Can nanotechnology improve the application of bioherbicides?

Bioherbicides are composed of microorganisms or natural compounds and are used for weed control; however, they have specific weaknesses and constraints that hinder their development and success under field conditions. Nanotechnology can help to overcome these limitations by providing a good starting point for the design of specific formulations and carriers that minimize the deficiencies of natural compounds and microorganisms, such as low solubility, short shelf life or a loss of viability. In addition, nanoformulations can help to improve the efficacy of bioherbicides by increasing their effectiveness and bioavailability, reducing the amount required for a treatment, and enhancing their ability to target specific weeds while preserving the crop. However, it is important to choose the right materials and nanodevices depending on specific needs and considering several factors inherent to nanomaterials such as production cost, safety or possible toxic effects. © 2023 Society of Chemical Industry.

Characterization of durum wheat resistance against leaf rust under climate change conditions of increasing temperature and [CO2].

Durum wheat cultivation in Mediterranean regions is threatened by abiotic factors, mainly related to the effects of climate change, and biotic factors such as the leaf rust disease. This situation requires an in-depth knowledge of how predicted elevated temperatures and [CO2] will affect durum wheat-leaf rust interactions. Therefore, we have characterised the response of one susceptible and two resistant durum wheat accessions against leaf rust under different environments in greenhouse assays, simulating the predicted conditions of elevated temperature and [CO2] in the far future period of 2070-2099 for the wheat growing region of Cordoba, Spain. Interestingly, high temperature alone or in combination with high [CO2] did not alter the external appearance of the rust lesions. However, through macro and microscopic evaluation, we found some host physiological and molecular responses to infection that would quantitatively reduce not only pustule formation and subsequent infection cycles of this pathogen, but also the host photosynthetic area under these predicted weather conditions, mainly expressed in the susceptible accession. Moreover, our results suggest that durum wheat responses to infection are mainly driven by temperature, being considered the most hampering abiotic stress. In contrast, leaf rust infection was greatly reduced when these weather conditions were also conducted during the inoculation process, resembling the effects of possible heat waves not only in disease development, but also in fungal germination and penetration success. Considering this lack of knowledge in plant-pathogen interactions combined with abiotic stresses, the present study is, to the best of our knowledge, the first to include the effects of the expected diurnal variation of maximum temperature and continuous elevated [CO2] in the durum wheat-leaf rust pathosystem.

Nanobiosensors and nanoformulations in agriculture: new advances and challenges for sustainable agriculture.

In the current scenario of climate change, global agricultural systems are facing remarkable challenges in order to increase production, while reducing the negative environmental impact. Nano-enabled technologies have the potential to revolutionise farming practices by increasing the efficiency of inputs and minimising losses, as well as contributing to sustainable agriculture. Two promising applications of nanotechnology in agriculture are nanobiosensors and nanoformulations (NFs). Nanobiosensors can help detect biotic and abiotic stresses in plants before they affect plant production, while NFs can make agrochemicals, more efficient and less polluting. NFs are becoming new-age materials with a wide variety of nanoparticle-based formulations such as fertilisers, herbicides, insecticides, and fungicides. They facilitate the site-targeted controlled delivery of agrochemicals enhancing their efficiency and reducing dosages. Smart farming aims to monitor and detect parameters related to plant health and environmental conditions in order to help sustainable agriculture. Nanobiosensors can provide real-time analytical data, including detection of nutrient levels, metabolites, pesticides, presence of pathogens, soil moisture, and temperature, aiding in precision farming practices, and optimising resource usage. In this review, we summarise recent innovative uses of NFs and nanobiosensors in agriculture that may boost crop protection and production, as well as reducing the negative environmental impact of agricultural activities. However, successful implementation of these smart technologies would require two special considerations: (i) educating farmers about appropriate use of nanotechnology, (ii) conducting field trials to ensure effectiveness under real conditions.

Reducing Nitrogen Dosage in Triticum durum Plants with Urea-Doped Nanofertilizers.

