Potato Berries as a Valuable Source of Compounds Potentially Applicable in Crop Protection and Pharmaceutical Sectors: A Review.

Potato (Solanum tuberosum) is a major agricultural crop cultivated worldwide. To meet market demand, breeding programs focus on enhancing important agricultural traits such as disease resistance and improvement of tuber palatability. However, while potato tubers get a lot of attention from research, potato berries are mostly overlooked due to their level of toxicity and lack of usefulness for the food production sector. Generally, they remain unused in the production fields after harvesting the tuber. These berries are toxic due to high levels of glycoalkaloids, which might confer some interesting bioactivities. Berries of various solanaceous species contain bioactive secondary metabolites, suggesting that potato berries might contain similarly valuable metabolites. Therefore, possible applications of potato berries, e.g., in the protection of plants against pests and pathogens, as well as the medical exploitation of their anti-inflammatory, anticarcinogenic, and antifungal properties, are plausible. The presence of valuable compounds in potato berries could also contribute to the bioeconomy by providing a novel use for otherwise discarded agricultural side streams. Here we review the potential use of these berries for the extraction of compounds that can be exploited to produce pharmaceuticals and plant protection products.

Exogenous Application of dsRNA in Plant Protection: Efficiency, Safety Concerns and Risk Assessment.

The use of double-stranded RNA (dsRNA) for plant protection shows great potential as a sustainable alternative to traditional pesticides. This review summarizes the current state of knowledge on using exogenous dsRNA in plant protection and includes the latest findings on the safety and efficiency of this strategy. The review also emphasizes the need for a cautious and comprehensive approach, considering safety considerations such as off-target effects and formulation challenges. The regulatory landscape in different regions is also discussed, underscoring the need for specific guidelines tailored to dsRNA-based pesticides. The review provides a crucial resource for researchers, regulators, and industry stakeholders, promoting a balanced approach incorporating innovation with thorough safety assessments. The continuous dialog emphasized in this review is essential for shaping the future of dsRNA-based plant protection. As the field advances, collaboration among scientists, regulators, and industry partners will play a vital role in establishing guidelines and ensuring the responsible, effective, and sustainable use of dsRNA in agriculture.

Smart and Sustainable Crop Protection: Design and Evaluation of a Novel α-Amylase-Responsive Nanopesticide for Effective Pest Control.

In this study, an α-amylase-responsive controlled-release formulation was developed by capping polydopamine onto ß-cyclodextrin-modified abamectin-loaded hollow mesoporous silica nanoparticles. The prepared Aba@HMS@CD@PDA were subjected to characterization using various analytical techniques. The findings revealed that Aba@HMS@CD@PDA, featuring a loading rate of 18.8 wt %, displayed noteworthy release behavior of abamectin in the presence of α-amylase. In comparison to abamectin EC, Aba@HMS@CD@PDA displayed a significantly foliar affinity and improved rainfastness on lotus leaves. The results of field trail demonstrated a significantly higher control efficacy against Spodoptera litura Fabricius compared to abamectin EC at all concentrations after 7, 14, and 21 days of spaying, showcasing the remarkable persistence of Aba@HMS@CD@PDA. These results underscore the potential of Aba@HMS@CD@PDA as a novel and persistently effective strategy for sustainable on-demand crop protection. The application of nanopesticides can enhance the effectiveness and efficiency of pesticide utilization, contributing to more sustainable agricultural practices.

Biophoton emission-based approach of the effects of systemic insecticides on the survival of Eurydema ventralis Kolenati, 1846 (Hemiptera: Pentatomidae) and on the photosynthetic activity of oilseed rape.

The choice of effective crop protection technologies is a key factors in the economical production of oilseed rape. Insecticides belonging to the group of active substances butenolides and diamides are active substances available as seed treatments in oilseed rape and promising control tools in the crop protection technologies. Our laboratory experiment demonstrated that the experimental insecticides flupyradifurone and cyantraniliprole are both effective against Eurydema ventralis (Hemiptera: Pentatomidae) when used as a seed and in-crop treatments, but there is a fundamental difference in their insect mortality inducing effects. Flupyradifurone was found to have a total mortality 96 h after application based on basipetal translocation. In the case of cyantraniliprole, the insecticidal effect of the same treatment was 27% less. The experiment showed that the acropetal translocation of the tested active substances after seed treatment did not induce efficacy comparable to that of the basipetal translocation. The study of the biophoton emission of the plants demonstrated a verifiable correlation between the different application methods of the insecticides and the photon emission intensity per unit plant surface area. In conclusion, the systematic insecticides tested, in addition to having the expected insecticidal effect, interfere with plant life processes by enhancing photosynthetic activity.

