Integrated Disease Management Strategies Research Articles

Integrated Management of Verticillium Wilt of Cacao

The vascular disease Verticillium wilt of cacao (Theobroma cacao), caused by the soilborne fungus Verticillium dahliae, is often qualified as a “minor” disease. However, it can cause severe losses locally, for example, in western Uganda and northeast of the Democratic Republic of Congo. This disease is difficult to control, notably due to an extended host range and prolonged survival in the soil. Therefore, Verticillium wilt must be addressed through an integrated disease management strategy as employed for other tree crops such as olive. Few studies, nonetheless, have focused on how to confront this disease in cacao. This paper aims to provide an overview of our knowledge on Verticillium wilt on cacao and the integrated disease management strategies for preventing and controlling it. Promising avenues based on findings in other crops that could be adapted to cacao are also explored. Good agricultural practices, genetic resistance, biological control, induced resistance, and the use of organic amendments with or without biocontrol agents are discussed. Moreover, the potential benefits of some of these solutions toward the resistance to other cacao diseases, abiotic stress, and nutrition improvement are presented.

An Overview of Chili Leaf Curl Disease: Molecular Mechanisms, Impact, Challenges, and Disease Management Strategies in Indian Subcontinent

Leaf curl disease in a chili plant is caused mainly by Chili leaf curl virus (ChiLCV) (Family: Geminiviridae, Genus: Begomovirus). ChiLCV shows a widespread occurrence in most of the chili (Capsicum spp.) growing regions. ChiLCV has a limited host range and infects tomatoes (Solanum lycopersicum), potatoes (S. tuberosum), and amaranth (Amaranthus tricolor). The virus genome is a monopartite circular single-stranded DNA molecule of 2.7 kb and associated with α and β-satellites of 1.3 and 1.4 kb, respectively. The virus genome is encapsulated in distinct twinned icosahedral particles of around 18–30 nm in size and transmitted by Bemisia tabaci (Family: Aleyrodidae, Order: Hemiptera). Recently, bipartite begomovirus has been found to be associated with leaf curl disease. The leaf curl disease has a widespread distribution in the major equatorial regions viz., Australia, Asia, Africa, Europe, and America. Besides the PCR, qPCR, and LAMP-based detection systems, recently, localized surface-plasmon-resonance (LPSR) based optical platform is used for ChiLCV detection in a 20–40 μl of sample volume using aluminum nanoparticles. Management of ChiLCV is more challenging due to the vector-borne nature of the virus, therefore integrated disease management strategies need to be followed to contain the spread and heavy crop loss. CRISPR/Cas-mediated virus resistance has gained importance in disease management of DNA and RNA viruses due to certain advantages over the conventional approaches. Therefore, CRISPR/Cas system-mediated resistance needs to be explored in chili against ChiLCV.

Plant Pathology: Advances in Disease Diagnosis and Management

Plant diseases pose significant threats to global agriculture, impacting crop yield, quality, and food security. Effective diagnosis and management of plant diseases are crucial for mitigating these threats and ensuring sustainable agricultural production. This review highlights recent advancements in plant pathology, focusing on innovative diagnostic techniques and integrated disease management strategies. We explore the development and application of molecular diagnostics, remote sensing technologies, and machine learning algorithms for early and accurate disease detection. Additionally, we discuss advances in disease management, including biocontrol agents, resistance breeding, and precision agriculture practices. Case studies demonstrating the successful implementation of these technologies and strategies are presented, showcasing their impact on reducing disease incidence and improving crop health. Finally, we address the challenges and future directions in plant pathology research, emphasizing the need for multidisciplinary approaches and enhanced collaboration between researchers, farmers, and policymakers.

