Genome-editing (GE) is having a tremendous influence around the globe in the life science community. Among its versatile uses, the desired modifications of genes, and more importantly the transgene (DNA)-free approach to develop genetically modified organism (GMO), are of special interest. The recent and rapid developments in genome-editing technology have given rise to hopes to achieve global food security in a sustainable manner. We here discuss recent developments in CRISPR-based genome-editing tools for crop improvement concerning adaptation, opportunities, and challenges. Some of the notable advances highlighted here include the development of transgene (DNA)-free genome plants, the availability of compatible nucleases, and the development of safe and effective CRISPR delivery vehicles for plant genome editing, multi-gene targeting and complex genome editing, base editing and prime editing to achieve more complex genetic engineering. Additionally, new avenues that facilitate fine-tuning plant gene regulation have also been addressed. In spite of the tremendous potential of CRISPR and other gene editing tools, major challenges remain. Some of the challenges are related to the practical advances required for the efficient delivery of CRISPR reagents and for precision genome editing, while others come from government policies and public acceptance. This review will therefore be helpful to gain insights into technological advances, its applications, and future challenges for crop improvement.
In the current era of rapid industrialization, the foremost challenge is the management of industrial wastes. Activities such as mining and industrialization spill over a large quantity of toxic waste that pollutes soil, water, and air. This poses a major environmental and health challenge. The toxic heavy metals present in the soil and water are entering the food chain, which in turn causes severe health hazards. Environmental clean‐up and reclamation of heavy metal contaminated soil and water are very important, and it necessitates efforts of environmentalists, industrialists, scientists, and policymakers. Phytoremediation is a plant‐based approach to remediate heavy metal/organic pollutant contaminated soil and water in an eco‐friendly, cost‐effective, and permanent way. This review covers the effect of heavy metal toxicity on plant growth and physiological process, the concept of heavy metal accumulation, detoxification, and the mechanisms of tolerance in plants. Based on plants' ability to uptake heavy metals and metabolize them within tissues, phytoremediation techniques have been classified into six types: phytoextraction, phytoimmobilization, phytovolatilization, phytodegradation, rhizofiltration, and rhizodegradation. The development of research in this area led to the identification of metal hyper‐accumulators, which could be utilized for reclamation of contaminated soil through phytomining. Concurrently, breeding and biotechnological approaches can enhance the remediation efficiency. Phytoremediation technology, combined with other reclamation technologies/practices, can provide clean soil and water to the ecosystem.
Plant pathology has been revolutionized by the emergence and intervention of next-generation sequencing technologies (NGS) which provide a fast, cost-effective, and reliable diagnostic for any class of pathogens. NGS has made tremendous advancements in the area of research and diagnostics of plant infecting viromes and has bridged plant virology with other advanced research fields like genome editing technologies. NGS in a broader perspective holds the potential for plant health improvement by diagnosing and mitigating the new or unusual symptoms caused by novel/unidentified viruses. CRISPR-based genome editing technologies can enable rapid engineering of efficient viral/viroid resistance by directly targeting specific nucleotide sites of plant viruses and viroids. Critical genes such as eIf (iso) 4E or eIF4E have been targeted via the CRISPR platform to produce plants resistant to single-stranded RNA (ssRNA) viruses. CRISPR/Cas-based multi-target DNA or RNA tests can be used for rapid and accurate diagnostic assays for plant viruses and viroids. Integrating NGS with CRISPR-based genome editing technologies may lead to a paradigm shift in combating deadly disease-causing plant viruses/viroids at the genomic level. Furthermore, the newly discovered CRISPR/Cas13 system has unprecedented potential in plant viroid diagnostics and interference. In this review, we have highlighted the application and importance of sequencing technologies on covering the viral genomes for precise modulations. This review also provides a snapshot vision of emerging developments in NGS technologies for the characterization of plant viruses and their potential utilities, advantages, and limitations in plant viral diagnostics. Furthermore, some of the notable advances like novel virus-inducible CRISPR/Cas9 system that confers virus resistance with no off-target effects have been discussed.