Entomopathogenic fungi in crops protection with an emphasis on bioactive metabolites and biological activities.

Plant pathogens with their abundance are harmful and cause huge damage to different agricultural crops and economy of a country as well as lead towards the shortage of food for humans. For their management, the utilization of entomopathogenic fungi is an eco-friendly technique, sustainable to the environment, safe for humans and has promising effect over chemical-based pesticides. This process requires a biochemical mechanism, including the production of enzymes, toxins, and other metabolites that facilitate host infection and invasion. Essential enzymes such as chitinase, proteinase, and lipase play a direct role in breaking down the host cuticle, the primary barrier to EPF (Entomopathogenic Fungi) infection. Additionally, secondary metabolites such as destruxins in Metarhizium, beauvericin in Beauveria, hirsutellides in Hirsutella, isarolides in Isaria, cordyols in Cordyceps, and vertihemipterins in Verticillium, among others, act both directly and indirectly to disable the defense mechanisms of insect hosts, thereby accelerating the EPF infection process. The chemical composition of these secondary metabolites varies, ranging from simple non-peptide pigments such as oosporine to highly complex piperazine derivatives such as vertihemiptellides. The biocontrol efficacy of EPF is extensively studied, with numerous fungal strains commercially available on a large scale for managing arthropod pests. This review emphasizes the role of proteins and enzymes against crop pathogens, detailing their mode of action, and describing the metabolites from entomopathogenic fungi and their biological activities. In doing so, these findings contribute to establishing a symbiotic equilibrium between agricultural productivity and environmental conservation.

New biologically active ionic liquids with benzethonium cation-efficient SAR inducers and antimicrobial agents.

BACKGROUND: An urgent need to find new methods for crop protection remains open due to the withdrawal from the market of the most toxic pesticides and increasing consumer awareness. One of the alternatives that can be used in modern agriculture is the use of bifunctional compounds whose actions towards plant protection are wider than those of conventional pesticides. RESULTS: In this study, we present the investigation of the biological efficacy of nine dual-functional salts containing a systemic acquired resistance (SAR)-inducing anion and the benzethonium cation. A significant result of the presented study is the discovery of the SAR induction activity of benzethonium chloride, which was previously reported only as an antimicrobial agent. Moreover, the concept of dual functionality was proven, as the application of presented compounds in a given concentrations resulted both in the control of human and plant bacteria species and induction of SAR. CONCLUSION: The strategy presented in this article shows the capabilities of derivatization of common biologically active compounds into their ionic derivatives to obtain bifunctional salts. This approach may be an example of the design of potential new compounds for modern agriculture. It provides plants with two complementary actions allowing to provide efficient protection to plants, if one mode of action is ineffective. © 2024 Society of Chemical Industry.

Improving RNA-based crop protection through nanotechnology and insights from cross-kingdom RNA trafficking.

Spray-induced gene silencing (SIGS) is a powerful and eco-friendly method for crop protection. Based off the discovery of RNA uptake ability in many fungal pathogens, the application of exogenous RNAs targeting pathogen/pest genes results in gene silencing and infection inhibition. However, SIGS remains hindered by the rapid degradation of RNA in the environment. As extracellular vesicles are used by plants, animals, and microbes in nature to transport RNAs for cross-kingdom/species RNA interference between hosts and microbes/pests, nanovesicles and other nanoparticles have been used to prevent RNA degradation. Efforts examining the effect of nanoparticles on RNA stability and internalization have identified key attributes that can inform better nanocarrier designs for SIGS. Understanding sRNA biogenesis, cross-kingdom/species RNAi, and how plants and pathogens/pests naturally interact are paramount for the design of SIGS strategies. Here, we focus on nanotechnology advancements for the engineering of innovative RNA-based disease control strategies against eukaryotic pathogens and pests.

Artificial nanovesicles for dsRNA delivery in spray-induced gene silencing for crop protection.

