Evaluating a naturally occurring baculovirus for extended biological control of the black cutworm (Lepidoptera: Noctuidae) in golf course habitats

Golf courses are a potential market for microbial insecticides, but how intensive management of such sites interacts with efficacy of entomopathogens is poorly known. We evaluated Agrotis ipsilon nucleopolyhedrovirus (AgipMNPV) for suppressing black cutworms, Agrotis ipsilon Hufnagel (Lepidoptera: Noctuidae), in turf representative of golf course habitats and on whole tees under actual play. In independent trials on sand- or soil-based putting greens and surrounds, or fairway-height creeping bentgrass (Agrostis stolonifera L.), < or = 1-wk-old AgipMNPV residues (10 x 10(8) occlusion bodies [OBs] per m2) typically gave 50-60% lethal infection of introduced third instars. In most cases, however, there was no residual control beyond 2-4 wk. Spraying fairway-height bentgrass with AgipMNPV alone (10 x 10(9) OBs per m2) gave 90, 85, and 7% infection of second instars introduced 4 d, 3 wk, or 5 wk later, but adjuvants (optical brightener, lignin, or both) intended to synergize and protect the virus from UV degradation did not extend infectivity. Fresh (< 1-wk-old) AgipMNPV residues killed 76-86% of neonates hatching from eggs on tees under play, but levels of control plummeted within a few weeks. Three species of braconids, an encyrtid Copidosoma bakeri (Howard), and a tachinid, Bonnetia comta (Fallen) collectively killed 24-31% of larvae recovered from those tees. AgipMNPV seems better suited for targeted control of early instars than for season-long control. Golf turf is a severe environment for baculoviruses so several applications per growing season would probably be needed to maintain high enough titers on grass foliage to effectively control caterpillar pests.     

Potential of Agrotis ipsilon nucleopolyhedrovirus for suppression of the black cutworm (Lepidoptera: Noctuidae) and effect of an optical brightener on virus efficacy.

Studies were performed in the laboratory, greenhouse and field to assess the potential of Agrotis ipsilon multicapsid nucleopolyhedrovirus (AgipMNPV) and a viral enhancing agent, M2R, for suppression of Agrotis ipsilon (Hufnagel). In laboratory droplet feeding bioassays, AgipMNPV was shown to be highly active against third-instar A. ipsilon. The optical brightener M2R significantly reduced LD50 estimates by approximately 160-fold, but had no direct effect on survival time estimates. In greenhouse trials, spray and bait formulations of AgipMNPV significantly reduced feeding damage to corn seedlings caused by third-instar A. ipsilon. In two sets of replicated field trials, bait formulations of AgipMNPV significantly reduced feeding damage to corn seedlings by third-instar A. ipsilon. However, there were no beneficial effects attributable to the inclusion of M2R in AgipMNPV formulations under greenhouse or field conditions. It seems likely that in an appropriately designed pest management program AgipMNPV could be used to suppress field populations of early and mid-instar A. ipsilon.     

Orientation Behavior and Regulatory Gene Expression Profiles in Migratory Agrotis ipsilon (Lepidoptera: Noctuidae)

Journal of Insect Behavior - Many insects such as the crop pest Agrotis ipsilon (black cutworm) migrate seasonally to exploit suitable breeding and winter habitats. The migration of a crop pest...     

Effect of tinopal LPW on the insecticidal properties and genetic stability of the nucleopolyhedrovirus of Spodoptera exigua (Lepidoptera: Noctuidae)

