Electrical synapse structure requires distinct isoforms of a postsynaptic scaffold.

Electrical synapses are neuronal gap junction (GJ) channels associated with a macromolecular complex called the electrical synapse density (ESD), which regulates development and dynamically modifies electrical transmission. However, the proteomic makeup and molecular mechanisms utilized by the ESD that direct electrical synapse formation are not well understood. Using the Mauthner cell of zebrafish as a model, we previously found that the intracellular scaffolding protein ZO1b is a member of the ESD, localizing postsynaptically, where it is required for GJ channel localization, electrical communication, neural network function, and behavior. Here, we show that the complexity of the ESD is further diversified by the genomic structure of the ZO1b gene locus. The ZO1b gene is alternatively initiated at three transcriptional start sites resulting in isoforms with unique N-termini that we call ZO1b-Alpha, -Beta, and -Gamma. We demonstrate that ZO1b-Beta and ZO1b-Gamma are broadly expressed throughout the nervous system and localize to electrical synapses. By contrast, ZO1b-Alpha is expressed mainly non-neuronally and is not found at synapses. We generate mutants in all individual isoforms, as well as double mutant combinations in cis on individual chromosomes, and find that ZO1b-Beta is necessary and sufficient for robust GJ channel localization. ZO1b-Gamma, despite its localization to the synapse, plays an auxiliary role in channel localization. This study expands the notion of molecular complexity at the ESD, revealing that an individual genomic locus can contribute distinct isoforms to the macromolecular complex at electrical synapses. Further, independent scaffold isoforms have differential contributions to developmental assembly of the interneuronal GJ channels. We propose that ESD molecular complexity arises both from the diversity of unique genes and from distinct isoforms encoded by single genes. Overall, ESD proteomic diversity is expected to have critical impacts on the development, structure, function, and plasticity of electrical transmission.

Reduced Sensitivity to Fluopyram in Meloidogyne graminis following Long-Term Exposure in Golf Turf.

In recent years, some golf course superintendents in Florida have reported that the turf health is no longer as great, and nematode responses to fluopyram have decreased. The objective of this research was to determine if the mechanism of the reported reduced efficacy was attributable to either: i) enhanced degradation accelerating its breakdown in the soil, or ii) reduced sensitivity to the nematicide in the nematode populations. In a field experiment, soil and nematodes were collected from small plots that had been treated multiple times over four years, for only one year, or never treated. Soil and nematodes were additionally collected from commercial turf sites where either multiple applications of fluopyram had been made for numerous years, or it had never been used. Bioassay experiments found no evidence of enhanced degradation. However, M. graminis collected from small field plots and commercial sites with long-term use of fluopyram were less sensitive to fluopyram in-vitro than those from small plots and commercial sites where fluopyram had not been used. These results indicate that nematicide resistance is a likely cause of reduced fluopyram efficacy on golf-course turf in Florida.

Morphological and Molecular Diversity among Pin Nematodes of the Genus Paratylenchus (Nematoda: Paratylenchidae) from Florida and Other Localities and Molecular Phylogeny of the Genus.

Pin nematodes (Paratylenchus spp.) are root parasites found worldwide. They have different life cycles and feeding habits and can damage a wide range of plants. A remarkable diversity of pin nematode species was found in soil samples from Florida and other states of the USA, Canada, and Spain. Using integrative taxonomy, two new species (Paratylenchus hawaiiensis sp. n. and P. roboris sp. n.), six valid species (Paratylenchulus acti, P. aquaticus, P. goldeni, P. paralatescens, P. minutus (=P. shenzhenensis syn. n.), and P. straeleni), and two undescribed species were identified from Florida; P. goldeni, P. hamatus, P. hamicaudatus, P. holdemani, and P. pedrami were found in California, P. minutus in Hawaii, P. goldeni in Oregon and Washington, and one new species, Paratylenchus borealis sp. n., in Alaska. Outside the USA, Paratylenchus projectus was detected in samples from Canada and Spain as well as P. holdemani and Paratylenchus sp. from Spain. The pin nematode species from Belgium and Russia identified in former studies as Paratylenchus sp. F was herein described as a new species with the name of P. borealis sp. n., using a population from Alaska. Previously reported molecular type A of P. aquaticus from Hawaii was reclassified as P. hawaiiensis sp. n., using a population from Florida. Paratylenchus roboris sp. n. from Florida has obese sedentary females with a stylet 63-71 µm long. The results of the molecular analysis of P. shenzhenensis from Florida and China indicated that it was conspecific with P. minutus from Hawaii and considered here as its junior synonym. New 26 D2-D3 expansion segments of 28S rRNA, 17 ITS rRNA, and 20 COI gene sequences were obtained in this study. Phylogenetic relationships of Paratylenchus are reconstructed using the D2-D3 of 28S rRNA, ITS rRNA, and COI gene sequences. Congruence of molecular and morphological evolution and species identification problems are discussed. Obese females were found in two major clades of Paratylenchus. The problem of reference materials is discussed, and it is proposed to make more efforts to collect topotype materials of known Paratylenchus species for molecular study.