Spray-induced gene silencing (SIGS) is an innovative and eco-friendly technology where topical application of pathogen gene-targeting RNAs to plant material can enable disease control. SIGS applications remain limited because of the instability of RNA, which can be rapidly degraded when exposed to various environmental conditions. Inspired by the natural mechanism of cross-kingdom RNAi through extracellular vesicle trafficking, we describe herein the use of artificial nanovesicles (AVs) for RNA encapsulation and control against the fungal pathogen, Botrytis cinerea. AVs were synthesized using three different cationic lipid formulations, DOTAP + PEG, DOTAP and DODMA, and examined for their ability to protect and deliver double stranded RNA (dsRNA). All three formulations enabled dsRNA delivery and uptake by B. cinerea. Further, encapsulating dsRNA in AVs provided strong protection from nuclease degradation and from removal by leaf washing. This improved stability led to prolonged RNAi-mediated protection against B. cinerea both on pre- and post-harvest plant material using AVs. Specifically, the AVs extended the protection duration conferred by dsRNA to 10 days on tomato and grape fruits and to 21 days on grape leaves. The results of this work demonstrate how AVs can be used as a new nanocarrier to overcome RNA instability in SIGS for crop protection.

Pest-removal services provided by birds on subsistence farms in south-eastern Nigeria.

To what extent birds provide the ecosystem service of pest control in subsistence farms, and how this service might depend on retained natural habitats near farmlands is unexplored in West Africa. To fill this knowledge gap, we placed plasticine mimics of insect pests on experimentally grown crops on the Mambilla Plateau, South Eastern Nigeria. We recorded bird attacks on the mimics and the proportion of mimics removed by birds. We also determined the influence of distance of crops from forest fragments on both attack and removal rates. We placed 90 potted plants of groundnut (Arachis hypogea) and bambara nut (Vigna subterranea) along 15 transects running 4.5 km from forest edge into open grassland. Each plant had six of the 540 mimics in total placed on their leaves. We inspected the potted plants weekly for 12 weeks to record (i) the presence of bird beak marks on mimics, and (ii) the number of missing mimics. Once a week we collected all the mimics from the plants and counted the number of assumed beak marks. After counting we replaced the mimics on the plants, mark free. We found a strong positive correlation between the abundance of insectivorous birds and the mean number of missing mimics and/or bird attack marks on mimics. However, this positive effect of insectivorous bird abundance on prey mimic attack/removal became less strong the farther they were from a forest fragment. We found increased predation rates and abundance of insectivorous birds closer to forest fragments. Our data suggest that pest predation may be a key ecosystem service provided by insectivorous birds on Nigerian farmlands. Farmlands that are closer to forest fragments may experience a higher rate of pest control by insectivorous birds than those further away, suggesting that retaining forest fragments in the landscape may enhance pest control services in sub-Saharan subsistence farms.

Tuning Beforehand: A Foresight on RNA Interference (RNAi) and In Vitro-Derived dsRNAs to Enhance Crop Resilience to Biotic and Abiotic Stresses.

Crop yield is severely affected by biotic and abiotic stresses. Plants adapt to these stresses mainly through gene expression reprogramming at the transcriptional and post-transcriptional levels. Recently, the exogenous application of double-stranded RNAs (dsRNAs) and RNA interference (RNAi) technology has emerged as a sustainable and publicly acceptable alternative to genetic transformation, hence, small RNAs (micro-RNAs and small interfering RNAs) have an important role in combating biotic and abiotic stresses in plants. RNAi limits the transcript level by either suppressing transcription (transcriptional gene silencing) or activating sequence-specific RNA degradation (post-transcriptional gene silencing). Using RNAi tools and their respective targets in abiotic stress responses in many crops is well documented. Many miRNAs families are reported in plant tolerance response or adaptation to drought, salinity, and temperature stresses. In biotic stress, the spray-induced gene silencing (SIGS) provides an intelligent method of using dsRNA as a trigger to silence target genes in pests and pathogens without producing side effects such as those caused by chemical pesticides. In this review, we focus on the potential of SIGS as the most recent application of RNAi in agriculture and point out the trends, challenges, and risks of production technologies. Additionally, we provide insights into the potential applications of exogenous RNAi against biotic stresses. We also review the current status of RNAi/miRNA tools and their respective targets on abiotic stress and the most common responsive miRNA families triggered by stress conditions in different crop species.