The influence of an optical brightener, Tinopal LPW, on the activity of a purified genotype of the nucleopolyhedrovirus (SeMNPV) of the beet armyworm, Spodoptera exigua (Hübner), was determined in second to fifth instar (L2-L5) S. exigua. When mixed with viral occlusion bodies (OB) 1% Tinopal LPW significantly reduced the median lethal dose (LD50) of the virus in all instars compared with insects treated with SeMNPV alone. Levels of enhancement, as determined by LD50 values, ranged from 2.6- to 580-fold, depending on the instar. The greatest enhancement occurred on the two later instars, L4 (70-fold) and L5 (580-fold), which show a much higher resistance to SeMNPV infection than earlier instars. The median time to death (MTD) values were not significantly different in any instar among larvae treated with SeMNPV + Tinopal LPW and those treated with SeMNPV alone. Larval development in SeMNPV + Tinopal LPW treated larvae was retarded, in second and fourth instars, compared with controls or larvae treated with SeMNPV alone. The OB yields from SeMNPV treated larvae were almost 1.6-fold greater in second instars (9.3 x 10(6) OBs/larvae), and 1.9-fold greater in fourth instars (1.9 x 10(8) OBs/larvae), than those obtained in larvae treated with SeMNPV + Tinopal LPW. The addition of 1% Tinopal LPW to the virus suspension did not alter the genotypic composition of viral progeny during four successive passages of the virus.     

Transgenic creeping bentgrass with delayed dollar spot symptoms

Creeping bentgrass (Agrostis palustris Huds) is animportant turfgrass used on golf course greens and fairways. It is susceptibleto a number of fungal pathogens and requires considerable fungicide use fordisease control. Transgenic approaches may be useful in improving the level ofdisease resistance. We have generated transgenic creeping bentgrass plantsexpressing PR5K from Arabidopsis thaliana (L.) Henyh. PR5Kis a receptor protein kinase whose extracellular domain is homologous to thePR5family of pathogenesis-related proteins. In a field test of plants inoculatedwith the fungal pathogen dollar spot (Sclerotiniahomoeocarpa F.T. Bennett) four of the eight transgenic lines showeddelays in disease expression of 29 to 45 days, relative to the control plants.     

Comparison of sticky wing and cone pheromone traps for monitoring seasonal abundance of black cutworm adults and larvae on golf courses

Black cutworm, Agrotis ipsilon (Hufnagel) (Lepidoptera: Noctuidae), flight activity was monitored on three golf courses in Wisconsin by using two types of pheromone traps: the Texas cone trap and sticky wing trap. The Texas cone trap caught significantly more black cutworm males compared with the sticky wing trap, capturing almost 12-fold more males. Black cutworm males were most abundant during mid-July in 2001 and 2002, between 700 and 800 cumulative degree-days. Flight activity also was detected in early May and mid-August, but these peaks were not as pronounced as in mid-July. No definitive relationship between black cutworm flight activity and subsequent larval infestations on golf course putting greens occurred.     

Characterization of 215 simple sequence repeat markers in creeping bentgrass (Agrostis stolonifera L.)

Creeping bentgrass (Agrostis stolonifera L.) is a versatile, cross-pollinated, temperate and perennial turfgrass species. It occurs naturally in a wide variety of habitats and is also cultivated on golf courses, bowling greens and tennis courts worldwide. Isozymes and amplified fragment length polymorphisms (AFLPs) have been used to determine genetic diversity, and restriction fragment length polymorphisms (RFLPs) and random amplified polymorphic DNA (RAPDs) were used to construct a genetic linkage map of this species. In the current report, we developed and characterized 215 unique genomic simple sequence repeat (SSR) markers in creeping bentgrass. The SSRs reported here are the first available markers in creeping bentgrass to date. Eight hundred and eighteen alleles were amplified by 215 SSR loci, an average of 3.72 alleles per locus. Fifty-nine per cent of those alleles segregated in a 1:1 Mendelian fashion (P > 0.05). Twenty-two per cent had a distorted segregation ratio (P ≤ 0.05). These SSR markers will be useful for assessing genetic diversity in creeping bentgrass and will be important for the development of genetic linkage maps and identifying quantitative trait loci. These markers could enhance breeding programmes by improving the efficiency of selection techniques.     