Urotensin II-related peptides, Urp1 and Urp2, control zebrafish spine morphology.

The spine provides structure and support to the body, yet how it develops its characteristic morphology as the organism grows is little understood. This is underscored by the commonality of conditions in which the spine curves abnormally such as scoliosis, kyphosis, and lordosis. Understanding the origin of these spinal curves has been challenging in part due to the lack of appropriate animal models. Recently, zebrafish have emerged as promising tools with which to understand the origin of spinal curves. Using zebrafish, we demonstrate that the urotensin II-related peptides (URPs), Urp1 and Urp2, are essential for maintaining spine morphology. Urp1 and Urp2 are 10-amino acid cyclic peptides expressed by neurons lining the central canal of the spinal cord. Upon combined genetic loss of Urp1 and Urp2, adolescent-onset planar curves manifested in the caudal region of the spine. Highly similar curves were caused by mutation of Uts2r3, an URP receptor. Quantitative comparisons revealed that urotensin-associated curves were distinct from other zebrafish spinal curve mutants in curve position and direction. Last, we found that the Reissner fiber, a proteinaceous thread that sits in the central canal and has been implicated in the control of spine morphology, breaks down prior to curve formation in mutants with perturbed cilia motility but was unaffected by loss of Uts2r3. This suggests a Reissner fiber-independent mechanism of curvature in urotensin-deficient mutants. Overall, our results show that Urp1 and Urp2 control zebrafish spine morphology and establish new animal models of spine deformity.

The backbone, or spine, is an integral part of the human body, providing support to our torsos so that we can sit, stand, bend and twist. If this structure does not form correctly, it can lead to pain, neurologic problems, and mobility issues. The spine normally has curves, but these can become deformed for many reasons, including genetic and muscular factors. There are also cases in which the cause of a spine distortion is unknown, such as in scoliosis (where the spine twists to the side), lordosis (where the lower part of the spine curves excessively), and kyphosis (where the upper part of the spine shows extreme curvature). The structure of the spine is laid out during embryonic development and maintained throughout life. Experiments in zebrafish have shown that a crucial element in preserving the shape of the spine is the flow of cerebrospinal fluid or CSF. Propelled by the movement of little 'hairs' at the surface of specialized cells, this liquid runs through our central nervous system along a cavity lined with neurons. These nerve cells produce Urp1 and Urp2, two short molecules (or peptides) built from the same components as proteins. In zebrafish embryos, lowering the levels of these peptides had previously been shown to cause early body deformities. But what role, if any, do Urp1 and Urp2 play in maintaining the shape of the spine in adult zebrafish? Bearce et al. set out to answer this question. First, they generated mutant zebrafish which did not carry either Urp1, Urp2 or both peptides. Contrary to previous findings, all three of these mutants developed normally as embryos. Once they were adults, zebrafish lacking Urp1 exhibited normal spines, while those lacking Urp2 had slightly deformed curves. However, zebrafish lacking both peptides had prominent curves in the tail-region of their spines, somewhat akin to lordosis in humans. This indicates that both peptides are necessary for adult spine structure, but work in a semi-redundant manner. Interestingly, the defects observed first appeared in adolescent fish and gradually worsened as they grew; many forms of human spinal abnormalities follow a similar trajectory. Bearce et al. also tested the role of the protein Uts2r3, a receptor for peptides which belong to the urotensin family (such as Urp1 and Urp2). Fish lacking this protein showed normal spine structure as embryos, but distorted spinal curves as adults, suggesting that Urp1 and Urp2 might control spine morphology by signaling via the Uts2r3 receptor. Together, Bearce et al.'s observations show that disturbing urotensin signaling leads to a lordosis-like condition in adult zebrafish, with evident deformities in the tail-region of the spine. Considering the broad similarities in structures between the zebrafish and the human spine, these results point to a possible involvement of urotensin signaling in spine distortion in humans. More studies using zebrafish will likely provide further insights into the principles that control the shape of the spine and what goes wrong when it breaks down.

