Foliar Symptoms Research Articles

Grapevine, peach, and plum response to simulated florpyrauxifen-benzyl drift

Abstract Off-target movement of rice herbicides is a concern in California, where sensitive crops are often grown nearby. Florpyrauxifen-benzyl and triclopyr are auxin-mimics commonly used in rice systems. To steward florpyrauxifen-benzyl around the time of its initial registration in the state, research was conducted to compare the onset of foliar symptoms from simulated florpyrauxifen-benzyl and triclopyr drift onto grapevine, peach, and plum. The rates were 1/200X, 1/100X, 1/33X, and 1/10X of the 29.4 g ai ha–1 florpyrauxifen-benzyl and 1/200X, 1/100X, and 1/33X of the 420.3 g ae ha–1 triclopyr rice use rates. Herbicides were applied on one side of one- to two-year-old peach and plum trees and one side of established grapevines in 2020 and 2021. The general symptoms for florpyrauxifen-benzyl and triclopyr were similar and included chlorosis, leaf curling, leaf distortion, leaf malformation, leaf crinkling, necrosis, and twisting on leaves. The symptoms from herbicides were observed on both sides of the grapevine canopy, whereas florpyrauxifen-benzyl symptoms on peach and plum were mostly observed on the treated side of the tree. Florpyrauxifen-benzyl and triclopyr symptoms were observed three days-after-treatment (DAT) for grapevines and seven DAT for peach and plum. In all crops, most symptoms persisted through 42 DAT. Some grape clusters showed deformation and dropping of berries. All treated crops gradually recovered during the season regardless of application rates. Because symptoms were relatively minor in peach and plum, this research suggested that application precautions that reduce off-site drift are likely to minimize the occurrence of significant injury. However, grapevines were more sensitive and showed injury symptoms up to 71% at 14 DAT with a 1/10X florpyrauxifen-benzyl simulated drift rate. Therefore, extra precautions such as using drift-management agents and proper wind speed conditions at the time of florpyrauxifen-benzyl applications may be necessary if there are nearby vineyards.

Sugarcane Yellow Leaf Virus – A Silent Threat?

Abstract Sugarcane yellow leaf virus (SCYLV) is the causal agent of yellow leaf disease, a significant threat to global sugarcane production. SCYLV is disseminated via infected planting material and transmitted by aphids, primarily by Melanaphis sacchari (Zehntner). The disease’s impact varies depending on the sugarcane cultivar, with some varieties showing little to no foliar symptoms, yet experiencing severe yield losses. Reliable diagnostic methods, including serological and molecular techniques, are crucial for accurate detection, though these methods vary in sensitivity. Advances in molecular techniques have revealed ten major SCYLV genotypes grouped into three phylogroups, further complicating detection efforts. Improved understanding of SCYLV’s distribution within the plant and its seasonal variability are yet to be understood. Management strategies focus on breeding SCYLV-resistant varieties, but this is challenging due to the virus’s variable impact on yield and the often asymptomatic nature of epidemics. Efforts to develop SCYLV-free propagative material have been implemented, but the virus can still spread via aphid vectors from neighbouring fields. Ongoing research aims to identify resistant cultivars, optimize diagnostic techniques, and better understand the epidemiology of SCYLV to mitigate its impact on sugarcane yield and quality. Information © The Authors 2024

Above‐ and belowground plant pathogens along elevational gradients: patterns and potential mechanisms

Plant pathogens are important for community assembly and ecosystem functioning and respond to a variety of abiotic and biotic factors, which may change along elevational gradients. Thus, elevational gradients are a valuable model system for exploring how environmental, plant community and soil factors influence pathogen communities. Yet, how these factors influence pathogens in natural ecosystems remains poorly understood. We examined the dynamics of plant fungal pathogens along elevational gradients, as well as the mechanisms shaping these dynamics, by combining a field survey on the Tibetan Plateau with a global meta‐analysis. In the field survey, increasing elevation was associated with a decrease in soil fungal pathogen richness but not in foliar fungal disease symptoms. Elevation was primarily related to soil fungal pathogen richness through abiotic factors, whereas no association was found between elevation and foliar fungal diseases. The meta‐analysis confirmed the generality of our field survey results: elevation was associated with a decrease in soil fungal pathogen richness, but it had no consistent relationship with foliar fungal diseases or pathogens. Thus, above‐ and belowground plant pathogen communities showed distinct elevational patterns, providing new insights into underlying mechanisms.

Prevalence of the Potato virus Y strain composition impacting Michigan seed potato production

Potato virus Y (PVY) is a complex of multiple strains that cause a variety of foliar and tuber symptoms. Several current strains have potential to result in variety-dependent yield and quality losses. Understanding the local PVY strain population and variety responses are critical for detection and management. From 2019-2023, PVY strain composition was monitored across seven Michigan seed potato-producing counties growing prevalent chip, russet, and tablestock market classes. Using multiplex reverse-transcriptase polymerase chain reaction methods, PVY strains were identified from potato tuber or leaf samples collected through the certification program (N = 78 positive lots). PVYN-Wi remained most prevalent in Michigan, however, tuber necrotic strains PVYNTN and PVYE also continued to be detected. These, as well as some PVYN-Wi isolates, are known to cause necrotic symptoms in tubers, causing yield and quality reductions for susceptible potato cultivars. As in other U.S. potato-growing regions, PVYN-Wi and PVYNTN were most prevalent in Michigan seed potato lots. Of note, mild to severe tuber necrosis was observed in diagnostic samples positive with local PVYN-Wi strains. Parental strain PVYO appeared to re-emerge in 2023 and should be considered in certification and management. Mild or asymptomatic reactions may still lead to yield or quality losses and may pose a risk to more susceptible cultivars. Due to the diversity of strains and varieties prevalent in the Great Lakes region, continued uniform certification is recommended. Information gathered from this and future survey efforts will be valuable for growers, inspectors, and future breeding and selection efforts.

