Prickly Pear Research Articles

Utilization of dried fruit peels from prickly pears as an eco-friendly bio-sorbent for adsorption and removing certain cationic dyes from water samples

Utilization of dried fruit peels from prickly pears as an eco-friendly bio-sorbent for adsorption and removing certain cationic dyes from water samples

Physical Methods as Pretreatment of Prickly Pear and Pineapple Peels for Producing Enzymes and Chemical Precursors

Physical Methods as Pretreatment of Prickly Pear and Pineapple Peels for Producing Enzymes and Chemical Precursors

MICRO-MORPHOLOGICAL AND PHYSICOCHEMICAL STANDARDISATION OF OPUNTIA ELATIOR MILL. CLADODE: AN UNEXPLORED TRADITIONAL PRICKLY PEAR CACTUS PLANT FROM MAHARASHTRA

Opuntia elatior Mill. (Cactaceae) is a significant medicinal herb In Maharashtra. In Ayurveda, it is referred to as Nagphani, and in English, prickly pear. Its fruits and cladode have long been used to remedy wounds, boils, coughs, fever, inflammation, and asthma. Despite its significant therapeutic potential, there isn't much information about standardised criteria for the species. Consequently, the current project was started to produce a comprehensive report on Opuntia elatior Mill's quality control and standardisation criteria. The current study aimed to highlight this traditional herbal crude, which would make it easier to identify fresh and dried samples using macro, microscopic, fluorescence, and physicochemical parameters in accordance with WHO guidelines. The present study's compilation of botanical characteristics can serve as a benchmark for the herbal medicine industry in identifying adulterations in market samples utilised for manufacturing different herbal remedies. These studies will provide reference information for accurately classifying Opuntia elatior Mill among closely related species within the same genus and family. They will also aid in identifying adulteration in market samples utilised to produce a range of herbal remedies.

Usefulness of Opuntia spp. on the Management of Obesity and Its Metabolic Co-Morbidities.

The plants of the Opuntia genus mainly grow in arid and semi-arid climates. Although the highest variety of wild species is found in Mexico, Opuntia spp. is widely distributed throughout the world. Extracts of these cacti have been described as important sources of bioactive substances that can have beneficial properties for the prevention and treatment of certain metabolic disorders. The objective of this review is to summarise the presently available knowledge regarding Opuntia ficus-indica (nopal or prickly pear), and some other species (O. streptacantha and O. robusta) on obesity and several metabolic complications. Current data show that Opuntia ficus-indica products used in preclinical studies have a significant capacity to prevent, at least partially, obesity and certain derived co-morbidities. On this subject, the potential beneficial effects of Opuntia are related to a reduction in oxidative stress and inflammation markers. Nevertheless, clinical studies have evidenced that the effects are highly contingent upon the experimental design. Moreover, the bioactive compound composition of nopal extracts has not been reported. As a result, there is a lack of information to elucidate the mechanisms of action responsible for the observed effects. Accordingly, further studies are needed to demonstrate whether Opuntia products can represent an effective tool to prevent and/or manage body weight and some metabolic disorders.

Response surface methodology‒guided color simulation of plant-based meat analogues before and after oven cooking: prickly pear cactus, mugwort, and sweet pumpkin pigments

This research aimed to find natural pigments that would give meat analogues the color of real meat, both before and after oven cooking. The natural pigments used were from the prickly pear cactus, mugwort, and sweet pumpkin. The key ingredient was the prickly pear cactus pigment, which consists mainly of betalains and loses its red color when exposed to heat. Yellow pigment derived from sweet pumpkin and green pigment derived from mugwort worked in synergy, and this mixture gave the analogues the characteristic brown hue of well-cooked meat. The optimal pigment ratios were established using response surface methodology, tailored for different meat types and cooking conditions. Quantitative findings revealed that the P3M1S1 combination at a 3% concentration provided the coloring most like that of real meat before and after cooking. In addition, sensory assessments confirmed that this mixture gave tasters an experience almost identical to that of eating actual meat. Response surface methodology validated the predictive accuracy, with deviations of less than 5% for color and less than 10% for sensory attributes. In addition, changes in important compounds responsible for color were identified when heat reduced the amounts of betalain in prickly pear cactus pigment and chlorophyll in mugwort pigment.

