Connecticut Agricultural Experiment Station Research Articles

First Report of Strawberry Anthracnose Crown Rot Caused by Colletotrichum siamense in New England.

In the summer of 2023, the Connecticut Agricultural Experiment Station was contacted by a farm in southern Connecticut due to reports of strawberry (Fragaria × ananassa) plants showing signs of severe wilting and crown rot across multiple fields, covering ~20 hectares. Cut crowns from diseased plants had marbled red and white lesions typically associated with anthracnose crown rot (ACR). Symptomatic plants were collected from five June-bearing cultivars (cvs. AC Valley Sunset, Lyla, Dickens, and Allstar) spanning four non-adjacent fields with incidence ranging from 5-90% and severity ranging mild wilting in low incidence fields to severe wilting/mortality in high incidence fields. Internal tissue from diseased crowns was surface sterilized in 0.6% NaOCL for 3 minutes, rinsed with sterile water, and plated on potato dextrose agar. After one-week, hyphal tips of fungi were transferred to fresh plates which formed dense mycelial mats of fluffy, greyish-white hyphae. Orange spore masses formed near the center of the colonies, each of which contained numerous cylindrical and fusiform straight conidia, matching spores within the genus Colletotrichum (De Silva et al. 2019). Average conidia (n=192) length was 15.7 ± 1.6 µm and width was 5.4 ± 0.7 µm. Fungi matching this morphology were isolated from 83% of the collected symptomatic crowns and hyphae were collected from two isolates, CT5-1 and CT23-1, for DNA extraction using the GeneJET Plant Genomic DNA Purification Kit. PCR was performed using primers targeting actin (ACT), calmodulin (CAL), β-tubulin (TUB2), GAPDH (gpdA), and ITS, followed by Sanger sequencing, which yielded identical sequences for both isolates (CT5-1 Accessions numbers: PP002078-81, OR999066)(Carbone and Kohn 1999; Hassan et al. 2018; Templeton et al. 1992). These were combined with sequences from fourteen Colletotrichum genomes, all of which were aligned, trimmed, and concatenated using Mega11 (Tamura, Stecher, and Kumar 2021). Model selection was conducted using IQ-TREE and selected parameters were used to generate maximum-likelihood trees from all five loci individually and the concatenated sequence, all of which placed the isolates in a high confidence cluster with Colletotrichum siamense (Nguyen et al. 2015). To confirm the pathogenicity of the pathogen, strawberry plants (cv. Jewel) (n=5) five weeks after bare root transplant were infected. The base of each crown was penetrated 5 mm deep with a sterile 20 µL pipette tip and then inoculated with 10 µL of spores at a concentration of 106 spores/mL. Control plants (n=5) were inoculated with 10 µL of sterile water. Plants were maintained at 30°C day (16-hour)/20°C night (8-hour) in a growth chamber and assessed after 14-days. Four of the five inoculated plants had visible wilt symptoms and bisected crowns revealed the marbled red and white lesions typifying ACR. Control plants had no clear wilting and bisected crowns were visibly healthy. C. siamense re-isolated from infected tissue presented with identical hyphal /spore morphology and ITS/Tub2 were re-amplified and sequenced, yielding identical sequences to CT5-1. Plant inoculations with the same variety were repeated, yielding identical symptom development and crown lesions. C. siamense has been a dominant source of ACR throughout the southeastern US but has not previously been a major problem in the Northeast. Given the extent of the field infection, it is likely that these isolates can survive the colder winter temperatures of New England, but further experimentation is necessary to determine the extent of the pathogen's winter hardiness.

First Report of Neopestalotiopsis spp. Causing Leaf Spot and Petiole Blight on Strawberry in New England.