Black Root Rot of Strawberry: A Disease Complex

Historically, root diseases have been a production-limiting problem for the strawberry industry worldwide. Even though this disease is of great economic importance, the etiology remains unresolved. However, soilborne fungal root pathogens, particularly Pythium and Rhizoctonia spp. have been implicated as major role players. Presence of nematodes in soil and favourable environmental conditions also play significant role in the disease development. Development of integrated disease management strategies is dependent upon a more complete understanding of the etiology, biology and ecology of the disease complex.

Enhanced suppression of soil-borne phytopathogenic bacteria Ralstonia solanacearum in soil and promotion of tomato plant growth by synergetic effect of green synthesized nanoparticles and plant extract.

Because of severe economic losses and food security concerns caused by plant pathogenic bacteria, Ralstonia solanacearum, there is a need to develop novel control methods. This study was aimed to green synthesize the zinc oxide nanoparticles (ZnO NPs) through Withania coagulans leaf extracts and checked their antibacterial potential alone or in combination with W. coagulans leaf extract for the management of R. solanacearum causing bacterial wilt disease in tomato. ZnO NPs were synthesized through an eco-friendly approach using leaves extract of W. coagulans and characterized through various spectroscopic approaches, that is Fourier transform infrared spectroscopic, UV-visible spectroscopy and energy dispersive spectroscopy. The antibacterial effect of W. coagulans leaf extract and ZnO NPs alone and in combination was tested in vitro and in vivo against bacterial wilt pathogen in tomato plants. The results showed that combine application of leaf extract and ZnO NPs inhibited in vitro growth of R. solanacearum more than applying alone. Three application times (0, 6 and 12days before transplantation) of leaf extract, ZnONPs and their combine application were tested in vivo. The combine treatment and longest application time (12days before transplantation) were more effective in suppressing soil population of R. solanacearum, reducing disease severity and enhancing plant growth than applying alone and smaller application time. It is concluded that W. coagulans leaf extract and ZnO NPs have strong antibacterial potential against R. solanacearum in vitro and in vivo. The results of this study suggest the potential application of leaf extract and ZnO nanoparticles for controlling R. solanacearum as safe, eco-friendly and less expensive integrated disease management strategy in tomato crop.

Actinoalloteichus cyanogriseus: a broad spectrum bio-agent against mulberry root rot pathogens

BackgroundIn India, severe yield and economic losses in sericulture were reported due to mulberry root rot disease. Since the disease was caused by complex soil-borne pathogens with wide host range, an attempt was made to develop actinobacteria based biological management strategy to mitigate it.ResultsAs a result of continuous screening for anti-fungal actinobacteria, 2 novel and potent isolates (M11 and M12) belong to the rare genus, Actinoalloteichus were obtained from mulberry rhizosphere. Both isolates were gram positive, grayish, aerobic and produced good amount of melanin pigment during sporulation. They exhibited excellent anti-fungal activity against mulberry root rot pathogens viz., Macrophomina phaseolina, Lasiodiplodia theobromae, Fusarium solani, Sclerotium rolfsii and Rhizoctonia solani. In addition, these melanogenic isolates grew in wide temperature and pH ranges. Their multi-stress tolerance ability/polyextremophilic nature was exposed when amending regular growth medium with excess concentration of salt and alkali. In addition, the potent isolates also showed tolerance to carbendazim (up to 1000 ppm) and UVB irradiation (up to 180 min at 10 cm distance).ConclusionThe present study focused on the identification and characterization of melanogenic actinobacteria, further made it potential candidate that could be incorporated in eco-friendly, integrated disease management strategies against complex mulberry root rot pathogens.