Optimizing prickly pear by-product valorization: formulating molasses with enhanced antioxidant capacities and sugar contents

ABSTRACT This study endeavoured to capitalize on prickly pear by-products for the optimization of molasses formulation, targeting elevated antioxidant capacities and superior sugar contents. Through robust statistical modelling, the optimal cooking parameters—temperature (70-80 °C) and duration (60–90 min)—were determined, guided by responses of antioxidant activity and Brix value. A D-Optimal mixture design further delineated the ideal proportions of molasses components (pulp, peel, and seeds). Characterization revealed that peel harboured the highest concentrations of total polyphenols (396.41 mg GAE/100g FW) and flavonoids (234.26 mg CE/100g FW), emphasizing its antioxidant potential (DPPH inhibition IC50: 12.72 µg/ml). The optimal cooking conditions were established at 78.35 °C for 79.70 min, with predictive equations guiding ingredient proportions (0.265 g pulp, 0.710 g peel, 0.025 g seed). Intriguingly, while peel inclusion enhanced total sugar content and antioxidant activity, seed incorporation exerted a contrasting effect by reducing total sugar content and limiting antioxidant activity.

Storage Conditions of Refrigerated Prickly Pears in Small Processing Industries

Prickly pears (Opuntia ficus-indica), which are well known for their beneficial properties for human health, are the subject of many studies due to their high content of bioactive compounds. However, the presence of spines on the fruit’s surface is a factor that limits consumption. Therefore, the viability and nutritional quality of peeled and packaged white, orange, and pink prickly pears were studied during storage. Refrigerated whole fruits stored at 8 °C and 85% RH for 0, 1, 2, 3, or 4 weeks were electrically peeled and packaged with micro-perforated film. At each time point, their microbial quality; physical parameters, such as hardness, texture, and color; and chemical parameters, including pH, titratable acidity, total soluble solids content, sugar content, ascorbic acid content, antioxidant capacity, and total phenol content, were analyzed. The mesophilic aerobic counts were lower than the values established by Spanish legislation (7 log (CFU/g f.w.)) until day 8 after packaging (or until day 6 after 4 weeks of storage). The hardness, texture, pH, sugar content, ascorbic acid content, and antioxidant capacity decreased significantly between day 0 and day 8 after packaging, independently of the number of weeks for which the whole fruits had been refrigerated previously. Furthermore, the changes in the parameters analyzed for whole fruits during storage were less remarkable. The evaluation of the sensory features was positive throughout the preserved period. The minimally processed prickly pears retained suitable microbial, nutritional, and sensory qualities when the whole fruits had been refrigerated for 1, 2, 3, or 4 weeks, facilitating their management in small processing companies.

Physicochemical characterization of four varieties of prickly pear (Opuntia spp.)

Objective: To characterize the physicochemical properties of the pulp from four varieties of prickly pear collected in three localities from the municipality of Salinas, San Luis Potosí. Design/Methodology/Approach: The following characteristics were recorded in 15 fruits of each variety: fruit size (equatorial and polar diameter), fresh biomass of total fruit, peel, and locule. Nine fruits per replicate were used (one fruit equals one replicate) to analyze moisture, ash content, total soluble solids, pH, titratable acidity, and total soluble sugars. Results: The physical differences of the fruits, total fresh weight, and equatorial and polar diameter match the typical characteristics of each variety. The largest fruits were (in descending order): Cristalina, Fafayuca, Naranjona, and Cardona. The highest moisture percentage (87.34%) was reported in Cristalina. Varieties such as Fafayuca from La Reforma and Naranjona from Salinas de Hidalgo had the highest and lowest concentration of total soluble sugars (25.84 and 11.46 g 100 g-1 FW, respectively). The fruits had a titratable acidity from 0.01 to 0.03% citric acid, while total soluble solids ranged from 11.60 to 15.84 °Brix. Study Limitations/Implications: Harvesting was based on the visual parameters established by the farmers. Findings/Conclusions: Fruits harvested from backyard orchards (<500 m2) had fruits of greater weight, which are used for self-consumption. The amount of total soluble solids and total soluble sugars per variety was different between localities, indicating that farmers from different localities have different harvesting criteria.

