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Is CRISPR/Cas9-based multi-trait enhancement of wheat forthcoming?
... CRISPR/Cas9-mediated genome editing can be used to target abiotic stressrelated genes for overexpression to enhance their function and improve abiotic stress tolerance such as; TaCER1-6A, TaHAG1, TaPGK which are related to drought (Overexpression increased wax alkane levels, reducing water loss and improving drought tolerance), salinity (Overexpression improved antioxidant activity, ionic balance, and salinity tolerance), and cold (Overexpression enhanced energy metabolism, reducing cellular damage under cold stress) receptively. It can also be used to knockout genes to remove their activity and study their role in stress responses or eliminate undesirable traits such as TaCER1-6A, TaHAG1, TaHSFA6e, TaHSP70, TaPGK, TaPHT1;9 that are related to drought (Knockout reduced wax alkane levels, leading to increased water loss and reduced drought tolerance), salinity (Knockout reduced antioxidant activity, increased Na + levels, and diminished salinity tolerance), heat (heat shock transcription factor Knockout reduced survival rates under heat stress), cold (Knockout reduced energy metabolism, increasing cellular damage and cold sensitivity) and heavy metals (phosphate transporter Knockout reduced arsenic uptake and improved growth under arsenic stress) respectively [53]. ...
Abiotic stress, including drought, heat, and salinity, is a major yield-limiting factor for wheat production, which is crucial for facing food scarcity. With the growing challenges posed by climate change, improving wheat's resilience to abiotic stresses is essential for ensuring food security. This chapter explores the damaging effects of these stresses on wheat and examines the genes, pathways, and mechanisms involved in tolerance, focusing on key stress-related genes and their regulatory networks, such as the TaDREB1 gene, which enhances drought tolerance by regulating water-use efficiency ; TaHKT1;5, which plays a pivotal role in salinity tolerance by maintaining ionic balance; and TaHSP17.4, which improves heat tolerance by reducing oxidative damage and stabilizing cellular functions. It also discusses the potential of genome editing, like CRISPR-Cas9 and TALENs, to improve wheat tolerance to these abiotic stresses, offering a sustainable approach to enhancing crop performance to meet future food demands.
... The advent of CRISPR-Cas9 technology has revolutionized the manipulation of wheat genomes (Wang et al., 2014;Haber et al., 2024), enabling precise integration of traits from WWRs. This transformative tool facilitates targeted gene edits to enhance disease resistance, stress tolerance, and nutritional content (Bortesi and Fischer, 2015;Zhang et al., 2019). ...
Modern agriculture faces increasing challenges from climate change and a rapidly growing global population, necessitating innovative strategies to ensure food security. Wheat wild relatives (WWR) represent a valuable genetic resource for improving wheat resilience and productivity. These species possess traits that confer resistance to pests and diseases, tolerance to environmental stresses such as drought and salinity, and enhanced nutritional quality. Recent advances in genomic sequencing and gene editing have facilitated the transfer of these beneficial traits into cultivated wheat. This review explores the potential of WWR in overcoming the limitations of current wheat varieties and enhancing climate resilience. Key topics include the genetic diversity and adaptability of WWR to harsh environments, recent breakthroughs in cross-breeding and genomics, and the emerging field of de novo domestication. Case studies showcase successful applications of wild wheat traits in modern agriculture. Harnessing WWR’s genetic resources presents a viable pathway to developing high-yielding, resilient crops that sustain future food supplies. Achieving this goal requires significant investment, interdisciplinary collaboration, and robust support for research, (pre-)breeding programs, and field trials.
... The gRNA sequence is complementary to the target DNA, guiding the Cas9 protein to accurately cleave the DNA, resulting in a double-strand break ). The cell then initiates repair through two main pathways: non-homologous end joining (NHEJ), which directly ligates DNA ends but often introduces mutations, making it suitable for gene knockout (Haber et al. 2024). The other is homology-directed repair (HDR), where a provided DNA template with the correct sequence guides precise repair or replacement of the DNA, commonly used for gene editing ). ...
Antibiotic resistance has become a public safety issue of the twenty-first century, posing a growing threat and drawing increased attention. Compared to traditional antibiotics, antimicrobial peptides (AMPs), as naturally produced small peptides, can target multiple pathways within pathogens and render them less prone to developing resistance. This makes them promising alternatives to antibiotics. However, traditional chemical synthesis methods face challenges, such as high costs, low yields, and poor stability, limiting the large-scale industrial production of AMPs. Despite extensive research to improve AMP production efficiency, issues such as low yields and complex extraction processes continue to pose significant barriers to commercial application. Therefore, there is an urgent need for new biosynthesis strategies and optimization methods to enhance AMP production efficiency and quality. This review summarizes the sources, classification, mechanisms of action and recent advances in the microbial synthesis of AMPs. It also explores innovative production methods, including recombinant microbial expression systems, fusion tags, codon optimization, tandem multimer expression, and hybrid peptide expression. Furthermore, we review the applications of gene editing technologies and artificial intelligence in AMP production, providing new perspectives and strategies for efficient, large-scale AMP production.
Graphical Abstract
... While the establishment of an efficient CRISPR-Cas9 multiplex system holds significant promise for shortening breeding times and enhancing wheat yields and quality, this potential has only been realized in a few specific cases. The technology is still in the early stages of application, and although it offers considerable possibilities, widespread implementation and consistent results across different wheat varieties have yet to be fully achieved [59][60][61]. These advancements underscore the transformative potential of genomics in addressing rust resistance challenges and advancing sustainable wheat agriculture. ...
Wheat rusts, including leaf, stripe, and stem rust, have been a threat to global food security due to their devastating impact on wheat yields. In recent years, significant strides have been made in understanding wheat rusts, focusing on disease spread mechanisms, the discovery of new host resistance genes, and the molecular basis of rust pathogenesis. This review summarizes the latest approaches and studies in wheat rust research that provide a comprehensive understanding of disease mechanisms and new insights into control strategies. Recent advances in genetic resistance using modern genomics techniques, as well as molecular mechanisms of rust pathogenesis and host resistance, are discussed. In addition, innovative management strategies, including the use of fungicides and biological control agents, are reviewed, highlighting their role in combating wheat rust. This review also emphasizes the impact of climate change on rust epidemiology and underscores the importance of developing resistant wheat varieties along with adaptive management practices. Finally, gaps in knowledge are identified and suggestions for future research are made. This review aims to inform researchers, agronomists, and policy makers, and to contribute to the development of more effective and sustainable wheat rust control strategies.
Herbicide resistance (HR) is fundamental for sustainable agriculture as global food security increasingly relies on efficient and eco-friendly weed management. Recent advances in CRISPR/dCas9-based epigenome editing offer a promising, non-genetic approach by precisely targeting regulatory regions of genes involved in herbicide sensitivity and detoxification. While CRISPR/Cas9 has successfully been used to develop HR crops, CRISPR/dCas9 remains underexplored in this field. We propose that CRISPR/dCas9-driven epigenome editing could enable time- and tissue-specific control of gene expression, allowing for the introduction of heritable HR traits without altering DNA sequences. This innovative approach could transform sustainable HR development, offering a powerful solution to enhance agricultural resilience and food security while aligning with eco-friendly weed management strategies.
The approach to halophytes remains a very important topic today, in a very fragile context represented by climate change and the global demographic explosion. Although the definition of halophytes itself raises a number of problems of biological interpretation, the knowledge accumulated so far has enabled a clear outline of some directions of biotechnological use of halophytes. Among the examples of biotechnological applications, we mention genetic and metabolic modifications, microbiome symbiosis, and high-precision technologies in agriculture.
Heat stress greatly threatens crop production. Plants have evolved multiple adaptive mechanisms, including alternative splicing, that allow them to withstand this stress. However, how alternative splicing contributes to heat stress responses in wheat (Triticum aestivum) is unclear.