SeedborneCercospora beticolaCan Initiate Cercospora Leaf Spot from Sugar Beet (Beta vulgaris) Fruit Tissue

Cercospora leaf spot (CLS) is a globally important disease of sugar beet (Beta vulgaris) caused by the fungus Cercospora beticola. Long-distance movement of C. beticola has been indirectly evidenced in recent population genetic studies, suggesting potential dispersal via seed. Commercial sugar beet "seed" consists of the reproductive fruit (true seed surrounded by maternal pericarp tissue) coated in artificial pellet material. In this study, we confirmed the presence of viable C. beticola in sugar beet fruit for 10 of 37 tested seed lots. All isolates harbored the G143A mutation associated with quinone outside inhibitor resistance, and 32 of 38 isolates had reduced demethylation inhibitor sensitivity (EC50 > 1 µg/ml). Planting of commercial sugar beet seed demonstrated the ability of seedborne inoculum to initiate CLS in sugar beet. C. beticola DNA was detected in DNA isolated from xylem sap, suggesting the vascular system is used to systemically colonize the host. We established nuclear ribosomal internal transcribed spacer region amplicon sequencing using the MinION platform to detect fungi in sugar beet fruit. Fungal sequences from 19 different genera were identified from 11 different sugar beet seed lots, but Fusarium, Alternaria, and Cercospora were consistently the three most dominant taxa, comprising an average of 93% relative read abundance over 11 seed lots. We also present evidence that C. beticola resides in the pericarp of sugar beet fruit rather than the true seed. The presence of seedborne inoculum should be considered when implementing integrated disease management strategies for CLS of sugar beet in the future.

Antagonistic activity of three species of bacteria against Fusarium oxysporum f. sp. albedinis, the causal agent of Bayoud disease on date palm (Phoenix dactylifera L.)

This work has the objective to study the antagonistic capacity of three bacte-rial species (Bacillus subtilus, B. amyloliquefasciens, and Pseudomonas pleco-glossicida), on mycelia growth of 20 isolates of Fusarium oxysporum f. sp. Albedinis (Foa), the causal agent of bayoud disease on date palm. We tested the inhibition of mycelial growth by dual confrontation method using these three bacteria species. A significant effect (P<0,05) of these microorganisms was observed on mycelial growth of 20 isolates of Foa, with the inhibition rates varied between 28,05 and 64,2%. The confirmation of this inhibition was observed by creation of inhibition zones in the Foa colonies confronted by these bacteria. The obtained results lead us to think about the possibility to apply these bacterial species as biofungicide in the control of Bayoud dis-ease in our contaminated groves and can be considerate as another alterna-tive fungicide in the efficient integrated disease management strategy.

Characterization and Control of Thielaviopsis punctulata on Date Palm in Saudi Arabia.

Date palm (Phoenix dactylifera L.) is the most important edible fruit crop in Saudi Arabia. Date palm cultivation and productivity are severely affected by various fungal diseases in date palm-producing countries. In recent years, black scorch disease has emerged as a devastating disease affecting date palm cultivation in the Arabian Peninsula. In the current survey, leaves and root samples were collected from deteriorated date palm trees showing variable symptoms of neck bending, leaf drying, tissue necrosis, wilting, and mortality of the entire tree in the Al-Ahsa region of Saudi Arabia. During microscopic examination, the fungus isolates growing on potato dextrose agar (PDA) media produced thick-walled chlamydospores and endoconidia. The morphological characterization confirmed the presence of Thielaviopsis punctulata in the date palm plant samples as the potential agent of black scorch disease. The results were further confirmed by polymerase chain reaction (PCR), sequencing, and phylogenetic dendrograms of partial regions of the ITS, TEF1-α, and β-tubulin genes. The nucleotide sequence comparison showed that the T. punctulata isolates were 99.9–100% identical to each other and to the T. punctulata isolate identified from Iraq-infecting date palm trees. The pathogenicity of the three selected T. punctulata isolates was also confirmed on date palm plants of Khalas cultivar. The morphological, molecular, and pathogenicity results confirmed that T. punctulata causes black scorch disease in symptomatic date palm plants in Saudi Arabia. Furthermore, seven commercially available fungicides were also tested for their potential efficacy to control black scorch disease. The in vitro application of the three fungicides Aliette, Score, and Tachigazole reduced the fungal growth zone by 86–100%, respectively, whereas the in vivo studies determined that the fungicides Aliette and Score significantly impeded the mycelial progression of T. punctulata with 40% and 73% efficiency, respectively. These fungicides can be used in integrated disease management (IDM) strategies to curb black scorch disease.