First Report of Curvularia trifolii Causing Leaf Spot on Zanthoxylum bungeanum in China.

Chinese prickly ash (Zanthoxylum bungeanum Maxim.), which is a Rutaceae plant as shrubs or small trees and indigenous to China, is widely grown in this country for its flavor, as well as its high economic and ecological value. So far, in China, the cultivated area and yield of Chinese prickly ash rank first in the world. In June 2023, a leaf spot disease with approximately 30% incidence was observed on Z. bungeanum in Zhenfeng County (25°44'21.38″ N, 105°56'47.15″ E, 1,083 m), Guizhou Province, China. Foliar symptoms appeared as irregularly shaped lesions, yellowish-brown with dark brown margins surrounded by yellow halos, which enlarged, resulting in the lesions dropping from the leaves and leaving holes. To isolate and identify the pathogen, symptomatic leaves were taken and cut into 5 mm × 5 mm pieces, surface sterilized with 2% NaClO for 3 min, 75% ethanol for 30 s, rinsed three times with sterile water, and incubated on PDA at 28°C. Ten isolates with identical morphology were obtained. After one week of incubation at 28℃, the colonies on PDA were brown, reverse dark brown, fluffy, reaching 7.0-7.5 cm in diameter. Conidia were straight or slightly curved, narrowly ellipsoidal or fusiform, 1-3 but mostly 3 septate, light or dark brown, with the middle cells usually darker than the terminal cells, smooth, 20.5-31.0 × 9.0-19.0 μm (x̄ = 26.0 × 14.0 μm, n = 30). The morphological features matched the description of Curvularia trifolii (Kauffman) Boedijn (Ellis 1971; Falloon 1976). Additionally, the internal transcribed spacer (ITS), large subunit (LSU) and glyceraldehyde-3-phosphate dehydrogenase (gapdh) genes were amplified by PCR with primers ITS5/ITS4 (White et al. 1990), LROR/LR5 (Vilgalys & Hester 1990) and GPD1/GPD2 (Berbee et al. 1999), and the ITS, LSU and gapdh sequences of the isolate GUCC 23-321 (PP837870, PP837881, PP855474) were deposited in GenBank. The BLAST showed 98.5% (ITS, HG779023, 598/709 bp), 99.87% (LSU, HG779077, 779/858 bp), and 97.79% (gapdh, HG779124, 543/498 bp) identities with C. trifolii (CBS 173.55). Furthermore, the phylogenetic tree of ML analysis based on the combined sequence data of ITS, LSU and gapdh revealed that GUCC 23-321 clustered with C. trifolii. Both morphology and phylogenetic analyses supported the identification of GUCC 23-321 as C. trifolii. Pathogenicity tests were carried out twice according to Koch's postulates. Five healthy 2-year-old Chinese prickly ash plants were sprayed with a conidial suspension (1 × 106 conidia/mL) of the isolate GUCC 23-321, while the controls (five other plants) were sprayed with sterile water. All plants were maintained in a greenhouse at 28°C, 80% relative humidity. After 8 days, the inoculated plants developed leaf spots similar to those showed in the field, but control plants were asymptomatic. Re-isolation of pathogenic fungi from the leaf lesions of the inoculated plants and according to molecular analysis and morphology, the fungi were identified as C. trifolii, fulfilling Koch's postulates. C. trifolii is a common fungal phytopathogen that has been reported to infect a variety of plants and cause leaf spot disease, such as Trifolium alexandrinum (Khadka 2016) and Nicotiana tabacum (Chen et al. 2017). This is the first worldwide report of C. trifolii causing Z. bungeanum leaf spot. The report will be beneficial for accurately diagnosing this disease, and proposing specific control measures.

First Report of Leaf Blight of Syringa oblata Caused by Botryosphaeria dothidea in China.