Fungal causal agents of the Black Spot of the cactus (Opuntia ficus-indica) in Colima, Mexico

Objective/Background: The prickly pear cactus (Opuntia ficus-indica) holds significant economic, social, and cultural importance in Mexico. However, it is recurrently affected by Black Spot disease (BS), caused by various phytopathogenic fungi. Identifying the causal agents of BS in commercial prickly pear crops is crucial for efficient agronomic management of the disease. The objective of this study was to identify the phytopathogenic fungi responsible for BS in prickly pear plantations in the Colima state, Mexico. Materials and Methods: Fifty cladodes from 50 plants exhibiting BS symptoms were collected from commercial plantations in Colima. The pathogenicity of the isolated fungi was verified using Koch’s postulates, and those causing the most severe BS symptoms were molecularly identified. Results: Thirty-five fungi were isolated from plants with BS symptoms, of which 20 exhibited distinct mycelial growth. Only six fungi induced BS symptoms; three of them were responsible for severe symptoms in cladodes: Alternaria alternata, Corynespora cassiicola, and Neoscytalidium dimidiatum. Conclusion: BS is caused by various phytopathogenic fungi, but this is the first report of C. cassiicola and N. dimidiatum as causal agents of BS in prickly pear cactus.

A Review of Non-wood Lignocellulose Waste Material Reinforced Concrete for Light-weight Construction Applications

In recent decades, non-wood lignocellulosic materials have gained significant attention, particularly in concrete applications for construction purposes. This study delves into utilising non-wood lignocellulosic materials for reinforcing concrete in construction applications. Lignocellulosic material emerges as a promising option for formulating new fibre cement compositions, thereby enhancing the sustainability, affordability, and performance of construction materials. Moreover, this research broadens the horizons of recycling agricultural waste by facilitating rational disposal and optimal utilisation. Through a comprehensive review, the study reveals that flax fibres, coir pith, prickly pear fibres, and rice husk ash waste exhibit superior workability compared to their counterparts. Furthermore, the strength of non-wood lignocellulosic reinforced concrete, incorporating bagasse ash, rice husk ash, and nutshell ash, peaked when fine aggregate replacement reached 15%, surpassing other types of non-wood lignocellulosic reinforced concrete. Adding a small quantity of prickly pear fibre to cement enhances the thermal conductivity of concrete, consequently improving compressive strength, flexural strength, tensile strength, and elastic modulus. This research is relevant to international research as it advances sustainable construction materials with desirable properties, benefiting society and various industries.

In vivo anti-Salmonella properties of aqueous extract of prickly pear (Opuntia ficus indica) cladode, hepatological and toxicological evaluation.

The present study aimed to evaluate antidiarrheal potential of prickly pear cladode and its hepatoprotective role in different groups of diarrhea-induced mice. Mice received cefixime (4 mg/kg of bw) and different concentrations of aqueous cladode extract (250 mg/kg of bw, 500 mg/kg of bw and 1000 mg/kg of bw). Feces Salmonella typhi ATCC 19430 was used to assess antidiarrheal potential and hematological, biochemical parameters, and histopathological analyses were carried out for 17 days. The results showed that administration of Salmonella typhi ATCC 19430 in mice produced liver injuries with histological damage, whereas 1000 mg/kg of bw cladode extract reduced the Salmonella typhi ATCC 19430 load of feces earlier as compared to the other groups during 17 days. Hematological parameters, like red blood cells (RBCs) (6.19 ± 1.85 × 106/mm3) and hemoglobin (Hb) (10.06 ± 2.03 g/dL), of negative control group decreased, while white blood cells (WBCs) (13.46 × 106/mm3) increased from reference values. In lipid profile, low-density lipoprotein (LDL) (9.0 ± 2.41 mg/dL), high-density lipoprotein (HDL) (6.07 ± 2.45 mg/dL) and total cholesterol (TC) (35.22 ± 8.29 mg/dL) of negative control group decreased, while triglycerides (TG) (168.35 ± 71.75 mg/dL) increased from reference values. Alanine transferase (ALT) (60.30 ± 20.33 IU/L), alkaline phosphatase (ALP) (359.9 ± 100.05 IU/L) and aspartate transferase (AST) (168.77 ± 66.61 IU/L) of negative control group increased from reference values. Urea (27.36 ± 10.54 mmol/L) and creatinine (35.29 ± 12.15 mmol/L) of the negative control group increased. Cefixime also ameliorated injuries due to the administration of Salmonella typhi ATCC 19430. Conclusively, these findings indicated that pure aqueous extract of cladode showed more promising results regarding antidiarrheal potential. Hence, cladode might be used in food and supplement formulations as a functional ingredient.

