Characterization of QoI-Fungicide Resistance in Cercospora Isolates Associated with Cercospora Leaf Blight of Soybean from the Southern United States.

Cercospora leaf blight (CLB) of soybean, caused by Cercospora cf. flagellaris, C. kikuchii, and C. cf. sigesbeckiae, is an economically important disease in the southern United States. Cultivar resistance to CLB is inconsistent; therefore, fungicides in the quinone outside inhibitor (QoI) class have been relied on to manage the disease. Approximately 620 isolates from plants exhibiting CLB were collected between 2018 and 2021 from 19 locations in eight southern states. A novel polymerase chain reaction-restriction fragment length polymorphism (PCR-RFLP) assay based on two genes, calmodulin and histone h3, was developed to differentiate between the dominant species of Cercospora, C. cf. flagellaris, and C. cf. sigesbeckiae. A multilocus phylogenetic analysis of actin, calmodulin, histone h3, ITS rDNA, and transcription elongation factor 1-α was used to confirm PCR-RFLP results and identify remaining isolates. Approximately 80% of the isolates collected were identified as C. cf. flagellaris, while 15% classified as C. cf. sigesbeckiae, 2% as C. kikuchii, and 3% as previously unreported Cercospora species associated with CLB in the United States. PCR-RFLP of cytochrome b (cytb) identified QoI-resistance conferred by the G143A substitution. Approximately 64 to 83% of isolates were determined to be QoI-resistant, and all contained the G143A substitution. Results of discriminatory dose assays using azoxystrobin (1 ppm) were 100% consistent with PCR-RFLP results. To our knowledge, this constitutes the first report of QoI resistance in CLB pathogen populations from Alabama, Arkansas, Kentucky, Mississippi, Missouri, Tennessee, and Texas. In areas where high frequencies of resistance have been identified, QoI fungicides should be avoided, and fungicide products with alternative modes-of-action should be utilized in the absence of CLB-resistant soybean cultivars.

De novo genome sequence of endophytic biocontrol strain Bacillus sp. ST24 isolated from rice seed.

In this study, we present the genome sequence of Bacillus sp. strain ST24, an endophytic bacterium isolated from rice seeds. The genome assembly comprises a total of 5,799,877 bp, with a GC content of 34.81%. Furthermore, our analysis revealed the presence of various genes associated with antibiotic production, as well as genes involved in polyketide biosynthesis and non-ribosomal polyketide-like clusters.

Multiple Biosynthetic Gene Pathways are Revealed in Whole-Genome Sequence of Endophytic Biocontrol Strain Pantoea stewartii subsp. indologenes ST25.

This report describes the draft genome sequence of Pantoea stewartii subsp. indologenes strain ST25, a biocontrol endophyte that was isolated from rice seed in Texas, USA. The genome assembly is 4,787,268 bp, with a GC content of 53.62%.

A High-Quality Genome Assembly for Cercospora janseana, Causal Agent of Narrow Brown Leaf Spot of Rice.

Cercospora janseana causes narrow brown leaf spot of rice. A nearly complete telomere-to-telomere reference genome was assembled with a combination of Oxford Nanopore and Illumina sequences. The genome assembly has a total length of 39,075,509 bp and consists of 15 chromosomes, 14 of which have telomeric repeats at both ends. The assembly N50 is 2.97 Mb and the L50 is five contigs. RNA-seq-mediated gene annotation identified 10,850 genes, including 955 predicted secreted proteins and 361 predicted effector proteins. This highly contiguous and almost complete C. janseana reference genome will be a vital resource for further investigation of host-pathogen interactions and genome evolution within this pathosystem. [Formula: see text] Copyright © 2023 The Author(s). This is an open access article distributed under the CC BY-NC-ND 4.0 International license.

Kernel Smut and False Smut: The Old-Emerging Diseases of Rice-A Review.

Kernel smut, caused by Tilletia horrida, is a disease characterized by the replacement of rice grains with black sooty masses of teliospores or chlamydospores. Kernel smut differs from rice false smut, caused by Ustilaginoidea virens, in the color of chlamydospores. False smut is characterized by globose, velvety spore balls ranging from orangish yellow to greenish black in color. Both kernel smut and false smut have been persistent but are considered minor diseases in many countries since they were discovered in the late 1870s to the 1980s due to their sporadic outbreaks and limited economic impacts. In recent years, however, kernel smut and false smut have emerged as two of the most economically important diseases in rice, including organic rice, in many countries, especially in the United States. The increased use of susceptible rice cultivars, especially hybrids, excessive use of nitrogen fertilizer, and short crop rotations have resulted in an increase in kernel smut and false smut, causing significant losses in grain yield and quality. In this article, we provide a review of the distribution and economic importance of kernel smut; our current understanding of the taxonomy, biology, and epidemiology of kernel smut; and the genomics of the kernel smut fungus as compared with false smut and its causal agent. We also provide an update on the current management strategies of pathogen exclusion, cultivar resistance, fungicides, biological control, and cultural practices for kernel smut and false smut of rice.