As an important landscaping plant, Syringa oblata is widely planted in northern China with high ornamental, medicinal, and edible value (Men et al. 2023). In September 2023, a new leaf blight disease on S. oblata was observed in Tianjin (39.0916°N, 117.1019°E), China. The onset of foliar symptoms was marked by the appearance of yellow-brown spots that originated from the tip and margin, subsequently evolving into irregularly shaped brown lesions. Finally, the lesions are distributed throughout the leaf surface, causing the leaf to wilt and seriously affecting photosynthesis. To identify the pathogen responsible for leaf blight of S. oblata, symptomatic leaves were collected and cut into square leaf blocks with a size of 0.3 cm², which were sterilized by immersion in 75% ethanol for 60 s and 5% NaClO for 30 s, and rinsed three times with sterile distilled water. The sterilized leaf pieces were then placed on potato dextrose agar (PDA) and incubated at 25 °C for 3-5 days. The peripheral hyphae of the fungal colony which developed from the infected tissues were isolated onto PDA plates. The fungal cultures on PDA plates were used for morphological observation and identification of the fungus. Colonies of B. dothidea on PDA medium were initially off-white, batting-shaped and gradually grayish-black. Aerial hyphae were well developed and could reach the tip of the petri dish. To induce sporulation, the hyphae were picked into medium containing sterilized pine needles. Conidia were found on pine needles after 30 d of incubation at 25°C. Conidia were hyaline, unicellular, oblong to spindle-shaped, and 17.5-23.0 µm in size × 6.1-8.5 µm(n=50). Based on these characteristics, the isolates were preliminarily identified as B. dothidea (Phillips et al. 2005). To provide additional evidence for the classification of the isolate, genomic DNA was extracted from the isolates of B. dothidea and used for Polymerase Chain Reaction (PCR). The internal transcribed spacer (ITS), glyceraldehyde-3-phosphate dehydrogenase (GPDH), and actin (ACT) were amplified with the primer pairs ITS1/ITS4, GDP1/GDP2, and ACT-512F/ACT-783R, respectively (Jiang et al. 2022). A BLAST search of sequences showed the ITS, GPDH, and ACT sequences had >99% identity with homologous sequences from B. dothidea isolates Bb158-4(HQ392696.1), PPO-46523(MG761771.1), and CMW7779 (AY972117.1), respectively. Phylogenetic analysis determined that the isolate was in the same clade position as B. dothidea, which confirmed the above morphological identification. To assess pathogenicity, the fungal cakes (6 mm diameter) were obtained from the edge of a fresh colony (cultured on a PDA plate for 7 days) using a sterile perforator. Ten surface-sterilized leaves of healthy S. oblata with uniform growth condition were collected and inoculated with fungal cakes after wounding. Ten leaves were inoculated with sterile PDA medium blocks as control. The test was repeated three times. All leaves were kept at 25°C and sterile H2O was sprayed daily to keep leaves surface moist. After five days, all vaccination sites showed lesions similar to those of the S. oblata diseased leaves in the field, while the controls were asymptomatic. B. dothidea was reisolated from symptomatic tissues, thus fulfilling Koch's postulates. B. dothidea is a member of Botryosphaeriaceae. Currently, B. dothidea infection of Syringa spp. plants has not been reported. However, in China, it has been reported to cause stem rot on Forsythia suspense (He et al. 2022) and leaf dieback on sweet osmanthus (Ling et al. 2010), demonstrating that B. dothidea can infect Oleaceae species. This study found that S. oblata could be infected by B. dothidea. To the best of our knowledge, this is the first report of B. dothidea causing leaf blight on S. oblata in China. Identifying the pathogen of S. oblata leaf blight is essential for the prevention and management of disease associated with S. oblata.

Isolation, identification and multi-locus sequence typing of Phytophthora capsici from capsicum fields and its cross-infectivity in different crop species

Phytophthora capsici is a major soil-borne oomycete plant pathogen, which severely affects the productivity of peppers including Capsicum annuum L., by causing fruit rot, root rot, and foliar blight diseases. It has a broad host range beyond Capsicum species and evolves rapidly to find new potential hosts for survival under different climatic conditions. In the present study, P. capsici was isolated from soil using castor seed baiting, and characterized through growth patterns and spore morphology. The species identification and molecular characterization were achieved through multi-locus sequence and phylogenetic analysis of three genomic loci (ITS, tef1 and cox), and also attributed given P. capsici isolate Pc_UHF into India-specific ITS-based evolutionary lineage Pc-X. Interestingly, dN/dS estimates revealed reasonably high positive selection in the tef1 gene, which could have possible implications on the pathogen's adaptation and/or virulence. Pathogenicity of this isolate was tested in three commercially growing bell pepper varieties and also in other crop species using detached leaf and/or in planta disease screening. Differential disease symptoms were induced by Pc_UHF in three bell pepper varieties, such as both collar rot and foliar blight symptoms observed in "Yolo Wonder" and "California Wonder", whereas only foliar blight symptoms were recorded in "Solan Bharpur". Furthermore, cross-pathogenicity was also recorded in tomato, citrus, cucumber and eggplant. This is the first report of molecular characterization and cross-pathogenically assessment of P. capsici isolate from farm soils in the North Western Himalayan region of India, which has a dynamic virulence profile and distinct host range.

First report of Rhizoctonia blight caused by Rhizoctonia solani on Chinese cabbage (Brassica rapa subsp. chinensis) in India.