In October 2023, a Connecticut grower contacted The Connecticut Agricultural Experiment Station about a field of strawberry plants (Fragaria × ananassa) (cv. Ruby June) showing symptoms of severe leaf spotting and visual wilting. Upon visiting the field, leaves had lesions with a diffuse black halo and a light brown center and wilting symptoms, which appeared driven by petiole lesions and presented as dark brown stripes with a reddish-purple halo. Symptoms were observed on 80 to 90% of plants within the block, nearly all of which (>90%) presented with both leaf spots and severe wilting. Diseased tissue was collected from 20 leaves and 25 petioles, sterilized in 0.6% NaOCL, and plated on potato dextrose agar. After hyphal tipping a morphologically identical fungus was isolated from 70% of leaves and 88% of petioles, which formed a dense white mycelial mat with moderate aerial mycelium and conidiomata that exuded dark brown conidial masses. The underside of the mycelial mat was yellowish. Conidia were fusoid, ellipsoid, straight to slightly curved, 4-septate with a single basal appendage and 2-5 apical, matching the description of species within the genus Neopestalotiopsis (Maharachchikumbura et al. 2014). The average conidia (n=74) length, not including appendages, was 29.9 ± 2.1 µm and the average width, at the widest point, was 7.5 ± 0.7 µm. Aerial hyphae were collected from two isolates, CT58-1 and CT62-2, and DNA was extracted for further molecular characterization. PCR was performed with primers targeting actin (ACT), β-tubulin (TUB2), and ITS prior to amplicon sequencing (Carbone and Kohn 1999; Hassan et al. 2018). Sequences were queried against the NCBI whole genome shotgun database, and aligned sequences from 13 species (including Neopestalotiopsis, Pestalotiopsis, and Pseudopestalotiopsis) were collected for each locus. Sequences were aligned, trimmed, and concatenated using Mega11, and IQ-TREE was employed for model selection (Nguyen et al. 2015; Tamura et al. 2021). A maximum-likelihood tree placed the isolates in a high-confidence cluster with Neopestalotiopsis rosae, confirming this placement of these isolates within the genus (CT58-1 Accession #: PP715979-89; PP707735). To confirm pathogenicity, CT58-1 was grown on autoclaved strawberry leaves to induce sporulation, and a suspension of 105 spores/ml was made. Five milliliters of this spore suspension was sprayed on six 6-week-old strawberries (cv. Jewel), and water was sprayed on the same number of control plants. Plants were at 100% humidity for two days and then kept in the greenhouse for 3 weeks to observe symptoms. Inoculated plants presented with identical leaf spot and petiole lesions to field samples and no visual symptoms were observed on control plants. New isolations were made from infected petioles, which produced morphologically identical spores to those described above, and ITS/ACT loci sequencing yielded sequences identical to those of CT58-1. Spore production and plant inoculations were repeated with this new isolate, and identical symptoms were observed. This is the first report of Neopestalotiopsis infecting strawberries in New England and given the high disease incidence in the initial infected field and relative lack of disease in a neighboring field, it is likely that this pathogen was introduced on bare root plants. As the plants were sourced from a nursery in Ontario, Canada, it is likely that the pathogen is capable of overwintering in the Northeastern United States.

Northern hydrilla (Hydrilla verticillata ssp. lithuanica): discovery and establishment outside the Connecticut River

AbstractHydrilla [Hydrilla verticillata (L. f.) Royle], an invasive aquatic weed, has had a rich introduction history into the United States, with multiple subspecies being introduced since the 1960s. The most recent occurred before 2016, when northern hydrilla (Hydrilla verticillata ssp. lithuanica) was discovered in the Connecticut River. By 2021, following a 3-yr survey from Agawam, MA, to the Long Island Sound by the Connecticut Agricultural Experiment Station Office of Aquatic Invasive Species, H. verticillata ssp. lithuanica was found in more than 113 km of the river, occupying 344 ha. Since this survey, there has been concern that H. verticillata ssp. lithuanica would spread to nearby waterbodies and have a significant negative impact. Here, we report the first documented spread and establishment of H. verticillata ssp. lithuanica from the Connecticut River to five waterbodies in Connecticut and one in Massachusetts. Of the eight sites where H. verticillata observations were made, 75% (n = 6) were confirmed to be H. verticillata ssp. lithuanica and 25% (n = 2) to be Hydrilla verticillata ssp. peregrina (wandering hydrilla). Except for one site, all six locations infested with H. verticillata ssp. lithuanica provide watercraft access through public or private boat ramps. The authors also postulate on the mechanisms facilitating the spread and establishment of this subspecies.