Wheat lodging extraction using Improved_Unet network.

The accurate extraction of wheat lodging areas can provide important technical support for post-disaster yield loss assessment and lodging-resistant wheat breeding. At present, wheat lodging assessment is facing the contradiction between timeliness and accuracy, and there is also a lack of effective lodging extraction methods. This study aims to propose a wheat lodging assessment method applicable to multiple Unmanned Aerial Vehicle (UAV) flight heights. The quadrotor UAV was used to collect high-definition images of wheat canopy at the grain filling and maturity stages, and the Unet network was evaluated and improved by introducing the Involution operator and Dense block module. The performance of the Improved_Unet was determined using the data collected from different flight heights, and the robustness of the improved network was verified with data from different years in two different geographical locations. The results of analyses show that (1) the Improved_Unet network was better than other networks (Segnet, Unet and DeeplabV3+ networks) evaluated in terms of segmentation accuracy, with the average improvement of each indicator being 3% and the maximum average improvement being 6%. The Improved_Unet network was more effective in extracting wheat lodging areas at the maturity stage. The four evaluation indicators, Precision, Dice, Recall, and Accuracy, were all the highest, which were 0.907, 0.929, 0.884, and 0.933, respectively; (2) the Improved_Unet network had the strongest robustness, and its Precision, Dice, Recall, and Accuracy reached 0.851, 0.892, 0.844, and 0.885, respectively, at the verification stage of using lodging data from other wheat production areas; and (3) the flight height had an influence on the lodging segmentation accuracy. The results of verification show that the 20-m flight height performed the best among the flight heights of 20, 40, 80 and 120 m evaluated, and the segmentation accuracy decreased with the increase of the flight height. The Precision, Dice, Recall, and Accuracy of the Improved_Unet changed from 0.907 to 0.845, from 0.929 to 0.864, from 0.884 to 0.841, and from 0.933 to 0.881, respectively. The results demonstrate the improved ability of the Improved-Unet to extract wheat lodging features. The proposed deep learning network can effectively extract the areas of wheat lodging, and the different height fusion models developed from this study can provide a more comprehensive reference for the automatic extraction of wheat lodging.

Multi-Locus Sequence Analysis Reveals Diversity of the Rice Kernel Smut Populations in the United States.

Rice (Oryza sativa) is the second leading cereal crop in the world and is one of the most important field crops in the US, valued at approximately $2.5 billion. Kernel smut (Tilletia horrida Tak.), once considered as a minor disease, is now an emerging economically important disease in the US. In this study, we used multi-locus sequence analysis to investigate the genetic diversity of 63 isolates of T. horrida collected from various rice-growing areas across in the US. Three different phylogeny analyses (maximum likelihood, neighbor-joining, and minimum evolution) were conducted based on the gene sequence sets, consisting of all four genes concatenated together, two rRNA regions concatenated together, and only ITS region sequences. The results of multi-gene analyses revealed the presence of four clades in the US populations, with 59% of the isolates clustering together. The populations collected from Mississippi and Louisiana were found to be the most diverse, whereas the populations from Arkansas and California were the least diverse. Similarly, ITS region-based analysis revealed that there were three clades in the T. horrida populations, with a majority (76%) of the isolates clustering together along with the 22 Tilletia spp. from eight different countries (Australia, China, India, Korea, Pakistan, Taiwan, The US, and Vietnam) that were grouped together. Two of the three clades in the ITS region-based phylogeny consisted of the isolates reported from multiple countries, suggesting potential multiple entries of T. horrida into the US. This is the first multi-locus analysis of T. horrida populations. The results will help develop effective management strategies, especially breeding for resistant cultivars, for the control of kernel smut in rice.

First Report of Fusarium Sheath Rot of Rice Caused by Fusarium incarnatum-equiseti Species Complex in the United States.