During January and February 2021, foliar blight symptoms were observed on the leaves of Chinese cabbage (Pak choi) at Lembucherra research farm, College of Agriculture, Tripura, India. The incidence of disease symptoms ranged from 5 to 10% of the plants observed in the field. The symptomatic leaves showed grayish colored water-soaked lesions with an irreguar shape and size. A total of 10 symptomatic leaves (1 leaf per plant) from Chinese cabbage infected plant were sampled, surface decontaminated with 1% NaOCl, washed twice in sterile water, plated on 2% water agar, and incubated at 25 ± 2°C. Hyphal tips from mycelium of 7-day old culture (2 isolates from two different plants) with right-angled branching were transferred to potato dextrose agar (PDA) media (SRL, India). Cream or light brown hyphae that branched at right angles, with septa near the point of the origin of hyphae, and a slight constriction at the base of the branch) were visible under a microscope. Olive-brown sclerotia were observed after 5 days of incubation. Multiple nuclei per cell were visible after staining with 4', 6-diamidino-2-phenylindole (Estandarte et al. 2016). Based on morphological characteristics (Parmeter et al. 1970) the isolates TP36 and TP37 were identified as Rhizoctonia solani. The internal transcribed spacer (ITS) region and glyceraldehyde-3-phosphate dehydrogenase gene (GAPDH) were amplified with ITS1& ITS4 (White et al. 1990) and (GAPDH F-5'- CAAGGAGAACCCAGGTGTTAAG-3' and GAPDH R- 5'-GGCGTCGAAGATAGAAGAGTGT-3') respectively for both isolates and sequenced (accession #. PP458158, PP458159, PP425343, PP425344). BLASTn analysis showed 99.26%( 668/673 nt) to 99.46% (659/664 nt) identity with R. solani sequences (GenBank MG397062.1 and KX674524.1) for ITS and 98.42% (552/562 nt) to 100% 540/540 nt)identity with R. solani sequences (GenBank HQ425709.1 and CP102644.1) for GAPDH. Isolates TP36 and TP37 were deposited in the Indian Type Culture Collection (ITCC), New Delhi as R. solani (nos. 9154 and 9319, respectively). Both isolates were amplified using (anastomosis group) AG1 subgroup specific primers (Matsumoto 2002; Prashantha et al. 2021) to identify their AG. The presence of a 265 bp amplicon for both isolates suggested that they belong to AG1-IA. A multilocus analysis of R. solani isolates from different host plants with concatenated sequences ITS and GAPH showed that TP36 and TP37 are closely related to rice isolate RS107. A pathogenicity test on five plants per treatment was conducted and repeated twice on one month old Chinese cabbage plants (hybrid, TOKITA, India) grown under glasshouse conditions in a sterilized mixture of soil and sand (3:1) at 27-28oC during January 2024 at ICAR-IARI, New Delhi. R. solani isolates TP36 and TP37 were grown on PDA and plants were inoculated by placing single sclerotia of 10-day old colony on different plant parts and covering it with moist cotton. After 7 day, typical lesions of R. solani infection were visible. No symptoms were observed on the control plants. The fungus was reisolated from the inoculated plants and identified as R. solani based on morphology. R. solani has previously been reported to cause disease on some members of Brassicaceae in different countries (Budge et al. 2009; Hua et al. 2014). Based on literature available this is the first report of R. solani infecting Chinese cabbage in India.

Unlocking Cassava Brown Streak Disease Resistance in Cassava: Insights from Genetic Variability and Combining Ability

Cassava brown streak disease (CBSD) threatens cassava production in sub-Saharan Africa despite the availability of resistant varieties. Extreme environmental factors weaken plant defenses, reducing CBSD resistance. This study examined CBSD inheritance in cassava populations, assessed genetic variability, and identified superior sources of resistance using F1, S1, and half-sib offspring populations derived from resistant sources. The offspring underwent field evaluation at two distinct sites from 2019 to 2021, and the symptom-free genotypes were analyzed using reverse transcription-quantitative polymerase chain reaction (RT-qPCR). Resistance to CBSD was categorized as most resistant, resistant, most tolerant, or tolerant based on symptoms and virus titers. The findings indicated that the resistance to CBSD is highly influenced by genotypes, F1/S1 types, and environmental conditions. An analysis of combining abilities revealed significant general combining abilities (GCAs) for CBSD, cassava mosaic disease (CMD), and traits associated with yield. The heritability estimates for resistance to CBSD varied between 43.4% and 63.2% for foliar symptoms and 14.6% and 57.9% for root necrosis across locations. The inheritance pattern involved a combination of additive and recessive genes with selfed (S1) populations displaying stronger and more effective resistance to the disease. The cassava brown streak virus (CBSV) was highly prevalent, and the Ugandan cassava brown streak virus (UCBSV) was not prevalent. Four genotypes were highly resistant to CBSD and could be key sources of resistance to this disease.

Identification of Sources of Resistance to Aphanomyces Root Rot in Pisum.

Aphanomyces root rot (ARR), caused by Aphanomyces euteiches, is one of the most devastating diseases that affect the production of peas. Several control strategies such as crop rotation, biocontrol, and fungicides have been proposed, but none provides a complete solution. Therefore, the deployment of resistant cultivars is fundamental. ARR resistance breeding is hampered by the moderate levels of resistance identified so far. The available screening protocols require post-inoculation root assessment, which is destructive, time-consuming, and tedious. In an attempt to address these limitations, we developed a non-destructive screening protocol based on foliar symptoms and used it to identify new sources of resistance in a Pisum spp. germplasm collection. Accessions were root inoculated separately with two A. euteiches isolates, and leaf symptoms were assessed at 5, 10, 14, 17, and 20 days after inoculation (DAI). Although the majority of accessions exhibited high levels of susceptibility, thirty of them exhibited moderate resistance. These thirty accessions were selected for a second experiment, in which they were inoculated with both A. euteiches isolates at two inoculum doses. The objective of this second trial was to confirm the resistance of these accessions by evaluating root and biomass loss, as well as foliar symptoms, and to compare root and foliar evaluations. As a result, a high correlation (R2 = 0.75) between foliar and root evaluations was observed, validating the foliar evaluation method. Notably, accessions from P.s. subsp. humile exhibited the lowest symptomatology across all evaluation methods, representing valuable genetic resources for breeding programs aimed at developing pea varieties resistant to ARR.

