Glycine max polygalacturonase inhibiting protein 11 (GmPGIP11) functions in the root to suppress Heterodera glycines parasitism.

Pathogen-secreted polygalacturonases (PGs) alter plant cell wall structure by cleaving the α-(1 â†’ 4) linkages between D-galacturonic acid residues in homogalacturonan (HG), macerating the cell wall, facilitating infection. Plant PG inhibiting proteins (PGIPs) disengage pathogen PGs, impairing infection. The soybean cyst nematode, Heterodera glycines, obligate root parasite produces secretions, generating a multinucleate nurse cell called a syncytium, a byproduct of the merged cytoplasm of 200-250 root cells, occurring through cell wall maceration. The common cytoplasmic pool, surrounded by an intact plasma membrane, provides a source from which H. glycines derives nourishment but without killing the parasitized cell during a susceptible reaction. The syncytium is also the site of a naturally-occurring defense response that happens in specific G. max genotypes. Transcriptomic analyses of RNA isolated from the syncytium undergoing the process of defense have identified that one of the 11 G. max PGIPs, GmPGIP11, is expressed during defense. Functional transgenic analyses show roots undergoing GmPGIP11 overexpression (OE) experience an increase in its relative transcript abundance (RTA) as compared to the ribosomal protein 21 (GmRPS21) control, leading to a decrease in H. glycines parasitism as compared to the overexpression control. The GmPGIP11 undergoing RNAi experiences a decrease in its RTA as compared to the GmRPS21 control with transgenic roots experiencing an increase in H. glycines parasitism as compared to the RNAi control. Pathogen associated molecular pattern (PAMP) triggered immunity (PTI) and effector triggered immunity (ETI) components are shown to influence GmPGIP11 expression while numerous agricultural crops are shown to have homologs.

Evaluation of genetic diversity, haplotype, and virulence of Fusarium oxysporum f. sp. vasinfectum field isolates from Alabama.

The United States is the third largest producer of cotton and the largest exporter of cotton globally. Fusarium wilt, caused by the soilborne fungal pathogen Fusarium oxysporum f. sp. vasinfectum (Fov), was estimated to cause a $21 million cotton yield loss in 2022. Historically, Alabama was an important producer of cotton in the southeastern United States and was the first state in which Fusarium wilt on cotton was described. To assess the genetic diversity of Fov field isolates in Alabama, 118 field isolates were collected from six counties across the state from 2014 to 2016. Phylogenetic analysis using TEF1 and RPB2 placed the Fov field isolates into 18 haplotypes. Upon profiling the Tfo1 transposon insertion in the NAT gene, it was determined that no race 4 isolates were recovered in Alabama. Representatives of all field isolate haplotypes caused disease on Upland cotton variety Rowden in a hydroponic test tube assay. Two haplotype A isolates were the most aggressive isolates recovered and haplotype A isolate TF1 was more aggressive than the race 4 isolate 89-1A on Upland cotton and had similar symptom severity on Pima cotton. Karyotype profiling indicted an abundance of small chromosomes characteristic of karyotypes that include accessory chromosomes, with considerable variability between isolates. Collectively, our study indicates that Fov isolates from Alabama are genetically diverse which may have been promoted by its persistence in cotton fields.

Characterizing the Diversity of Oomycetes Associated with Diseased Cotton Seedlings in Alabama.

Many oomycete species are associated with the seedlings of crops, including upland cotton (Gossypium hirsutum L.), which leads to annual threats. The diversity of oomycete species in Alabama needs to be better understood since the last survey of oomycetes associated with cotton in Alabama was 20 years ago-before significant updates to taxonomy and improvements in identification of oomycetes using molecular tools. Our current study aimed to identify oomycetes associated with Alabama cotton seedlings, correlate diversity with soil edaphic factors, and assess virulence toward cotton seed. Thirty symptomatic cotton seedlings were collected independently from 25 fields in 2021 and 2022 2 to 4 weeks after planting. Oomycetes were isolated by plating root sections onto a semiselective medium. The internal transcribed spacer (ITS) region was sequenced to identify the resulting isolates. A seed virulence assay was conducted in vitro to verify pathogenicity, and 347 oomycete isolates were obtained representing 36 species. Northern Alabama soils had the richest oomycete communities and a greater silt and clay concentration than sandier soils in the central and southern coastal plains. Globisporangium irregulare and Phytophthora nicotianae were consistently recovered from cotton roots in both years. Globisporangium irregulare was pathogenic and recovered from all Alabama regions, whereas P. nicotianae was pathogenic but recovered primarily in areas with lower sand content in northern Alabama. Many oomycete species have not been previously reported in Alabama or the southeastern United States. Altogether, this knowledge will help facilitate effective management strategies for cotton seedling diseases caused by oomycetes in Alabama and the United States.