We reveal that the heat shock transcription factor gene TaHSFA6e is alternatively spliced in response to heat stress. TaHSFA6e generates two major functional transcripts: TaHSFA6e‐II and TaHSFA6e‐III. TaHSFA6e‐III enhances the transcriptional activity of three downstream heat shock protein 70 (TaHSP70) genes to a greater extent than does TaHSFA6e‐II. Further investigation reveals that the enhanced transcriptional activity of TaHSFA6e‐III is due to a 14‐amino acid peptide at its C‐terminus, which arises from alternative splicing and is predicted to form an amphipathic helix.
Results show that knockout of TaHSFA6e or TaHSP70s increases heat sensitivity in wheat. Moreover, TaHSP70s are localized in stress granule following exposure to heat stress and are involved in regulating stress granule disassembly and translation re‐initiation upon stress relief. Polysome profiling analysis confirms that the translational efficiency of stress granule stored mRNAs is lower at the recovery stage in Tahsp70s mutants than in the wild types.
Our finding provides insight into the molecular mechanisms by which alternative splicing improves the thermotolerance in wheat.
The discovery of the CRISPR/Cas genome-editing system has revolutionized our understanding of the plant genome. CRISPR/Cas has been used for over a decade to modify plant genomes for the study of specific genes and biosynthetic pathways as well as to speed up breeding in many plant species, including both model and non-model crops. Although the CRISPR/Cas system is very efficient for genome editing, many bottlenecks and challenges slow down further improvement and applications. In this review we discuss the challenges that can occur during tissue culture, transformation, regeneration, and mutant detection. We also review the opportunities provided by new CRISPR platforms and specific applications related to gene regulation, abiotic and biotic stress response improvement, and de novo domestication of plants.
Here, we reported the complete profiling of the crotonylation proteome in common wheat. Through a combination of crotonylation and multi-omics analysis, we identified a TaPGK associated with wheat cold stress. Then, we confirmed the positive role of TaPGK-modulating wheat cold tolerance. Meanwhile, we found that cold stress induced lysine crotonylation of TaPGK. Moreover, we screened a lysine decrotonylase TaSRT1 interacting with TaPGK and found that TaSRT1 negatively regulated wheat cold tolerance. We subsequently demonstrated TaSRT1 inhibiting the accumulation of TaPGK protein, and this inhibition was possibly resulted from decrotonylation of TaPGK by TaSRT1. Transcriptome sequencing indicated that overexpression of TaPGK activated glycolytic key genes and thereby increased pyruvate content. Moreover, we found that exogenous application of pyruvate sharply enhanced wheat cold tolerance. These findings suggest that the TaSRT1-TaPGK model regulating wheat cold tolerance is possibly through mediating pyruvate. This study provided two valuable cold tolerance genes and dissected diverse mechanism of glycolytic pathway involving in wheat cold stress.
In the smallholder, low-input farming systems widespread in sub-Saharan Africa, farmers select and propagate crop varieties based on their traditional knowledge and experience. A data-driven integration of their knowledge into breeding pipelines may support the sustainable intensification of local farming. Here, we combine genomics with participatory research to tap into traditional knowledge in smallholder farming systems, using durum wheat (Triticum durum Desf.) in Ethiopia as a case study. We developed and genotyped a large multiparental population, called the Ethiopian NAM (EtNAM), that recombines an elite international breeding line with Ethiopian traditional varieties maintained by local farmers. A total of 1,200 EtNAM lines were evaluated for agronomic performance and farmers' appreciation in three locations in Ethiopia, finding that women and men farmers could skillfully identify the worth of wheat genotypes and their potential for local adaptation. We then trained a genomic selection (GS) model using farmer appreciation scores and found that its prediction accuracy over grain yield (GY) was higher than that of a benchmark GS model trained on GY. Finally, we used forward genetics approaches to identify marker-trait associations for agronomic traits and farmer appreciation scores. We produced genetic maps for individual EtNAM families and used them to support the characterization of genomic loci of breeding relevance with pleiotropic effects on phenology, yield, and farmer preference. Our data show that farmers' traditional knowledge can be integrated in genomics-driven breeding to support the selection of best allelic combinations for local adaptation.
Plants compete for light partly by over-producing chlorophyll in leaves. The resulting high light absorption is an effective strategy for out competing neighbors in mixed communities, but it prevents light transmission to lower leaves and limits photosynthesis in dense agricultural canopies. We used a CRISPR/Cas9-mediated approach to engineer rice plants with truncated light-harvesting antenna (TLA) via knockout mutations to individual antenna assembly component genes CpSRP43, CpSRP54a, and its paralog, CpSRP54b. We compared the photosynthetic contributions of these components in rice by studying the growth rates of whole plants, quantum yield of photosynthesis, chlorophyll density and distribution, and phenotypic abnormalities. Additionally, we investigated a Poales-specific duplication of CpSRP54. The Poales are an important family that includes staple crops such as rice, wheat, corn, millet, and sorghum. Mutations in any of these three genes involved in antenna assembly decreased chlorophyll content and light absorption and increased photosynthesis per photon absorbed (quantum yield). These results have significant implications for the improvement of high leaf-area-index crop monocultures.
Summary Increase of grain yield is always a major objective of wheat genetic improvement. The SQUAMOSA‐promoter binding protein‐like (SPL) genes, coding for a small family of diverse plant‐specific transcription factors, represent important targets for improving grain yield and other major agronomic traits in rice. The function of the SPL genes in wheat remains to be investigated in this respect. In this study, we identified 56 wheat orthologues of rice SPL genes belonging to 19 homoeologous groups. Like in rice, nine orthologous TaSPL genes harbors the microRNA156 recognition elements (MRE) in their last exons except for TaSPL13, which harbors the MRE in its 3´‐untranslated region (3´UTR). We modified the MRE of TaSPL13 using CRISPR‐Cas9 and generated 12 mutations in the three homoeologous genes. As expected, the MRE mutations led to an approximate two‐fold increase in the TaSPL13 mutant transcripts. The phenotypic evaluation showed that the MRE mutations in TaSPL13 resulted in decrease in flowering time, tiller number, and plant height and concomitantly increase in grain size and number. The results show that the TaSPL13 mutants exhibit a combination of different phenotypes observed in Arabidopsis AtSPL3/4/5 mutants and rice OsSPL13/14/16 mutants and hold great potential in improving wheat yield by simultaneously increasing grain size and number and by refining plant architecture. The novel TaSPL13 mutations generated can be utilized in wheat breeding programs to improve these agronomic traits.
Wheat stripe rust caused by the fungus Puccinia striiformis f. sp. tritici (Pst) is one of the most destructive wheat diseases resulting in significant losses to wheat production worldwide. The development of disease‐resistant varieties is the most economical and effective measure to control diseases. Altering the susceptibility genes that promote pathogen compatibility via CRISPR/Cas9‐mediated gene editing technology has become a new strategy for developing disease‐resistant wheat varieties. Calcineurin B‐like protein (CBL)‐interacting protein kinases (CIPKs) has been demonstrated to be involved in defense responses during plant‐pathogen interactions. However, whether wheat CIPK functions as susceptibility factor is still unclear. Here, we isolated a CIPK homologue gene TaCIPK14 from wheat. Knockdown of TaCIPK14 significantly increased wheat resistance to Pst, whereas overexpression of TaCIPK14 resulted in enhanced wheat susceptibility to Pst by decreasing different aspects of the defense response, including accumulation of ROS and expression of pathogenesis‐relative genes. We generated wheat Tacipk14 mutant plants by simultaneous modification of the three homoeologs of wheat TaCIPK14 via CRISPR/Cas9 technology. The Tacipk14 mutant lines expressed race‐nonspecific (RNS) broad‐spectrum resistance (BSR) to Pst. Moreover, no significant difference was found in agronomic yield traits between Tacipk14 mutant plants and Fielder control plants under greenhouse and field conditions. These results demonstrate that TaCIPK14 acts as an important susceptibility factor in wheat response to Pst, and knock‐out of TaCIPK14 represents a powerful strategy for generating new disease‐resistant wheat varieties with BSR to Pst.