Fungal diseases, including sheath rot (Sarocladium oryzae), cause significant losses of yield and milling quality of rice (Oryza sativa). In August 2021, symptoms like sheath rot were observed on 20% of rice plants (cv. Presidio) in 1-hectare field in Eagle Lake, Texas. Initial lesions occurred on the upper flag leaf sheaths and were oblong or irregular oval, with gray to light brown centers, and a dark reddish-brown diffuse margin. Lesions enlarged, coalesced, and covered a large area of the sheath. Infection led to panicle rot with kernels turning dark brown. Unlike sheath rot, sheath infection also led to inside culm infection with irregular dark brown lesions. Infected tissue pieces were sterilized with 1% NaOCl for 2 min, followed by 75% ethanol for 30 s, washed in sterile H2O three times, air dried and incubated on PDA at 27℃. Fungal isolates were obtained from 15 diseased plant samples and their singled-spored fungal colonies were whitish, loosely floccose and produced light yellow pigmentation. On carnation leaf agar, macroconidia were slightly curved and tapered at the ends, with 3 to 5 septa, and measured 17.5 to 34.3 × 3.1 to 5.0 µm. Microconidia were ovoid, usually with 0 to 1 septum and were 4.0 to 15.5 × 2.5 to 4.5 µm. Spherical shaped chlamydospores were produced in chain. These morphological characteristics were consistent to those described for Fusarium incarnatum-equiseti species complex (O'Donnell et al. 2009), including F. incarnatum (Wang et al. 2021) and F. equiseti (Avila et al. 2019). For molecular identification, DNA of a representative isolate was extracted and ITS, LSU, and EF1 of the fungus were amplified using the primers of ITS1/ITS4 (Wang et al. 2014), D1/D2 domain region of LSU (Fell et al. 2000), and EF1 (Wang et al. 2014), respectively, and sequenced. The ITS sequence (OL344049) was 99.61% identical to F. incarnatum-equiseti species complex (FD_01692) in Fusarium-ID database and 99.61% identical to F. equiseti (LC514690, KY523100, MW016539) and F. incarnatum (MH979697) in NCBI database. The LSU sequence (OK559512) was 98.77% similar to F. equiseti (MN877913, MN368509) and F. incarnatum (MH877332, MH877326); the EF1 sequence (OK570044) was 99.27% similar to F. equiseti (MK278902) in NCBI database. A phylogenetic analysis based on the concatenated nucleotide sequences grouped this isolate in the F. incarnatum-equiseti species complex clade at 100% bootstrap support. To evaluate pathogenicity, a conidial suspension of 1 x 106 conidia/ml or sterilized water (the controls) was injected into the sheaths and young panicles of three rice plants (cv. Presidio) at boot. Treated plants were maintained in a greenhouse at 25 to 30℃. After 3 weeks, typical symptoms, like those observed in the field, developed on the inoculated plants but not on the controls. The same fungus was consistently re-isolated from the diseased plants. To our knowledge, this is the first report of Fusarium sheath rot caused by F. incarnatum-equiseti species complex in rice in the U. S. F. incarnatum-equiseti species complex has been reported to be associated with panicle infection in wild rice (O. latifolia) in Brazil (Tralamazza et al. 2021). F. incarnatum has also been reported to cause panicle rot in China (Wang et al. 2021). F. proliferatum has been reported to cause Fusarium sheath rot in India (Prabhukarthikeyan et al. 2021) and the U. S. (Cartwright et al. 1995). This research demonstrates the potential of different pathogens being involved in causing sheath rot of rice.

First Report of Leaf Spot of Rice Caused by Epicoccum sorghinum in the United States.