New Approaches to Plant Pathogen Detection and Disease Diagnosis.

Detecting plant pathogens and diagnosing diseases are critical components of successful pest management. These key areas have undergone significant advancements driven by breakthroughs in molecular biology and remote sensing technologies within the realm of precision agriculture. Notably, nucleic acid amplification techniques, with recent emphasis on sequencing procedures, particularly next-generation sequencing, have enabled improved DNA or RNA amplification detection protocols that now enable previously unthinkable strategies aimed at dissecting plant microbiota, including the disease-causing components. Simultaneously, the domain of remote sensing has seen the emergence of cutting-edge imaging sensor technologies and the integration of powerful computational tools, such as machine learning. These innovations enable spectral analysis of foliar symptoms and specific pathogen-induced alterations, making imaging spectroscopy and thermal imaging fundamental tools for large-scale disease surveillance and monitoring. These technologies contribute significantly to understanding the temporal and spatial dynamics of plant diseases.

First report of Triticum mosaic virus infecting oat crop in the United States.

Triticum mosaic virus (TriMV, genus Poacevirus, family Potyviridae) was first reported in 2006 (Seifers et al. 2008) to infect wheat, and since then, it has been established as a constraint for US wheat production (Byamukama et al. 2013). In the field, TriMV often exists as a coinfection with wheat streak mosaic virus (WSMV), and these two viruses interact synergistically to produce severe symptoms and greater yield loss (Byamukama et al. 2012; Tatineni et al. 2022). Both TriMV and WSMV are transmitted by wheat curl mites (Aceria tosichella Keifer) (McMechan et al. 2014). Wheat is the primary host reported for TriMV in the field, but Seifers et al. (2010) established oat, rye, barley, and several other cereals and grasses as hosts under controlled conditions. However, there are no documented cases of TriMV infecting oats in the field. Between 10-25 June, 2023, a total of 273 field oat plants showing foliar yellowing, yellow flecking, and streaking symptoms were collected from four different fields in Nebraska (Big Springs: 41.1029° N, 102.1451° W; Mead: 41.2292° N, 96.4938° W; Odell: 40.0459° N, 96.7984° W; Stumf: 40.5048° N, 101.4223° W). Total RNA was extracted using the MagMax Plant RNA Isolation kit (Thermo Fisher Scientific) and the KingFisher Flex Magnetic Particle Processor (Thermo Fisher Scientific) (Mondal et al. 2023). Sample RNA was assayed with a single-step multiplex reverse transcription polymerase chain reaction (RT-PCR) to determine presence of WSMV and TriMV. Out of 273 symptomatic oat plants, 254 (93.04%) tested positive for at least one virus. Out of total positive samples, 238 were positive for WSMV (93.70 %), 12 plants tested positive for both TriMV and WSMV (4.70%), and 4 plants were infected with TriMV alone (1.60%). As a secondary confirmation, amplified fragments from the TriMV single infection were gel purified using a gel extraction kit (QIAquick) and sequenced (Eurofins Genomics). The nucleotide sequences were analysed using the BlastN program, compiled, and edited in the BioEdit software (Hall 1999). Sequences were deposited in the NCBI GenBank database (accession number PP475806). Nucleotide BLAST searches of the target coat protein (CP) gene showed > 98% identity to the corresponding sequences in TriMV accession MK318274. For further validation, virus inoculum was prepared by grinding field-collected plant material from plants with only TriMV present in 20 mM sodium phosphate buffer, pH 7.0, and then mechanically inoculating two-week-old oats (cv. Shaw n=8) and wheat (cv. Sattler, n=8) plants. Three weeks post-inoculation, all the eight wheat plants exhibited mild yellowing and streaking symptoms, while oat plants did not show obvious foliar symptoms. All wheat and oat plants were further tested positive with DAC-ELISA (antibodies produced against TriMV CP at the USDA-ARS facility in Lincoln, NE) and with RT-PCR. The specific attribution of these symptoms to TriMV in oats is not possible as none produced prominent symptoms. Asymptomatic oat infection from symptomatic field-collected oat samples could be due to oat cultivar differences. Although the prevalence of TriMV in wheat has been established across the Great Plains of the United States, to our knowledge, this is the first report of TriMV infection in US oat fields. Our finding warrant further investigation into the incidence and impact of the virus in oat crop and its potential for serving as a asymptomatic virus reservoir.