Isolation and Characterization of Barley (Hordeum vulgare) Extracellular Vesicles to Assess Their Role in RNA Spray-Based Crop Protection.

The demonstration that spray-induced gene silencing (SIGS) can confer strong disease resistance, bypassing the laborious and time-consuming transgenic expression of double-stranded (ds)RNA to induce the gene silencing of pathogenic targets, was ground-breaking. However, future field applications will require fundamental mechanistic knowledge of dsRNA uptake, processing, and transfer. There is increasing evidence that extracellular vesicles (EVs) mediate the transfer of transgene-derived small interfering (si)RNAs in host-induced gene silencing (HIGS) applications. In this study, we establish a protocol for barley EV isolation and assess the possibilities for EVs regarding the translocation of sprayed dsRNA from barley (Hordeum vulgare) to its interacting fungal pathogens. We found barley EVs that were 156 nm in size, containing predominantly 21 and 19 nucleotide (nts) siRNAs, starting with a 5'-terminal Adenine. Although a direct comparison of the RNA cargo between HIGS and SIGS EV isolates is improper given their underlying mechanistic differences, we identified sequence-identical siRNAs in both systems. Overall, the number of siRNAs isolated from the EVs of dsRNA-sprayed barley plants with sequence complementarity to the sprayed dsRNA precursor was low. However, whether these few siRNAs are sufficient to induce the SIGS of pathogenic target genes requires further research. Taken together, our results raise the possibility that EVs may not be mandatory for the spray-delivered siRNA uptake and induction of SIGS.

Eco-Friendly Nanoplatforms for Crop Quality Control, Protection, and Nutrition.

Agricultural chemicals have been widely utilized to manage pests, weeds, and plant pathogens for maximizing crop yields. However, the excessive use of these organic substances to compensate their instability in the environment has caused severe environmental consequences, threatened human health, and consumed enormous economic costs. In order to improve the utilization efficiency of these agricultural chemicals, one strategy that attracted researchers is to design novel eco-friendly nanoplatforms. To date, numerous advanced nanoplatforms with functional components have been applied in the agricultural field, such as silica-based materials for pesticides delivery, metal/metal oxide nanoparticles for pesticides/mycotoxins detection, and carbon nanoparticles for fertilizers delivery. In this review, the synthesis, applications, and mechanisms of recent eco-friendly nanoplatforms in the agricultural field, including pesticides and mycotoxins on-site detection, phytopathogen inactivation, pest control, and crops growth regulation for guaranteeing food security, enhancing the utilization efficiency of agricultural chemicals and increasing crop yields are highlighted. The review also stimulates new thinking for improving the existing agricultural technologies, protecting crops from biotic and abiotic stress, alleviating the global food crisis, and ensuring food security. In addition, the challenges to overcome the constrained applications of functional nanoplatforms in the agricultural field are also discussed.

Insights into the multitrophic interactions between the biocontrol agent Bacillus subtilis MBI 600, the pathogen Botrytis cinerea and their plant host.

Botrytis cinerea is a plant pathogen causing the gray mold disease in a plethora of host plants. The control of the disease is based mostly on chemical pesticides, which are responsible for environmental pollution, while they also pose risks for human health. Furthermore, B. cinerea resistant isolates have been identified against many fungicide groups, making the control of this disease challenging. The application of biocontrol agents can be a possible solution, but requires deep understanding of the molecular mechanisms in order to be effective. In this study, we investigated the multitrophic interactions between the biocontrol agent Bacillus subtilis MBI 600, a new commercialized biopesticide, the pathogen B. cinerea and their plant host. Our analysis showed that this biocontrol agent reduced B. cinerea mycelial growth in vitro, and was able to suppress the disease incidence on cucumber plants. Moreover, treatment with B. subtilis led to induction of genes involved in plant immunity. RNA-seq analysis of B. cinerea transcriptome upon exposure to bacterial secretome, showed that genes coding for MFS and ABC transporters were highly induced. Deletion of the Bcmfs1 MFS transporter gene, using a CRISP/Cas9 editing method, affected its virulence and the tolerance of B. cinerea to bacterial secondary metabolites. These findings suggest that specific detoxification transporters are involved in these interactions, with crucial role in different aspects of B. cinerea physiology.

Evaluation of seed associated endophytic bacteria from tolerant chilli cv. Firingi Jolokia for their biocontrol potential against bacterial wilt disease.