Photosynthetic Response of Blueberries Grown in Containers.

Recently, there has been increased interest in container blueberry production as a viable alternative to open-field blueberry planting. Container production of blueberries offers numerous advantages, among these, a lack of limitation by suboptimal soil conditions in the open field and the ability to control substrate pH, drainage, and organic matter. The photosynthetic response for three container-grown Southern highbush blueberry (interspecific Vaccinium hybrids) cultivars including 'Jewel', 'Meadowlark', and 'Victoria' and a rabbiteye blueberry (Vaccinium virgatum) 'Baldwin', were measured during the spring and summer of 2022. It was hypothesized that the three cultivars evaluated would have different photosynthetic responses. The objective of this study was to determine the photosynthetic activity of different blueberry cultivars during the first year of crop establishment. A series of measurements were conducted every 2 h throughout the day and for different dates using a gas exchange data analyzer on newly matured fully expanded leaves located in the top middle section of the canopy for each cultivar. The response curves showed that net photosynthesis (A) became saturated at moderate light, with saturation occurring at a photosynthetic photon flux density (PPFD) of 1932 µmol m-2 s-1. At this point, the rate of CO2 assimilation was approximately 16.84 µmol CO2 m-2 s-1. No differences in (A) were found among cultivars. Overall, the attained values of photosynthesis provide a strong conceptual basis for understanding the cultivar variation response when grown in containers; therefore, the containerized system may serve as a production system for early fruiting blueberries in Alabama, USA.

Evaluation of Meloidogyne Incognita and Rotylenchulus Reniformis Nematode-resistant Cotton Cultivars with Supplemental Corteva Agriscience Nematicides.

Meloidogyne incognita- and Rotylenchulus reniformis-resistant new cotton cultivars have recently become available, giving growers a new option in nematode management. The objectives of this study were: (i) to determine the yield potential of the new cultivars PHY 360 W3FE (M. incognita-resistant) and PHY 332 W3FE (R. reniformis-resistant) in nematode-infested fields and (ii) to evaluate the effects of combining the nematicides Reklemel (fluazaindolizine), Vydate C-LV (oxamyl), and the seed treatment BIOST Nematicide 100 (heat killed Burkholderia rinojenses and its non-living spent fermentation media) with resistant cotton cultivars on nematode population levels and lint yield. Field experiments in 2020 and 2021 indicated M. incognita population levels were 73% lower on PHY 360 W3FE (R) and 80% lower for R. reniformis on the PHY 332 W3FE (R) at 40 days after planting. Nematode eggs per gram of root were further reduced an average of 86% after the addition of Reklemel and Vydate C-LV when averaging both cultivars over the two years. Tests with BIOST Nematicide 100 + Reklemel + Vydate C-LV (0.56 + 2.5 L/ha) in both M. incognita and R. reniformis fields produced higher lint yields. Overall, planting PHY 360 W3FE (R) and PHY 332 W3FE (R) improved yields an average of 364 kg/ha while limiting nematode population increases. The addition of the nematicides further increased yields 152 kg/ha of the nematode-resistant cultivars.

Two Intra-Individual ITS1 rDNA Sequence Variants Identified in the Female and Male Rotylenchulus reniformis Populations of Alabama.