Comparative study on the susceptibility of cutworms (Lepidoptera: Noctuidae) to Agrotis segetum nucleopolyhedrovirus and Agrotis ipsilon nucleopolyhedrovirus

The common cutworm (Agrotis segetum) and the black cutworm (Agrotis ipsilon) are serious soil pests of many vegetable and field crops all over the world. We have demonstrated the cross-infectivity of two baculoviruses, A. segetum nucleopolyhedrovirus (AgseNPV) and A. ipsilon nucleopolyhedrovirus (AgipNPV) for these two insect pests. The susceptibility of A. segetum to AgipNPV was confirmed by DNA restriction endonuclease analyses of DNA isolated from virus harvested from infected A. segetum larvae. For an initial comparison of both viruses, partial polyhedrin sequences were amplified by PCR, cloned, and sequenced. Both viruses shared a very similar polyhedrin gene sequence resulting in only three amino acid substitutions. Phylogenetic analyses clearly demonstrated that both viruses belong to NPV group II and are most closely related to a clade consisting of Spodoptera exigua NPV, Spodoptera frugiperda NPV, and Spodoptera littoralis NPV. Since AgipNPV shows high virulence for both cutworm species, it appears to be a suitable candidate as a single biological control agent of A. segetum and A. ipsilon.     

Enhanced toxicity of Bacillus thuringiensis subspecies kurstaki and aizawai to black cutworm larvae (Lepidoptera: Noctuidae) with Bacillus sp. NFD2 and Pseudomonas sp. FNFD1

Bacillus thuringiensis subspecies kurstaki and aizawai are important control agents for lepidopteran pests. Bioassays were designed to test B. t. kurstaki and aizawai against second- and-fourth instar black cutworm larvae with and without Bacillus sp. NFD2 and Pseudomonas sp. FNFD1 bacteria. B. thuringiensis subsp. aizawai (XenTari) was more toxic to both second- and fourth-instar black cutworm, Agrotis ipsilon (Hufnagel) (Lepidoptera: Noctuidae), larvae than B. t. kurstaki (DiPel) at 7 d after treatment (DAT). When DiPel was combined with NFD2 or FNFD1 versus second instars, the LC50s were 5.0X and 4.7X lower, respectively, than with DiPel alone. DiPel combined with both NFD2 and FNFD1 versus second instars resulted in an LC50 value 7.7X lower than with DiPel alone. When XenTari was combined with NFD2 or FNFD1 versus second instars, the LC50s were 5.2X and 3.8X lower, respectively, than with XenTari alone. XenTari combined with both NFD2 and FNFD1 versus second instars resulted in an LC50 9.7X lower than with XenTari alone. When DiPel was combined with NFD2 or FNFD1 versus fourth instars, the LC50s were 4.4X and 3.4X lower, respectively, than with DiPel alone. DiPel combined with both NFD2 and FNFD1 versus fourth instars resulted in an LC50 5.0X lower than with DiPel alone. When XenTari was combined with NFD2 or FNFD1 versus fourth instars, the LC50s were 5.7X and 3.3X lower, respectively, than with XenTari alone. XenTari combined with both NFD2 and FNFD1 versus fourth instars resulted in an LC50 6.7X lower than with XenTari alone.     

Efficacy of botanical insecticides from Piper species (Piperaceae) extracts for control of Ruropean chafer (Coleoptera: Scarabaeidae)

Biopesticides, including botanicals, can offer a safe and effective alternative to conventional insecticides for controlling major insect pests within an integrated pest management program. The current study highlights the practical application of a botanical insecticide for controlling a major insect pest of turfgrass: European chafer, Rhizotrogus majalis (Razoumowsky). Greenhouse and field trials were conducted to test the efficacy of a botanical formulation based on black pepper, Piper nigrum L. (Piperaceae), seed extracts to R. majalis larvae. The 7-d P. nigrum extract LC50 for R. majalis third instars was 2.5%. Successful treatment in the field was accomplished with the application of a 2% P. nigrum formulation to turfgrass infested with R. majalis second and third instars, whereas 4% extract was required in a second field trial with older third instars. The 2% pepper extract activity was comparable with the conventional insecticide diazinon in the first field trial. However, the 4% pepper extracts significantly affected the earthworm populations in treated plots compared with diazinon in the second field trail. The analysis of soil residues for piperamides in the P. nigrum extract determined a half-life of 1 - 2.6 d in the first and second field trials, respectively. This confirmed the expectation that under field conditions the residual activity would be less than conventional insecticides, thereby reducing the environmental risk associated with pesticide use. We recommend the pepper formulation for spot treatment applications when population densities reveal an epicenter of infestation rather than broadcasting over large areas, thus helping to minimize cost and negative affects on nontarget invertebrates.     