Integrated disease management strategies against leaf spot and root rot diseases of mungbean

Integrated disease management strategies against leaf spot and root rot diseases of mungbean

Stemphylium Leaf Blight: A Re-Emerging Threat to Onion Production in Eastern North America.

Stemphylium leaf blight (SLB), caused by Stemphylium vesicarium, is a foliar disease of onion worldwide, and has recently become an important disease in the northeastern United States and Ontario, Canada. The symptoms begin as small, tan to brown lesions on the leaves that can progress to defoliate plants. Crop loss occurs through reduced photosynthetic area, resulting in smaller, lower-quality bulbs. Leaf necrosis caused by SLB also can compromise bulb storage, as green leaves are required for the uptake of sprout inhibitors applied prior to harvest. The pathogen can overwinter on infested onion residue and infected volunteer plants. Asymptomatic weedy hosts near onion fields may also be a source of inoculum. Production of ascospores of the teleomorph (Pleospora allii) peaks in early spring in northeastern North America, often before the crop is planted, and declines rapidly as daily mean air temperatures rise. Conidia are usually present throughout the growing season. Application of fungicides is a standard practice for management of the complex of fungi that can cause foliar diseases of onion in this region. Recent assessments have shown that populations of S. vesicarium in New York and Ontario are resistant to at least three single-site mode-of-action fungicides. Three disease prediction systems have been developed and evaluated that may enable growers to reduce the frequency and/or number of fungicide applications, but the loss of efficacious fungicides due to resistance development within S. vesicarium populations threatens sustainability. The lack of commercially acceptable onion cultivars with sufficient resistance to reduce the number of fungicides for SLB also limits the ability to manage SLB effectively. Integrated disease management strategies for SLB are essential to maintain profitable, sustainable onion production across eastern North America.

Biofumigation: A Cover Crop Option 12 Months of the Year to Manage Three Soilborne Pathogens Ailing the Australian Vegetable Industry

Brassica biofumigant cover crops are being increasingly considered in vegetable crop rotations as part of an integrated disease management strategy and simply as a cover cropping choice. Nine biofumigant varieties were assessed to see if they could be grown year-round in the Lockyer Valley South East Queensland region, for yield, days to incorporation and glucosinolate concentrations, as well as efficacy against 3 soilborne pathogens; Sclerotinia sclerotiorum, Sclerotium rolfsii and Macrophomina phaseolina. The fastest growing brassica biofumigant was BQ Mulch which reached 25% flowering in 36 and 59 days from planting to incorporation with a summer and winter planting respectively. Nemcon and Nemclear took the longest to incorporation when planted in summer, 101 days and failed to flower, while Caliente, Tillage Radish and Biofum reached 25% flowering and incorporation in 98 days when planted in winter. BQ Mulch produced the least amount of biomass, 30.93 t/ha fresh weight and 2.92 t/ha dry weight with a summer planting. Biofum producing the greatest amount of biomass, 185.76 t/ha fresh weight and 17.34 t/ha dry weight with a summer and winter planting respectively. Most varieties produced more total glucosinolates during summer compared to winter. Caliente produced the highest levels of Total GSLs with 53.47 µmol/g DW in summer compared to 23.78 µmol/g DW in winter. This was reflected in their efficacy against the soilborne pathogens. Caliente and Mustclean were more efficacious at controlling Macrophomina and Sclerotinia in summer compared to winter while all varieties were more efficacious at controlling Sclerotinia with a summer planting compared to a winter planting.