Harnessing Plant Bioelectricity through Prickly Pear as Botanical Batteries

Plants which are essential for life, have the potential to become a renewable energy source in the Future. They can generate electricity, reducing greenhouse gas emissions and being environmentally friendly. This study aims to explore the untapped potential of botanical batteries and contribute to green energy technology. Plants' capacity to convert sunlight into chemical energy could be a viable and environmentally friendly source for electrical power generation, offering a sustainable solution to the world's growing energy demands while mitigating climate change impacts. To achieve the goal of the study, pure experimental research was applied. And the researchers used cactus (prickly pear), copper nails, zinc nails, copper wire, alligator clips, and disposable plastic containers. The study was conducted at Bayugan National Comprehensive High School, Bayugan City. Throughout the analysis of the data obtained after the three tests conducted, it has been found that like an electrochemical cell, copper and zinc electrodes inserted into prickly pear leaves can generate energy. With the highest current magnitude and maximum value at the highest contact area, the prickly pear plant has the highest potential for energy harvesting. Cut-off or partially leaved leaves can be harvested for their energy, which can then be utilized to charge batteries or power low-power devices.

Phytochemical characterization and biological activities evaluation of Opuntia sp. cladodes

The prickly pear is a plant belonging to the Cactaceae family, and it is the best-known species that has gained significant recognition in recent years. Opuntia sp. cladodes have been traditionally utilized as a functional food and in traditional medicine for treating chronic diseases. Therefore, this study aims to evaluate the antioxidant, antimicrobial, and pharmacological activities of cladode extracts from Opuntia microdasys (Cl1) and Opuntia macrorhiza (Cl2), as well as to identify their phytochemical compounds. The chemical composition of the cladode extracts was analyzed using spectrophotometric methods, while the antioxidant activity was determined through DNA nicking assays. Microdilution assays were employed to assess the antimicrobial activity. Analgesic and anti-inflammatory activities were determined using the acetic acid writhing test in mice and the carrageenan-induced Wistar rat paw edema test. The results revealed that the two extracts of cladodes from Cl1 and Cl2 were characterized by a high phenolic content (58.13 and 44.58 GAE/g of extract, respectively). All the tested extracts exhibited antimicrobial activity against the strains tested, with Minimum Inhibitory Concentration (MIC) values ranging from 0.312 to 5 mg/mL and Minimum Bactericidal Concentration (MBC) values ranging from 1.25 to 10 mg/mL. The cladode extracts of Cl1 and Cl2 at a dose of 300 mg/kg showed a dose-dependent inhibition of acetic acid-induced writhing in mice, with respective values of 64.76 % and 61.91 %. The maximum anti-inflammatory activity of all doses of cladode extracts was observed 5 h after carrageenan-induced paw edema, and all tested doses significantly inhibited the induced inflammation.