Ovipositional preferences and larval survival of annual bluegrass weevil,Listronotus maculicollis,on Poa annua and selected bentgrasses (Agrostis spp.)

The annual bluegrass weevil (ABW), Listronotus maculicollis Kirby (Coleoptera: Curculionidae), is a serious and expanding pest of short‐cut turfgrass on golf courses in eastern North America. Increasing problems with the development of insecticide resistance in this pest highlights the need for more sustainable management approaches. Plant resistance is one of the most promising alternative strategies. Bentgrasses are the dominant grass species on golf course fairways, tees, and putting greens in the areas affected by ABW. But Poa annua L. (Poaceae), a highly invasive weed, often constitutes a large percentage of turf stands in short‐mown golf courses and is thought to be particularly susceptible to ABW. We studied resistance to ABW in four cultivars of creeping bentgrass, Agrostis stolonifera L., and two cultivars each of colonial bentgrass, Agrostis capillaris L., and velvet bentgrass, Agrostis canina L. (Poaceae), in comparison with P. annua by addressing the three major components of resistance: antixenosis (adult ovipositional and feeding preferences), antibiosis (larval survival and growth), and grass tolerance (grass damage). Our findings suggest that antixenosis/non‐preference is at least partially involved in bentgrass resistance to ABW. Even though oviposition was observed in all tested grasses, females laid significantly fewer eggs in Agrostis spp. than in P. annua. Compared to P. annua, Agrostis spp. were also less suitable for larval development with lower numbers of ABW immatures recovered and larvae weighing less and being less advanced in development. Resistance levels to ABW larvae varied significantly among Agrostis spp. and cultivars. Agrostis canina was least preferred by females for oviposition and A. stolonifera was the least suitable for larval survival and development. Agrostis spp., especially A. stolonifera, were more tolerant to ABW feeding than P. annua. Our findings suggest that reduction in P. annua and replacement with Agrostis spp., especially A. stolonifera, wherever feasible should be integral to more sustainable approaches to ABW management.     

Greenhouse evaluation of fitness-related reproductive traits in roundup<Superscript>®</Superscript>-tolerant transgenic creeping bentgrass (<Emphasis Type="Italic">Agrostis Stolonifera</Emphasis> L.)

Summary Creeping bentgrass is a very important turfgrass species used extensively on golf course greens, fairways, and tees. One of the challenges of creeping bentgrass management is the control of grassy weeds, most of which respond to herbicides in a similar manner to that of creeping bentgrass. As part of a weed management program for golf courses, Roundup^?-tolerant creeping bentgrass will be simple to employ and more effective in controlling problem weeds than currently available methods. The goal of this research was to evaluate fitness-related reproductive traits in four transgenic creeping bentgrass events modified to express a Roundup^?-tolerant gene, cp4 epsps, to determine if these creeping bentgrass events had gained an unexpected reproductive fitness advantage. We compared transgenic events ASR 333, ASR801 with their nontransformed tissue culture line, C99056L and transgenic events ASR365, ASR368 with their non-transformed tissue culture line, B99061R. Populations of plants from three conventional cultivars were also included for comparison to determine whether significant variations, if present in transgenic events, were novel to the non-transformed organism, Agrostis stolonifera L. Our results showed that none of the four transgenic events surveyed were significantly different from the respective non-transformed tissue culture line plants for the following characteristics: first heading date, anthesis duration, inflorescence length, number of florets per inflorescence, pollen size, and seed-set capacity through open-pollination. One of the transgenic events, ASR333, needed significantly more days for anthesis initiation than the nontransformed tissue culture line, C99056L; while another transgenic event, ASR801, exhibited significantly shorter pollen longevity than plants of the tissue culture line, C99056L. However, ASR801 was not significantly different from the conventional cultivars ‘Penn A-4’ and ‘Penncross’ for pollen longevity. Plants of both transgenic events ASR365 and ASR368 did not differ significantly from plants of the tissue culture line, B99061R, for all characters measured.     