1373. Using Geospatial Analysis to Describe the Association between Active Tick Surveillance Data and Clinical Cases of Anaplasmosis

Abstract Background Anaplasmosis is a vector-borne disease spread by Ixodes scapularis ticks mainly in the Northeastern United States. The Connecticut Department of Public Health (CT DPH) made anaplasmosis a state-wide reportable disease in 2008 and a large increase in reportable cases was seen in Connecticut between 2014 and 2019. Methods This study used clinical cases of anaplasmosis reported to the CT DPH and tick infection prevalence data to understand emerging geographic hotspots for the disease in the state and to evaluate the association between the human and tick infections. Human incidence rates (IR) were calculated per 100,000 in the population by county. Tick infection prevalence was calculated as an acarological risk index (ARI) using active tick surveillance data from the Connecticut Agricultural Experiment Station (CAES). These two calculations were analyzed with a Spearman rank correlation in SAS version 9.4 and overlaid on ArcMap. Results During the 2019-2020 period, 420 human cases of anaplasmosis were included and 148 infected ticks were identified. There was a significant positive correlation between IR and ARI identified (Spearman correlation 0.54, p = 0.03). Litchfield County had the highest number of human cases (128) and Fairfield County had the highest proportion of infected ticks (11.25%). The area with the greatest increase in both ARI and IR in Connecticut from 2019-2020 was New London County (ARI increased from 0.073 to 0.108, IR increased from 2.64 to 8.57). Conclusion This study supports the use of active tick surveillance in identifying geographic areas high incidence of human anaplasmosis and provides a strategy to evaluate tick surveillance for anaplasma infection. Active tick surveillance can be useful in guiding public health interventions and inform public health and clinician education on anaplasmosis. Disclosures All Authors: No reported disclosures.

From nanotoxicology to nano-enabled agriculture: Following the science at the Connecticut Agricultural Experiment Station (CAES)

The Connecticut Agricultural Experiment Station was founded in 1875 and has a 147-year history of “Putting Science to Work for Society.” No program exemplifies that core mission more than the Station’s 12-year effort on nanotechnology. This work first started in the realm of nanotoxicology, evaluating the negative effects of these potential emerging contaminants on crop species and in agricultural systems. However, as evidence began increasing that select nanomaterials at appropriate doses and under certain conditions could actually convey some benefit to plant species, work in nano-enabled agriculture began. With an increasing population and a changing climate driving food insecurity, CAES has developed a robust program on the safe and sustainable application of nanotechnology for food production and preservation. Initiatives include novel strategies for crop protection against a range of pathogens; enhancing crop tolerance abiotic stresses such as drought, salinity, and toxic metal contamination; elevating rates of photosynthesis; developing biopolymer-based delivery systems for agrochemicals with much greater efficiency and efficacy; and novel food nanoscale strategies for food preservation. By collaborating with over 30 domestic and international institutions, CAES has and will continue to lead and participate in efforts to combat global food insecurity.

Yale School of Public Health Symposium: An overview of the challenges and opportunities associated with per- and polyfluoroalkyl substances (PFAS).