Brown spot (Cochliobolus miyabeanus), blast (Magnaporthe oryzae) and stackburn (Alternaria padwickii) are common diseases in rice with similar leaf spot symptoms. In August 2021, a leaf spot disease, with symptoms dissimilar to these diseases, occurred on almost 100% of the leaves and sheaths of rice plants (cv. Presidio) in a 1-hectare field in Eagle Lake, Texas. Lesions started as small dark brown spots on lower leaves and sheaths. The spots enlarged to become round or oval (1.5 to 5.0 mm) spots having round ends with gray centers, dark-brown borders or rings, and slight gold halos. The spots on the sheaths were similar to those on the leaf blades, with lesion size ranging from 2 to 5 mm. Pieces of infected tissue were cut from the margin of necrotic lesions, surface disinfected with 1% NaOCl for 2 min followed by 75% ethanol for 30 s and rinsed with sterile distilled water three times. The tissues were then dried on sterilized filter paper, placed on potato dextrose agar (PDA), and incubated at 25℃ for 7 days. Two isolates (LS36 and LS37) were obtained, and their colonies were initially villose, gray at the center and pale at the margin, and then turned dark gray, with the reverse side becoming scarlet. Chlamydospores were unicellular or multicellular and massively produced in nearly spherical shape (11 to 26 × 10 to 22 µm, n=100). Pycnidia were dark and mostly spheroid (105 to 171 × 76 to 128 µm, n=100). Conidia were unicellular, hyaline, ellipsoidal, with the size of 3.6 to 5.8× 1.9 to 2.8 µm (n=100). These morphological characteristics were similar to those described for Epicoccum sorghinum (Zhou et al. 2018). The rDNA internal transcribed spacer (ITS), rRNA large subunit (LSU), and translation elongation factor 1 alpha (EF1) gene of an representative isolate (LS37) were amplified (Fell et al. 2000; Wang et al. 2014) and sequenced. The ITS sequence (OK189534) of the isolate was 96.95% identical to E. sorghinum (KX758542); the EF1 sequence (OK236518) was 98.37% identical to E. sorghinum (MN461167); and the LSU sequence (OK189535) was 99.29% identical to E. sorghinum (MK817520, MK817521, and MK817522). Rice plants (cv. Presidio) at heading were inoculated with the two isolates individually by placing a drop of conidial suspension of 1 x 106 conidia/ml or a 2-mm PDA plug of 7-day-old cultures on the wounded or unwounded leaves and sheaths (3 sites/leaf or sheath, 3 plants/treatment). The wound was made by penetrating the epidermis using a 0.5-mm-diameter pin. The plants inoculated with sterilized water or PDA-only plugs served as the controls. The treated plants were placed in a dew chamber at 26℃ for 2 days and then transferred in a greenhouse (25 to 30℃). After 5 days, typical symptoms, similar to those observed in the field, developed on all of the inoculated leaves and sheaths, with the wound inoculation inducing more rapid development of symptoms than the unwounded inoculation. No symptoms developed on the controls. The two isolates produced similar symptoms and the fungus was consistently re-isolated from the infected plants and confirmed to be E. sorghinum based on morphological characteristics. The pathogenicity test was repeated twice with similar results. To our knowledge, this is the first report of leaf spot caused by E. sorghinum in rice in the United States. This disease was first reported on rice in China in 2020 (Liu et al. 2020). This research will help identify this new disease from other leaf spot-like diseases and develop management strategies for control of this disease.

First Report of Brown Leaf Spot of Rice Caused by Curvularia hawaiiensis in the United States.

Multiple diseases, including brown spot (Cochliobolus miyabeanus), leaf spot (Epicoccum sorghimum), and blast (Magnaporthe oryzae), can cause spot-like symptoms on the leaves of rice. In July 2021, a disease showing symptoms like brown spot was observed in an 8-hectare field of rice, with disease incidence of >30%, in Beaumont, Texas. Lesions started as small pinhead-size blackish spots on leaf tips or from the edges of leaf blades. The spots enlarged to become irregular (most) or oval brown spots with a slight chlorotic halo. Diseased leaves were collected, washed in running tap water and cut into small pieces. Pieces of the tissue were surface sterilized with 1%NaOCl for 2 min followed by 75% ethanol for 30 s and then washed in sterile distilled water three times with each time lasting for 1 min. The disinfected tissue pieces were air dried, placed on potato dextrose agar (PDA) medium and incubated at 25℃. Initially fungal colonies were hairy in texture with light dark brown center and whitish edge and dark brown pigmentation at the reverse side. Mature colonies turned to black in the center and dark brown toward the edge, with black at the reverse side after 2 or more weeks of incubation. Conidia were oval to narrowly oblong, rounded at the ends, with 2 to 6 distoseptate, and 15 to 35 × 6 to 10 µm in size. These morphological characteristics were similar to those described for Curvularia hawaiiensis (Aslam et al. 2019; Ellis 1971; Kusai et al. 2015). For molecular identification, DNA was extracted and the two different rRNA regions internal transcribed spacer (ITS) and large subunit (LSU), and the two genes RNA Polymerase II (RPB1) and translation elongation factor 1 alpha (EF1) of the fungus were amplified using the primers of ITS1/ITS4 (Wang et al. 2014), D1/D2 domain region of LSU (Fell et al. 200), and RPB1 and EF1 (Wang et al. 2014), respectively, and sequenced. The ITS sequence (OK397200) was 98.27% identical to C. hawaiiensis (KP131943); the EF1 sequence (OK492159) was 99.78% identical to C. hawaiiensis (KC503942); the LSU sequence (OK397295) was 98.96% identical to multiple C. hawaiiensis (MN055715, MH160813, MH875853, etc.); the RPB1 sequence (OK492160) was 97.41% identical to C. hawaiiensis (JN992363). To evaluate pathogenicity, three rice plants (cv. Presidio) at the 3-leaf stage were spray inoculated with a conidial suspension of 1 x 106 conidia/ml. Another set of three plants that were sprayed with sterilized distilled water served as the controls. Treated plants were maintained in a greenhouse with temperature ranging from 25 to 30℃. After 2 weeks, typical symptoms, like those observed in the field, developed on the inoculated plants while no symptoms developed on the control plants. The same fungus was consistently re-isolated from the diseased plants. The pathogenicity test was conducted three times with similar results. To our knowledge, this is the first report of brown leaf spot caused by C. hawaiiensis in rice in the United States. Curvularia species are frequently associated with rice grain and cause blackish discoloration symptoms on grain kernels. Recently, however, C. hawaiiensis has also been reported to cause brown leaf spot in Malaysia (Kusai et al. 2015) and Pakistan (Aslam et al. 2019). This research will help identify this disease from other leaf spot-like diseases and develop effective management strategies.