The effects of fungicides on the phylloplane yeast populations of creeping bentgrass

The effects of fungicides on population size and the development of fungicide resistance in the phylloplane yeast flora of bentgrass was investigated. In the spring of 2001, azoxystrobin, chlorothalonil, flutolanil, and propiconazole were applied separately over a 6-week period to creeping bentgrass (Agrostis palustris Huds.). Total and fungicide-resistant yeast populations were assessed by dilution plating onto either potato dextrose agar or potato dextrose agar amended with the test fungicides. Total yeast populations in the fungicide-treated plots were significantly lower than the check plots on three out of four sample dates. In the fall, azoxystrobin or propiconazole were applied twice to the bentgrass over 3 weeks. Significantly larger total yeast populations were observed compared with resistant or highly resistant populations for each treatment on every sample date. Total yeast populations were significantly higher in the check plots compared with either the propiconazole- or azoxystrobin-treated plots on the first three of five sample dates. A collection of yeasts (N = 114) with no prior exposure to fungicides were more sensitive to chlorothalonil, propiconazole, flutolanil, and iprodione than a second group (N = 115) isolated from fungicide-treated turfgrass. These results suggest that fungicide resistance among phylloplane yeasts is widespread and could be an important factor in the development of biological control agents for turfgrass diseases.     

Efficacy and persistence of entomopathogenic nematodes for black cutworm control in turfgrass

Field experiments were conducted in turf maintained under golf course fairway conditions in May, June, and August 2009 and in August and September 2010 to evaluate the ability of entomopathogenic nematodes to control larval populations of the black cutworm, Agrotis ipsilon, on golf courses. Commercial products containing the entomopathogenic nematodes Heterorhabditis bacteriophora, Steinernema carpocapsae, S. feltiae, and S. riobrave were applied at 1.0 or 2.5×10⁹ infective juveniles per ha against fourth-instar black cutworms. Larval mortality and turf damage were evaluated at 4 and/or 7 days after treatment (DAT). Steinernema carpocapsae was the best performing species due to a combination of high control rates (average 83%), most consistent results (70–90% range), high speed of kill (average 68% at 4 DAT), and prevention of significant turf damage despite very high larval densities at 0 DAT. Efficacy of S. feltiae and H. bacteriophora was often similar to that of S. carpocapsae but overall less consistent. Short-term persistence of the nematodes was evaluated in four turfgrass sites maintained under golf course putting green, fairway, or rough conditions in June and August 2009 by baiting soil samples at 0, 4, 7, and 14 DAT. Relative to recovery immediately after application, at least 50% of S. feltiae and 25% of S. carpocapsae consistently persisted up to 4 days in one of the greens and up to 7 days in some trials. Our finding suggests that S. carpocapsae and S. feltiae may provide adequate black cutworm control in golf course turf under moderate summer temperatures.     

Activity of wild-type and hybrid<Emphasis Type="Italic"> Bacillus thuringiensis</Emphasis> δ-endotoxins against<Emphasis Type="Italic"> Agrotis ipsilon</Emphasis>

Twelve Cry1 and two Cry9 delta-endotoxins from Bacillus thuringiensis were tested for their activity against black cutworm ( Agrotis ipsilon). A. ipsilon was not susceptible to many toxins, but three toxins had significant activity. Cry9Ca was the most toxic, followed by Cry1Aa and Cry1Fb. Hybrids between these three active proteins were made by in vivo recombination and analyzed for activity against A. ipsilon. Analysis of hybrids between Cry1Aa and Cry1Fb indicated that domain I of Cry1Aa protein was involved in its higher activity.     