Advances in proteome research have opened the gateway to understanding numerous metabolic pathways and fundamental mechanisms involved in abiotic stress tolerance. In the present study, the antioxidant capacity of four tomato genotypes i.e., Kashi Amrit, Kashi Anupam, EC-317-6-1, and WIR-4360 was determined under drought stress to ascertain the scavenging potential for reactive oxygen species (ROS). A significant increase in the superoxide dismutase (SOD), Ascorbate peroxidase (APX), and catalase (CAT) activities in all the four genotypes under drought stress was observed, which seemed to be associated with a protective role against ROS (p < 0.001). Based on the antioxidant enzyme activities, a proteomic approach was applied to study differential protein expression in two selected genotypes from different species i.e., EC-317-6-1 (Solanum pimpinellifolium) and Kashi Amrit (Solanum lycopersicum) grown under irrigated, drought, and re-watering conditions. To reveal the protein network regulated under these conditions, two-dimensional gel electrophoresis was employed to identify and quantify the number of proteins in drought-sensitive (Kashi Amrit) and tolerant (EC-317-6-1) genotypes. Matrix-assisted laser desorption/ionization-time of flight analysis (MALDI-TOF) revealed a total of 453 spots after fine-tuning factors i.e., smoothness, saliency, and minimum area that responded to drought. Out of 453 total spots, 93 spots were identified in Kashi Amrit and 154 in EC-317-6-1 under irrigated conditions, whereas 4 spots were identified in Kashi Amrit and 77 spots in EC-317-6-1 under drought conditions. Furthermore, differentially expressed proteins were distinguished according to the fold change of their expression. Information provided in this report will be useful for the selection of proteins or genes in analyzing or improving drought tolerance in tomato cultivars. These findings may assist in the construction of a complete proteome database encompassing various divergent species which could be a valuable source for the improvement of crops under drought-stress conditions in the future.
A research trial was conducted at Agronomy Farm (SKUAST-K, Wadura, Jammu & Kashmir), during kharif 2017 and 2018 to evaluate nutrient removal in rice under various rice establishment methods and weed control measures. The study comprised of two factors: rice establishment techniques {(Transplanting (TPR); Direct seeding (DSR) and System of rice intensification (SRI)} as main plot treatments and weed control measures {(Butachlor @ 1500 g a.i ha−1 (B); Penoxsulam @ 22.5 g a.i ha−1 (P); Pyrazosulfuron ethyl + Pretilachlor @ 15 and 600 g a.i ha−1 (PP); Bensulfuron methyl + Pretilachlor @ 60 and 600 g a.i ha−1 (BP); 2 Conoweeding/Hand Weeding (CW/HW); Weed free (WF) and weedy check (WC)} as sub-plot treatments meant to evaluate the best establishment method and weed management practice for rice. Over DSR and transplanted rice, the SRI technique yielded a significant increase in dry biomass accumulation (17.04 and 17.20 t ha−1) and grain (7.92 and 8.17 t ha−1) and straw (9.60 and 10.17 t ha−1) yields. Penoxsulam herbicide significantly showed higher grain and straw yield of 8.19 and 8.28 t ha−1 and 10.13 and 10.44 t ha−1, respectively, than other weed management measures by comparing the means using critical difference. TPR excelled in reducing dry weed biomass more than other established methods. All herbicides considerably reduced dry weed biomass, but Penoxsulam herbicide showed the greatest reduction in dry weed biomass and proved superior against complex weed flora. Weeds showed maximum contribution towards total Biomass under DSR, among rice establishment techniques. In contrast, among different weed control measures, it was maximum in weedy check treatment (Untreated Control) and minimum in penoxsulam treatment. SRI significantly excelled in crop (grain and straw) nutrient uptake compared to the DSR and TPR method, although different crop establishment techniques non-significantly influenced nutrient concentrations. Furthermore, penoxsulam treatment demonstrated higher crop (grain and straw) nutrient uptake among the various weed management measures. However, available soil nutrients were observed among establishment techniques, highest in DSR and lowest in SRI. Moreover, direct-seeded rice excelled SRI and transplanted rice in weed nutrient uptake, and among the different herbicidal treatments, penoxsulam recorded the lowest uptake in weeds. Nutrient budgeting demonstrated that DSR showed the maximum percentage of nutrient removal by weeds, and the minimum ratio was in TPR. In contrast, the lowest rate of nutrients removed via weeds were seen in penoxsulam application under various weed management measures.
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