Around 300 different plant species are infected by the plant-parasitic reniform nematode (Rotylenchulus reniformis), including cotton. This is a devasting nematode with a preference for cotton; it is commonly found in Alabama farms and causes severe reduction in yields. Its first internal transcribed spacer (ITS1) region can be sequenced, and potential mutations can be found in order to study the population dynamics of the reniform nematode. The goal of our study was to sequence the ITS1 rDNA region in male and female RNs that were collected from BelleMina, Hamilton, and Lamons locations in Alabama. After separating the single male and female RNs from the samples collected from the three selected listed sites above, the ITS1 region was amplified selectively using specific primers, and the resulting products were cloned and sequenced. Two distinct bands were observed after DNA amplification of male and female nematodes at 550 bp and 730 bp, respectively. The analysis of sequenced fragments among the three populations showed variation in average nucleotide frequencies of female and male RNs. Singletons within the female and male Hamilton populations ranged from 7.8% to 10%, and the variable sites ranged from 13.4% to 26%. However, female and male BelleMina populations had singletons ranging from 7.1% to 19.7% and variable regions in the range of 13.9% to 49.3%. The female and male Lamons populations had singletons ranging from 2.5% to 8.7% and variable regions in the range of 2.9% to 14.2%. Phylogenetic (neighbor-joining) analysis for the two ITS1 fragments (ITS-550 and ITS-730) showed relatively high intra-nematode variability. Different clone sequences from an individual nematode often had greater similarity with other nematodes than with their own sequences. RNA fold analysis of the ITS1 sequences revealed varied stem and loop structures, suggesting both conserved and variable regions in the variants identified from female and male RNs, thus underscoring the presence of significant intra- and inter-nematodal variation among RN populations in Alabama.

The heterologous expression of conserved Glycine max (soybean) mitogen activated protein kinase 3 (MAPK3) paralogs suppresses Meloidogyne incognita parasitism in Gossypium hirsutum (upland cotton).

Two conserved Glycine max (soybean) mitogen activated protein kinase 3 (MAPK3) paralogs function in defense to the parasitic soybean cyst nematode Heterodera glycines. Gene Ontology analyses of RNA seq data obtained from MAPK3-1-overexpressing (OE) and MAPK3-2-OE roots compared to their control, as well as MAPK3-1-RNA interference (RNAi) and MAPK3-2-RNAi compared to their control, hierarchically orders the induced and suppressed genes, strengthening the hypothesis that their heterologous expression in Gossypium hirsutum (upland cotton) would impair parasitism by the root knot nematode (RKN) Meloidogyne incognita. MAPK3-1 expression (E) in G. hirsutum suppresses the production of M. incognita root galls, egg masses, and second stage juveniles (J2s) by 80.32%, 82.37%, and 88.21%, respectfully. Unexpectedly, egg number increases by 28.99% but J2s are inviable. MAPK3-2-E effects are identical, statistically. MAPK3-1-E and MAPK3-2-E decreases root mass 1.49-fold and 1.55-fold, respectively, as compared to the pRAP15-ccdB-E control. The reproductive factor (RF) of M. incognita for G. hirsutum roots expressing MAPK3-1-E or MAPK3-2-E decreases 60.39% and 50.46%, respectively, compared to controls. The results are consistent with upstream pathogen activated molecular pattern (PAMP) triggered immunity (PTI) and effector triggered immunity (ETI) functioning in defense to H. glycines. The experiments showcase the feasibility of employing MAPK3, through heterologous expression, to combat M. incognita parasitism, possibly overcoming impediments otherwise making G. hirsutum's defense platform deficient. MAPK homologs are identified in other important crop species for future functional analyses.

Glycine max Homologs of DOESN'T MAKE INFECTIONS 1, 2, and 3 Function to Impair Heterodera glycines Parasitism While Also Regulating Mitogen Activated Protein Kinase Expression.