Selection of a nucleopolyhedrovirus for control of Spodoptera frugiperda (Lepidoptera: Noctuidae): structural, genetic, and biological comparison of four isolates from the Americas

Spodoptera frugiperda (J. E. Smith) (Lepidoptera: Noctuidae) is the principal pest of maize in tropical and subtropical regions of the Americas. Larvae of this species are susceptible to a nucleopolyhedrovirus (NPV) which has attracted interest as a potential biocontrol agent. Four strains of NPV isolated from infected S. frugiperda larvae in the United States, Nicaragua, and Argentina were subjected to a structural, genetic, and biological comparison to select a candidate isolate for use in biocontrol experiments in Mexico and Honduras. All isolates had an occlusion body polyhedrin protein of 32 kDa, but the virions of each isolate differed subtly in the pattern and abundance of certain structural polypeptides revealed by SDS-PAGE analysis. Restriction endonuclease analysis of viral DNA confirmed that these isolates were strains of a single virus species but showed that they were not genetically homogeneous; each isolate could be differentiated from the others using common restriction enzymes. Droplet feeding bioassays indicated that an isolate from Nicaragua (Sf-NIC) and an isolate from the United States (Sf-US) had the highest infectivity when tested against 2nd instars originating from a Honduran S. frugiperda colony. No significant differences were detected in the speed of kill of Sf-NIC (102.7 h), Sf-US (102.3 h) and Sf-AR (103.4 h), whereas that of Sf-2 (97.3 h) was significantly shorter. Additional bioassays of the Sf-NIC isolate against 2nd to 6th instars demonstrated that LC50 values increased with larval stage from 2.03 x 10(5) OBs/ml for 2nd instars to 1.84 x 10(8) OBs/ml for 5th instars. The concentration required to elicit a lethal infection of 6th instars was so high that a reliable estimate of LC50 could not be obtained. The mean time to death for each stage challenged with the Sf-NIC isolate increased with instar from an average of 102.7 h in 2nd instars to 136.9 h in 5th instars.     

Gene Action of Dollar Spot Resistance in Creeping Bentgrass

The dollar spot disease, incited by Sclerotinia homoeocarpa F.T. Bennet, is one of the most important diseases of creeping bentgrass (Agrostis stolonifera L.) on golf courses. An understanding of the inheritance of dollar spot resistance could enhance genetic improvement efforts in creeping bentgrass. The objectives of this study were to evaluate the response of two creeping bentgrass crosses to two different isolates of S. homoeocarpa, determine gene action and identify number of loci involved in resistance to individual fungal isolates. Parental clones, pseudo F₂, pseudo F₃, BC₁ and BC₂ progenies from two crosses were established in a field trial in a randomized complete block split‐plot design in the fall of 2002. Progeny of each generation (subplots) were inoculated with each of two isolates of S. homoeocarpa (main plots) applied at a rate of 0.25 g/m² of prepared inoculum and evaluated for dollar spot disease. Minimum loci calculations averaged 1.0–2.6. Midparent heterosis calculations were not significant. Backcross population means were closest to the recurrent parent. Generation mean analysis supports a simple additive‐dominance model for both crosses and both isolates, although there was also some evidence of epistatic gene action depending on the cross and the isolate. These results confirm previous research that dollar spot disease is quantitatively inherited and indicate that there may be a few genes interacting in a mainly additive fashion to confer dollar spot disease resistance in creeping bentgrass.     