Detection of fusarium wilt-induced physiological impairment in strawberry plants using hyperspectral imaging and machine learning

AbstractStrawberry (Fragraria x ananassa) is a crop affected by various soil-borne fungal pathogens with mostly non-specific foliar symptoms and often requiring laboratory isolation for correct diagnosis. Moreover, these nonspecific foliar symptoms, appreciated by the human eye, appear after some time following infection by the pathogen. Early detection of plant diseases is one of the primary objectives in agriculture because it may contribute to identifying more tolerant cultivars in breeding programs and optimise pesticide use in agricultural production with earlier applications in emerging disease foci. New technologies, such as remote sensing and machine learning (ML) algorithms, have arisen as potential tools to improve the ability to detect and classify different crop diseases. The combined use of hyperspectral imagery and ML algorithms were investigated to detect and classify the physiological stress caused by early infections of Fusarium wilt in strawberry plants. Six ML models, namely artificial neural network, decision tree, K-nearest neighbour, support vector machine, multinomial logistic regression and Naïve Bayes were developed to estimate physiological stress associated with Fusarium wilt disease. The results showed that stomatal conductance (gs) and photosynthesis (A) declined even without visual symptoms of the disease. Among the six ML models evaluated, the artificial neural network model showed the highest classification performance with an overall accuracy of 81%, regardless of the physiological parameter utilized for model training. Moreover, the artificial neural network accurately predicted the absolute values of both physiological parameters (gs and A) based on the complete spectral signature from visually healthy foliar tissue, achieving coefficients of determination of 84% and 81%, respectively. Consequently, ML models utilizing physiological response data and hyperspectral imaging exhibited remarkable robustness, facilitating the estimation of Fusarium wilt severity in strawberry plants even without visual symptoms.

A single viral amino acid shapes the root system architecture of a plant host upon virus infection

BackgroundGrapevine fanleaf virus (GFLV) is one of the most detrimental viral pathogens of grapevines worldwide but no information is available on its effect on the root system architecture (RSA) of plant hosts. We used two wildtype GFLV strains and their single amino acid mutants to assess RSA traits in infected Nicotiana benthamiana and evaluate transcriptomic changes in host root gene expression in replicated time course 3’RNA-Seq experiments. Mutations targeted the multi-functional GFLV-encoded protein 1EPol*/Sd, a putative RNA-dependent RNA polymerase and determinant of foliar symptoms in N. benthamiana plants.ResultsPlant infection with wildtype GFLV strain GHu and mutant GFLV strain F13 1EPolG802K, both carrying a lysine in position 802 of protein 1EPol*/Sd, resulted in a significantly lower number of root tips (-30%), and a significantly increased average root diameter (+ 20%) at 17 days post inoculation (dpi) in comparison with roots of mock inoculated plants. In contrast, the RSA of plants infected with wildtype GFLV strain F13 and mutant GFLV strain GHu 1EPolK802G, both carrying a glycine in position 802 of protein 1EPol*/Sd, resembled that of mock inoculated plants. Modifications of RSA traits were not associated with GFLV titer. Root tissue transcriptome analysis at 17 dpi indicated dysregulation of pattern recognition receptors, plant hormones, RNA silencing, and genes related to the production of reactive oxygen species (ROS). For wildtype GFLV strain GHu, RSA modifications were correlated with an abundant accumulation of ROS in the pericycle of primary roots at 7 dpi and the duration of vein clearing symptom expression in apical leaves. Dysegulation of a hypersensitive response was an overarching gene ontology found through enrichment analyses of 3’RNA-Seq data.ConclusionsOur findings revealed the causative role of lysine in position 802 of protein 1EPol*/Sd in a novel RSA phenotype during viral infection and documented GFLV-N. benthamiana interactions at the root level based on (i) antiviral response, (ii) receptor mediated production of ROS, and (iii) hormone regulation. A correlation between above and below ground symptoms was reported for the first time in plants infected with wildtype GFLV strain GHu. Further work is warranted to test whether the modified RSA of a plant host might impact GFLV acquisition and transmission by the ectoparasitic dagger nematode Xiphinema index.

Late-season decline, a bacterial disease of corn caused by Pantoea ananatis, identified in sorghum

Foliar symptoms resembling those associated with late-season decline (LSD) disease of corn, caused by the bacteria Pantoea ananatis, were observed in commercial fields of sorghum plants in 2023 in the Texas Panhandle. Diagnosis of symptomatic tissues consistently recovered bacterial isolates that were subsequently identified, based on partial sequences of their respective 16S rRNA gene, as P. ananatis. The pathogenicity of a representative isolate on sorghum was investigated using the method of Koch’s postulate and resulted in the successful reproduction of LSD-like foliar symptoms identical to those described for corn, as well as those observed in affected sorghum fields.

First Report of Watermelon Crinkle Leaf-Associated Virus-2 (WCLaV-2) in Watermelon (Citrullus lanatus) in Georgia, USA.