Improve nutritive value of silage based on prickly pear peel by-products

Prickly pear (Opuntia ficus-indica L. Mill) is widely cultivated in arid regions of every continent for its nutritious fruits and various commercial applications, cactus pear cladodes are used as forage in the most arid regions, while recently prickly pear by-products are studied as ruminant feed. They are characterised by a high moisture and sugar content; therefore, it is necessary to study suitable conservation methods. The aim of this study was to verify the effectiveness of silage to preserve prickly pear peels (PP) in a mixture with other by-products at different inclusion levels (5% straw and 6% or 12% wheat bran). For each treatment, six under vacuum micro-silos were prepared and, after 40 d of storage, the state of preservation was evaluated. Subsequently, the aliquots were analysed for chemical composition and incubated with sheep rumen fluid to evaluate the fermentation kinetics. The use of wheat bran and straw added to PP resulted in an increase in dry matter (DM); PP silage with wheat bran was found to have higher protein content and nutritional value. The greater presence of lactic acid was found in PP, such as acetic acid, therefore the lactic/acetic ratio was 2.33, significantly lower than silages with the addition of wheat bran. Silage with 5% straw showed the lowest level of organic matter disappearance (OMD) and the cumulative volume of gas (OMCV) produced during the incubation. The PP showed the fastest fermentation rate achieved after 12 h of fermentation, while those containing wheat bran, presented a higher maximum fermentation rate (Rmax), which need longer time to be achieved. Canonical discriminant analysis (CDA) showed clear discrimination of different silages based on their chemical characteristics and parameters detected during in vitro incubation. Overall, the silages with the addition of wheat bran can be considered the best and, of the two, those produced with the addition of 12% bran. Finally, ensilage represents a conservation technique suitable for preserving the nutritional characteristics of PP.

Discrimination of different prickly pear (Opuntia spp.) accessions by NMR-based metabolomics

Mexico is one of the main producers of prickly pear worldwide and has around 50 registered accessions. Despite its great nutritional value, there are no reports on its metabolite profile. The objective of this study was to analyze 12 accessions of prickly pear (TAC, TNL, TCB, TCO, TV, TZ, TRI, TRP, TRR, TS, TSH and TT) of different colors using nuclear magnetic resonance (NMR) spectroscopy to identify their metabolic profiles. Principal component analysis (PCA) and orthogonal partial least squares discriminant analysis (OPLS-DA) were used to compare the metabolome. In total, 34 primary metabolites were identified in the 1H NMR spectra. The PCA model used to compare the 12 accessions did not show a clear separation between the groups. The OPLS-DA model made it possible to discriminate the TRI and TCB accessions. The heatmap showed that these accessions have a higher concentration of some metabolites. The TCB accession has high concentrations of sugars and amino acids while the TRI accession has high concentrations of organic acids. In addition, The TCO accession has a lower concentration of primary metabolites. The OPLS-DA model made it possible to classify the prickly pears by color. For orange prickly pears, the TNL accession has a higher concentration of primary metabolites than the TAC accession. In green prickly pears, the accession with the highest concentration of primary metabolites is the TCB accession, while it is the TRI accession in red prickly pears. This work demonstrated that NMR-based metabolomics is a highly reproducible and robust methodology to classify prickly pear accessions.

A REVIEW: OPUNTIA FICUS-INDICA AS A SOURCE OF BIOACTIVE COMPOUND INGREDIENTS FOR FUNCTIONAL FOODS, NUTRITION, HUMAN DISEASE AND HEALTH