Sublethal effects of Cry 1F Bt corn and clothianidin on black cutworm (Lepidoptera: Noctuidae) larval development

Black cutworm, Agrotis ipsilon (Hufnagel) (Lepidoptera: Noctuidae), is an occasional pest of maize (corn), Zea mays L., that may cause severe stand losses and injury to corn seedlings. The efficacy of the neonicotinoid seed treatment clothianidin at two commercially available rates and their interaction with a transgenic corn hybrid (Bt corn), trait expressing the Bacillus thuringiensis variety aizawai insecticidal toxin Cry 1Fa2, against black cutworm larvae was investigated. Clothianidin at a rate of 25 mg kernel(-1) on Bt corn increased larval mortality and reduced larval weight gains additively. In contrast, weights of larvae fed non-Bt corn seedlings treated with clothianidin at a rate of 25 mg kernel(-1) increased significantly, suggesting either compensatory overconsumption, hormesis, or hormoligosis. Both Bt corn alone and clothianidin at a rate of 125 mg kernel(-1) applied to non-Bt corn seedlings caused increased mortality and reduced larval weight gains. In two field trials, plots planted with Bt corn hybrids consistently had the highest plant populations and yields, regardless of whether they were treated with clothianidin at the lower commercial rate of 25 mg kernel(-1) The use of Bt corn alone or in combination with the low rate of clothianidin (25 mg kernel(-1)) seems suitable as a means of suppressing black cutworm in no-tillage cornfields, although rescue treatments may still be necessary under severe infestations. Clothianidin alone at the low rate of 25 mg kernel(-1) is not recommended for black cutworm control until further studies of its effects on larval physiology and field performance have been completed.     

A PCR-based linkage map of Agrostis stolonifera and identification of QTL markers for dollar spot resistance

Creeping bentgrass (Agrostis stolonifera L.) is the most widely utilized cool-season turf species for intensively managed sports playing surfaces, including bowling greens and golf course putting greens, tees, and fairways. One of the biggest disease problems affecting creeping bentgrass is dollar spot disease caused by Sclerotinia homoeocarpa F.T. Bennett. Relative to traditional food crops, little attention has been paid to applying molecular technology to traditional creeping bentgrass breeding programs. The objective of this study was to develop a PCR-based linkage map of creeping bentgrass and identify quantitative trait loci (QTLs) associated with dollar spot resistance. Mapping populations segregating for dollar spot resistance were created, phenotyped for disease resistance, and genotyped for simple sequence repeat, conserved intron scanning primer, intron length polymorphism, and amplified fragment length polymorphism markers. As expected, 14 linkage groups (LGs) were detected for each parental map, covering a total of 1,424 and 1,374 cM for the 7418-3 and the L93-10 parental maps, respectively. A total of eight QTL regions (23 markers) for dollar spot resistance were observed for three isolates (Crenshaw, PRG, and UMass1) in our creeping bentgrass mapping populations. LGs 1, 4, and 5 contained at least two overlapping QTL regions to different isolates, indicating that these regions may play a significant role in dollar spot resistance. Identification of QTLs associated with disease resistance will help to facilitate marker-assisted selection in traditional creeping bentgrass breeding programs.     

Biology of Microplitis kewleyi (Hymenoptera: Braconidae): A parasite of Agrotis ipsilon (Lepidoptera: Noctuidae) larvae

Microplitis kewleyi Muesebeck is a gregarious internal parasite of larvae of the black cutworm Agrotis ipsilon (Hufnagel). Studies of the biology of the parasite revealed that there was an inverse relationship between host instar and parasite preference. Duration of development from egg to pupa ranged from 18 days at 27°C to 68.7 days at 16°C. Development from egg to pupa took 13.5–21.6 days when fourth and first instar host larvae, respectively, were parasitized. A larger number of parasites emerged from hosts parasitized in the fourth instar (22.4) than the first instar (11.5). Parasite pupation occurred when the host was in the fifth/sixth instar, depending on the instar parasitized. Thirty‐nine per cent of host larvae exposed as first instars to parasites died before parasite emergence. This decreased to 0% for host larvae exposed as fourth instars. The sex ratio was 1:1.2 (M:F). Thirty‐seven per cent of hosts exposed diurnally were stung, compared to 24% exposed nocturnally. Mean daily progeny was highest (12) on the first day, decreasing to zero after 20 days. Percent host parasitism was also highest on the first day (35%) decreasing to nearly 0% after 18 days. There appear to be three parasite larval instars. Host larvae often remained alive after parasite emergence.