Glycine max root cells developing into syncytia through the parasitic activities of the pathogenic nematode Heterodera glycines underwent isolation by laser microdissection (LM). Microarray analyses have identified the expression of a G. max DOESN'T MAKE INFECTIONS3 (DMI3) homolog in syncytia undergoing parasitism but during a defense response. DMI3 encodes part of the common symbiosis pathway (CSP) involving DMI1, DMI2, and other CSP genes. The identified DMI gene expression, and symbiosis role, suggests the possible existence of commonalities between symbiosis and defense. G. max has 3 DMI1, 12 DMI2, and 2 DMI3 paralogs. LM-assisted gene expression experiments of isolated syncytia under further examination here show G. max DMI1-3, DMI2-7, and DMI3-2 expression occurring during the defense response in the H. glycines-resistant genotypes G.max [Peking/PI548402] and G.max [PI88788] indicating a broad and consistent level of expression of the genes. Transgenic overexpression (OE) of G. max DMI1-3, DMI2-7, and DMI3-2 impairs H. glycines parasitism. RNA interference (RNAi) of G. max DMI1-3, DMI2-7, and DMI3-2 increases H. glycines parasitism. The combined opposite outcomes reveal a defense function for these genes. Prior functional transgenic analyses of the 32-member G. max mitogen activated protein kinase (MAPK) gene family has determined that 9 of them act in the defense response to H. glycines parasitism, referred to as defense MAPKs. RNA-seq analyses of root RNA isolated from the 9 G. max defense MAPKs undergoing OE or RNAi reveal they alter the relative transcript abundances (RTAs) of specific DMI1, DMI2, and DMI3 paralogs. In contrast, transgenically-manipulated DMI1-3, DMI2-7, and DMI3-2 expression influences MAPK3-1 and MAPK3-2 RTAs under certain circumstances. The results show G. max homologs of the CSP, and defense pathway are linked, apparently involving co-regulated gene expression.

Additional fertilizer and nematicide combinations on upland cotton to manage Rotylenchulus reniformis and Meloidogyne incognita in Alabama.

Plant parasitic nematodes are major pests on upland cotton worldwide and in the United States. The reniform nematode, Rotylenchulus reniformis and the southern root-knot nematode Meloidogyne incognita are some of the most damaging nematodes on cotton in the United States. Current management strategies focus on reducing nematode populations with nematicides. The objective of this research was to integrate additional fertilizer and nematicide combinations into current practices to establish economical nematode management strategies while promoting cotton yield and profit. Microplot and field trials were run to evaluate fertilizer and nematicide combinations applied at the pinhead square (PHS) and first bloom (FB) plant growth stages to reduce nematode population density and promote plant growth and yield. Cost efficiency was evaluated based on profit from lint yields and chemical input costs. Data combined from 2019 and 2020 suggested a nematicide seed treatment (ST) ST + (NH4)2SO4 + Vydate® C-LV + Max-In® Sulfur was the most effective in increasing seed cotton yields in the R. reniformis microplot trials. In R. reniformis field trials, a nematicide ST + (NH4)2SO4 + Vydate® C-LV at PHS supported the largest lint yield and profit per hectare at $1176. In M. incognita field trials, a nematicide ST + 28-0-0-5 + Vydate® C-LV + Max-In® Sulfur at PHS and FB supported the largest lint yields and profit per hectare at $784. These results suggest that combinations utilizing fertilizers and nematicides applied together across the season in addition to current fertility management show potential to promote yield and profit in R. reniformis and M. incognita infested cotton fields.

Meta-Analysis of the Field Efficacy of Seed- and Soil-Applied Nematicides on Meloidogyne incognita and Rotylenchulus reniformis Across the U.S. Cotton Belt.

Meta-analysis was used to compare yield protection and nematode suppression provided by two seed-applied and two soil-applied nematicides against Meloidogyne incognita and Rotylenchulus reniformis on cotton across 3 years and several trial locations in the U.S. Cotton Belt. Nematicides consisted of thiodicarb- and fluopyram-treated seed, aldicarb and fluopyram applied in furrow, and combinations of the seed treatments and soil-applied fluopyram. The nematicides had no effect on nematode reproduction or root infection but had a significant impact on seed cotton yield response ([Formula: see text]), with an average increase of 176 and 197 kg/ha relative to the nontreated control in M. incognita and R. reniformis infested fields, respectively. However, because of significant variation in yield protection and nematode suppression by nematicides, five or six moderator variables (cultivar resistance [M. incognita only], nematode infestation level, nematicide treatment, application method, trial location, and growing season) were used depending on nematode species. In M. incognita-infested fields, greater yield protection was observed with nematicides applied in furrow and with seed-applied + in-furrow than with solo seed-applied nematicide applications. Most notable of these in-furrow nematicides were aldicarb and fluopyram (>131 g/ha) with or without a seed-applied nematicide compared with thiodicarb. In R. reniformis-infested fields, moderator variables provided no further explanation of the variation in yield response produced by nematicides. Furthermore, moderator variables provided little explanation of the variation in nematode suppression by nematicides in M. incognita- and R. reniformis-infested fields. The limited explanation by the moderator variables on the field efficacy of nematicides in M. incognita- and R. reniformis-infested fields demonstrates the difficulty of managing these pathogens with nonfumigant nematicides across the U.S. Cotton Belt.