Assessment of in vitro antimicrobial efficacy of biologically synthesized metal nanoparticles against pathogenic bacteria

The microbial infections due to biofilm forming bacterial pathogens are very common in human subjects. The intensive application of antibiotics in integrated disease management strategy has led to increased multidrug resistance incommon pathogens. Thus, indicating need of developing an alternative method for the control of these multidrug resistant pathogens. Present study involves the Moringa oleifera aqueous extract mediated biological synthesis of silver (Ag nanoparticles (NPs)- Avg. size 82.5 nm; zeta potential = −27.9 mV), copper oxide (CuONPs- Avg. size 61 nm; zeta potential = −19.3 mV), iron oxide (FeONPs- Avg. size 83.3 nm; zeta potential = −9.37 mV) and alumina (AlONPs- Avg. size 87.3 nm; zeta potential = −10.9 mV) nanoparticles. Biological nanoparticles were detected by visual observation, spectrophotometric detection followed by zeta potential analysis, nanoparticle tracking analysis, Fourier transform infrared spectrometry and X-ray diffraction analysis. Nanoparticles were further evaluated for their in vitro antimicrobial potential, membrane damage effectiveness, biofilm inhibition activity by MTT assay. Nanoparticles were assessed against human pathogens viz. two Gram-positive (Bacillus subtilis MTCC 441 and Staphylococcus haemolyticus MTCC 3383) and two Gram-negative bacteria (Enterobacter aerogenes MTCC 111 and Salmonella enterica ser. Typhi MTCC 8767). The nanoparticles exhibited akin activity pattern against all pathogens studied i.e. AgNPs > CuONPs > AlONPs > FeONPs. Tested nanoparticles registered lower MIC values and more intensified growth inhibition against Gram-negative bacteria compared to their Gram-positive counterparts. These results pointed out that the M. oleifera mediated nanoparticles can be prospectivelyutilized in the development of alternative antimicrobials against diverse bacterial infections.

Feeding the Microbes: A Strategy to Control Verticillium Wilt

Verticillium dahliae is one of the most devastating soilborne pathogens for horticulture production. The pathogen has a broad host range and currently there is no effective chemical disease management, therefore, novel sustainable integrated disease management strategies should be considered. In this respect, we questioned whether the soil application of common microbiological growth media can influence the plant—microbe interactions and, subsequently, offer protection against V. dahliae. Indeed, the addition of Nutrient Broth (NB) and Potato Dextrose (PD) in non-sterilized soil reduced Verticillium wilt symptoms in eggplants. The addition of NB in sterilized soil did not reduce the disease symptoms compared to controls, however the addition of PD enhanced plant protection against V. dahliae. Following the results of a split root bioassay in eggplants, the possibility that NB and PD triggered the plant defense mechanisms against V. dahliae was excluded, since NB and PD did not reduce wilt symptom. Therefore, PD may be used as an easy food source for V. dahliae, detouring the pathogen from the root system of plants, while NB may affect the soil microbiome by enhancing antagonism in rhizosphere, or antagonistic interaction between V. dahliae and rhizospheric microbiome. Subsequently, several bacterial strains were isolated from the NB-treated rhizosphere and examined for their biocontrol activity against V. dahliae. Among the examined strains, a Pseudomonas putida strain, Z13, significantly reduced Veticillium severity and incidence under greenhouse conditions.

Ecology of pitaya stem rot caused by Gilbertella persicaria and its biological control by phyllosphere bacteria