Heavy metal (HM) contamination is a serious concern across the globe, and in recent times, HMs’ intensity has significantly increased, posing a serious threat to crop growth and productivity. Heavy metals pose serious health issues in humans by entering the human food chains. Therefore, it is direly needed to reduce the effects of HMs on plants and humans by adapting appropriate practices. In this context, application of micronutrients can be an essential practice to mitigate the toxic effects of HMs. Zinc (Zn) is a crucial nutrient needed for plant growth, and Zn application reduced the HM-induced toxicity in plants. This review highlights Zn’s role in mitigating the HMs toxicity in plants. We have systematically described the potential mechanisms mediated by Zn to mitigate HMs in plants. Zinc application reduced the HMs uptake and translocation plants, which is considered an essential mechanism of HM stress tolerance. Zn application also improves membrane stability, plant water relationship, nutrient uptake, photosynthetic performance, osmolytes accumulation, anti-oxidant activities, and gene expression. In addition to this, the Zn application substantially improves photosynthesis by enhancing the synthesis of photosynthetic pigments, photosystem activities, enzymatic activities, and maintaining photosynthetic apparatus structure, ensuring better growth under HM stress. Therefore, Zn nutrition could improve the plant performance under HM stress by modulating the plant’s physiological and biochemical functioning, anti-oxidant activities, osmolytes accumulation, and gene expression.
Plants have evolved a two‐branched innate immune system to detect and cope with pathogen attack, which are initiated by cell‐surface and intracellular immune receptors leading to pattern‐triggered immunity (PTI) and effector‐triggered immunity (ETI), respectively. A core transducer including PAD4‐EDS1 node is proposed as the convergence point for a two‐tiered immune system in conferring pathogen immunity. However, the transcriptional regulatory mechanisms controlling expression of these key transducers remain largely unknown.
Here, we identified histone acetyltransferase TaHAG1 as a positive regulator of powdery mildew resistance in wheat. TaHAG1 regulates expression of key transducer gene TaPAD4 and promotes SA and reactive oxygen species accumulation to accomplish resistance to Bgt infection.
Moreover, overexpression and CRISPR‐mediated knockout of TaPAD4 validate its role in wheat powdery mildew resistance. Furthermore, TaHAG1 physically interacts with TaPLATZ5, a plant‐specific zinc‐binding protein. TaPLATZ5 directly binds to promoter of TaPAD4 and together with TaHAG1 to potentiate the expression of TaPAD4 by increasing the levels of H3 acetylation.
Our study revealed a key transcription regulatory node in which TaHAG1 acts as an epigenetic modulator and interacts with TaPLATZ5 that confers powdery mildew resistance in wheat through activating a convergence point gene between PTI and ETI, which could be effective for genetic improvement of disease resistance in wheat and other crops.
Pulses and whole grains are considered staple foods that provide a significant amount of calories, fibre and protein, making them key food sources in a nutritionally balanced diet. Additionally, pulses and whole grains contain many bioactive compounds such as dietary fibre, resistant starch, phenolic compounds and mono- and polyunsaturated fatty acids that are known to combat chronic disease. Notably, recent research has demonstrated that protein derived from pulse and whole grain sources contains bioactive peptides that also possess disease-fighting properties. Mechanisms of action include inhibition or alteration of enzyme activities, vasodilatation, modulation of lipid metabolism and gut microbiome and oxidative stress reduction. Consumer demand for plant-based proteins has skyrocketed primarily based on the perceived health benefits and lower carbon footprint of consuming foods from plant sources versus animal. Therefore, more research should be invested in discovering the health-promoting effects that pulse and whole grain proteins have to offer.
During the last century, the progressive substitution of landraces with modern, high yielding varieties, led to a dramatic reduction of in situ conserved crop diversity in Europe. Nowadays there is limited and scattered information on where landraces are cultivated. To fill this gap and lay the groundwork for a regional landrace in situ conservation strategy, information on more than 19,335 geo-referenced landrace cultivation sites were collated from 14 European countries. According to collected data, landraces of 141 herbaceous and 48 tree species are cultivated across Europe: Italy (107 species), Greece (93), Portugal (45) and Spain (44) hold the highest numbers. Common bean, onion, tomato, potato and apple are the species of main interest in the covered countries. As from collected data, about 19.8% of landrace cultivation sites are in protected areas of the Natura 2000 network. We also got evidence that 16.7% and 19.3% of conservation varieties of agricultural species and vegetables are currently cultivated, respectively. Results of the GIS analysis allowed the identification of 1261 cells (25 km × 25 km) including all the cultivation sites, distributed across all European biogeographical regions. Data of this study constitute the largest ever produced database of in situ-maintained landraces and the first 2 attempt to create an inventory for the entire Europe. The availability of such resource will serve for better planning of actions and development of policies to protect landraces and foster their use.
Tree mortality during global‐change‐type drought is usually attributed to xylem dysfunction, but as climate change increases the frequency of extreme heat events, it is necessary to better understand the interactive role of heat stress. We hypothesized that some drought‐stressed plants paradoxically open stomata in heatwaves to prevent leaves from critically overheating. We experimentally imposed heat (>40°C) and drought stress onto 20 broadleaf evergreen tree/shrub species in a glasshouse study. Most well‐watered plants avoided lethal overheating, but drought exacerbated thermal damage during heatwaves. Thermal safety margins (TSM) quantifying the difference between leaf surface temperature and leaf critical temperature, where photosynthesis is disrupted, identified species vulnerability to heatwaves. Several mechanisms contributed to high heat tolerance and avoidance of damaging leaf temperatures—small leaf size, low leaf osmotic potential, high leaf mass per area (i.e., thick, dense leaves), high transpirational capacity, and access to water. Water‐stressed plants had smaller TSM, greater crown dieback, and a fundamentally different stomatal heatwave response relative to well‐watered plants. On average, well‐watered plants closed stomata and decreased stomatal conductance (gs) during the heatwave, but droughted plants did not. Plant species with low gs, either due to isohydric stomatal behavior under water deficit or inherently low transpirational capacity, opened stomata and increased gs under high temperatures. The current paradigm maintains that stomata close before hydraulic thresholds are surpassed, but our results suggest that isohydric species may dramatically increase gs (over sixfold increases) even past their leaf turgor loss point. By actively increasing water loss at high temperatures, plants can be driven toward mortality thresholds more rapidly than has been previously recognized. The inclusion of TSM and responses to heat stress could improve our ability to predict the vulnerability of different tree species to future droughts.
Multiplex genome-editing (MGE) technologies are recently developed versatile bioengineering tools for modifying two or more specific DNA loci in a genome with high precision. These genome-editing tools have greatly increased the feasibility of introducing desired changes at multiple nucleotide levels into a target genome. In particular, clustered regularly interspaced short palindromic repeats (CRISPR)/CRISPR-associated protein (Cas) [CRISPR/Cas] system-based MGE tools allow the simultaneous generation of direct mutations precisely at multiple loci in a gene or multiple genes. MGE is enhancing the field of plant molecular biology and providing capabilities for revolutionizing modern crop-breeding methods as it was virtually impossible to edit genomes so precisely at the single base-pair level with prior genome-editing tools, such as zinc-finger nucleases (ZFNs) and transcription activator-like effector nucleases (TALENs). Recently, researchers have not only started using MGE tools to advance genome-editing applications in certain plant science fields but also have attempted to decipher and answer basic questions related to plant biology. In this review, we discuss the current progress that has been made toward the development and utilization of MGE tools with an emphasis on the improvements in plant biology after the discovery of CRISPR/Cas9. Furthermore, the most recent advancements involving CRISPR/Cas applications for editing multiple loci or genes are described. Finally, insights into the strengths and importance of MGE technology in advancing crop-improvement programs are presented.