Supplemental Fumigant Placement Improves Root Knot and Fusarium Wilt Management for Tomatoes Produced on a Raised-Bed Plasticulture System in Florida's Myakka Fine Sand.

Fresh-market tomatoes are produced on a raised-bed plasticulture system that relies heavily on soil-applied preplant fumigants for the management of soilborne pathogens, nematodes, and weeds. Since the transition from methyl bromide to alternative fumigants, growers have experienced a resurgence of several soilborne pests and pathogens, including root-knot nematode caused by Meloidogyne spp. and Fusarium wilt caused by Fusarium oxysporum f. sp. lycopersici race 3. This resurgence is attributed to the inability of the alternative fumigants to effectively disperse through the soil in the same manner as methyl bromide. Two supplemental fumigation strategies, the application of chloropicrin (PIC) below bed edges (herein "supplemental PIC") and broadcast deep-shank applications of 1,3-dichloropropene (1,3-D), were evaluated in conjunction with standard raised-bed applications of Pic-Clor 60, Pic-Clor 80, and Pic 100 covered with a virtually impermeable film or a totally impermeable film. Large-plot replicated studies were conducted in two separate commercial tomato fields with a history of production losses caused by root-knot nematode and Fusarium wilt. Deep-shank 1,3-D applications significantly reduced the recovery of root-knot and total parasitic nematodes across field sites before the preparation of raised beds. Both supplemental PIC and deep-shank 1,3-D reduced root-knot galling and Fusarium wilt incidence, but the latter supplemental treatment statistically had the greatest impact. Fumigant applied within raised beds or plastic film had no significant effect on root-knot galling or Fusarium wilt. Although both supplemental fumigation strategies had a significant effect on pest and disease pressure, neither statistically improved tomato yields based on small subplot harvests. Controlled laboratory experiments confirmed the fungicidal activity of 1,3-D against F. oxysporum f. sp. lycopersici, with 75, 90, 95, and 99% lethal doses corresponding to estimated field application rates of 56.1, 93.5, 121.6, and 184.7 liters/ha, respectively. The results demonstrate how fumigant placement can improve pest and disease control activity with current fumigant alternatives to methyl bromide and further support the broader pesticidal activity of some chemical fumigants.

Evaluation of the mechanism of action of Bacillus spp. to manage Meloidogyne incognita with split root assay, RT-qPCR and qPCR.

The goal of this research is to determine the mechanism of action of two Bacillus spp. that can manage Meloidogyne incognita population density in cotton. The overall objectives are 1) determine the efficacy and direct antagonistic capabilities of the Bacillus spp. and 2) determine the systemic capabilities of the Bacillus spp. The greenhouse in planta assay indicated B. amyloliquefaciens QST713 and B. firmus I-1582 could manage M. incognita similarly to the chemical standard fluopyram. An in vitro assay determined that B. firmus I-1582 and its extracted metabolites were able to directly manage M. incognita second stage juveniles by increasing mortality rate above 75%. A split root assay, used to determine systemic capabilities of the bacteria, indicated B. amyloliquefaciens QST713 and B. firmus I-1582 could indirectly decrease the nematode population density. Another species, B. mojavensis strain 2, also demonstrated systemic capabilities but was not a successful biological control agent because it supported a high population density in greenhouse in planta assay and in the split root assay. A RT-qPCR assay was used to confirm any systemic activity observed in the split root assay. At 24 hours both B. amyloliquefaciens QST713 and B. firmus I-1582 upregulated one gene involved in the initial stages of JA synthesis pathway but not another gene involved in the later stages of JA synthesis. These results point to a JA intermediate molecule, most likely OPDA, stimulated by the bacteria rather than JA in a short-term systemic response. After 1 week, the Bacillus spp. stimulated a SA-responsive defense related gene. The long-term systemic response to the Bacillus spp. indicates salicylic acid also plays a role in defense conferred by these bacteria. The final assay was a qPCR to determine the concentration of the bacteria on the cotton roots after 24 days. Bacillus amyloliquefaciens QST713 and B. firmus I-43 1582 were able to colonize the root successfully, with the concentration after 24 days not significantly differing from the concentration at inoculation. This study identifies two bacteria that work via systemic resistance and will help aid in implementing these species in an integrated management system.