A Symbiotic Approach to Generating Stress Tolerant Crops.

Studies were undertaken to determine if fungal endophytes from plants in stressful habitats could be commercialized to generate climate resilient crop plants. Fungal endophytes were isolated from weedy rice plants and grasses from South Korea and the USA, respectively. Endophytes (Curvularia brachyspora and Fusarium asiaticum) from weedy rice plants from high salt or drought stressed habitats in South Korea conferred salt and drought stress tolerance to weedy rice and commercial varieties reflective of the habitats from which they were isolated. Fungal endophytes isolated from grasses in arid habitats of the USA were identified as Trichoderma harzianum and conferred drought and heat stress tolerance to monocots and eudicots. Two T. harzianum isolates were exposed to UV mutagenesis to derive strains resistant to fungicides in seed treatment plant protection packages. Three strains that collectively had resistance to commonly used fungicides were used for field testing. The three-strain mixture (ThSM3a) increased crop yields proportionally to the level of stress plants experienced with average yields up to 52% under high and 3-5% in low stress conditions. This study demonstrates fungal endophytes can be developed as viable commercial tools for rapidly generating climate resilient crops to enhance agricultural sustainability.

Hybrid Rice Outperforms Inbred Rice in Resistance to Sheath Blight and Narrow Brown Leaf Spot.

Sheath blight (ShB; caused by Rhizoctonia solani AG1-1A) and narrow brown leaf spot (NBLS; Cercospora janseana) are among the most important diseases affecting rice production in Texas and other southern parts of the United States. Because of the high yield potential, hybrid rice acreage is continually increasing. Understanding the relative levels of resistance to ShB and NBLS in hybrids compared to those of inbreds is important for effective disease management; however, this information remains largely unknown. A comparison of the performance of hybrid rice and inbred rice was performed involving 173 hybrid and 155 inbred genotypes (cultivars and elite breeding lines) over the course of five crop seasons (2016 to 2020) and two locations in Texas. The results showed that genotype, cultivar type (hybrid or inbred), location, and their interactions had significant effects on the severity of ShB and NBLS. The ShB severity in hybrid genotypes was significantly lower than that in inbred genotypes, with an average reduction of 27% in disease severity during the 5-year, two-location evaluation. Most (53%) of the hybrid genotypes were rated moderately resistant, whereas almost all (97%) of the inbred genotypes ranged from very susceptible to moderately susceptible. Similarly, NBLS severity in hybrid genotypes is significantly lower than that in inbred genotypes. All but four hybrid genotypes exhibit immune reactions to NBLS. In contrast, 77% of inbred genotypes exhibit NBLS symptoms, with disease resistance reactions ranging from susceptible to resistant. The results demonstrate that hybrid rice is generally less susceptible to sheath blight and has a higher level of resistance against NBLS compared with inbred rice.

Combined Use of PGPRs and Reduced Rates of Azoxystrobin to Improve Management of Sheath Blight of Rice.