First Report of Belonolaimus Longicaudatus Infecting Soybean in Indiana.

Soybeans (Glycine max) are an important crop for Indiana, playing a major role in the state's economy. In June 2021, symptomatic soybean plants were submitted to Purdue University's Plant and Pest Diagnostic Laboratory for diagnosis. Sting nematodes were observed on the surface of the washed roots using stereo- and brightfield compound microscopy. A total of 76 sting nematodes per 100 cm3 soil were recovered from a composite soil sample. Morphological features and measurements of adult females and males of the sting nematode population were similar to those described for Belonolaimus longicaudatus. Molecular analysis confirmed the morphological identification using D2-D3 expansion segment of the 28S large subunit ribosomal DNA and internal transcribed spacer (ITS) regions. The consensus sequences were submitted to the National Center for Biotechnology Information (NCBI) database with accession numbers OM632679 and OM632681 for the D2-D3 and ITS regions, respectively. Parasitism of the sting nematode population to soybean plants was confirmed as its average population increased from 27 to 40 nematodes per pot 4 wk after inoculation under greenhouse conditions. To the best of our knowledge, this represents the first report of the sting nematode B. longicaudatus in Indiana.

First Report of Meloidogyne Incognita Infecting Mitragyna Speciosa in the United States.

Kratom (Mitragyna speciosa) belongs to the coffee family of Rubiaceae. The tree is native to Southeast Asia and primarily grown in Malaysia, Thailand, and Indonesia. Recently, it has been introduced and cultivated in other countries including the United States. The leaves and extracts of the leaves are used for medicinal and recreational purposes. In February 2022, kratom root and soil samples were submitted to the University of Florida Nematode Assay Laboratory for diagnosis by a commercial grower in Florida. Root galls were observed on the roots. On examination of soil and root samples, it is revealed that high numbers of root-knot nematodes (Meloidogyne sp.) are present. Molecular species identification was performed by a combination of the mitochondria haplotyping and species-specific primer techniques using TRNAH/MHR106 and MORF/MTHIS primer sets and Meloidogyne incognita-specific primers (MIF/MIR). The root-knot nematode infecting kratom is identified as M. incognita by molecular analysis. To our knowledge, this paper is the first report of M. incognita infecting kratom in the United States.

Bermudagrass Cultivars with Different Tolerance to Nematode Damage Are Characterized by Distinct Fungal but Similar Bacterial and Archaeal Microbiomes.

Turfgrass landscapes have expanded rapidly in recent decades and are a major vegetation type in urbanizing ecosystems. While turfgrass areas provide numerous ecosystem services in urban environments, ecological side effects from intensive management are raising concerns regarding their sustainability. One potentially promising approach to ameliorate the ecological impact and decrease the use of agricultural chemicals is to take advantage of naturally evolved turfgrass-associated microbes by harnessing beneficial services provided by microbiomes. Unfortunately, especially compared to agricultural crops, the microbiomes of turfgrasses are not well understood. Here, we analyzed microbial communities inhabiting the leaf and root endospheres as well as soil in two bermudagrass cultivars, 'Latitude 36' and 'TifTuf', which exhibit distinct tolerance to nematode damage, with the goal of identifying potential differences in the microbiomes that might explain their distinct phenotype. We used 16S rRNA gene V4 and ITS2 amplicon sequencing to characterize the microbiomes in combination with microbial cultivation efforts to identify potentially beneficial endophytic fungi and bacteria. Our results show that Latitude 36 and TifTuf showed markedly different fungal microbiomes, each harboring unique taxa from Ascomycota and Glomeromycota, respectively. In contrast, less difference was observed from bacterial and archaeal microbiomes, which were dominated by Bacteroidetes and Thaumarchaeota, respectively. The TifTuf microbiomes exhibited lower microbial diversity compared to Latitude 36. Many sequences could not be classified to a higher taxonomic resolution, indicating a relatively high abundance of hitherto undescribed microorganisms. Our results provide new insights into the structure and composition of turfgrass microbiomes but also raise important questions regarding the functional attributes of key taxa.