On December 13, 2019, the Yale School of Public Health hosted a symposium titled "Per- and Polyfluoroalkyl Substances (PFAS): Challenges and Opportunities" in New Haven, Connecticut. The meeting focused on the current state of the science on these chemicals, highlighted the challenges unique to PFAS, and explored promising opportunities for addressing them. It brought together participants from Yale University, the National Institute of Environmental Health Sciences, the University of Massachusetts Amherst, the University of Connecticut, the Connecticut Agricultural Experiment Station, the Connecticut Departments of Public Health and Energy and Environmental Protection, and the public and private sectors. Presentations during the symposium centered around several primary themes. The first reviewed the current state of the science on the health effects associated with PFAS exposure and noted key areas that warranted future research. As research in this field relies on specialized laboratory analyses, the second theme considered commercially available methods for PFAS analysis as well as several emerging analytical approaches that support health studies and facilitate the investigation of a broader range of PFAS. Since mitigation of PFAS exposure requires prevention and cleanup of contamination, the third theme highlighted new nanotechnology-enabled PFAS remediation technologies and explored the potential of green chemistry to develop safer alternatives to PFAS. The fourth theme covered collaborative efforts to assess the vulnerability of in-state private wells and small public water supplies to PFAS contamination by adjacent landfills, and the fifth focused on strategies that promote successful community engagement. This symposium supported a unique interdisciplinary coalition established during the development of Connecticut's PFAS Action Plan, and discussions occurring throughout the symposium revealed opportunities for collaborations among Connecticut scientists, state and local officials, and community advocates. In doing so, it bolstered the State of Connecticut's efforts to implement the ambitious initiatives that its PFAS Action Plan recommends.

Old and new plants from the Americas to Europe: potatoes, corn and the genetics of double hybrid corn (1800–1940)

AbstractWe discuss the extension of corn and potatoes in Galician Atlantic agriculture during the nineteenth and twentieth centuries as an innovation process that facilitated rapid circulation of a new cattle feed from the Americas to Europe. Specifically, we focus on Galicia from 1890 to 1940, a time of significant scientific interest with regard to genetic improvements. This new science made it possible to develop double hybrid corn plants that became widespread after the 1920s. In this article we will describe the conditions accompanying the introduction and spread of these American crops, as recorded by modernist historiography, then analyse the institutional and social framework – knowledge networks, innovation systems and institutional and social tools – that enabled genetic advances in the twentieth century. To accomplish this, we must trace the journey of seeds and knowledge across the Atlantic from places such as the Connecticut Agricultural Experiment Station (1918) to the Galician Biological Mission (1921), among others.

Prevalence of Infection and Co-Infection and Presence of Rickettsial Endosymbionts in Ixodes scapularis (Acari: Ixodidae) in Connecticut, USA

Ixodes scapularis is currently known to transmit 7 pathogens responsible for Lyme disease, anaplasmosis, babesiosis, tick-borne relapsing fever, ehrlichiosis, and Powassan encephalitis. Ixodes scapularis can also be colonized by endosymbiotic bacteria including those in the genus of Rickettsia. We screened 459 I. scapularis ticks submitted to the Connecticut Agricultural Experiment Station Tick Testing Laboratory with the objectives to (1) examine differences in infection prevalence of Borrelia burgdorferi, Anaplasma phagocytophilum, Babesia microti, and Borrelia miyamotoi, (2) evaluate whether prevalence of co-infections occur at the same frequency that would be expected based on single infection, and (3) determine the presence of rickettsial endosymbionts in I. scapularis. The prevalence of infection in I. scapularis was highest with Bo. burgdorferi sensu lato (nymph = 45.8%; female = 47.0%), followed by A. phagocytophilum (nymph = 4.0%; female = 6.9%), Ba. microti (nymph = 5.7%; female = 4.7%), and Bo. miyamotoi (nymph = 0%; female = 7.3%). We also identified rickettsial endosymbionts in 93.3% of I. scapularis. Nymphs were significantly more likely to be infected with Bo. burgdorferi if they were infected with Ba. microti, whereas adult females were significantly more likely to be infected with Bo. burgdorferi if they were infected with A. phagocytophilum. Our study suggests that the infection prevalence of Bo. burgdorferi is not independent of other co-circulating pathogens and that there is a substantially higher infection of Bo. miyamotoi in I. scapularis females compared with nymphs in this study. High prevalence of infection and co-infection with multiple pathogens in I. scapularis highlights the public health consequences in Connecticut, a state endemic for Lyme and other tick-borne diseases.