Diversity and temporal distribution of Fusarium oxysporum f. sp. vasinfectum races and genotypes as influenced by Gossypium cultivar.

This study assess the population diversity and temporal variability of caused by Fusarium oxysporum f. sp. vasinfectum (FOV) races/genotypes infecting cotton cultivars with either FOV or Meloidogyne incognita resistance. All plants sampled demonstrated typical symptoms of FOV including wilting, chlorosis and necrosis of the leaves, and discoloration of the vascular tissue in the stem. A diverse population of FOV was characterized. Eight races/genotypes of FOV were collected throughout the three site years. FOV race 1 was the most predominant in all tests (AUDPC=101.1); statistically higher numbers of isolates from LA-108 (AUDPC=59.9), race 8 (AUDPC=47.5), and race 2 (AUDPC=38.6) were also found compared to other races and genotypes collected. FOV race 1, race 2, race 8, and 108 were the most virulent races identified. The genotypes MDS-12, LA-110, and LA-127/140 were found in all tests but at a low incidence, and LA-112 was only found in trace amounts. MDS-12, LA-110, LA-112, and LA-127/140 produced less disease pressure. FOV race 4 which is highly virulent and present in California and Texas was not found in Alabama. A positive correlation was observed between the accumulation of growing degree days and FOV race 1, race 2, race 8, LA-108, and LA-110. Later symptom expression influenced by seasonal heat partially mitigates damage allowing cotton to produce bolls though they may be reduced in number and lint quality. Plant resistance to the FOV as expressed in these cultivars appears to provide better protection than M. incognita resistance. PhytoGen 72, which is resistant to FOV races/genotypes had low levels of FOV infection even though it sustained a high level of M. incognita root population density. The M. incognita resistant cultivars Deltapine 1558NR B2RF and PhytoGen 480 W3FE supported a lower nematode population density, however, FOV disease incidence was not reduced. FOV races/genotypes did not vary significantly between the nematode resistant and nematode susceptible cultivars.

Cotton Leafroll Dwarf Virus US Genomes Comprise Divergent Subpopulations and Harbor Extensive Variability.

Cotton leafroll dwarf virus (CLRDV) was first reported in the United States (US) in 2017 from cotton plants in Alabama (AL) and has become widespread in cotton-growing states of the southern US. To investigate the genomic variability among CLRDV isolates in the US, complete genomes of the virus were obtained from infected cotton plants displaying mild to severe symptoms from AL, Florida, and Texas. Eight CLRDV genomes were determined, ranging in size from 5865 to 5867 bp, and shared highest nucleotide identity with other CLRDV isolates in the US, at 95.9-98.7%. Open reading frame (ORF) 0, encoding the P0 silencing suppressor, was the most variable gene, sharing 88.5-99.6% and 81.2-89.3% amino acid similarity with CLRDV isolates reported in cotton growing states in the US and in Argentina and Brazil in South America, respectively. Based on Bayesian analysis, the complete CLRDV genomes from cotton in the US formed a monophyletic group comprising three relatively divergent sister clades, whereas CLRDV genotypes from South America clustered as closely related sister-groups, separate from US isolates, patterns reminiscent of phylogeographical structuring. The CLRDV isolates exhibited a complex pattern of recombination, with most breakpoints evident in ORFs 2 and 3, and ORF5. Despite extensive nucleotide diversity among all available CLRDV genomes, purifying selection (dN/dS < 1) was implicated as the primary selective force acting on viral protein evolution.