Mist chamber extraction for improved diagnosis of Meloidogyne spp. from golf course bermudagrass.

Meloidogyne spp. are among the most damaging plant-parasitic nematodes to golf course bermudagrass in the southern United States. Diagnostic samples processed by centrifugal flotation often recovered only low numbers of vermiform Meloidogyne spp. life stages (J2 and males) from soil, while roots were found to be heavily infested by sedentary life stages. Therefore, the University of Florida Nematode Assay Lab (NAL) evaluated mist extraction from turf plugs as a method for diagnosis of Meloidogyne spp. from golf course bermudagrass. Soil and turf plugs were obtained from 596 golf course bermudagrass small plots from multiple locations and cultivars over several years, and vermiform Meloidogyne spp. extracted from 100 cm3 of soil by centrifugal flotation and by mist chamber extraction from four 3.8-cm-diam. turf plugs were compared. Additionally, both extraction methods were performed on 431 golf course bermudagrass diagnostic samples received by the NAL from Florida, 36 golf course bermudagrass diagnostic samples from Texas, and 34 golf course bentgrass/bluegrass samples from California. In the small plots, and the bermudagrass samples from Florida and Texas, mist extraction had higher detection and recovery rates of vermiform Meloidogyne spp. than did centrifugal flotation. However, centrifugal flotation had higher detection and recovery rates than mist extraction from bentgrass/bluegrass samples from California. Mist extraction from turf plugs is superior to centrifugal flotation from soil for diagnosis of Meloidogyne spp. on golf course bermudagrass, but not on golf course bentgrass and bluegrass.

Yellow and purple nutsedge and coffee senna as hosts of common plant nematodes in Florida.

Yellow (Cyperus esculentus) and purple (C. rotundus) nutsedges, and coffee senna (Senna occidentalis) are common weeds in the southern USA and each have been reported as alternative hosts for plant-parasitic nematodes. Our objective was to determine the host suitability of these weeds to plant-parasitic nematodes common in Florida agriculture and turfgrass systems. The root-knot nematode (RKN) species tested included Meloidogyne arenaria, M. enterolobii, M. floridensis, M. graminis, M. hapla, M. incognita, and M. javanica. The host status of sting nematode, Belonolaimus longicaudatus, was also evaluated, but only on the nutsedge species. All RKN species evaluated reproduced on both nutsedge species and had a reproductive factor greater than one, except for M. graminis on yellow nutsedge. However, only M. hapla, M. javanica, and M. graminis induced visual galls on yellow nutsedge and only M. graminis caused galling on purple nutsedge. Meloidogyne arenaria and M. graminis reproduced at a greater rate on purple nutsedge than on yellow nutsedge. Both nutsedge species were good hosts to B. longicaudatus. Coffee senna was a host to M. enterolobii, a poor host to M. incognita, and nonhost to the other RKN species evaluated.

Nematicide effects on non-target nematodes in bermudagrass.