Predicting spatiotemporal patterns of Lyme disease incidence from passively collected surveillance data for Borrelia burgdorferi sensu lato-infected Ixodes scapularis ticks

Lyme disease is the most prevalent vector-borne disease in the United States. Ixodes scapularis, commonly referred to as the blacklegged tick, is the primary vector of Lyme disease spirochetes, Borrelia burgdorferi sensu lato (s.l.), in the eastern United States. Connecticut has pervasive populations of I. scapularis and remains a hotspot for Lyme disease. A primary aim of this study was to determine if passively collected data on human-biting I. scapularis ticks in Connecticut could serve as a useful proxy for Lyme disease incidence based on the cases reported by the Connecticut Department of Public Health (CDPH). Data for human-biting I. scapularis ticks submitted to the Tick Testing Laboratory at the Connecticut Agricultural Experiment Station (CAES-TTL), and tested for infection with B. burgdorferi s.l., were used to estimate the rate of submitted nymphs, nymphal infection prevalence, and the rate of submitted infected nymphs. We assessed spatiotemporal patterns in tick-based measures and Lyme disease incidence with generalized linear and spatial models. In conjunction with land cover and household income data, we used generalized linear mixed effects models to examine the association between tick-based risk estimates and Lyme disease incidence. Between 2007 and 2017, the CAES-TTL received 26,116 I. scapularis tick submissions and the CDPH reported 23,423 Lyme disease cases. The rate of submitted nymphs, nymphal infection prevalence, the rate of submitted infected nymphs, and Lyme disease incidence all decreased over time during this eleven-year period. The rate of submitted nymphs, the rate of submitted infected nymphs, and Lyme disease incidence were spatially correlated, but nymphal infection prevalence was not. Using a mixed modeling approach to predict Lyme disease incidence and account for spatiotemporal structuring of the data, we found the best fitting tested model included a strong, positive association with the rate of submitted infected nymphs and a negative association with the percent of developed land for each county. We show that within counties, submissions of B. burgdorferi s.l. infected nymphs were strongly and positively associated with inter-annual variation in reported Lyme disease cases. Tick-based passive surveillance programs may be useful in providing independent measures of entomological risk, particularly in settings where Lyme disease case reporting practices change substantially over time.

Detection of Borrelia in Ixodes scapularis ticks by silver stain, immunohistochemical and direct immunofluorescent methods.

Lyme disease (LD) is one of the most common tick-borne diseases caused by several Borrelia species of spirochetes. Ixodes scapularis is responsible for the transmission of LD in the northeastern United States. The rate of infection varies with the duration of tick attachment to the host; however, this information may be unknown. In skin biopsies, it is often difficult to identify spirochetes. Testing of ticks is not routinely performed. Treatment is established by clinical presentation. To test paraffin-embedded I. scapularis ticks for Borrelia by different methods. We examined 20 paraffin-embedded ticks by silver stain, immunohistochemical (IHC) and direct immunofluorescent (DIF) methods and compared the percentage of positivity with DIF results from the Connecticut Agricultural Experiment Station. The results were similar by DIF, which proved to be the most sensitive method, followed by the silver stain and IHC. We found that the identification of spirochetes in paraffin-embedded ticks was less difficult than in tissue with a comparable turnaround time to that of a routine biopsy. Timely identification of Borrelia in ticks may influence the clinical management of the patients.

53rd North American Chemical Residue Workshop.