In this study, the seed endosphere of a bacterial wilt tolerant chilli cv. Firingi Jolokia was explored in order to find effective agents for bacterial wilt disease biocontrol. A total of 32 endophytic bacteria were isolated from freshly collected seeds and six isolates were selected based on R. solanacearum inhibition assay. These isolates were identified as Bacillus subtilis (KJ-2), Bacillus velezensis (KJ-4), Leuconostoc mesenteroides (KP-1), Lactococcus lactis (LB-3), Bacillus amyloliquefaciens (WK-2), and Bacillus subtilis (WK-3) by 16S rRNA gene sequencing. In the in planta R. solanacearum inhibition assay carried out by seedling root bacterization method, Bacillus subtilis (KJ-2) exhibited highest biocontrol efficacy of 86.6 % on 7th day post R. solanacearum inoculation and a minimum biocontrol efficacy of 52.9 % was noted for Leuconostoc mesenteroides (KP-1). GC-HRMS analysis detected several known antimicrobial compounds in the extract of the culture supernatant of Bacillus subtilis (KJ-2); which may contribute to inhibition of R. solanacearum. In the growth promotion assay conducted using these isolates, only two of them namely Bacillus subtilis (KJ-2) and Bacillus amyloliquefaciens (WK-2) showed growth promotion in true leafed tomato plants. All the selected seed endophytic isolates were able to control bacterial wilt of tomato at the seedling stage and Bacillus subtilis (KJ-2) was found to be most effective in controlling the disease. The results of the present study highlighted that seed endosphere of bacterial wilt tolerant cultivar is a rich source of R. solanacearum antagonizing bacterial isolates.

Genomics assisted functional characterization of Paenibacillus polymyxa HK4 as a biocontrol and plant growth promoting bacterium.

The diseases caused by phytopathogens account for huge economic losses in the agricultural sector. Paenibacillus polymyxa is one of the agriculturally important biocontrol agents and plant growth promoting bacterium. This study describes the antifungal potential of P. polymyxa HK4 against an array of fungal phytopathogens and its ability to stimulate seed germination of cumin and groundnut under in vitro conditions. The cumin and groundnut seeds bacterized with HK4 exhibited enhanced germination efficiency in comparison to controls. The use of HK4 as a soil inoculant significantly promoted the shoot length and fresh weight of groundnut plants in pot studies. The draft genome analysis of HK4 revealed the genetic attributes for motility, root colonization, antagonism, phosphate solubilization, siderophore production and production of volatile organic compounds. The bacterium HK4 harnessed several hydrolytic enzymes that may assist its competence in the rhizosphere. The PCR amplification and sequence analysis of the conserved region of the fusA gene amplicon revealed the ability of HK4 to produce fusaricidin. Furthermore, the LC-ESI-MS/MS of crude cell pellet extract of HK4 confirmed the presence of fusaricidin as a major antifungal metabolite. This study demonstrated the potential of HK4 as a biocontrol agent and a plant growth promoter.

Compatibility of Bt biopesticides and adjuvants for Spodoptera frugiperda control.

Spodoptera frugiperda is a pest of economic importance for several crops with resistance reports to Bt crops and pesticides. Eco-friendly Bt biopesticides may be an alternative to chemical insecticides due to their selectivity and specificity. However, the efficacy of Bt biopesticides may be influenced by the association with other chemicals, such as adjuvants. This study evaluated the compatibility and toxicity of Bt biopesticides mixed with adjuvants for the control of S. frugiperda. The treatments included the association of Dipel SC and Dipel PM with adjuvants. Compatibility tests were used to evaluate the Bt mixture. Bt suspensions obtained from mixtures of Bt and adjuvants at 106 and 3 × 108 spores/mL-1 were used to evaluate S. frugiperda mortality and distilled water was used as the control. The addition of the adjuvant LI increased growth and sporulation, indicating compatibility with Bt biopesticides. The other adjuvants were toxic to reducing Bt growth and sporulation. Only the mixture of Bt with LI and Bt alone was effective to S. frugiperda. The addition of adjuvants to Bt biopesticide affect the Bt sporulation, growth and mortality.

Livestock integration into soybean systems improves long-term system stability and profits without compromising crop yields.