In turfgrass systems, nematicides are a valuable tool for managing plant-parasitic nematode populations, but few studies have examined nematicide effects on non-target nematodes. The study evaluated effects of turfgrass nematicide formulations of abamectin (Divanem SC), fluopyram (Indemnify), furfural (MultiGuard Protect EC), and fluensulfone (Nimitz Pro G) on non-target nematode populations in bermudagrass (Cynodon spp.). Nematicides were applied at labeled rates every four weeks as a summer treatment program from June 7, 2016 to August 30, 2016 and April 24, 2017 to July 18, 2017. Samples were collected before the initial treatment and 2 d, 14 d, 56 d, and 238 d after the final treatment in both years for nematode community analysis. Data from each nematicide treatment were compared to the untreated at each sample date using analysis of covariance with initial population counts serving as the covariate. Abamectin had moderate impact and fluopyram had substantial impact on the non-target nematodes. Furfural and fluensulfone had minimal impact on non-target nematodes. The results of this study suggest nematicides can impact non-target nematode densities in bermudagrass.In turfgrass systems, nematicides are a valuable tool for managing plant-parasitic nematode populations, but few studies have examined nematicide effects on non-target nematodes. The study evaluated effects of turfgrass nematicide formulations of abamectin (Divanem SC), fluopyram (Indemnify), furfural (MultiGuard Protect EC), and fluensulfone (Nimitz Pro G) on non-target nematode populations in bermudagrass (Cynodon spp.). Nematicides were applied at labeled rates every four weeks as a summer treatment program from June 7, 2016 to August 30, 2016 and April 24, 2017 to July 18, 2017. Samples were collected before the initial treatment and 2 d, 14 d, 56 d, and 238 d after the final treatment in both years for nematode community analysis. Data from each nematicide treatment were compared to the untreated at each sample date using analysis of covariance with initial population counts serving as the covariate. Abamectin had moderate impact and fluopyram had substantial impact on the non-target nematodes. Furfural and fluensulfone had minimal impact on non-target nematodes. The results of this study suggest nematicides can impact non-target nematode densities in bermudagrass.

Developing a One-Step Multiplex PCR Assay for Rapid Detection of Four Stubby-Root Nematode Species, Paratrichodorus allius, P. minor, P. porosus, and Trichodorus obtusus.

Four trichodorid species, Paratrichodorus allius, P. minor, P. porosus, and Trichodorus obtusus, were found in multiple states in the United States. Traditional diagnosis based on morphology and morphometrics is laborious and requires an experienced taxonomist. Additionally, end-point diagnosis using PCR was only available for P. allius. To increase diagnostic efficiency and reduce costs, a one-step multiplex PCR assay was developed to simultaneously identify these four species using one PCR reaction. Available sequences of 18S ribosomal DNA and internal transcribed spacer 1 (ITS1) region of these species were aligned and five primers were designed. The conserved forward primer located in the 18S region, in combination with the species-specific antisense primer in the ITS1 region, amplified a single distinctive PCR fragment for each species (421/425 bp for P. allius, 190 bp for P. minor, 513 bp for P. porosus, and 353 bp for T. obtusus). In silico analysis with 10 other trichodorid species and experimental analysis using samples with these four species, 20 other plant-parasitic and three non-plant-parasitic nematodes demonstrated high specificity with the primers designed. The multiplex PCR amplified desirable fragments using a set of artificially mixed templates containing one, two, three, or four targeted species. The reliability of multiplex PCR results was demonstrated by using nematode populations isolated from infested fields from diverse geographic regions in eight states. The multiplex PCR-based tool developed in this study for the first time provides a simple, rapid, and cost-friendly assay for accurate diagnosis of the four major trichodorid nematodes in the United States.

Responses of Anastrepha suspensa, Diachasmimorpha longicaudata, and Sensitivity of Guava Production to Heterorhabditis bacteriophora in Fruit Fly Integrated Pest Management.