Rice (Oryza sativa L.) is an important food crop species in China. Cultivating high-yielding rice varieties that have a high photosynthetic efficiency is an important goal of rice breeding in China. In recent years, due to the continual innovation of molecular breeding methods, many excellent genes have been applied in rice breeding, which is highly important for increasing rice yields. In this paper, the hexokinase gene OsHXK1 was knocked out via the CRISPR/Cas9 gene-editing method in the indica rice varieties Huanghuazhan, Meixiangzhan, and Wushansimiao, and OsHXK1-CRISPR/Cas9 lines were obtained. According to the results of a phenotypic analysis and agronomic trait statistics, the OsHXK1-CRISPR/Cas9 plants presented increased light saturation points, stomatal conductance, light tolerance, photosynthetic products, and rice yields. Moreover, transcriptome analysis showed that the expression of photosynthesis-related genes significantly increased. Taken together, our results revealed that knocking out OsHXK1 via the CRISPR/Cas9 gene-editing method could effectively lead to the cultivation of high-photosynthetic efficiency and high-yielding rice varieties. They also revealed the important roles of OsHXK1 in the regulation of rice yield and photosynthesis.
The molecular responses of plants to the important abiotic stress, heat stress (HS), have been extensively studied at the transcriptional level. Alternative splicing (AS) is a post-transcriptional regulatory process in which an intron-containing gene can generate more than one mRNA variant. The impact of HS on the pre-mRNA splicing process has been reported in various eukaryotes but seldom discussed in-depth, especially in plants. Here, we review AS regulation in response to HS in different plant species. We discuss potential molecular mechanisms controlling heat-inducible AS regulation in plants and hypothesize that AS regulation participates in heat-priming establishment and HS memory maintenance. We propose that the pre-mRNA splicing variation is an important regulator of plant HS responses (HSRs).
Introduction
Phytic acid (PA) is an important antinutrient agent present in cereal grains which reduces the bioavailability of iron and zinc in human body, causing malnutrition. Inositol pentakisphosphate 2- kinase 1 (IPK1) gene has been reported to be an important gene for PA biosynthesis.
Objective
A recent genome editing tool CRISPR/Cas9 has been successfully applied to develop biofortified rice by disrupting IPK1 gene, however, it remained a challenge in wheat. The aim of this study was to biofortify wheat using CRISPR/Cas9.
Methodology
In this study, we isolated 3 TaIPK1 homeologs in wheat designated as TaIPK1.A, TaIPK1.B and TaIPK1.D and found that the expression abundance of TaIPK1.A was stronger in early stages of grain filling. Using CRISPR/Cas9, we have disrupted TaIPK1.A gene in cv. Borlaug-2016 with two guide RNAs targeting the 1st and 2nd exons.
Results
We got several genome-edited lines in the T0 generation at frequencies of 12.7% and 10.8%. Sequencing analysis revealed deletion of 1-23 nucleotides and even an addition of 1 nucleotide in various lines. Analysis of the genome-edited lines revealed a significant decrease in the PA content and an increase in iron and zinc accumulation in grains compared with control plants.
Conclusion
Our study demonstrates the potential application of CRISPR/Cas9 technique for the rapid generation of biofortified wheat cultivars.
Wheat (Triticum aestivum L.) is a staple food crop consumed by more than 30% of world population. Nitrogen (N) fertilizer has been applied broadly in agriculture practice to improve wheat yield to meet the growing demands for food production. However, undue N fertilizer application and the low N use efficiency (NUE) of modern wheat varieties are aggravating environmental pollution and ecological deterioration. Under nitrogen‐limiting conditions, the rice (Oryza sativa) abnormal cytokinin response1 repressor1 (are1) mutant exhibits increased NUE, delayed senescence and consequently, increased grain yield. However, the function of ARE1 ortholog in wheat remains unknown. Here, we isolated and characterized three TaARE1 homoeologs from the elite Chinese winter wheat cultivar ZhengMai 7698. We then used CRISPR/Cas9‐mediated targeted mutagenesis to generate a series of transgene‐free mutant lines either with partial or triple‐null taare1 alleles. All transgene‐free mutant lines showed enhanced tolerance to N starvation, and showed delayed senescence and increased grain yield in field conditions. In particular, the AABBdd and aabbDD mutant lines exhibited delayed senescence and significantly increased grain yield without growth defects compared to the wild‐type control. Together, our results underscore the potential to manipulate ARE1 orthologs through gene editing for breeding of high‐yield wheat as well as other cereal crops with improved NUE.
Wheat (Triticum aestivum, 2n = 6x = 42, AABBDD) is one of the most important staple food crops in the world. Despite that wheat production has significantly increased over the past decades, future wheat production will face unprecedented challenges following global climate changes, increasing world population, and water shortages in arid and semi-arid lands. Furthermore, undue applications of diverse fertilizers and pesticides are exacerbating environmental pollution and ecological deterioration. To ensure global food and ecosystem security, it is essential to enhance resilience of wheat production while minimizing environmental pollution through using cutting-edge technologies. However, the hexaploid genome and gene redundancy complicate foreword genetic research and precision gene modifications in wheat improvement, thus impeding the breeding of elite wheat cultivars. In this review, we first introduce the state-of-art genome editing technologies in crop plants, especially in wheat, for both functional genomics and genetic improvement. We then outline applications of other technologies, such as GWAS, high throughput genotyping and phenotyping, speed breeding, and synthetic biology in wheat, respectively. Lastly, we discuss the existing challenges in wheat genome editing and propose future prospective of precision gene modifications using advanced genome editing technologies. We conclude that the combination of genome editing and other molecular breeding strategies will greatly facilitate wheat genetic improvement for sustainable global production.
Plants are attacked by a large diversity of pathogens. These pathogens can affect plant growth and fitness directly but also indirectly by inducing changes in the host plant that affect interactions with beneficial and antagonistic insects. Yet, we lack insights into the relative importance of direct and indirect effects of pathogens on their host plants, and how these effects differ among pathogen species.
In this study, we examined four fungal pathogens on the wood anemone Anemone nemorosa. We used field observations to record the impacts of each pathogen species on plant growth and fitness throughout the season, and experimental hand pollination and insect feeding trials to assess whether fitness impacts were mediated by pathogen‐induced changes in plant–pollinator and plant–herbivore interactions.
Three out of four pathogens negatively affected plant size, and pathogens differed strongly in their effect on plant architecture. Infected plants had lower fitness, but this effect was not mediated by pollinators or herbivores. Even so, two out of four pathogens reduced herbivory on anemones in the field, and we found negative effects of pathogen infection on herbivore preference and performance in feeding trials.
Synthesis. Our results are of broader significance in two main respects. First, we demonstrated that pathogens negatively affected plant growth and fitness, and that the magnitude of these effects varied among pathogen species, suggesting that pathogens constitute important selective agents that differ in strength. Second, direct effects on plant fitness were more important than effects mediated by beneficial and antagonistic insects. In addition, although we did not detect insect‐mediated effects on plant fitness, the negative effects of some pathogens on herbivore preference and performance indicate that pathogen communities influence the distribution and abundance of herbivores.
