Recent innovation in the field of genome engineering encompasses numerous levels of plant genome engineering which attract the substantial excitement of plant biologist worldwide. RNA-guided CRISPR Cas9 system has appeared a promising tool in site-directed mutagenesis due to its innovative utilization in different branches of biology. CRISPR-Cas9 nuclease system have supersedes all previously existed strategies and their associated pitfalls encountered with site-specific mutagenesis. Here we demonstrated an efficient sequence specific integration/mutation of FAD2–2 gene in soybean using CRISPR-Cas9 nuclease system. A single guided RNA sequence was designed with the help of a number of bioinformatics tools aimed to target distinct sites of FAD2–2 loci in soybean. The binary vector (pCas9-AtU6-sgRNA) has been successfully transformed into soybean cotyledon using Agrobacterium tumafacien. Taken together our findings complies soybean transgenic mutants subjected to targeted mutation were surprisingly detected in our target gene. Furthermore, the detection of Cas9 gene, BAR gene, and NOS terminator were carried out respectively. Southern blot analysis confirmed the stable transformation of Cas9 gene into soybean. Real time expression with qRT-PCR and Sanger sequencing analysis confirmed the efficient CRISPR-Cas9/sgRNA induced mutation within the target sequence of FAD2–2 loci. The integration of FAD2–2 target region in the form of substitution, deletions and insertions were achieved with notably high frequency and rare off-target mutagenesis. High frequent mutation efficiency was recorded as 21% out of all transgenic soybean plants subjected to targeted mutagenesis. Furthermore, Near-infrared spectroscopy (NIR) indicates the entire fatty acid profiling obtained from the mutants seeds of soybean. A considerable modulation in oleic acid content up to (65.58%) whereas the least level of linoleic acid is (16.08%) were recorded. Based on these finding CRISPR-Cas9 system can possibly sum up recent development and future challenges in producing agronomically important crops.

Fifteen transcription factors in the CAMTA (calmodulin binding transcription activator) family of soybean were reported to differentially regulate in multiple stresses; however, their functional analyses had not yet been attempted. To characterize their role in stresses, we first comprehensively analyzed the GmCAMTA family in silico and thereafter determined their expression pattern under drought. The bioinformatics analysis revealed multiple stress-related cis-regulatory elements including ABRE, SARE, G-box and W-box, 10 unique miRNA (microRNA) targets in GmCAMTA transcripts and 48 proteins in GmCAMTAs’ interaction network. We then cloned the 2769 bp CDS (coding sequence) of GmCAMTA12 in an expression vector and overexpressed in soybean and Arabidopsis through Agrobacterium-mediated transformation. The T3 (Transgenic generation 3) stably transformed homozygous lines of Arabidopsis exhibited enhanced tolerance to drought in soil as well as on MS (Murashige and Skoog) media containing mannitol. In their drought assay, the average survival rate of transgenic Arabidopsis lines OE5 and OE12 (Overexpression Line 5 and Line 12) was 83.66% and 87.87%, respectively, which was ~30% higher than that of wild type. In addition, the germination and root length assays as well as physiological indexes such as proline and malondialdehyde contents, catalase activity and leakage of electrolytes affirmed the better performance of OE lines. Similarly, GmCAMTA12 overexpression in soybean promoted drought-efficient hairy roots in OE chimeric plants as compare to that of VC (Vector control). In parallel, the improved growth performance of OE in Hoagland-PEG (polyethylene glycol) and on MS-mannitol was revealed by their phenotypic, physiological and molecular measures. Furthermore, with the overexpression of GmCAMTA12, the downstream genes including AtAnnexin5, AtCaMHSP, At2G433110 and AtWRKY14 were upregulated in Arabidopsis. Likewise, in soybean hairy roots, GmELO, GmNAB and GmPLA1-IId were significantly upregulated as a result of GmCAMTA12 overexpression and majority of these upregulated genes in both plants possess CAMTA binding CGCG/CGTG motif in their promoters. Taken together, we report that GmCAMTA12 plays substantial role in tolerance of soybean against drought stress and could prove to be a novel candidate for engineering soybean and other plants against drought stress. Some research gaps were also identified for future studies to extend our comprehension of Ca-CaM-CAMTA-mediated stress regulatory mechanisms.

This review delves into the mesmerizing technology of nano-agrochemicals, specifically pesticides and herbicides, and their potential to aid in the achievement of UN SDG 17, which aims to reduce hunger and poverty globally. The global market for conventional pesticides and herbicides is expected to reach USD 82.9 billion by 2027, growing 2.7% annually, with North America, Europe, and the Asia–Pacific region being the biggest markets. However, the extensive use of chemical pesticides has proven adverse effects on human health as well as the ecosystem. Therefore, the efficacy, mechanisms, and environmental impacts of conventional pesticides require sustainable alternatives for effective pest management. Undoubtedly, nano-agrochemicals have the potential to completely transform agriculture by increasing crop yields with reduced environmental contamination. The present review discusses the effectiveness and environmental impact of nanopesticides as promising strategies for sustainable agriculture. It provides a concise overview of green nano-agrochemical synthesis and agricultural applications, and the efficacy of nano-agrochemicals against pests including insects and weeds. Nano-agrochemical pesticides are investigated due to their unique size and exceptional performance advantages over conventional ones. Here, we have focused on the environmental risks and current state of nano-agrochemicals, emphasizing the need for further investigations. The review also draws the attention of agriculturists and stakeholders to the current trends of nanomaterial use in agriculture especially for reducing plant diseases and pests. A discussion of the pros and cons of nano-agrochemicals is paramount for their application in sustainable agriculture.

The recent millennium has witnessed a notable shift in consumer focus towards natural products for addressing lifestyle-related disorders, driven by their safety and cost-effectiveness. Nutraceuticals and functional foods play an imperative role by meeting nutritional needs and offering medicinal benefits. With increased scientific knowledge and awareness, the significance of a healthy lifestyle, including diet, in reducing disease risk is widely acknowledged, facilitating access to a diverse and safer diet for longevity. Plant-based foods rich in phytochemicals are increasingly popular and effectively utilized in disease management. Agricultural waste from plant-based foods is being recognized as a valuable source of nutraceuticals for dietary interventions. Citrus peels, known for their diverse flavonoids, are emerging as a promising health-promoting ingredient. Globally, citrus production yields approximately 15 million tons of by-products annually, highlighting the substantial potential for utilizing citrus waste in phyto-therapeutic and nutraceutical applications. Citrus peels are a rich source of flavonoids, with concentrations ranging from 2.5 to 5.5 g/100 g dry weight, depending on the citrus variety. The most abundant flavonoids in citrus peel include hesperidin and naringin, as well as essential oils rich in monoterpenes like limonene. The peel extracts exhibit high antioxidant capacity, with DPPH radical scavenging activities ranging from 70 to 90%, comparable to synthetic antioxidants like BHA and BHT. Additionally, the flavonoids present in citrus peel have been found to have antioxidant properties, which can help reduce oxidative stress by 30% and cardiovascular disease by 25%. Potent anti-inflammatory effects have also been demonstrated, reducing inflammatory markers such as IL-6 and TNF-α by up to 40% in cell culture studies. These findings highlight the potential of citrus peel as a valuable source of nutraceuticals in diet-based therapies.