Caribbean fruit fly, also known as Caribfly or Anastrepha suspensa , is a major tephritid pest of guavas. A virulent entomopathogenic nematode (EPN) species was investigated to suppress the fruit-to-soil stages of Caribflies, which are also attacked by the koinobiont parasitoid Diachasmimorpha longicaudata in south Florida. The main objective was to develop a feasible and cost-effective EPN-application method for integrated pest management (IPM) of Caribfly to improve guava production. Naturally infested guavas were treated with increasing Heterorhabditis bacteriophora infective juvenile (IJ) concentration or rate (0, 25, 50, …, 1,600 IJs cm -2 ) in field trials to measure the optimum IJ rate and then examine sensitivity of producing guavas to inclusion of Heterorhabditis bacteriophora in Caribfly IPM plans. Relative survival of Caribfly in treatments significantly decreased with increasing IJ rate from 0 to 100 IJs cm -2 . Similarly, probability of observing large numbers of parasitoid wasps ( Diachasmimorpha longicaudata ) in EPN treatments significantly declined with increasing IJ rate (0-100 IJs cm -2 ), even though the non-target effects of Heterorhabditis bacteriophora on relative survival of Diachasmimorpha longicaudata could not be determined because of few emerging parasitoid wasps. Optimum suppression (⩾ 60%) of Caribfly was consistently achieved at 100 IJs cm -2 or 17,500 IJs fruit -1 . Profitability analysis showed that Heterorhabditis bacteriophora can be included in Caribfly IPM tactics to produce guavas. Costs of EPNs in Caribfly IPM are minimized if Heterorhabditis bacteriophora is strategically applied by spot treatment of fruit. Repayment of costs of EPN-augmentation by spot treatments appears achievable by recovering 5.71% of the annual yield losses (⩾1,963 kg ha -1 ≈ US$ 8,650 ha -1 ), which are largely due to Caribfly infestation. Hectare-wide EPN-augmentation (or broadcasting) method requires more fruit recovery than the total annual yield losses to repay its high costs. Profitability of guava production in south Florida will not be very sensitive to marginal costs of the spot treatment method, when compared to the field-wide broadcasting of Heterorhabditis bacteriophora .

Molecular Characterization and Identification of Stubby Root Nematode Species From Multiple States in the United States.

Stubby root nematodes (SRN) are important plant parasites infecting many crops and widely distributed in many regions of the United States. SRN transmit Tobacco rattle virus, which causes potato corky ringspot disease, thereby having a significant economic impact on the potato industry. In 2015 to 2017, 184 soil samples and 16 nematode suspensions from North Dakota, Minnesota, Idaho, Oregon, Washington, South Carolina, North Carolina, and Florida were assayed for the presence of SRN. SRN were found in 106 soil samples with population densities of 10 to 320 SRN per 200 g of soil and in eight of the nematode suspensions. Sequencing of ribosomal DNA (rDNA) or species-specific polymerase chain reaction assays revealed the presence of four SRN species, including Paratrichodorus allius, P. minor, P. porosus, and Trichodorus obtusus. Accordingly, their rDNA sequences were characterized by analyzing D2-D3 of 28S rDNA, 18S rDNA, and internal transcribed spacer (ITS) rDNA obtained in this study and retrieved from GenBank. Both intra- and interspecies variations were higher in ITS rDNA than 18S rDNA and D2-D3 of 28S rDNA. Based on phylogenetic analysis, the four SRN species formed a monophyletic group, with P. allius more closely related to P. porosus than P. minor and T. obtusus. Indel variation of ITS2 rDNA was present in P. allius populations from the same geographic regions. This study documented the occurrence of SRN species across multiple states. The intra- and interspecies genetic diversity of rDNA in this study will provide more information for understanding the evolutionary relationships of SRN and will be valuable for future studies of SRN species identification and management.

Vertical Distribution of Pasteuria penetrans Parasitizing Meloidogyne incognita on Pittosporum tobira in Florida.

Pasteuria penetrans is considered as the primary agent responsible for soil suppressiveness to root-knot nematodes widely distributed in many agricultural fields. A preliminary survey on a Pittosporum tobira field where the grower had experienced a continuous decline in productivity caused by Meloidogyne incognita showed that the nematode was infected with Pasteuria penetrans. For effective control of the nematode, the bacterium and the host must coexist in the same root zone. The vertical distribution of Pasteuria penetrans and its relationship with the nematode host in the soil was investigated to identify (i) the vertical distribution of P. penetrans endospores in an irrigated P. tobira field and (ii) the relationship among P. penetrans endospore density, M. incognita J2 population density, and host plant root distribution over time. Soil bioassays revealed that endospore density was greater in the upper 18 cm of the top soil compared with the underlying depths. A correlation analysis showed that the endospore density was positively related to the J2 population density and host plant root distribution. Thus, the vertical distribution of P. penetrans was largely dependent on its nematode host which in turn was determined by the distribution of the host plant roots. The Pasteuria was predominant mostly in the upper layers of the soil where their nematode host and the plant host roots are abundant, a factor which may be a critical consideration when using P. penetrans as a nematode biological control agent.