AbstractGilbertella persicaria stem rot is a disease that causes rotting in the stem nodules and fruits of pitaya. Here, we aimed to study the ecology of G. persicaria and its biological control using several Bacillus subtilis strains isolated from the phyllosphere. Sporangiospores of G. persicaria were detected on different parts of the plant, with maximum density (c.239.9 propagules/cm2) found on the petals after flowering, and spore germination was observed to be considerably affected by humidity. Furthermore, the study revealed that an incision wound of at least 2 mm depth was necessary for infection by the pathogen to the stem nodule, and exposure to high temperatures (50°C) accelerated disease incidence, by disrupting resistance to the spread of the pathogen. Additionally, biological control of G. persicaria was effectively achieved by inoculation with B. subtilis strains isolated from pitaya stem nodules. Intriguingly, treatment with antagonistic bacteria reduced stem rot incidence to zero (bacterial suspension: PNB‐5, PNB‐7, and PNB‐93; culture filtrate: PNB‐7, PNB‐93, and PNB‐95), equivalent to that achieved by fungicide application in both prophylactic and simultaneous treatments. Collectively, this is the first study to demonstrate the ecology of G. persicaria and to show the potential applicability of phyllosphere bacterial strains for biological control of G. persicaria in pitaya, which could help develop effective integrated disease management strategies for the control of pitaya stem rot.

Monitoring of Brazilian wheat blast field populations reveals resistance to QoI, DMI, and SDHI fungicides

AbstractWheat blast is one of the most important and devastating fungal diseases of wheat in South America, South‐east Asia, and now in southern Africa. The disease can reduce grain yield by up to 70% and is best controlled using integrated disease management strategies. The difficulty in disease management is compounded by the lack of durable host resistance and the ineffectiveness of fungicide sprays. New succinate dehydrogenase inhibitor (SDHI) fungicides were recently introduced for the management of wheat diseases. Brazilian field populations of the wheat blast pathogen Pyricularia oryzae Triticum lineage (PoTl) sampled from different geographical regions in 2012 and 2018 were shown to be resistant to both QoI (strobilurin) and DMI (azole) fungicides. The main objective of the current study was to determine the SDHI baseline sensitivity in these populations. Moderate levels of SDHI resistance were detected in five out of the six field populations sampled in 2012 and in most of the strains isolated in 2018. No association was found between target site mutations in the sdhB, sdhC, and sdhD genes and the levels of SDHI resistance, indicating that a pre‐existing resistance mechanism not associated with target site mutations is probably present in Brazilian wheat blast populations.

Evaluation of 20 enset (Ensete ventricosum) landraces for response to Xanthomonas vasicola pv. Musacearum infection

Xanthomonas wilt, caused by Xanthomonas vasicola pv. musacearum (Xvm), formerly X. campestris pv. musacearum, is the most threatening and economically important disease of enset (Ensete ventricosum), the multipurpose food security crop orphan in south and southwestern Ethiopia. Xvm has also had a major impact on banana and plantain production in east Africa following its detection in Uganda in 2001 and subsequent spread. The only current effective control of this disease relies on integrated disease management strategies including minimizing field pathogen inoculum and deployment of wilt-resistant enset landraces. Identifying landraces with stable and durable Xvm resistance will greatly accelerate breeding programmes. While previous reports have identified landraces with varying degrees of tolerance to Xvm, no systematic study has collectively assessed their relative resistance. Here we undertook a detailed “common garden” analysis of 20 enset landraces previously reported to exhibit lower susceptibility to Xvm using an aggressive Xvm inoculum isolated from a disease hotspot area. Detailed longitudinal and survival analyses were applied to each landrace, using a combination of area-under-disease progress stairs, disease index and apparent infection rate to capture disease metrics as well as disease progression symptoms. Considerable variation was observed among the 20 landraces; however, none exhibited full immunity to Xvm infection. Landraces Haella, Mazia and Lemat showed the lowest susceptibility to Xvm as evidenced by reduced disease units and higher survival rates compared to the susceptible control landrace Arkiya, which exhibited the highest infection level and lowest survival rate, consistent with a high degree of susceptibility to Xvm. Thus, we have in this controlled experiment identified new material suitable for incorporation into future breeding programmes to develop Xvm-resistant enset varieties.

Real-Time PCR Assay for the Diagnosis and Quantification of Co-infections by Diaporthe batatas and Diaporthe destruens in Sweet Potato.