Climate models project greater weather variability over the coming decades. High yielding systems that can maintain stable crop yields under variable environmental scenarios are critical to enhance food security. However, the effect of adding a trophic level (i.e. herbivores) on the long-term stability of agricultural systems is not well understood. We used a 16-year dataset from an integrated soybean-beef cattle experiment to measure the impacts of grazing on the stability of key crop, pasture, animal and whole-system outcomes. Treatments consisted of four grazing intensities (10, 20, 30 and 40 cm sward height) on mixed black oat (Avena strigosa) and Italian ryegrass (Lolium multiflorum) pastures and an ungrazed control. Stability of both human-digestible protein production and profitability increased at moderate to light grazing intensities, while over-intensification or absence of grazing decreased system stability. Grazing did not affect subsequent soybean yields but reduced the chance of crop failure and financial loss in unfavorable years. At both lighter and heavier grazing intensities, tradeoffs occurred between the stability of herbage production and animal live weight gains. We show that ecological intensification of specialized soybean systems using livestock integration can increase system stability and profitability, but the probability of win-win outcomes depends on management.

Lethal and antifeedant effects of Bordeaux mixture on the marsh slug (Deroceras laeve).

The marsh slug, Deroceras laeve (Müller), is a generalist pest of agricultural crops. Slugs are mainly managed with synthetic pesticides that can also have high toxicity toward vertebrates and nontarget arthropods. Besides, they are not labeled for use in organic crop systems. Bordeaux mixture (BM) is an alternative product often used in organic crops for controlling plant diseases. The molluscicidal activity of BM has been reported; however, to our knowledge, no study has determined its efficacy to control D. laeve. This study aimed to determine the lethal (concentration- and time-mortality curves, and foliar persistence) and antifeedant (reduction in leaf consumption) effects of BM-treated cabbage leaves on D. laeve. The LC50 of BM was 28.15%, and the LT50 was 8.83 h. The BM LC25 reduced D. laeve leaf consumption by 3.31-fold. Furthermore, high control effectiveness (mortality > 90%) was attained until the 7th day after spraying. These findings reveal that BM reduces damage by killing D. laeve in a fast faction and by reducing foliar consumption. Therefore, BM can be an alternative to D. laeve management in both conventional and organic systems.

Searching for New Beneficial Bacterial Isolates of Wild Raspberries for Biocontrol of Phytopathogens-Antagonistic Properties and Functional Characterization.

The threat caused by plants fungal and fungal-like pathogens is a serious problem in the organic farming of soft fruits. The European Commission regulations prohibit some commercially available chemical plant protection products, and instead recommend the use of natural methods for improving the microbial soil status and thus increasing resistance to biotic stresses caused by phytopathogens. The solution to this problem may be biopreparations based on, e.g., bacteria, especially those isolated from native local environments. To select proper bacterial candidates for biopreparation, research was provided to preliminarily ensure that those isolates are able not only to inhibit the growth of pathogens, but also to be metabolically effective. In the presented research sixty-five isolates were acquired and identified. Potentially pathogenic isolates were excluded from further research, and beneficial bacterial isolates were tested against the following plant pathogens: Botrytis spp., Colletotrichum spp., Phytophthora spp., and Verticillium spp. The eight most effective antagonists belonging to Arthrobacter, Bacillus, Pseudomonas, and Rhodococcus genera were subjected to metabolic and enzymatic analyses and a resistance to chemical stress survey, indicating to their potential as components of biopreparations for agroecology.

A simple PCR-based method for the rapid and accurate identification of spider mites (Tetranychidae) on cassava.

The morphological identification of mites entails great challenges. Characteristics such as dorsal setae and aedeagus are widely used, but they show variations between populations, and the technique is time consuming and demands specialized taxonomic expertise that is difficult to access. A successful alternative has been to exploit a region of the mitochondrial cytochrome oxidase I (COI) gene to classify specimens to the species level. We analyzed the COI sequences of four mite species associated with cassava and classified them definitively by detailed morphological examinations. We then developed an identification kit based on the restriction fragment length polymorphism-polymerase chain reaction of subunit I of the COI gene focused on the three restriction enzymes AseI, MboII, and ApoI. This set of enzymes permitted the simple, accurate identification of Mononychellus caribbeanae, M. tanajoa, M. mcgregori, and Tetranychus urticae, rapidly and with few resources. This kit could be a vital tool for the surveillance and monitoring of mite pests in cassava crop protection programs in Africa, Asia, and Latin America.