Management of Root-knot Nematode (Meloidogyne incognita) on Pittosporum tobira Under Greenhouse, Field, and On-farm Conditions in Florida.

Root-knot nematodes are important pests of cut foliage crops in Florida. Currently, effective nematicides for control of these nematodes on cut foliage crops are lacking. Hence, research was conducted at the University of Florida to identify pesticides or biopesticides that could be used to manage these nematodes. The research comprised on-farm, field, and greenhouse trials. Nematicide treatments evaluated include commercial formulations of spirotetramat, furfural, and Purpureocillium lilacinum (=Paecilomyces lilacinus) strain 251. Treatment applications were made during the spring and fall seasons according to manufacturer's specifications. Efficacy was evaluated based on J2/100 cm3 of soil, J2/g of root, and crop yield (kg/plot). Unlike spirotetramat, which did not demonstrate any measurable effects on Meloidogyne incognita J2 in the soil, furfural and P. lilacinum were marginally effective in reducing the population density of M. incognita on Pittosporum tobira. However, nematode reduction did not affect yield significantly. Although furfural and P. lilacinum have some potential for management of M. incognita on cut foliage crops, their use as a lone management option would likely not provide the needed level of control. Early treatment application following infestation provided greater J2 suppression compared to late application, suggesting the need for growers to avoid infested fields.

Mitochondrial Haplotype-based Identification of Root-knot Nematodes (Meloidogyne spp.) on Cut Foliage Crops in Florida.

Florida accounts for more than 75% of the national cut foliage production. Unfortunately, root-knot nematodes (RKN) (Meloidogyne spp.) are a serious problem on these crops, rendering many farms unproductive. Currently, information on the Meloidogyne spp. occurring on most commonly cultivated cut foliage crops in Florida, and tools for their rapid identification are lacking. The objectives of this study were to (i) identify specific RKN infecting common ornamental cut foliage crops in Florida and (ii) evaluate the feasibility of using the mtDNA haplotype as a molecular diagnostic tool for rapid identification of large samples of RKN. A total of 200 Meloidogyne females were collected from cut foliage plant roots. Meloidogyne spp. were identified by PCR and RFLP of mitochondrial DNA. PCR and RFLP of mitochondrial DNA were effective in discriminating the Meloidogyne spp. present. Meloidogyne incognita is the most dominant RKN on cut foliage crops in Florida and must be a high target for making management decisions. Other Meloidogyne spp. identified include M. javanica, M. hapla, Meloidogyne sp. 1, and Meloidogyne sp. 2. The results for this study demonstrate the usefulness of the mtDNA haplotype-based designation as a valuable molecular tool for identification of Meloidogyne spp.

Effects of Infection by Belonolaimus longicaudatus on Rooting Dynamics among St. Augustinegrass and Bermudagrass Genotypes.

Understanding rooting dynamics using the minirhizotron technique is useful for cultivar selection and to quantify nematode damage to roots. A 2-yr microplot study including five bermudagrass ('Tifway', Belonolaimus longicaudatus susceptible; two commercial cultivars [TifSport and Celebration] and two genotypes ['BA132' and 'PI 291590'], which have been reported to be tolerant to B. longicaudatus) and two St. Augustinegrass ('FX 313', susceptible, and 'Floratam' that was reported as tolerant to B. longicaudatus) genotypes in a 5 x 2 and 2 x 2 factorial design with four replications, respectively, was initiated in 2012. Two treatments included were uninoculated and B. longicaudatus inoculated. In situ root images were captured each month using a minirhizotron camera system from April to September of 2013 and 2014. Mixed models analysis and comparison of least squares means indicated significant differences in root parameters studied across the genotypes and soil depths of both grass species. 'Celebration', 'TifSport' and 'PI 291590' bermudagrass, and 'Floratam' St. Augustinegrass had significantly different root parameters compared to the corresponding susceptible genotypes (P ≤ 0.05). Only 'TifSport' had no significant root loss when infested with B. longicaudatus compared to non-infested. 'Celebration' and 'PI 291590' had significant root loss but retained significantly greater root densities than 'Tifway' in B. longicaudatus-infested conditions (P ≤ 0.05). Root lengths were greater at the 0 to 5 cm depth followed by 5 to 10 and 10 to 15 cm of vertical soil depth for both grass species (P ≤ 0.05). 'Celebration', 'TifSport', and 'PI 291590' had better root vigor against B. longicaudatus compared to Tifway.