Farmers rely heavily on the use of strobilurin fungicides to manage sheath blight (ShB) caused by Rhizoctonia solani AG1-IA, the most important disease in rice in the southern United States. Greenhouse and field studies were conducted to evaluate the potential use of plant growth-promoting rhizobacteria (PGPRs) in combination with a reduced rate of azoxystrobin application as a strategy to improve the current fungicide-reliant management. Of the nine antagonistic PGPR strains screened in the greenhouse, Bacillus subtilis strain MBI600 provided the most significant and consistent suppression of ShB. Efficacy of strain MBI600 was further evaluated at the concentrations of 0, 103, 106, 109, and 1011 CFU/ml alone or in combinations with 0, 17, 33, 50, 67, 83, and 100% of the recommended application rate (0.16 kg a.i./ha) of azoxystrobin. Strain MBI600 applied at 106,109, and 1011 CFU/ml alone was effective in reducing ShB severity. Combinations of this strain at these rates with ≥33% of the recommended application rate of azoxystrobin further reduced ShB severity. A dose-response model defining the relationships between strain MBI600, azoxystrobin, and ShB severity was established. Estimates of the effective concentrations (EC50 and EC90) of strain MBI600 when applied in combination with 50% of the recommended application rate of azoxystrobin were 104 and 109 CFU/ml, respectively. A field trial was conducted over 4 years to verify the efficacy of their combinations. Strain MBI600 alone, when applied at 109 CFU/ml at the boot stage, reduced ShB severity but did not significantly increase grain yields each year. Combination of strain MBI600 with azoxystrobin at half of the recommended application rate improved efficacy of strain MBI600, reducing ShB severity to a level comparable to that of azoxystrobin applied at the full rate in all 4 years. The combined treatment also increased grain yield by 14 to 19%, comparable to the fungicide applied at the full rate in 3 of 4 years. Combined use of PGPR strain MBI600 with a reduced rate of azoxystrobin application can be a viable management option for control of ShB while allowing producers to use less fungicide on rice.

Sterile White Basidiomycete Fungus Marasmius graminum: A New Pathogen Causing Seedling Blight in Rice.

In April 2018, damping-off of rice (Oryza sativa L.) seedlings at the 2-to-3-leaf stage was observed in three fields in the counties of Wharton and Matagorda of Texas and Jefferson-Davis Parish of Louisiana. All affected areas were 1 ha or greater, with 10 to 20% of the seedlings showing the symptoms. Infected seedlings showed dark-brown necrotic lesions on the roots and/or mesocotyls where white superficial mycelium was usually present. Symptomatic tissues excised from 10 diseased seedlings of each field were surface sterilized with 1% NaOCl, double rinsed in sterilized distilled water, and plated on potato dextrose agar (PDA). The plates were incubated at 25°C with a 12-h photoperiod in a growth chamber. After 48 h, hyphal tips of fungal colonies were transferred onto PDA and 12 isolates were obtained. Clamp connections and dolipore septa were observed in young hyphae, indicating that these isolates were a basidiomycete fungus. Young hyphal cells were binucleate based on safranin O stain (Bandoni 1979). No fruiting bodies or sclerotia produced on PDA after one month of incubation. Based on these morphological characteristics, these isolates were identified as belonging to sterile white basidiomycetes (SWB) (Howard et al. 1977). To further identify the isolates into the species level, the internal transcribed spacer (ITS) region of ribosomal DNA (rDNA) of a representative isolate was sequenced with primer ITS1 and ITS4 (Vinnere et al. 2005). The ITS sequence (GenBank acc. no. MT524457) had more than 97% sequence similarity with known Marasmius graminum strains from Denmark (JN943595) (Schoch et al. 2012) and Sweden (MH857692) (Vu et al. 2019). Pathogenicity was tested with three representative isolates in a growth chamber using a modified method (Carling and Leiner 1990). Pots (6.5 cm in diameter x 7.5 cm height) were filled with 100 g of sterilized sand and watered to field capacity. Five PDA plugs (4 mm in diameter) from 5-day-old growing culture were placed on the sand surface of each pot. Pots inoculated with PDA plugs without fungus served as the controls. Five seeds of rice cv. Presidio were planted into each pot and covered with 10 g of sterilized sand. Pots were maintained at 25±2°C in a growth chamber with a 12-h photoperiod for 14 days. There were four replicated pots for each treatment and the experiment repeated twice. After 2 weeks, severe damping-off and associated symptoms similar to those observed in the field appeared in the inoculated pots. No symptoms developed in the control pots. The same fungus was consistently re-isolated from infected plants. Based on morphological characteristics and rDNA-ITS sequencing, these isolates were identified as M. graminum. The SWB fungus was first reported as a causal agent of stem rot of snap bean in Florida (Howard et al. 1977) and Nebraska (Harveson 2002), root or hypocotyl rot of corn, snap bean, squash and peanut in Georgia (Sumner et al. 1979; Bell and Sumner 1984), and crown rot of pigeon pea (Cajanus cajan) in Puerto Rico (Kaiser et al. 1987). Later, the SWB strain (ATCC 28344) causing stem rot of snap bean in Florida was further identified as M. graminum based on nuclear large subunit rRNA gene (Vinnere et al. 2005). Comparing the ITS region of this isolate (AY445120) with our isolate revealed a 99% similarity. To our knowledge, this is the first report that the SWB fungus M. graminum causes seedling blight in rice. Identification of this new disease will help to develop management strategies for control of stand loss in rice.