Heterakis gallinarum and Histomonas meleagridis DNA persists in chicken houses years after depopulation.

The poultry pathogen Histomonas meleagridis is transmitted by chicken cecal worms (Heterakis gallinarum) and is potentially transmitted by second order insect vectors and paratenic hosts. Darkling beetles (Alphitobius diaperinus) are poultry farm pests that infest barns. An outstanding question is the degree to which darkling beetles transmit both Heterakis and Histomonas. In this study we monitored populations of darkling beetles and assessed their positivity for both Heterakis and Histomonas by PCR. Uniquely, this study was conducted during the scheduled deconstruction of Auburn University's Poultry Research Farm. Therefore, we were able to monitor beetle and litter infection status months and years after bird depopulation. The duration of our monitoring continued through three seasons. We show that environmental DNA from both Heterakis and Histomonas persist in the environment long after prior infections, even in the absence of living Heterakis and its hosts. Finally, in an intensive search for live Heterakis, we discovered reniform nematodes (plant parasitic nematodes) residing in the soil floor of poultry farms.

Conserved oligomeric Golgi (COG) complex genes functioning in defense are expressed in root cells undergoing a defense response to a pathogenic infection and exhibit regulation my MAPKs.

The conserved oligomeric Golgi (COG) complex maintains correct Golgi structure and function during retrograde trafficking. Glycine max has 2 paralogs of each COG gene, with one paralog of each gene family having a defense function to the parasitic nematode Heterodera glycines. Experiments presented here show G. max COG paralogs functioning in defense are expressed specifically in the root cells (syncytia) undergoing the defense response. The expressed defense COG gene COG7-2-b is an alternate splice variant, indicating specific COG variants are important to defense. Transcriptomic experiments examining RNA isolated from COG overexpressing and RNAi roots show some COG genes co-regulate the expression of other COG complex genes. Examining signaling events responsible for COG expression, transcriptomic experiments probing MAPK overexpressing roots show their expression influences the relative transcript abundance of COG genes as compared to controls. COG complex paralogs are shown to be found in plants that are agriculturally relevant on a world-wide scale including Manihot esculenta, Zea mays, Oryza sativa, Triticum aestivum, Hordeum vulgare, Sorghum bicolor, Brassica rapa, Elaes guineensis and Saccharum officinalis and in additional crops significant to U.S. agriculture including Beta vulgaris, Solanum tuberosum, Solanum lycopersicum and Gossypium hirsutum. The analyses provide basic information on COG complex biology, including the coregulation of some COG genes and that MAPKs functioning in defense influence their expression. Furthermore, it appears in G. max and likely other crops that some level of neofunctionalization of the duplicated genes is occurring. The analysis has identified important avenues for future research broadly in plants.

Enhanced biological control of root-knot nematode, Meloidogyne incognita, by combined inoculation of cotton or soybean seeds with a plant growth-promoting rhizobacterium and pectin-rich orange peel.

LC-MS analysis of plant growth-promoting rhizobacterium (PGPR) Bacillus velezensis AP203 supernatants indicated the presence of nematode-inhibiting compounds that increased in abundance when B. velezensis AP203 was grown on orange peel. Meloidogyne incognita J2 were incubated with B. velezensis AP203 spores and orange peel, spores alone, orange peel alone, or with a non-inoculated control, and the combination of B. velezensis AP203 with orange peel resulted in 94% mortality of M. incognita juveniles (p ≤ 0.05). The J2 mortality rate for B. velezensis alone was 53%, compared to 59% mortality with orange peel, and the non-inoculated control exhibited 7% mortality. When tested on soybeans raised in a greenhouse, it was observed that when grown in the presence of orange peel, B. velezensis AP203 culture broth, cell suspension or supernatant reduced the numbers of M. incognita eggs per g of root at 45 days after planting (DAP) compared to inoculated controls in soybean and cotton (p ≤ 0.05). Likewise, soybean root length and fresh root weight significantly increased after inoculation with B. velezensis AP203 amended with orange peel. In cotton, shoot and root length significantly increased after inoculation with cell pellets of B. velezensis AP203 amended with orange peel compared to the M. incognita inoculated control. These data indicate that B. velezensis AP203 responds to growth on pectin-rich orange peel by production of biologically active secondary metabolites that can promote plant growth and inhibit root-knot nematode viability.