Nanotechnology is emerging as a very promising tool towards more efficient and sustainable practices in agriculture. In this work, we propose the use of non-toxic calcium phosphate nanoparticles doped with urea (U-ACP) for the fertilization of Triticum durum plants. U-ACP nanoparticles present very similar morphology, structure, and composition than the amorphous precursor of bone mineral, but contain a considerable amount of nitrogen as adsorbed urea (up to ca. 6 wt % urea). Tests on Triticum durum plants indicated that yields and quality of the crops treated with the nanoparticles at reduced nitrogen dosages (by 40%) were unaltered in comparison to positive control plants, which were given the minimum N dosages to obtain the highest values of yield and quality in fields. In addition, optical microscopy inspections showed that Alizarin Red S stained nanoparticles were able to penetrate through the epidermis of the roots or the stomata of the leaves. We observed that the uptake through the roots occurs much faster than through the leaves (1 h vs. 2 days, respectively). Our results highlight the potential of engineering nanoparticles to provide a considerable efficiency of nitrogen uptake by durum wheat and open the door to design more sustainable practices for the fertilization of wheat in fields.

Engineering Biomimetic Calcium Phosphate Nanoparticles: A Green Synthesis of Slow-Release Multinutrient (NPK) Nanofertilizers.

Biomimetic calcium phosphate nanoparticles (CaP) have been actively used in biomedicine, due to their high biodegradability and biocompatibility. However, much less progress has been made regarding their application in precision agriculture, i.e., for the controlled delivery of active species to plants. Herein, we report a straightforward and green synthetic method to dope CaP with potassium (K) and nitrogen (N, as nitrate and urea). By modulating the synthetic conditions in terms of maturation time (at 37 °C) and doping, we prepared K- and N-doped nanoparticles in the form of either amorphous calcium phosphate (ACP) or nanocrystalline apatite (Ap) and studied the impact of the dopants on the ACP-to-Ap transformation pathways. Importantly, we found out that ACP, isolated at low maturation times, incorporates nitrogen (in the form of nitrate and urea) to a larger extent than Ap (2.6 vs 1.1 wt %, respectively). Multinutrient nanofertilizers (so-called nanoU-NPK) with the following composition (wt %) were obtained: Ca (23.3), P (10.1), K (1.5), NO3 (2.9), and urea (4.8). The nanoU-NPK provides a slow and gradual release of the most important plant macronutrients (NPK), with N in two chemical forms, and different kinetics. The concentration of nutrients supplied by 10 g L-1 of nanoU-NPK to the media after 1 week (in mg L-1) was Ca (27.0), P (6.2), K (41.0), NO3 (134.0), and urea (315.0). Preliminary tests on durum wheat have shown that the application of nanoU-NPK allows reducing the amount of nitrogen supplied to the plants by 40% with respect to a conventional treatment, without affecting the final kernel weight per plant. The application of these slow-release NPK nanofertilizers is a promising strategy toward enhancing the efficiency of the fertilization, complying with the concept of precision agriculture.

Assessment of functional and structural changes of soil fungal and oomycete communities in holm oak declined dehesas through metabarcoding analysis.

Forest decline is nowadays a major challenge for ecosystem sustainability. Dehesas, which consists of savannah-like mediterranean ecosystems, are threatened by the holm oak decline in the south-west of Iberian Peninsula. Phytophthora cinnamomi is considered the main agent of holm oak root rot, but little is known about the relationship between diversity of soilborne microbial community and the decline syndrome of holm oak. It would be hypothesized that the changes in the structure and functionality of the soil microbiome might influence tree health status through changes in richness and diversity of beneficial organisms such as mycorrhizal species, or fungal plant pathogens such as Fusarium spp. or Alternaria spp. Total DNA of soil samples from declined oak dehesas was extracted and analyzed through metabarcoding techniques, to evaluate the specific composition and diversity of the fungal and oomycete communities and their relationship with the disease symptoms. The fungal community included a wide range of pathogens and abundance of ectomycorrhizal key taxa related with low defoliation degree. Phytophthora cinnamomi and Pythium spiculum did not appear among the most abundant oomycetes, nor were they related directly to defoliation levels. Moreover, a particular taxon belonging to the genus Trichoderma was strongly correlated with the scarcity of pathogenic Phytophthora spp. The diversity and composition of fungal and oomycete communities were related to the severity of the decline symptoms. The metabarcoding study of microbiome represents a powerful tool to develop biocontrol strategies for the management of the holm oak root rot.

Safe nanotechnologies for increasing the effectiveness of environmentally friendly natural agrochemicals.