Watermelon (Citrullus lanatus) is a key horticultural crop in Georgia with farmgate value of 142 million USD (2022 Farm Gate Value Report), yet faces challenges from whitefly-transmitted viruses, especially during periods of elevated whitefly populations in the fall. Foliar symptoms on watermelon plants including yellow mottling and chlorosis, wrinkling, bunching, and upward curling, were observed in experimental fields at UGA Tifton and commercial fields in Colquitt County in the fall of 2023. These were similar to those described for watermelon crinkle leaf-associated viruses (WCLaV-1 and WCLaV-2) from Florida (Hendrick et al, 2021) and Texas (Hernandez et al., 2021). The disease incidence reached 100% in both locations. WCLaV-1 was previously identified in Georgia (Adeleke et al., 2022a); however, WCLaV-2 remained undetected in further surveys (Adeleke et al., 2022b). Total nucleic acid was extracted from symptomatic leaf tissues with the MagMAXTM 96 Viral RNA isolation kit (ThermoFisher Scientific, USA), following the manufacturer's guidelines, with the omission of DNAse treatment. The presence of WCLaV-1 was identified through reverse transcription-polymerase chain reaction (RT-PCR) (Hernandez et al., 2021) in 17 out of 24 samples in Tift County, and 12 out of 15 samples from Colquitt County. Within the same set of samples, WCLaV-2 was identified in three samples from Tift County and four samples from Colquitt County by RT-PCR directed at the RdRp gene (Hernandez et al., 2021). WCLaV-1 was not detected in the three samples from Tift County that had WCLaV-2, while mixed infections of WCLaV-2 and WCLaV-1 were observed in the samples form Colquitt County. Two whitefly transmitted viruses, previously reported in Georgia were also identified as mixed infection in these samples (Table S1). The presence of WCLaV-2 was further confirmed by amplifying the movement protein (MP) gene of WCLaV-2 by RT-PCR assays (Hernandez et al., 2021). The amplicons, with expected sizes of 968bp for the RdRp gene and 562bp for the MP gene of WCLaV-2, located on RNA 1 and RNA 2 segments respectively, were directly sequenced from both directions (Genewiz, USA) from a sample collected in Tift County. The resulting data were analyzed via BLASTn search. The MP gene fragment (PP178543) shared 100% identity with isolates from Brazil (LC636074.1), Texas (MW559086.1), and Florida (MZ325858.1). RdRp gene (PP178542) shared >99.7% identity with isolates from Brazil (LC636073.1), Texas (MW559083.1) and Florida (MZ325855.1). WCLaV-1 and WCLaV-2, initially discovered in Asia (Xin et al., 2017), have been assigned to the genus Coguvirus, in the family Phenuiviridae (Walker et al., 2022). Subsequent reports from the USA (Hendrick et al., 2021; Hernandez et al., 2021), Australia (Mulholland et al., 2023), and Brazil (Maeda et al., 2022) indicate the global spread of these viruses. Watermelon is the primary host of WCLaV-2. Despite these findings, biological information, including vector relations, for both viruses and other members of the genus Coguvirus remains elusive. The impact of these viruses on watermelon production and yield in the identified regions remains largely unknown, underscoring the need for further investigations.

Standard area diagrams for severity assessment of eucalypt bacterial leaf blight caused by Xanthomonas spp.

AbstractBacterial leaf blight (BLB) caused by Xanthomonas spp. is an important threat for eucalypt production. Planting of resistant genotypes is the best strategy for eucalypt BLB control. However, selection of resistant clones is challenging since the disease is caused by at least three Xanthomonas species that can be sympatric and cause a range of foliar symptoms. So far, standard area diagrams (SADs) to assess eucalypt BLB severity caused by different Xanthomonas species have not been reported. Here, SADs for eucalypt BLB severity assessment were developed. Plants of five eucalypt genotypes were individually inoculated with different Xanthomonas species and leaves exhibiting lesions varying in colour, shape, and size were digitally scanned for disease assessment. Among 150 scanned leaves, the actual leaf affected area ranged from 0.01% to 55%. Based on these minimum and maximum values and the frequency of severity distribution, the 10 leaves that best represented the distribution of affected leaf area were selected to assemble the SADs. Groups of experienced and inexperienced raters estimated BLB severity on a subset of 50 out of the 150 original leaves without and then with the use of the SADs. Correlation concordance analyses revealed that SADs provided a significant improvement in accuracy, precision and reliability of disease estimates by both rater groups, with higher gains observed for the inexperienced group. The SADs developed in this study will be useful for selection of eucalypt genotypes resistant to BLB under controlled conditions when knowledge on the Xanthomonas species composition is lacking or when local populations are comprised of several species.

Large gradient of susceptibility to esca disease revealed by long-term monitoring of 46 grapevine cultivars in a common garden vineyard

Grapevine (Vitis vinifera L.) is prone to many fungal diseases, including esca, a severe vascular disease threatening the wine sector and for which there is no cost-effective cure. Susceptibility to esca varies between cultivars in different infection conditions. It may therefore be possible to use the genetic diversity of grapevine cultivars to mitigate disease impact. However, the genetic component of esca susceptibility has rarely been investigated in the vineyard, and the specific mechanisms and varietal traits underlying esca susceptibility remain unknown.In this study, we monitored the incidence and severity of esca foliar symptoms and plant dieback (apoplexy and mortality) at plant level for seven years, on 46 cultivars planted in an experimental common garden, to separate the genetic component of esca susceptibility from the effects of environment and cropping practices. We observed a broad gradient of varietal susceptibility, with a mean incidence of 0 to 26 % of vines expressing esca foliar symptoms depending on the variety. This gradient remained similar across years and, unlike the severity of foliar symptoms, the incidence of grapevine dieback was significantly correlated with that of foliar symptoms. We detected a significant but weak and very localised phylogenetic signal for the incidence of esca foliar symptoms in this panel of cultivars.We then explored the relationships between epidemiological metrics and ecophysiological and phenological traits phenotyped on the same plot. Esca disease incidence was negatively correlated with δ13C across cultivars, suggesting that varieties with higher water use efficiency are less prone to the expression of esca symptoms on leaves. Moreover, the least vigorous cultivars were among the least susceptible, although this relationship was not significant. By contrast, neither phenological stages nor nitrogen status were significantly predictive of cultivar susceptibility to the disease.Together, these results provide new insight into the potential of genetic resources for use in the sustainable management of grapevine trunk diseases and open up new perspectives for studying the pathological and physiological determinants of their incidence.