Effects of a Commercial Formulation of Bacillus firmus I-1582 on Golf Course Bermudagrass Infested with Belonolaimus longicaudatus.

One of the primary pests of bermudagrass (Cynodon spp.) on golf courses in the southeastern United States is Belonolaimus longicaudatus (sting nematode). In 2011, a commercial formulation of Bacillus firmus I-1582, Nortica 5WG, was launched in the United States for management of plant-parasitic nematodes on turfgrasses. To test the efficacy of late winter/early spring application of this biopesticide on B. longicaudatus, two field trials in 2009 compared B. firmus with fenamiphos and untreated control treatments. In 2011, two additional field trials compared treatment with B. firmus with untreated control only. These trials measured treatment effects on the population density of B. longicaudatus, turf root length, and turf percent green cover. In all four trials, treatment with B. firmus improved root length and decreased numbers of B. longicaudatus in contrast to the untreated. These results indicate that late winter/early spring application of B. firmus is an effective biopesticide treatment for management of B. longicaudatus on golf course bermudagrass.

Effects of a Commercial Formulation of Paecilomyces lilacinus Strain 251 on Overseeded Bermudagrass Infested with Belonolaimus longicaudatus.

Belonolaimus longicaudatus is an important parasite of both warm-season bermudagrass and winter overseed grasses used on golf courses in the southeastern United States. Field trials were conducted to study the effects of a commercial formulation of Paecilomyces lilacinus strain 251 applied to overseed grasses during the winter and early spring on population density of B. longicaudatus and bermudagrass health in late spring after bermudagrass broke dormancy. These studies found that P. lilacinus reduced numbers of B. longicaudatus in most cases, but not below damaging levels. Multiple applications of 1 × 10(10) spores/m(2) were generally more effective than 2 × 10(10) spores/m(2) in reducing nematode numbers and improving turf roots. These results indicate that application of this formulation of P. lilacinus strain 251 to overseeded turf in the spring may be a useful integrated pest management tool for B. longicaudatus on bermudagrass, but is not sufficient as a stand-alone nematode management tactic.

Interaction Between Belonolaimus longicaudatus and Helicotylenchus pseudorobustus on Bermudagrass and Seashore Paspalum Hosts.

Belonolaimus longicaudatus and Helicotylenchus pseudorobustus are among the most common nematode parasites of turfgrasses in Florida. Bermudagrass (Cynodon dactylon × C. transvaalensis) and seashore paspalum (Paspalum vaginatum) are the two turf species most commonly used on Florida golf courses. This paper explores the interactions between B. longicaudatus and H. pseudorobustus on bermudagrass and seashore paspalum hosts. Data collected from thousands of nematode samples submitted to the Florida Nematode Assay Lab over a 8-yr period revealed a negative relationship between B. longicaudatus and H. pseudorobustus on bermudagrass, but not seashore paspalum. In a multi-year field plot experiment using multiple cultivars of bermudagrass, and seashore paspalum B. longicaudatus and H. pseudorobustus were negatively related on both turf species. Greenhouse trials where multiple cultivars of both turf species were inoculated with different combinations of B. longicaudatus and H. pseudorobustus found that each nematode species was inhibitory to the other on both host species. Belonolaimus longicaudatus and H. pseudorobustus clearly impact each other on turfgrass hosts, although the mechanism of the nematode-nematode interactions is unknown.