Natural compounds and living organisms continue to play a limited role in crop protection, and few of them have reached the market, despite their attractiveness and the efforts made in research. Very often these products have negative characteristics compared to synthetic compounds, e.g., higher costs of production, lower effectiveness, lack of persistence, and inability to reach and penetrate the target plant. Conversely, nanotechnologies are having an enormous impact on all human activities, including agriculture, even if the production of some nanomaterials is not environmentally friendly or could have adverse effects on agriculture and the environment. Thus, certain nanomaterials could facilitate the development of formulated natural pesticides, making them more effective and more environmentally friendly. Nanoformulations can improve efficacy, reduce effective doses, and increase shelf-life and persistence. Such controlled-release products can improve delivery to the target pest. This review considers certain available nanomaterials and nanotechnologies for use in agriculture, discussing their properties and the feasibility of their use in sustainable crop protection, in particular, in improving the effectiveness of natural bio-based agrochemicals. © 2019 Society of Chemical Industry.

Physical and Chemical Barriers in Root Tissues Contribute to Quantitative Resistance to Fusarium oxysporum f. sp. pisi in Pea.

Fusarium wilt caused by Fusarium oxysporum f. sp. pisi (Fop) is one of the most destructive diseases of pea worldwide. Control of this disease is difficult and it is mainly based on the use of resistant cultivars. While monogenic resistance has been successfully used in the field, it is at risk of breakdown by the constant evolution of the pathogen. New sources of quantitative resistance have been recently identified from a wild relative Pisum spp. collection. Here, we characterize histologically the resistance mechanisms occurring in these sources of quantitative resistance. Detailed comparison, of the reaction at cellular level, of eight pea accessions with differential responses to Fop race 2, showed that resistant accessions established several barriers at the epidermis, exodermis, cortex, endodermis and vascular stele efficiently impeding fungal progression. The main components of these different barriers were carbohydrates and phenolic compounds including lignin. We found that these barriers were mainly based on three defense mechanisms including cell wall strengthening, formation of papilla-like structures at penetration sites and accumulation of different substances within and between cells. These defense reactions varied in intensity and localization between resistant accessions. Our results also clarify some steps of the infection process of F. oxysporum in plant and support the important role of cell wall-degrading enzymes in F. oxysporum pathogenicity.

The interactive effects of arbuscular mycorrhiza and plant growth-promoting rhizobacteria synergistically enhance host plant defences against pathogens.

Belowground interactions between plant roots, mycorrhizal fungi and plant growth-promoting rhizobacteria (PGPR) can improve plant health via enhanced nutrient acquisition and priming of the plant immune system. Two wheat cultivars differing in their ability to form mycorrhiza were (co)inoculated with the mycorrhizal fungus Rhizophagus irregularis and the rhizobacterial strain Pseudomonas putida KT2440. The cultivar with high mycorrhizal compatibility supported higher levels of rhizobacterial colonization than the low compatibility cultivar. Those levels were augmented by mycorrhizal infection. Conversely, rhizobacterial colonization of the low compatibility cultivar was reduced by mycorrhizal arbuscule formation. Single inoculations with R. irregularis or P. putida had differential growth effects on both cultivars. Furthermore, while both cultivars developed systemic priming of chitosan-induced callose after single inoculations with R. irregularis or P. putida, only the cultivar with high mycorrhizal compatibility showed a synergistic increase in callose responsiveness following co-inoculation with both microbes. Our results show that multilateral interactions between roots, mycorrhizal fungi and PGPR can have synergistic effects on growth and systemic priming of wheat.

Differences in Crenate Broomrape Parasitism Dynamics on Three Legume Crops Using a Thermal Time Model.

Root parasitic weeds are a major limiting production factor in a number of crops, and control is difficult. Genetic resistance and chemical control lead the fight, but without unequivocal success. Models that help to describe and even predict the evolution of parasitism underground are a valuable tool for herbicide applications, and even could help in breeding programs. Legumes are heavily affected by Orobanche crenata (crenate broomrape) in the Mediterranean basin. This work presents a descriptive model based on thermal time and correlating growing degree days (GDD) with the different developmental stages of the parasite. The model was developed in three different legume crops (faba bean, grass pea and lentil) attacked by crenate broomrape. The developmental stages of the parasite strongly correlated with the GDD and differences were found depending on the host crop.

Quantum dot and superparamagnetic nanoparticle interaction with pathogenic fungi: internalization and toxicity profile.