First report of the new Neopestalotiopsis species causing strawberry leaf spot and fruit rot in Georgia.

In April 2023, symptomatic strawberry (Fragaria × ananassa) plants (cv. 'Camarosa' and 'Florida Brilliance') were observed at a commercial farm in Worth County, GA (USA). Symptoms included foliar, irregularly distributed, and different-sized spots (dark brown with light brown centers) and dark brown V-shaped necrotic areas starting at the leaf edge. By the time of sample collection, ~50% incidence was reported in the field. Leaf samples were collected and shipped overnight to the laboratory. Black acervuli were observed readily on old necrotic foliar lesions. Conidial morphology was consistent with that observed with Neopestalotiopsis species (Maharachchikumbura et al. 2014). Conidia were ellipsoid to fusiform, five-celled, with three light brown median cells and one hyaline apical and basal cell. Apical cells had two-to-four flexuous appendages, and the basal cell had one non-flexuous appendage (Fig 1). The average (n = 20) conidia length, not including the appendages, was 26.6 µm (SD: 2.8), and width was 6.3 µm (SD: 0.94). Fungal isolation was conducted on acidified PDA and incubated at 25°C for 6 days. Dense, white mycelia were observed on the upper plate surface, while a pale pink/orange coloration was observed on the underside (Fig 1). Black acervuli formed on the surface of the white mycelial mat. Six isolates were purified and selected to confirm the species identity. DNA was extracted from 6-day-old cultures and PCR was conducted following Kaur et al. (2023). Amplified DNA was digested with the restriction enzyme BsaWI and two bands were clearly visualized (~130 and ~290 bp), along with a faint band of 20-bp (Fig 2). Four of the six isolates were selected for sequencing of the β-tubulin gene. BLAST queries using the consensus sequence showed that all isolates had 100% identity to strain N21002 from Florida (FL), characterized as Neopestalotiopsis sp. (Kaur et al. 2023). One representative isolate (AJ07-2023) was deposited in GenBank (accession No. PP316103). Pathogenicity tests were performed on 27-day-old transplants of Sensation 'Florida127' provided by Natalia Peres from the UF. Plants were grown in 10.5 cm pots in the greenhouse. Isolate AJ07-2023 was grown on PDA for 30 days at 25°C, and the spore suspension was adjusted at 106 spore/ml. Five strawberry plants were sprayed with 5 ml of inoculum using a Preval sprayer with a CO2 canister, and 5 plants were sprayed with sterile distilled water. Plants were placed in a growth chamber for 6 days and covered with plastic bags after the sixth day to maintain ~85% relative humidity and 25°C. Foliar symptoms, including dark-brown circular lesions occurring towards the edge of leaves with light-brown center and light-yellow halo, developed 13 days after inoculation. No symptoms were observed on control plants. Neopestalotiopsis sp. was reisolated from inoculated plants as described above. Colony, conidial morphology, and PCR results were consistent with the original isolates. Neopestalotiopsis disease has been reported on strawberry in FL (Baggio et al. 2021), OH (Rotondo et al. 2022) and IN (Guan et al. 2023). Although the disease has been observed sporadically in GA since 2020 (Brannen, personal communication), to our knowledge, this is the first official report of the new Neopestalotiopsis sp. in GA. It has been reported that this new strain is more aggressive on fruits and leaves than other Neopestalotiopsis spp. (Baggio et al. 2021), therefore, accurate identification is critical for proper management.

Thiosulfinate Tolerance Gene Clusters Are Common Features of Burkholderia Onion Pathogens.

Burkholderia gladioli pv. alliicola, B. cepacia, and B. orbicola are common bacterial pathogens of onion. Onions produce organosulfur thiosulfinate defensive compounds after cellular decompartmentalization. Using whole-genome sequencing and in silico analysis, we identified putative thiosulfinate tolerance gene (TTG) clusters in multiple onion-associated Burkholderia species similar to those characterized in other Allium-associated bacterial endophytes and pathogens. Sequence analysis revealed the presence of three Burkholderia TTG cluster types, with both Type A and Type B being broadly distributed in B. gladioli, B. cepacia, and B. orbicola in both the chromosome and plasmids. Based on isolate natural variation and generation of isogenic strains, we determined the in vitro and in vivo contribution of TTG clusters in B. gladioli, B. cepacia, and B. orbicola. The Burkholderia TTG clusters contributed to enhanced allicin tolerance and improved growth in filtered onion extracts by all three species. TTG clusters also made clear contributions to B. gladioli foliar necrosis symptoms and bacterial populations. Surprisingly, the TTG cluster did not contribute to bacterial populations in onion bulb scales by these three species. Based on our findings, we hypothesize onion-associated Burkholderia may evade or inhibit the production of thiosulfinates in onion bulb tissues. [Formula: see text] Copyright © 2024 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.