Polyploidy occurs prevalently and plays an important role during plant speciation and evolution. This phenomenon suggests polyploidy could develop novel features that enable them to adapt wider range of environmental conditions compared with diploid progenitors. Bread wheat (Triticum aestivum L., BBAADD) is a typical allohexaploid species and generally exhibits greater salt tolerance than its tetraploid wheat progenitor (BBAA). However, little is known about the underlying molecular basis and the regulatory pathway of this trait. Here, we show that the histone acetyltransferase TaHAG1 acts as a crucial regulator to strengthen salt tolerance of hexaploid wheat. Salinity-induced TaHAG1 expression was associated with tolerance variation in polyploidy wheat. Overexpression, silencing and CRISPR-mediated knockout of TaHAG1 validated the role of TaHAG1 in salinity tolerance of wheat. TaHAG1 contributed to salt tolerance by modulating ROS production and signal specificity. Moreover, TaHAG1 directly targeted a subset of genes that are responsible for hydrogen peroxide production, and enrichment of TaHAG1 triggered increased H3 acetylation and transcriptional upregulation of these loci under salt stress. In addition, we found the salinity-induced TaHAG1-mediated ROS production pathway is involved in salt tolerance difference of wheat accessions with varying ploidy. Our findings provide insight into the molecular mechanism of how an epigenetic regulatory factor facilitates adaptability of polyploidy wheat and highlights this epigenetic modulator as a strategy for salt tolerance breeding in bread wheat.
Ensuring the sustainability of agriculture under climate change has led to a surge in alternative strategies for crop improvement. Advances in integrated crop breeding, social acceptance, and farm-level adoption are crucial to address future challenges to food security. Societal acceptance can be slow when consumers do not see the need for innovation or immediate benefits. We consider how best to address the issue of social licence and harmonised governance for novel gene technologies in plant breeding. In addition, we highlight optimised breeding strategies that will enable long-term genetic gains to be achieved. Promoted by harmonised global policy change, innovative plant breeding can realise high and sustainable productivity together with enhanced nutritional traits.
Common wheat (Triticum aestivum L.) is one of the three major food crops in the world; thus, wheat breeding programs are important for world food security. Characterizing the genes that control important agronomic traits and finding new ways to alter them are necessary to improve wheat breeding. Functional genomics and breeding in polyploid wheat has been greatly accelerated by the advent of several powerful tools, especially CRISPR/Cas9 genome editing technology, which allows multiplex genome engineering. Here, we describe the development of CRISPR/Cas9, which has revolutionized the field of genome editing. In addition, we emphasize technological breakthroughs (e.g., base editing and prime editing) based on CRISPR/Cas9. We also summarize recent applications and advances in the functional annotation and breeding of wheat, and we introduce the production of CRISPR-edited DNA-free wheat. Combined with other achievements, CRISPR and CRISPR-based genome editing will speed progress in wheat biology and promote sustainable agriculture.
Environmental problems have always received immense attention from scientists. Toxicants pollution is a critical environmental concern that has posed serious threats to human health and agricultural production. Heavy metals and pesticides are top of the list of environmental toxicants endangering nature. This review focuses on the toxic effect of heavy metals (cadmium (Cd), lead (Pb), copper (Cu), and zinc (Zn)) and pesticides (insecticides, herbicides, and fungicides) adversely influencing the agricultural ecosystem (plant and soil) and human health. Furthermore, heavy metals accumulation and pesticide residues in soils and plants have been discussed in detail. In addition, the characteristics of contaminated soil and plant physiological parameters have been reviewed. Moreover, human diseases caused by exposure to heavy metals and pesticides were also reported. The bioaccumulation, mechanism of action, and transmission pathways of both heavy metals and pesticides are emphasized. In addition, the bioavailability in soil and plant uptake of these contaminants has also been considered. Meanwhile, the synergistic and antagonistic interactions between heavy metals and pesticides and their combined toxic effects have been discussed. Previous relevant studies are included to cover all aspects of this review. The information in this review provides deep insights into the understanding of environmental toxicants and their hazardous effects.
The changing climate scenarios harshen the biotic stresses including boosting up the population of insect/pest and disease, uplifting weed growth, declining soil beneficial microbes, threaten pollinator, and boosting up abiotic stresses including harsh drought/waterlogging, extremisms in temperature, salinity/alkalinity, abrupt rainfall pattern)) and ulitamtely affect the plant in multiple ways. This nexus review paper will cover four significant points viz (1) the possible impacts of climate change; as the world already facing the problem of food security, in such crucial period, climatic change severely affects all four dimensions of food security (from production to consumption) and will lead to malnutrition/malnourishment faced by low-income peoples. (2) How some major crops (wheat, cotton, rice, maize, and sugarcane) are affected by stress and their consequent loss. (3) How to develop a strategic work to limit crucial factors, like their significant role in climate-smart breeding, developing resilience to stresses, and idiotypic breeding. Additionally, there is an essence of improving food security, as much of our food is wasted before consumption for instance post-harvest losses. (4) Role of biotechnology and genetic engineering in adaptive introgression of the gene or developing plant transgenic against pests. As millions of dollars are invested in innovation and research to cope with future climate change stresses on a plant, hence community base adaptation of innovation is also considered an important factor in crop improvements. Because of such crucial predictions about the future impacts of climate change on agriculture, we must adopt measures to evolve crop.
Background
Low-temperature severely affects the growth and development of chrysanthemum which is one kind of ornamental plant well-known and widely used in the world. Lysine crotonylation is a recently identified post-translational modification (PTM) with multiple cellular functions. However, lysine crotonylation under low-temperature stress has not been studied.
Results
Proteome-wide and lysine crotonylation of chrysanthemum at low-temperature was analyzed using TMT (Tandem Mass Tag) labeling, sensitive immuno-precipitation, and high-resolution LC-MS/MS. The results showed that 2017 crotonylation sites were identified in 1199 proteins. Treatment at 4 °C for 24 h and − 4 °C for 4 h resulted in 393 upregulated proteins and 500 downregulated proteins (1.2-fold threshold and P < 0.05). Analysis of biological information showed that lysine crotonylation was involved in photosynthesis, ribosomes, and antioxidant systems. The crotonylated proteins and motifs in chrysanthemum were compared with other plants to obtain orthologous proteins and conserved motifs. To further understand how lysine crotonylation at K136 affected APX (ascorbate peroxidase), we performed a site-directed mutation at K136 in APX. Site-directed crotonylation showed that lysine decrotonylation at K136 reduced APX activity, and lysine complete crotonylation at K136 increased APX activity.
Conclusion
In summary, our study comparatively analyzed proteome-wide and crotonylation in chrysanthemum under low-temperature stress and provided insights into the mechanisms of crotonylation in positively regulated APX activity to reduce the oxidative damage caused by low-temperature stress. These data provided an important basis for studying crotonylation to regulate antioxidant enzyme activity in response to low-temperature stress and a new research ideas for chilling-tolerance and freezing-tolerance chrysanthemum molecular breeding.
Most common food grains contain gluten proteins and can cause adverse medical conditions generally known as gluten-related disorders. Celiac disease is an immune-mediated enteropathy triggered by gluten in individuals carrying a specific genetic make-up. The presence of the human leukocyte antigens (HLA)-DQ2 and HLA-DQ8 haplotypes together with gluten intake is a necessary, although not sufficient, condition, to develop celiac disease. Fine mapping of the human genome has revealed numerous genetic variants important in the development of this disease. Most of the genetic variants are small nucleotide polymorphisms located within promoters and transcriptional enhancer sequences. Their importance is underlined by an increased risk in DQ2/DQ8 carriers who also have these non-HLA alleles. In addition, several immune-mediated diseases share susceptibility loci with celiac disease, shedding light on the reasons for co-occurrence between these diseases. Finally, most of the genes potentially involved in celiac disease by fine genetic mapping of non-HLA loci were confirmed in gene expression studies. In contrast to celiac disease, very little is known about the genetic make-up of non-celiac wheat sensitivity (NCWS), a clinically defined pathology that shares symptoms and gluten dependence with the celiac disease. We recently identified differentially expressed genes and miRNAs in the intestinal mucosa of these patients. Remarkably, the differentially expressed genes were long non-coding RNAs possibly involved in the regulation of cell functions. Thus, we can speculate that important aspects of these diseases depend on alteration of regulatory genetic circuits. Furthermore, our finding suggests that innate immune response is involved in the pathogenic mechanism of NCWS. This review is intended to convey the idea that in order to fully understand celiac disease and its relationship with other gluten-related disorders, it is worth learning more about non-HLA variants.