For several years now, nanoscaled materials have been implemented in biotechnological applications related to animal (in particular human) cells and related pathologies. However, the use of nanomaterials in plant biology is far less widespread, although their application in this field could lead to the future development of plant biotechnology applications. For any practical use, it is crucial to elucidate the relationship between the nanomaterials and the target cells. In this work we have evaluated the behavior of two types of nanomaterials, quantum dots and superparamagnetic nanoparticles, on Fusarium oxysporum, a fungal species that infects an enormous range of crops causing important economic losses and is also an opportunistic human pathogen. Our results indicated that both nanomaterials rapidly interacted with the fungal hypha labeling the presence of the pathogenic fungus, although they showed differential behavior with respect to internalization. Thus, whereas magnetic nanoparticles appeared to be on the cell surface, quantum dots were significantly taken up by the fungal hyphae showing their potential for the development of novel control approaches of F. oxysporum and related pathogenic fungi following appropriate functionalization. In addition, the fungal germination and growth, accumulation of ROS, indicative of cell stress, and fungal viability have been evaluated at different nanomaterial concentrations showing the low toxicity of both types of nanomaterials to the fungus. This work represents the first study on the behavior of quantum dots and superparamagnetic particles on fungal cells, and constitutes the first and essential step to address the feasibility of new nanotechnology-based systems for early detection and eventual control of pathogenic fungi.

A method to quantify infection and colonization of holm oak (Quercus ilex) roots by Phytophthora cinnamomi.

Phytophthora cinnamomi Rands. is an important root rot pathogen widely distributed in the north hemisphere, with a large host range. Among others diseases, it is known to be a principal factor in the decline of holm oak and cork oak, the most important tree species in the "dehesa" ecosystem of south-western Spain. Previously, the focus of studies on P. cinnamomi and holm oak have been on molecular tools for identification, functional responses of the host, together with other physiological and morphological host variables. However, a microscopic index to describe the degree of infection and colonization in the plant tissues has not yet been developed. A colonization or infection index would be a useful tool for studies that examine differences between individuals subjected to different treatments or to individuals belonging to different breeding accessions, together with their specific responses to the pathogen. This work presents a methodology based on the capture and digital treatment of microscopic images, using simple and accessible software, together with a range of variables that quantify the infection and colonization process.

Synthesis, application, and tracking of magnetic carbon-coated nanoparticles in plants.

The behavior of nanoparticles inside plants is gaining importance for its implications in research about putative applications and toxicology. Magnetic carbon-coated nanoparticles can be easily traced through plant tissues using simple and affordable histological techniques. Here we present a methodology for the synthesis of such nanoparticles. We also describe methods for growing plants using rhizotrons (in order to observe the roots), procedures for applying the nanoparticles, taking and processing the samples.

Effect of amphotericin B nanodisks on plant fungal diseases.

BACKGROUND: The development of water-soluble nanodevices extends the potential use of compounds developed for other purposes (e.g. antifungal drugs or antibiotics) for applications in agriculture. For example, the broad-spectrum, water-insoluble, macrolide polyene antibiotic amphotericin B (AMB) could be used to inhibit phytopathogenic fungi. A new formulation embedding AMB in nanodisks (NDs) enhances antibiotic solubility and confers protection against environmental damage. In the present study, AMB-NDs were tested for efficacy against several phytopathogenic fungi in vitro and on infected living plants (chickpea and wheat). RESULTS: Compared with AMB in dimethylsulfoxide (DMSO), AMB-NDs increased the sensitivity of several fungal species to this antimycotic in vitro. Sensitivity varied with fungal species as well as with the forma specialis. Phytophthora cinnamomi, previously reported as insensitive to other polyene antimycotics, remained unaffected at the doses examined. Some effect against disease symptoms were obtained with AMB-NDs against fusarium wilt in chickpea, whereas the results were highly variable in wheat, depending on both the species and treatment regimen. CONCLUSION: The results confirm that formulation of AMB into ND increases its effectiveness against phytopathogenic fungi in vitro, opening the possibility for its use on infected plants in the field.

Absorption and translocation to the aerial part of magnetic carbon-coated nanoparticles through the root of different crop plants.

The development of nanodevices for agriculture and plant research will allow several new applications, ranging from treatments with agrochemicals to delivery of nucleic acids for genetic transformation. But a long way for research is still in front of us until such nanodevices could be widely used. Their behaviour inside the plants is not yet well known and the putative toxic effects for both, the plants directly exposed and/or the animals and humans, if the nanodevices reach the food chain, remain uncertain. In this work we show that magnetic carbon-coated nanoparticles forming a biocompatible magnetic fluid (bioferrofluid) can easily penetrate through the root in four different crop plants (pea, sunflower, tomato and wheat). They reach the vascular cylinder, move using the transpiration stream in the xylem vessels and spread through the aerial part of the plants in less than 24 hours. Accumulation of nanoparticles was detected in wheat leaf trichomes, suggesting a way for excretion/detoxification. This kind of studies is of great interest in order to unveil the movement and accumulation of nanoparticles in plant tissues for assessing further applications in the field or laboratory.

