and confirmed through colony PCR, restriction analysis, and sequencing. Confirmed constructs were moved into Agrobacterium and subsequently used for transformation. Agroinfilteration in N. benthamiana revealed delayed symptoms (3–5 days) with improved resistance against CLCuD. In addition, viral titer was also low (20–40%) in infected plants co-infiltrated with Cas9-gRNA, compared to control plants (infected with virus only). Similar results were obtained in cotton seedlings. The results of transient expression in N. benthamiana and cotton seedlings demonstrate the potential of multiplex CRISPR/Cas to develop resistance against CLCuD. Five transgenic plants developed from three experiments showed resistance (60−70%) to CLCuV, out of which two were selected best during evaluation and screening. The technology will help breeding CLCuD-resistant cotton varieties for sustainable cotton production.

We developed alginate-based floating microbeads of clarithromycin with therapeutic oils for the possible eradication of Helicobacter pylori (H. pylori) infections by enhancing the residence time of the carrier at the site of infection. In pursuit of this endeavor, the alginate was blended with hydroxy propyl methyl cellulose (HPMC) as an interpenetrating polymer to develop beads by ionotropic gelation using calcium carbonate as a gas generating agent. The developed microbeads remained buoyant under gastric conditions for 24 h. These microbeads initially swelled and afterwards decreased in size, possibly due to the erosion of the polymer. Furthermore, swelling was also affected by the type of encapsulated oil, i.e., swelling decreased with increasing concentrations of eucalyptus oil and increased with increasing concentrations of oleic acid. Antibacterial assays of the formulations showed significant antibacterial activity against Staphylococcus aureus (S. aureus) and Escherichia coli (E. coli); these assays also showed synergistic activity between clarithromycin and therapeutic oils as evident from the higher zone of inhibition of the microbeads as compared to the pure drug and oils. Scanning electron microscopy (SEM) images revealed a smoother surface for oleic acid containing the formulation as compared to eucalyptus oil containing the formulation. Differential scanning calorimetry (DSC) and thermogravimetric analysis (TGA) revealed the development of a stable formulation, while Fourier transform infrared spectrophotometry (FTIR) studies did not reveal any interaction between the polymers and the active ingredients. Optimized formulations (CLM3 and CLM6) were designed to release the drug in a controlled manner in gastric media by Fickian diffusion. Conclusively, the developed microbeads are a promising carrier to overcome the narrow therapeutic index and low bioavailability of clarithromycin, while the presence of therapeutic oils will produce synergistic effects with the drug to eradicate infection effectively.

Understanding the natural resources of native flora in a particular area is essential to be able to identify, record, and update existing records concerning the flora of that area, especially medicinal plants. Until recently, there has been very little scientific documentation on the biological diversity of Aljumum flora. The current study aimed to document medicinal plants among the flora of this region and determine the traditional usages that are documented in the literature. In the flowering season from November 2019 to May 2020, we conducted more than 80 field trips to the study area. The results reported 90 species belonging to 79 genera and 34 families in the Aljumum region, which constitute 82 species of medicinal plants from a total of 2253 known species in Saudi Arabia. The most distributed species were Calotropis procera, Panicum turgidum, and Aerva javanica (5.31%); within four endemic families, we found Fabaceae (32.35%), Poaceae (20.58%), and Asteraceae and Brassicaceae (17.64%). The present study reviews a collection of medicinal plants in Aljumum used in ethnomedicine. Additionally, these natural resources should be preserved, and therefore, conservation programs should be established to protect the natural diversity of the plant species in this region with sustainable environmental management.