Anthropogenic activities in the past and present eras have created global warming and consequently a storm of drought stress, affecting both plants and animals. Being sessile, plants are more vulnerable to drought stress and consequently reduce plant growth and yield. To mitigate the effects of drought stress on plants, it is very crucial to determine the plant response mechanisms against drought stress. Drought response mechanism includes morph-physiological, biochemical, cellular and molecular processes takes place in plants underlying drought stress. These processes include improvement in root system, leaf structure, osmotic adjustment, relative water content and stomata regulation. In addition, calcium and phytohormone (Abscisic acid, Jasmonic acid, Salicylic acid, Auxins, Gibberellins, Ethylene etc.) signaling pathways and scavenging of reactive oxygen species are the key mechanisms to cope with drought stress. Moreover, microorganisms such as bacteria and fungi also have an important role in drought tolerance enhancement. To further elucidate and improve drought tolerance in plants, quantitative trait loci, transgenic approach and application of exogenous substances (nitric oxide, 24-epibrassinoide, glycine betaine and proline) are very crucial. Hereby, the present study integrates various mechanisms of drought tolerance in plants.
In the past, there have been drought events in different parts of the world, which have negatively influenced the productivity and production of various crops including wheat (Triticum aestivum L.), one of the world’s three important cereal crops. Breeding new high yielding drought-tolerant wheat varieties is a research priority specifically in regions where climate change is predicted to result in more drought conditions. Commonly in breeding for drought tolerance, grain yield is the basis for selection, but it is a complex, late-stage trait, affected by many factors aside from drought. A strategy that evaluates genotypes for physiological responses to drought at earlier growth stages may be more targeted to drought and time efficient. Such an approach may be enabled by recent advances in high-throughput phenotyping platforms (HTPPs). In addition, the success of new genomic and molecular approaches rely on the quality of phenotypic data which is utilized to dissect the genetics of complex traits such as drought tolerance. Therefore, the first objective of this review is to describe the growth-stage based physio-morphological traits that could be targeted by breeders to develop drought-tolerant wheat genotypes. The second objective is to describe recent advances in high throughput phenotyping of drought tolerance related physio-morphological traits primarily under field conditions. We discuss how these strategies can be integrated into a comprehensive breeding program to mitigate the impacts of climate change. The review concludes that there is a need for comprehensive high throughput phenotyping of physio-morphological traits that is growth stage-based to improve the efficiency of breeding drought-tolerant wheat.
The rusts of wheat, caused by three species of Puccinia, are very devastating diseases and are major biotic constraints in efforts to sustain wheat production worldwide. Their capacity to spread aerially over long distances, rapid production of infectious uredospores, and abilities to evolve new pathotypes, makes the management of wheat pathogens a very challenging task. The development and deployment of resistant wheat varieties has proven to be the most economic, effective and efficient means of managing rust diseases. Rust resistance used in wheat improvement has included sources from the primary gene pool as well as from species distantly related to wheat. The 1BL/1RS translocation from cereal rye was used widely in wheat breeding, and for some time provided resistance to the wheat leaf rust, stripe rust, and stem rust pathogens conferred by genes Lr26, Yr9, and Sr31, respectively. However, the emergence of virulence for all three genes, and stripe rust resistance gene Yr27, has posed major threats to the cultivation of wheat globally. To overcome this threat, efforts are going on worldwide to monitor rust diseases, identify rust pathotypes, and to evaluate wheat germplasm for rust resistance. Anticipatory breeding and the responsible deployment of rust resistant cultivars have proven to be effective strategies to manage wheat rusts. Efforts are still however being made to decipher the recurrence of wheat rusts, their epidemiologies, and new genomic approaches are being used to break the yield barriers and manage biotic stresses such as the rusts. Efficient monitoring of pathotypes of Puccinia species on wheat, identification of resistance sources, pre-emptive breeding, and strategic deployment of rust resistant wheat cultivars have been the key factors to effective management of wheat rusts in India. The success in containing wheat rusts in India can be gauged by the fact that we had no wheat rust epiphytotic for nearly last five decades. This publication provides a comprehensive overview of the wheat rust research conducted in India.
CRISPR/Cas9 gene-editing technologies have been very effective in editing target genes in all major crop plants and offer unprecedented potentials in crop improvement. A major challenge in using CRISPR gene-editing technology for agricultural applications is that the target gene-edited crop plants need to be transgene free to maintain trait stability and to gain regulatory approval for commercial production. In this article, we present various strategies for generating transgene-free and target gene-edited crop plants. The CRISPR transgenes can be removed by genetic segregation if the crop plants are reproduced sexually. Marker-assisted tracking and eliminating transgenes greatly decrease the time and labor needed for identifying the ideal transgene-free plants. Transgenes can be programed to undergo self-elimination when CRISPR genes and suicide genes are sequentially activated, greatly accelerating the isolation of transgene-free and target gene-edited plants. Transgene-free plants can also be generated using approaches that are considered non-transgenic such as ribonucleoprotein transfection, transient expression of transgenes without DNA integration, and nano-biotechnology. Here, we discuss the advantages and disadvantages of the various strategies in generating transgene-free plants and provide guidance for adopting the best strategies in editing a crop plant.
Bacterial blight of rice is an important disease in Asia and Africa. The pathogen, Xanthomonas oryzae pv. oryzae (Xoo), secretes one or more of six known transcription-activator-like effectors (TALes) that bind specific promoter sequences and induce, at minimum, one of the three host sucrose transporter genes SWEET11, SWEET13 and SWEET14, the expression of which is required for disease susceptibility. We used CRISPR–Cas9-mediated genome editing to introduce mutations in all three SWEET gene promoters. Editing was further informed by sequence analyses of TALe genes in 63 Xoo strains, which revealed multiple TALe variants for SWEET13 alleles. Mutations were also created in SWEET14, which is also targeted by two TALes from an African Xoo lineage. A total of five promoter mutations were simultaneously introduced into the rice line Kitaake and the elite mega varieties IR64 and Ciherang-Sub1. Paddy trials showed that genome-edited SWEET promoters endow rice lines with robust, broad-spectrum resistance.
Most genetic variants that contribute to disease¹ are challenging to correct efficiently and without excess byproducts2, 3, 4–5. Here we describe prime editing, a versatile and precise genome editing method that directly writes new genetic information into a specified DNA site using a catalytically impaired Cas9 endonuclease fused to an engineered reverse transcriptase, programmed with a prime editing guide RNA (pegRNA) that both specifies the target site and encodes the desired edit. We performed more than 175 edits in human cells, including targeted insertions, deletions, and all 12 types of point mutation, without requiring double-strand breaks or donor DNA templates. We used prime editing in human cells to correct, efficiently and with few byproducts, the primary genetic causes of sickle cell disease (requiring a transversion in HBB) and Tay–Sachs disease (requiring a deletion in HEXA); to install a protective transversion in PRNP; and to insert various tags and epitopes precisely into target loci. Four human cell lines and primary post-mitotic mouse cortical neurons support prime editing with varying efficiencies. Prime editing shows higher or similar efficiency and fewer byproducts than homology-directed repair, has complementary strengths and weaknesses compared to base editing, and induces much lower off-target editing than Cas9 nuclease at known Cas9 off-target sites. Prime editing substantially expands the scope and capabilities of genome editing, and in principle could correct up to 89% of known genetic variants associated with human diseases.
Drought is considered as one of the major limiting factors affecting growth and productivity of crop plants. It severely affects the morphological and physiological activities of the plants and hampers the seed germination, root proliferation, biomass accumulation and final yield of field crops. Drought stress disrupts the biosynthesis of chlorophyll contents, carotene and decreases photosynthesis in plants. It gradually reduces CO2 assimilation rates owing to decrease in stomatal conductance. In addition, drought affects cell membrane stability and disrupts water relations of a plant by reducing water use efficiency. To cope with these situations, plants adopt different mechanisms such as drought tolerance, avoidance and escape. In this review, we discussed about the effects of drought on morphological and physiological characteristics of plants and suggested the different agronomic practices to overcome the deleterious effects of drought stress.