ADVERTISEMENT RETURN TO ISSUESymposium Introducti...Symposium IntroductionNEXT53rd North American Chemical Residue WorkshopBrian D. Eitzer*†, Sherry T. Garris‡, and Joan M. Stevens§View Author Information† The Connecticut Agricultural Experiment Station, 123 Huntington Street, New Haven, Connecticut 06511, United States‡ The South Carolina Department of Agriculture, 123 Ballard Court, West Columbia, South Carolina 29172, United States§ Agilent Technologies, Incorporated, 2850 Centerville Road, Wilmington, Delaware 19808, United States*E-mail: [email protected]Cite this: J. Agric. Food Chem. 2017, 65, 34, 7251Publication Date (Web):July 20, 2017Publication History Published online20 July 2017Published inissue 30 August 2017https://doi.org/10.1021/acs.jafc.7b00453Copyright © 2017 American Chemical SocietyRIGHTS & PERMISSIONSArticle Views75Altmetric-Citations-LEARN ABOUT THESE METRICSArticle Views are the COUNTER-compliant sum of full text article downloads since November 2008 (both PDF and HTML) across all institutions and individuals. These metrics are regularly updated to reflect usage leading up to the last few days.Citations are the number of other articles citing this article, calculated by Crossref and updated daily. Find more information about Crossref citation counts.The Altmetric Attention Score is a quantitative measure of the attention that a research article has received online. Clicking on the donut icon will load a page at altmetric.com with additional details about the score and the social media presence for the given article. Find more information on the Altmetric Attention Score and how the score is calculated. Share Add toView InAdd Full Text with ReferenceAdd Description ExportRISCitationCitation and abstractCitation and referencesMore Options Share onFacebookTwitterWechatLinked InReddit Read OnlinePDF (164 KB) Get e-AlertsSUBJECTS:Chromatography,Mass spectrometry,Sample preparation,Toxins Get e-Alerts

Origins and Evolution of the National Nutrient Databank Conference

Food composition tables are largely a twentieth century product. The early tables in America developed by W. O. Atwater in the 1890's at the Storrs Connecticut Agricultural Experiment Station grew exponentially during the first half of the 20th century as one after another vitamin and mineral was found to be essential for life. As tables became more complex the user required better tools for calculating food composition. The rapid growth of computer applications in the mid to late 20th century allowed nutritionists to access these complex tables. This confluence of data and computer applications spawned an organization created specifically to study food composition databases.

A PRELIMINARY REPORT ON ASIAN CHESTNUT GALL WASP ON SPECIES AND HYBRIDS OF CHESTNUT IN CONNECTICUT

Seven species of chestnut ( spp.) and most of the possible hybrid combinations of these species are growing in plantings maintained by The Connecticut Agricultural Experiment Station (CAES). In 2011, Asian chestnut gall wasp (Dryocosmus kuriphilus) was found for the first time in these plantings, and had clearly been present for at least two years. Some of the individual trees of the species have not been infested, nor have very few galls. Some of the hybrids between species also have few or no galls. Thus, breeding for resistance to this insect may be possible. Selective crosses have been made and progeny have been planted to examine the genetics of the resistance.