New chemical clues for broomrape-sunflower host-parasite interactions: synthesis of guaianestrigolactones.

A comparative structure-activity relationship (SAR) study has been conducted with several guaianolide sesquiterpene lactones (SLs) as inducers of the germination of sunflower broomrape (Orobanche cumana) seeds. Compounds were selected and synthesized to study the influence of the lactone-enol-gamma-lactone moiety on the selectivity of SLs toward the stimulation of sunflower broomrape germination. The results clearly illustrate that SLs are recognized only by O. cumana, while the introduction of a strigol-like second lactone moiety in the guaianolide backbone results in the loss of specificity and hence the germination of other broomrape species. We have named this new class of compounds guaianestrigolactones (GELs).

Nanoparticle penetration and transport in living pumpkin plants: in situ subcellular identification.

BACKGROUND: In recent years, the application of nanotechnology in several fields of bioscience and biomedicine has been studied. The use of nanoparticles for the targeted delivery of substances has been given special attention and is of particular interest in the treatment of plant diseases. In this work both the penetration and the movement of iron-carbon nanoparticles in plant cells have been analyzed in living plants of Cucurbita pepo. RESULTS: The nanoparticles were applied in planta using two different application methods, injection and spraying, and magnets were used to retain the particles in movement in specific areas of the plant. The main experimental approach, using correlative light and electron microscopy provided evidence of intracellular localization of nanoparticles and their displacement from the application point. Long range movement of the particles through the plant body was also detected, particles having been found near the magnets used to immobilize and concentrate them. Furthermore, cell response to the nanoparticle presence was detected. CONCLUSION: Nanoparticles were capable of penetrating living plant tissues and migrating to different regions of the plant, although movements over short distances seemed to be favoured. These findings show that the use of carbon coated magnetic particles for directed delivery of substances into plant cells is a feasible application.

Nanotechnology for parasitic plant control.

The field of nanotechnology opens up novel potential applications for agriculture. Nanotechnology applications are already being explored and used in medicine and pharmacology, but interest for use in crop protection is just starting. The development of nanodevices as smart delivery systems to target specific sites and nanocarriers for controlled chemical release is discussed. Some nanotechnologies can improve existing crop management techniques in the short to medium term. Nanocapsules would help to avoid phytotoxicity on the crop by using systemic herbicides against parasitic weeds. Nanoencapsulation can also improve herbicide application, providing better penetration through cuticles and tissues, and allowing slow and constant release of the active substances. On the other hand, new crop management tools could be developed on the basis of medical applications. Nanoparticles have a great potential as 'magic bullets', loaded with herbicides, chemicals or nucleic acids, and targeting specific plant tissues or areas to release their charge. Viral capsids can be altered by mutagenesis to achieve different configurations and deliver specific nucleic acids, enzymes or antimicrobial peptides acting against the parasites. Many issues are still to be addressed, such as increasing the scale of production processes and lowering costs, as well as toxicological issues, but the foundations of a new plant treatment concept have been laid, and applications in the field of parasitic plant control can be started.

Breeding approaches for crenate broomrape (Orobanche crenata Forsk.) management in pea (Pisum sativum L.).

BACKGROUND: Pea cultivation is strongly hampered in Mediterranean and Middle East farming systems by the occurrence of Orobanche crenata Forsk. Strategies of control have been developed, but only marginal successes have been achieved. Most control methods are either unfeasible, uneconomical, hard to achieve or result in incomplete protection. The integration of several control measures is the most desirable strategy. RESULTS: [corrected] Recent developments in control are presented and re-evaluated in light of recent developments in crop breeding and molecular genetics. These developments are placed within a framework that is compatible with current agronomic practices. CONCLUSION: The current focus in applied breeding is leveraging biotechnological tools to develop more and better markers to speed up the delivery of improved cultivars to the farmer. To date, however, progress in marker development and delivery of useful markers has been slow. The application of knowledge gained from basic genomic research and genetic engineering will contribute to more rapid pea improvement for resistance against O. crenata and/or the herbicide.