The application of clustered regularly interspaced short palindromic repeats (CRISPR) for genetic manipulation has revolutionized life science over the past few years. CRISPR was first discovered as an adaptive immune system in bacteria and archaea, and then engineered to generate targeted DNA breaks in living cells and organisms. During the cellular DNA repair process, various DNA changes can be introduced. The diverse and expanding CRISPR toolbox allows programmable genome editing, epigenome editing and transcriptome regulation in plants. However, challenges in plant genome editing need to be fully appreciated and solutions explored. This Review intends to provide an informative summary of the latest developments and breakthroughs of CRISPR technology, with a focus on achievements and potential utility in plant biology. Ultimately, CRISPR will not only facilitate basic research, but also accelerate plant breeding and germplasm development. The application of CRISPR to improve germplasm is particularly important in the context of global climate change as well as in the face of current agricultural, environmental and ecological challenges.
Early sowing has been extensively used in high-latitude areas to avoid drought stress during sowing; however, cold damage has become the key limiting factor of early sowing. To relieve cold stress, plants develop a series of physiological and biochemical changes and sophisticated molecular regulatory mechanisms. The biomembrane is the barrier that protects cells from injury as well as the primary place for sensing cold signals. Chilling tolerance is closely related to the composition, structure, and metabolic process of membrane lipids. This review focuses on membrane lipid metabolism and its molecular mechanism, as well as lipid signal transduction in peanut (Arachis hypogaea L.) under cold stress to build a foundation for explicating lipid metabolism regulation patterns and physiological and molecular response mechanisms during cold stress and to promote the genetic improvement of peanut cold tolerance.
Grain size and weight are important components of a suite of yield‐related traits in crops. Here, we showed that the CRISPR‐Cas9 gene editing of TaGW7, a homolog of rice OsGW7 encoding a TONNEAU1‐recruiting motif (TRM) protein, affects grain shape and weight in allohexaploid wheat. By editing the TaGW7 homoeologs in the B and D genomes, we showed that mutations in either of the two or both genomes increased the grain width and weight but reduced the grain length. The effect sizes of mutations in the TaGW7 gene homoeologs coincided with the relative levels of their expression in the B and D genomes. The effects of gene editing on grain morphology and weight traits were dosage dependent with the double‐copy mutant showing larger effect than the respective single copy mutants. The TaGW7‐centered gene co‐expression network indicated that this gene is involved in the pathways regulating cell division and organ growth, also confirmed by the cellular co‐localization of TaGW7 with α‐ and β‐tubulin proteins, the building blocks of microtubule arrays. The analyses of exome capture data in tetraploid domesticated and wild emmer, and hexaploid wheat revealed the loss of diversity around TaGW7‐associated with domestication selection, suggesting that TaGW7 is likely to play an important role in the evolution of yield component traits in wheat. Our study showed how integrating CRISPR‐Cas9 system with cross‐species comparison can help to uncover the function of a gene fixed in wheat for allelic variants targeted by domestication selection and select targets for engineering new gene variants for crop improvement.
It has long been recognized that stomatal movement modulates CO2 availability and as a consequence the photosynthetic rate of plants, and that this process is feedback-regulated by photoassimilates. However, the genetic components and mechanisms underlying this regulatory loop remain poorly understood, especially in monocot crop species. Here, we report the cloning and functional characterization of a maize (Zea mays) mutant named closed stomata1 (cst1). Map-based cloning of cst1 followed by confirmation with the clustered regularly interspaced short palindromic repeats (CRISPR)/ CRISPR associated protein 9 system identified the causal mutation in a Clade I Sugars Will Eventually be Exported Transporters (SWEET) family gene, which leads to the E81K mutation in the CST1 protein. CST1 encodes a functional glucose transporter expressed in subsidiary cells, and the E81K mutation strongly impairs the oligomerization and glucose transporter activity of CST1. Mutation of CST1 results in reduced stomatal opening, carbon starvation, and early senescence in leaves, suggesting that CST1 functions as a positive regulator of stomatal opening. Moreover, CST1 expression is induced by carbon starvation and suppressed by photoassimilate accumulation. Our study thus defines CST1 as a missing link in the feedback-regulation of stomatal movement and photosynthesis by photoassimilates in maize.
CRISPR technology has vividly increased its applications in last fve years for genome editing in a wide range of organisms
from bacteria to plants. It is mostly applied in the feld of mammalian research. This emerging versatile tool can be utilized in
crop improvement by targeting various traits to increase economic value and adaptability of the crop species under changing
climate. In plants, Arabidopsis and rice are the most studied plant species in genome editing through CRISPR technology.
Wheat is lagging behind in the utilization of CRISPR based genome modifcations. The hexaploid, large genome size and the
recalcitrant nature in terms of tissue culture are the major obstacles for CRISPR utilization in wheat. Recently, the IWGSC
released the high quality of reference genome for wheat which will greatly accelerate the application of CRISPR-based
genome engineering in wheat and helps to resolve the global issue of food security in coming decades. The exogenous DNAfree improved mutants with CRISPR technology having desired traits will increase the productivity under biotic and abiotic
stress conditions. To address complex traits involving multigene, recently developed multiplex genome editing toolkits can
be used. This is a frst review of its kind in which the practical utilization and updates on CRISPR validation in wheat along
with its future prospects for use of this technology in wheat improvement are comprehensively discussed. Thus, the compiled
information will immensely beneft the researchers for utilization of CRISPR system in wheat improvement across the globe.
Arsenate (AsV) is one of the most common forms of arsenic (As) in environment and plant high-affinity phosphate transporters (PHT1s) are the primary plant AsV transporters. However, few PHT1s involved in AsV absorption have been identified in crops. In our previous study, TaPHT1;3, TaPHT1;6 and TaPHT1;9 were identified to function in phosphate absorption. Here, their AsV absorption capacities were evaluated using several experiments. Ectopic expression in yeast mutants indicated that TaPHT1;9 had the highest AsV absorption rates, followed by TaPHT1;6, while not for TaPHT1;3. Under AsV stress, further, BSMV-VIGS-mediated TaPHT1;9-silencing wheat plants exhibited higher AsV tolerance and lower As concentrations than TaPHT1;6-silenced plants, whereas TaPHT1;3-silencing plants had similar phenotype and AsV concentrations to control. These suggested that TaPHT1;9 and TaPHT1;6 possessed AsV absorption capacity with the former showing higher activities. Under hydroponic condition, furthermore, CRISPR-edited TaPHT1;9 wheat mutants showed the enhanced tolerance to AsV with decreased As distributions and concentrations, whereas TaPHT1;9 ectopic expression transgenic rice plants had the opposite results. Also, under AsV-contaminated soil condition, TaPHT1;9 transgenic rice plants exhibited depressed AsV tolerance with increased As concentrations in roots, straws and grains. Moreover, Pi addition alleviated the AsV toxicity. These suggested that TaPHT1;9 should be a candidate target gene for AsV phytoremediation.
The advent of clustered regularly interspaced short palindromic repeat (CRISPR) genome editing, coupled with advances in computing and imaging capabilities, has initiated a new era in which genetic diseases and individual disease susceptibilities are both predictable and actionable. Likewise, genes responsible for plant traits can be identified and altered quickly, transforming the pace of agricultural research and plant breeding. In this Review, we discuss the current state of CRISPR-mediated genetic manipulation in human cells, animals, and plants along with relevant successes and challenges and present a roadmap for the future of this technology.