Foot rot disease caused by Diaporthe destruens (formerly Plenodomus destruens) has become a major concern for the production of sweet potato [Ipomoea batatas (L.) Lam.] in Japan. A related fungus Diaporthe batatas, which causes dry rot disease of sweet potato, is native and is widespread in fields in Japan. The similar characteristics of these two pathogens pose a challenge for conventional disease diagnosis. Currently, there are no effective molecular measures for identifying and distinguishing D. destruens and D. batatas. Here, we demonstrate a real-time PCR assay that distinguishes and quantifies D. batatas and D. destruens from co-infected sweet potato. The assay was performed with various simulated DNA combinations of D. batatas and D. destruens ranging from 1:1 to 1:100000. The assay was also used with the ratios of D. batatas: D. destruens: sweet potato DNA ranging from 1:1:1 to 1:1:100000. These assays produced a specific amplification product for each of the pathogens, and quantified the fungal biomass over the entire range tested without detecting false positives. The assay was validated by using infected sweet potato collected from various fields; it showed sufficient sensitivity and specificity to quantify and distinguish D. batatas and D. destruens from these field samples. Thus, our real-time PCR assay would be a useful tool for diagnosis of D. batatas and D. destruens and is expected to provide the foundation for the design of integrated disease management strategies for foot rot disease in sweet potato.

Trends in Molecular Diagnosis and Diversity Studies for Phytosanitary Regulated Xanthomonas.

Bacteria in the genus Xanthomonas infect a wide range of crops and wild plants, with most species responsible for plant diseases that have a global economic and environmental impact on the seed, plant, and food trade. Infections by Xanthomonas spp. cause a wide variety of non-specific symptoms, making their identification difficult. The coexistence of phylogenetically close strains, but drastically different in their phenotype, poses an added challenge to diagnosis. Data on future climate change scenarios predict an increase in the severity of epidemics and a geographical expansion of pathogens, increasing pressure on plant health services. In this context, the effectiveness of integrated disease management strategies strongly depends on the availability of rapid, sensitive, and specific diagnostic methods. The accumulation of genomic information in recent years has facilitated the identification of new DNA markers, a cornerstone for the development of more sensitive and specific methods. Nevertheless, the challenges that the taxonomic complexity of this genus represents in terms of diagnosis together with the fact that within the same bacterial species, groups of strains may interact with distinct host species demonstrate that there is still a long way to go. In this review, we describe and discuss the current molecular-based methods for the diagnosis and detection of regulated Xanthomonas, taxonomic and diversity studies in Xanthomonas and genomic approaches for molecular diagnosis.

Induction of Plant Resistance in Tobacco (Nicotiana tabacum) against Tomato Spotted Wilt Orthotospovirus through Foliar Application of dsRNA.

Thrips-transmitted tomato spotted wilt orthotospovirus (TSWV) continues to be a constraint to peanut, pepper, tobacco, and tomato production in Georgia and elsewhere. TSWV is being managed by an integrated disease management strategy that includes a combination of cultural practices, vector management, and growing virus-resistant varieties where available. We used a non-transgenic strategy to induce RNA interference (RNAi)-mediated resistance in tobacco (Nicotiana tabacum) plants against TSWV. Double-stranded RNA (dsRNA) molecules for the NSs (silencing suppressor) and N (nucleoprotein) genes were produced by a two-step PCR approach followed by in vitro transcription. When topically applied to tobacco leaves, both molecules elicited a resistance response. Host response to the treatments was measured by determining the time to symptom expression, and the level of resistance by absolute quantification of the virus. We also show the systemic movement of dsRNA_N from the inoculated leaves to younger, non-inoculated leaves. Post-application, viral siRNAs were detected for up to nine days in inoculated leaves and up to six days in non-inoculated leaves. The topical application of dsRNAs to induce RNAi represents an environmentally safe and efficient way to manage TSWV in tobacco crops and could be applicable to other TSWV-susceptible crops.