Warming increases Bacterial Panicle Blight (Burkholderia glumae) occurrences and impacts on USA rice production.

Bacterial Panicle Blight (BPB), caused by Burkholderia glumae, is a bacterial disease in rice (Oryza sativa) that reduces rice yield and quality for producers and consequently creates higher market prices for consumers. BPB is caused by the simultaneous occurrence of high daily minimum temperatures (~22°C) and relative humidity (~77%), which may increase under the current scenario of global warming. This study hypothesized that the economic damage from warming may cause an increase in economic losses, though at a decreasing rate per degree. Thus, this study estimates the yield losses associated with BPB occurrences at the county level in the Mid-South United States (US) for annual rice production in 2003-2013 and under +1-3°C warming scenarios using daily weather information with appropriate thresholds. From the estimated losses, the total production potential of a BPB-resistant rice was quantified using a spatial equilibrium trade model to further estimate market welfare changes with the counterfactual scenario that all US county-level rice production were BPB resistant. Results from the study indicate that the alleviation of BPB would represent a $69 million USD increase in consumer surplus in the US and a concomitant increase in rice production that would feed an additional 1.46 million people annually assuming a global average consumption of 54 Kg per person. Under the 1°C warming scenario, BPB occurrences and production losses would cause price increases for rice and subsequently result in a $112 million USD annual decrease in consumer surplus in the US and a loss of production equivalent to feeding 2.17 million people. Under a 3°C warming scenario, production losses due to BPB cause an annual reduction of $204 million USD in consumer surplus in the US, and a loss in production sufficient to feed 3.98 million people a year. As global warming intensifies, BPB could become a more common and formidable rice disease to combat, and breeding for BPB resistance would be the primary line-of-defense as currently no effective chemical options are available. The results of this study inform agriculturalists, policymakers, and economists about the value of BPB-resistance in the international rice market and also help support efforts to focus future breeding toward climate change impact resilience.

Plant-Based Culture Media for Improved Growth and Sporulation of Cercospora janseana.

Cercospora janseana is the causal agent of narrow brown leaf spot, one of the major diseases of rice in the United States. In vitro studies were conducted to evaluate various plant-based agar media in comparison with potato dextrose agar (PDA) for their effects on radial growth and sporulation of C. janseana. The agar media were evaluated individually with the following plant-extract ingredients: fresh rice leaves, dried rice leaves, dried rice stems, dried rice roots, dried rice hulls, dried rice straw, rice bran, dried sorghum leaves, and dried barnyard grass leaves. Plant-based extracts obtained from 25, 50, 100, or 200 g/liter (plant material/water) were tested either alone or in combination with 10 and 20% clarified V8 juice. PDA served as the control medium. Media with 10 and 20% V8 juice were also included for comparison. Media were inoculated with a disc of 1-week-old actively growing C. janseana at the center of Petri plates and incubated at 28°C with a 12/12-h (light/dark) cycle for 2 weeks. Radial growth was measured at 7 and 14 days after inoculation (DAI), whereas sporulation was measured at 14 DAI. Radial growth and sporulation differed among various media and their amendment concentrations. Media with extracts of fresh rice leaves, dried rice leaves, dried rice stems, dried barnyard grass leaves, and clarified V8 juice enhanced radial growth compared with the PDA control. Of all the agar media evaluated, fresh rice leaf extract amendment at 25 g/liter with no V8 supplementation consistently supported the greatest radial growth, and 10% V8-only medium resulted in greater sporulation of C. janseana than other media evaluated.

Screening Brassicaceous Plants as Biofumigants for Management of Rhizoctonia solani AG1-IA.