In the context of climate change, the magnitude and frequency of temperature extremes (low and high temperatures) are increasing worldwide. Changes to the lower extremes of temperature, known as cold stress (CS), are one of the recurrent stressors in many parts of the world, severely limiting agricultural production. A series of plant reactions to CS could be generalized into morphological, physiological, and biochemical responses based on commonalities among crop plants. However, the differing originality of crops revealed varying degrees of sensitivity to cold and, therefore, exhibited large differences in these responses among the crops. This review discusses the vegetative and reproductive growth effects of CS and highlights the species-specific aspect of each growth stage whereby the reproductive growth CS appears more detrimental in rice and wheat, with marginal yield losses. To mitigate CS negative effects, crop plants have evolved cold-acclimation mechanisms (with differing capability), characterized by specific protein accumulation, membrane modification, regulation of signaling pathways, osmotic regulation, and induction of endogenous hormones. In addition, we reviewed a comprehensive account of management strategies for regulating tolerance mechanisms of crop plants under CS.
Cuticular waxes cover the aerial surfaces of land plants and protect them from various environmental stresses. Alkanes are major wax components and contribute to plant drought tolerance, but the biosynthesis and regulation of alkanes remain largely unknown in wheat (Triticum aestivum L.). Here, we identified and functionally characterized a key alkane biosynthesis gene ECERIFERUM1-6A (TaCER1-6A) from wheat. The CRISPR/Cas9-mediated knockout mutation in TaCER1-6A greatly reduced the contents of C27, C29, C31, and C33 alkanes in wheat leaves, while TaCER1-6A overexpression significantly increased the contents of these alkanes in wheat leaves, suggesting that TaCER1-6A is specifically involved in the biosynthesis of C27, C29, C31, and C33 alkanes on wheat leaf surfaces. TaCER1-6A knockout lines exhibited increased cuticle permeability and reduced drought tolerance, whereas TaCER1-6A overexpression lines displayed reduced cuticle permeability and enhanced drought tolerance. TaCER1-6A was highly expressed in flag leaf blades and seedling leaf blades and could respond to abiotic stresses and abscisic acid (ABA). TaCER1-6A was located in the endoplasmic reticulum (ER), which is the subcellular compartment responsible for wax biosynthesis. A total of three haplotypes (HapI/II/III) of TaCER1-6A were identified in 43 wheat accessions, and HapI was the dominant haplotype (95%) in these wheat varieties. Additionally, we identified two R2R3-MYB transcription factors TaMYB96-2D and TaMYB96-5D that bound directly to the conserved motif CAACCA in promoters of the cuticular wax biosynthesis genes TaCER1-6A, TaCER1-1A, and fatty acyl-CoA reductase4 (TaFAR4). Collectively, these results suggest that TaCER1-6A is required for C27, C29, C31, and C33 alkanes biosynthesis and improves drought tolerance in wheat.
As the current crisis unfolds, international attention is focused on the many supply-related effects that compound existing failings and inequalities of the global food system, which were amplified by the COVID-19 pandemic. We may again see civil unrest as witnessed during
the 2008 food crisis. The war in Ukraine and trade sanctions are triggering a level of volatility that could easily overwhelm
existing mitigation mechanisms. The top priority must be to mitigate near-term food security crises through increased production and demand-side interventions supported by appropriate policy incentives (such as price guarantees and subsidized agricultural inputs). Demand-side interventions that conserve grain stocks for human consumption and shift to lower-cost flour blends can also improve food insecurity in the short term. Beyond this, we see opportunities for expanding
wheat production into new locations that have comparative advantages, for optimizing seed systems and other requirements of local wheat production and supply, and for establishing wheat-production
monitoring capacities. In parallel, a transition to agri-food system resilience will require balancing food supply needs with imperatives for climate change mitigation and adaptation, gender equity, nutritional
sufficiency and livelihood security.
The emergence of CRISPR-Cas systems has accelerated the development of gene editing technologies, which are widely used in the life sciences. To improve the performance of these systems, workers have engineered and developed a variety of CRISPR-Cas tools with a broader range of targets, higher efficiency and specificity, and greater precision. Moreover, CRISPR-Cas-related technologies have also been expanded beyond making cuts in DNA by introducing functional elements that permit precise gene modification, control gene expression, make epigenetic changes, and so on. In this review, we introduce and summarize the characteristics and applications of different types of CRISPR-Cas tools. We discuss certain limitations of current approaches and future prospects for optimizing CRISPR-Cas systems.
As a critical second messenger in plants, Ca²⁺ is involved in numerous biological processes including biotic and abiotic stress responses. The CBL-interacting protein kinases, known as CIPKs, are essential components in Ca²⁺-mediated signal transduction pathways. Here, we found that CIPK14 plays a role in the process of regulating immune response in Arabidopsis. The CIPK14 loss-of-function mutants exhibited enhanced resistance to the P. syringae, whereas CIPK14 overexpression plants were more susceptible to bacterial pathogen. Enhanced resistance in cipk14 mutants were accompanied by increased accumulation of SA and elevated expression of defense marker genes (PR1, EDS1, EDS5, ICS1). Overexpression of CIPK14 suppressed Pst DC3000, Pst DC3000 hrcC and flg22 induced generation of ROS and callose deposition. As compared with wild type plants, the expression levels of MPK3/6-dependent PTI marker genes (FRK1, CYP81F2, WAK2, FOX) were up-regulated in cipk14 mutants but down-regulated in CIPK14 overexpression plants after flg22 and elf18 treatment. Additionally, both loss-of-function and gain-of-function of CIPK14 significantly altered the phosphorylation status of MPK3/6 under flg22 treatment, suggesting that CIPK14 is a general modulator of plant immunity at both transcriptional and post-transcriptional level. Taken together, our results uncover that CIPK14 acts as a negative regulator in plant immune response.
CRISPR technology, which is widely used for plant genome editing, will accelerate the breeding of food crops beyond what was imaginable before its development. Here we provide a brief overview of CRISPR technology, its most important applications for crop improvement and several technological breakthroughs. We also make predictions of the applications of CRISPR technology to food crops, which we believe would provide the potential for synthetic biology and domestication of crops. We also discuss the implications of regulatory policy for deployment of the technology in the developing world.
Eukaryotic cells deploy overlapping repair pathways to resolve DNA damage. Advancements in genome editing take advantage of these pathways to produce permanent genetic changes. Despite recent improvements, genome editing can produce diverse outcomes that can introduce risks in clinical applications. Although homology-directed repair is attractive for its ability to encode precise edits, it is particularly difficult in human cells. Here we discuss the DNA repair pathways that underlie genome editing and strategies to favour various outcomes. Harnessing DNA repair pathways in genome editing In this Review, Yeh, Richardson and Corn discuss the DNA repair pathways that underlie genome editing and recent improvements and strategies to yield desired genomic alterations in cells and organisms.
The current trajectory for crop yields is insufficient to nourish the world’s population by 2050¹. Greater and more consistent crop production must be achieved against a backdrop of climatic stress that limits yields, owing to shifts in pests and pathogens, precipitation, heat-waves and other weather extremes. Here we consider the potential of plant sciences to address post-Green Revolution challenges in agriculture and explore emerging strategies for enhancing sustainable crop production and resilience in a changing climate. Accelerated crop improvement must leverage naturally evolved traits and transformative engineering driven by mechanistic understanding, to yield the resilient production systems that are needed to ensure future harvests.
Multiplex genome editing involves the simultaneous targeting of multiple related or unrelated targets. The latter is most straightforward using the CRISPR/Cas9 system because multiple gRNAs can be delivered either as independent expression cassettes with their own promoters or as polycistronic transcripts processed into mature gRNAs by endogenous or introduced nucleases. Multiplex genome editing in plants initially focused on input traits such as herbicide resistance, but has recently expanded to include hormone biosynthesis and perception, metabolic engineering, plant development and molecular farming, with more than 100 simultaneous targeting events reported. Usually the coding region is targeted but recent examples also include promoter modifications to generate mutants with varying levels of gene expression.