Originally from Mexico, today, prickly pears (Opuntia ficus-indica (L.) Mill.) can be found worldwide, but they are most common in Africa., Australia, and the countries of the Mediterranean. The Cactaceae family, which has more than 1500 species of cactus, includes prickly pears. Additionally, consumers are becoming more and more interested in it because it contains bioactive substances like carotenoids, sterols, polyunsaturated fatty acids, polyphenols, and vitamins that are linked to important biological and functional activities such as effects on antioxidants, inflammation, and hypoglycemia as it prospers in dry and semiarid areas, this plant is very effective in preventing desertification and can be exploited in fields and gardens as a fence. Although it is frequently used today to make culinary products like jams and juices, it is also eaten as a fruit. If one wishes to create novel products with health-promoting properties for food, cosmetics, or pharmaceutical fields, As a feasible wellspring of phytochemicals for practical food sources, natural additions, or nutritional supplements, prickly pears are affordable. Peer Review History: Received 3 December 2023; Revised 25 January 2024; Accepted 6 March; Available online 15 March 2024 Academic Editor: Dr. Sally A. El-Zahaby, Pharos University in Alexandria, Egypt, sally.elzahaby@yahoo.com Average Peer review marks at initial stage: 6.0/10 Average Peer review marks at publication stage: 7.0/10 Reviewers: Dr. Gehan Fawzy Abdel Raoof Kandeel, Pharmacognosy Department, National Research Centre, Dokki, 12622, Giza, Egypt, gehankandeel9@yahoo.com Dr. Evren Alğin Yapar, Turkish Medicines and Medical Devices Agency, Turkiye, evren.yapar@yahoo.com

Exotic predators can sequester and use novel toxins from exotic non-coevolved prey.

Defensive chemicals of prey can be sequestered by some coevolved predators, which take advantage of prey toxins for their own defence. The increase in the number of invasive species in the Anthropocene has resulted in new interactions among non-coevolved predator and prey species. While novelty in chemical defence may provide a benefit for invasive prey against non-coevolved predators, resident predators with the right evolutionary pre-adaptations might benefit from sequestering these novel defences. Here, we chose a well-known system of invasive species to test whether non-coevolved predators can sequester and use toxins from exotic prey. Together with the invasive prickly pear plants, cochineal bugs (Dactylopius spp.) are spreading worldwide from their native range in the Americas. These insects produce carminic acid, a defensive anthraquinone that some specialized predators sequester for their own defence. Using this system, we first determined whether coccinellids that prey on cochineal bugs in the Mediterranean region tolerated, sequestered, and released carminic acid in reflex bleeding. Then, we quantified the deterrent effect of carminic acid against antagonistic ants. Our results demonstrate that the Australian coccinellid Cryptolaemus montrouzieri sequestered carminic acid, a substance absent in its coevolved prey, from exotic cochineal bugs. When attacked, the predator released this substance through reflex bleeding at concentrations that were deterrent against antagonistic ants. These findings reveal that non-coevolved predators can sequester and use novel toxins from exotic prey and highlights the surprising outcomes of novel interactions that arise from species invasions.

Selenium-Induced Enhancement in Growth and Rhizosphere Soil Methane Oxidation of Prickly Pear.

As an essential element for plants, animals, and humans, selenium (Se) has been shown to participate in microbial methane oxidation. We studied the growth response and rhizosphere methane oxidation of an economic crop (prickly pear, Rosa roxburghii Tratt) through three treatments (Se0.6 mg/kg, Se2.0 mg/kg, and Se10 mg/kg) and a control (Se0 mg/kg) in a two-month pot experiment. The results showed that the height, total biomass, root biomass, and leaf biomass of prickly pear were significantly increased in the Se0.6 and Se2.0 treatments. The root-to-shoot ratio of prickly pear reached a maximum value in the Se2 treatment. The leaf carotenoid contents significantly increased in the three treatments. Antioxidant activities significantly increased in the Se0.6 and Se2 treatments. Low Se contents (0.6, 2 mg/kg) promoted root growth, including dry weight, length, surface area, volume, and root activity. There was a significant linear relationship between root and aboveground Se contents. The Se translocation factor increased as the soil Se content increased, ranging from 0.173 to 0.288. The application of Se can improve the state of rhizosphere soil's organic C and soil nutrients (N, P, and K). Se significantly promoted the methane oxidation rate in rhizosphere soils, and the Se10 treatment showed the highest methane oxidation rate. The soil Se gradients led to differentiation in the growth, rhizosphere soil properties, and methane oxidation capacity of prickly pear. The root Se content and Se translocation factor were significantly positively correlated with the methane oxidation rate. Prickly pear can accumulate Se when grown in Se-enriched soil. The 2 mg/kg Se soil treatment enhanced growth and methane oxidation in the rhizosphere soil of prickly pear.