Brassicaceae plants rich in glucosinolates have been used as biofumigants for management of Rhizoctonia solani (teleomorph: Thanatephorus cucumeris) and other soilborne pathogens. Efficacy of brassica plant tissue has mainly been attributed to toxic isothiocyanates released upon hydrolysis of glucosinolates. Management of R. solani AG 1-IA, the causal agent of sheath blight in rice (Oryza sativa), using biofumigation, is promising but needs more validation. Biofumigation activity of nine Brassicaceae plants and two other related species were evaluated in vitro with soils from Texas, Arkansas, or Mississippi. All plants evaluated significantly suppressed the mycelium growth of R. solani AG 1-IA. Mustard (Brassica juncea) cultivars ('Brand 199', 'Ruby Streak', 'Florida Broadleaf', and 'Green Wave') consistently provided the greatest (>90%) mycelial inhibition, while sunn hemp (Crotalaria juncea) and Chinese cabbage (B. rapa) had the least suppressive effect. B. juncea 'Red Giant' and 'Sheali Hong', turnip (B. rapa), kale (B. oleracea), and arugula (Eruca sativa) showed intermediate efficacy or were inconsistent. Effects of soil pasteurization and plant tissue amendment rates were examined with B. juncea Brand 199 and Texas soil. Inhibition of mycelial growth became greater with increasing plant amendment rates up to 3.2% (wt/wt) in the soil. Soil pasteurization almost completely suppressed the release of allyl isothiocyanate (AITC). The nonpasteurized soil amended with 0.5% (wt/wt) of the plant material released 96% more AITC than the soil amended with 0.25% (wt/wt) of the plant material. The highest levels of AITC release were observed at 12 and 24 h after soil amendment, with 0.25 and 0.5% (wt/wt) of the plant material, respectively. Antifungal effects of B. juncea are attributed to dose-dependent production of volatile AITC and could be used for managing rice sheath blight caused by R. solani AG 1-IA.

Assessment of Aerial Agrichemical Spraying Effect Using Moderate-Resolution Satellite Imagery.

Remote sensing technique can be used to examine the effects of agrichemical application on the performance of field crops at a large scale in an effort to develop precision agricultural aerial spraying technology. In this study, an airplane M-18B at the 4-m flight height was used to spray a mix of agrichemicals (a fungicide and a plant growth regulator) to control rice leaf blast disease and improve the growth vigor of rice plants in the field. After the aerial spraying, satellite imagery of tested area was acquired and processed to calculate vegetation indices (VIs). Ground agrichemical concentration data were also collected. The relationships between droplets deposition and VIs were analyzed. The results indicated that the highest correlation coefficient between single phase spectral feature (NDVI) and droplets deposition points density (DDPD, points·cm-2) was 0.315 with P-value of 0.035 while the highest correlation coefficient between temporal change characteristic (MSAVI) and droplets deposition volume density (DDVD, µL·cm-2) was 0.312 with P-value of 0.038). Rice plants with the greatest growth vigor were all detected within the spraying swath, with a gradual decrease in the vigor of rice plants with the increase of droplets drift distance. There were similar trend patterns in the changes of the spraying effects based on the spatial interpolation maps of droplets deposition data and spectral characteristics. Therefore, vegetation indexes, NDVI and MSAVI calculated from satellite imagery can be used to determine the aerial spraying effects in the field on a large scale.

Integration of Brassica Cover Crop with Host Resistance and Azoxystrobin for Management of Rice Sheath Blight.

Sheath blight caused by Rhizoctonia solani is the most important rice disease that can cause significant losses in grain yield and quality in the southern United States. Current management options for sheath blight primarily consist of fungicides, tolerant cultivars, and cultural practices. These options are not always very effective. Brassica plants have been used for soil fumigation to manage a variety of different soilborne pathogens. In this field study, the efficacy of a Brassica juncea cover crop integrated with use of a tolerant rice cultivar and fungicide application was evaluated in 2011, 2012, and 2013. The B. juncea cover crop significantly lowered sheath blight severity in all 3 years and led to a significantly higher grain yield in 2013 as compared with the fallow control. 'Presidio' rice had lower sheath blight severity and higher yield than 'Cocodrie' in 2012 and 2013. Fungicide applications with azoxystrobin at the label rate (0.16 kg a.i./ha) or half the label rate (0.08 kg a.i./ha) significantly reduced sheath blight severity in all 3 years, resulting in a yield increase in 2 of the 3 years. B. juncea along with use of a tolerant rice cultivar and half the label rate of azoxystrobin can be an effective approach for management of sheath blight in rice.