Entomopathogenic nematode management of small hive beetles (Aethina tumida) in three native Alabama soils under low moisture conditions.

The goal was to determine the efficacy of entomopathogenic nematodes (EPNs) on Aethina tumida small hive beetle (SHB) in Alabama soils. The objectives were to (i) determine the pupation success of SHB wandering larvae; (ii) determine the efficacy of EPNs on SHB wandering larvae in natural and autoclaved soil; and (iii) determine the efficacy of EPNs on SHB wandering larvae in three Alabama soil types at typical low moisture levels. The Alabama soils were Kalmia loamy sand (KLS), Benndale fine sandy loam (BFSL), and Decatur silt loam (DSL). Heterorhabditis bacteriophora, H. indica, Steinernema carpocapsae, S. feltiae, S. kraussei, and S. riobrave were tested at population densities of 5, 10, 20, 40, and 80 third-stage infective EPN juveniles (IJ3) per 130 cm3 soil. Pupation success in SHB population densities of 5, 10, and 20 wandering larvae per Petri dish were similar. Of the six EPN species, S. carpocapsae achieved the highest efficacy across all EPN population densities in both natural and autoclaved soil. Steinernema riobrave and H. indica achieved the next highest efficacies; however, they were significantly less effective than S. carpocapsae. Steinernema carpocapsae parasitized 87% SHB wandering larvae across all population densities tested. Steinernema carpocapsae achieved the best efficacy colonizing 94% of the SHB in the KLS soil, 80% in the BFSL soil, and 47% in the DSL soil. In conclusions, S. carpocapsae is be a promising biological control EPN to implement into a management system on SHB.

First report of Meloidogyne incognita infecting Cannabis sativa in Alabama.

Hemp (Cannabis sativa L.) is a new crop in Alabama. In 2019, symptomatic plants with stunted growth, poor root development, and numerous galls were observed in hemp plants grown in Geneva County, AL. After harvest, soil samples were collected from areas with the symptomatic plants and root-knot nematode (Meloidogyne spp.) were found in the soil. Based on morphological features and the polymerase chain reactions using species-specific primers, it was identified as Meloidogyne incognita. Further, a host differential test in a greenhouse assay confirmed it to be M. incognita race 3. The pathogenicity of the nematode to the hemp was confirmed by a modified version of Koch's postulates. To our knowledge, this is the first report of M. incognita infecting Cannabis sativa in Alabama.

Evaluation of a new chemical nematicide, fluazaindolizine (ReklemelTM active), for plant-parasitic nematode management in bermudagrass.

Plant-parasitic nematodes are a major pest of turfgrass in the United States, yet there are few options for successful management. Most current management strategies rely on the use of a limited number of chemical nematicides, so finding a new management option for nematode suppression would be extremely valuable for turfgrass managers. The aim of this study is to evaluate a new nematicide, fluazaindolizine (Reklemel™ active), for its ability to reduce plant-parasitic nematode population density and improve turfgrass quality. Separate research trials were conducted on bermudagrass infested with Belonolaimus longicaudatus and Meloidogyne incognita in greenhouse, microplot, and field settings over 2018 and 2019. Both greenhouse evaluations demonstrated multiple rates of fluazaindolizine reduced B. longicaudatus population density, and one of the two M. incognita trials showed multiple rates of fluazaindolizine reduced nematode population density. Fluazaindolizine was also effective at reducing population density of both B. longicaudatus and M. incognita in microplot settings for both 2018 and 2019, and a significant improvement in turf quality was observed for both visual turfgrass ratings and NDVI. Field trials demonstrated a significant reduction for both B. longicaudatus and M. incognita population density by multiple rates of fluazaindolizine, but no significant differences in turf quality ratings were observed. Overall, fluazaindolizine shows promise as a chemical nematicide for plant-parasitic nematode management on turfgrass.