Chemistry and functional properties of bioactive compounds present in Prickly pear fruits

Chemistry and functional properties of bioactive compounds present in Prickly pear fruits

First Report of rattail cactus necrosis-associated virus Infecting Prickly pear (Opuntia macrocentra) in the United States.

Black-spined prickly pear (Opuntia macrocentra Engelmann; Cactaceae) is a cactus native to Arizona, New Mexico, Texas, and northwest Mexico. The plant is often grown for ornamental purposes in the United States. In February 2023, virus-like symptoms such as concentric ringspots and chlorotic spots were observed on O. macrocentra plants grown at the vicinity of Maricopa County Cooperative extension, University of Arizona, Phoenix, AZ (33°24'24.6"N, 111°59'15.3"W). Total RNA was extracted from two samples (YPHC-60-A and YPHC-60-B), following the protocol by Tzanetakis et al. (2007). Reverse transcription polymerase chain reaction (RT-PCR) was performed with degenerate tobamovirus, TobamodF/TobamodR (Li et al. 2018) and potexvirus, 1RC, Potex 2RC, and Potex 5 (van der Vlugt and Berendsen 2002) primers. An expected amplicon of ~880 bp was obtained from both samples using TobamodF/TobamodR primers, while no amplification was observed with potexvirus primers. Further, RT-PCR was carried out using species-specific primers to detect cacti related tobamoviruses: cactus mild mottle virus (CMMoV), rattail cactus necrosis-associated virus (RCNaV) (Park et al. 2018) and Opuntia virus 2 (Salgado-Ortiz et al. 2020). Amplicons of ~540 bp were amplified from both samples using RCNaV specific primers, whereas no amplification was obtained using CMMoV and Opuntia virus 2 specific primers. Then, the amplicons from both YPHC-60 (A-B) isolates (~540 bp) were Sanger sequenced and shared 99.22% nucleotide identity to each other. A BLAST search revealed 93% nucleotide identity with RCNaV CP sequences (KY581586.1, JF729471, and MT130378.1). The sequences were submitted in the GenBank (accessions no. OQ914798 and OR828526). Furthermore, complete RCNaV- RNA dependent RNA polymerase (RdRP) gene was amplified using primers 3490-s-5'GTAGGTGGTACCGCATAGCA-3'; 3490as 5'AAACGCAAGTCMRYGACYGA-3' (designed in this study from accession no. JF729471.1, position 3490-3509 and 4905-4925). The expected amplicons of ~1,500 bp were obtained from both YPHC-60 (A-B) samples and sequenced (GenBank: OQ914799 and OR823954) showing 87.5 % identity with RCNaV sequences (JF729471.1 and NC_016442.1). The maximum-likelihood phylogenetic tree clustered YPHC-60 (A-B) isolates in a single clade with other RCNaV isolates. RCNaV virus particles were isolated from YPHC-60 (A-B) and submitted for RNA extraction, testing positive for RCNaV by RT-PCR. Sap extract of YPHC-60 (A-B) prepared in 0.01 M phosphate buffer (pH =7.0) was used to mechanically inoculate 3 indicator plant species (n=10): Phaseolus vulgaris, Medicago sativa, and Cucumis melo. Also, infected tissue was used to graft Opuntia sp. plants. Symptoms such as local lesions were observed on M. sativa and vein thickening on P. vulgaris 14 days post-inoculation, while Opuntia sp. showed chlorosis 30 days after grafting. RCNaV infection in mechanically inoculated P. vulgaris, M. sativa, and Opuntia sp. was also confirmed through RT-PCR. C. melo and non-inoculated control plants did not show any symptoms, nor tested positive through RT-PCR. RCNaV has been reported earlier to infect cactus species in South Korea (Park et al. 2018) and O. albicarpa in Mexico (De La Torre-Almaráz et al. 2016), where it was found in several orchards. To the best of our knowledge, this is the first report of RCNaV infecting O. macrocentra in the United States. This study highlights that RCNaV is easily transmitted mechanically or by grafting, which could impact the nursery industry as most cacti are clonally propagated.