THE BREEDING PROGRAM OF THE AMERICAN CHESTNUT FOUNDATION

Charles Burnham hypothesized in 1981 that the blight resistance of Castanea mollissima could be backcrossed to C. dentate to produce a tree that would perform in the forest like the formerly dominant American chestnut. The American Chestnut Foundation (TACF) was formed in 1983 to test this hypothesis using a traditional plant breeding program. Two B1 chestnut trees, ‘Clapper’ and ‘Graves,’ produced by former breeding programs of the U.S. Department of Agriculture and the Connecticut Agricultural Experiment Station, were used to jump-start the program. These former breeding programs identified Chinese chestnut as the most promising source of blight resistance and developed most of the methods used in the current program. ‘Clapper’ and ‘Graves’ each were backcrossed into approximately thirty American chestnut from the vicinity of Sugar Grove, Virginia, backcrossed again then intercrossed twice. Prior to each cross, trees were selected for blight resistance and recurrent morphological traits. The resultant B3F3 seedlings were planted into the forest for testing beginning in 2009. Tests of the parents of these B3F3s suggested that their blight resistance will fall somewhere between an intermediate and a high level. We are midway through selection in B3F2 seed orchards and resistance among B3F3 progeny should increase as more trees are culled. The ultimate blight resistance of these trees in the forest will be influenced by their adaptation and also may be affected by hypoviruses and other factors; it may take decades for a clear picture to emerge. In addition to ‘Clapper’ and ‘Graves,’ ‘Nanking’ Chinese chestnut has been backcrossed into twenty American backgrounds. More than twenty other sources of blight resistance from Chinese chestnut have been backcrossed, to varying degrees, into American chestnut but into only a few backgrounds; by themselves they do not constitute a viable breeding population. Other Castanea species, C. crenata, C. henryi and C. sequinii have been used to a lesser extent, as well as large, surviving American chestnut trees. This breeding program has been extended throughout the range of the American chestnut in TACF’s network of seventeen state chapters. Through the efforts of those chapters, the effective size of our core population (Ne) should exceed five hundred. The chapters also plan to develop viable breeding populations from the additional sources of blight resistance mentioned above.

Registration of ‘B2’ Connecticut Broadleaf Cigar‐Wrapper Tobacco Resistant to Fusarium Wilt, Tobacco Mosaic Virus, Cyst Nematodes, and Blue Mold

‘B2’ Connecticut broadleaf cigar‐wrapper tobacco (Nicotiana tabacum L.) (Reg. No. CV‐124, PI 664300) is a male‐sterile hybrid released by the Connecticut Agricultural Experiment Station and is resistant to Fusarium wilt, Tobacco mosaic virus (TMV), the tobacco cyst nematode [TCN; Globodera tabacum tabacum (Lownsbery and Lownsbery 1954) Stone 1973], and blue mold [caused by Peronospora tabacina de Bary Adam 1933 (= Peronospora hyoscyami de Bary 1863 f. sp. tabacina Skalicky 1964)]. Initial crosses were made in 1987 between VA‐81 and PD4 flue‐cured tobacco and three broadleaf inbreds resistant to Fusarium wilt [caused by Fusarium oxysporum Schlechtend.:Fr. f. sp. nicotianae (J. Johnson) W.C. Snyder & H.N. Hans.] and TMV. F2 selections were backcrossed twice to broadleaf lines, inbred and selected across 12 generations using bulk modified single‐seed descent with 2% of plants (20 plants of 1000) selected most years; selection for TMV and TCN resistance was conducted in greenhouse screens with fewer plants. Lines were field selected for reduced ozone weather fleck. One F12 selection was backcrossed again to the broadleaf cultivar ‘Scantic’ and inbred via pedigree selection across 10 generations with field and greenhouse selection. Progeny testing was used to select F8 plants homozygous for resistance to TCN and for hypersensitive TMV resistance from Nicotiana glutinosa L. A superior male‐fertile inbred (MF‐2) resulting from 27 generations of selection for broadleaf characteristics with 8 cycles of selection each for Fusarium wilt, TMV and TCN resistance was identified and crossed with a male‐sterile CT broadleaf line resistant to TMV and Fusarium wilt (MS‐1) to produce the F1 male‐sterile hybrid B2.

First Report of Black Leg of Hydroponic Basil in the United States Caused by Plectosporium tabacinum.

During August 2007 and again in January 2008, compact sweet basil (Ocimum basilicum 'Genovese') plants grown hydroponically in Indiana displayed dark, irregular, stem lesions extending 2 to 3 cm above the interface of the nutrient solution. These necrotic stem lesions (black leg), observed on 20 to 30% of the basil plants caused very weak, brittle stems so that they could not be marketed fresh. Although no wilting was noted, reduced plant height was observed. Similar symptoms of blackleg and poor growth have been reported from Italy on greenhouse-grown basil infected with Microdochium tabacinum (1,2). Diseased plant samples were sent to diagnostic clinics at Purdue University and the University of Massachusetts. Stem samples were surface sterilized and plated on potato dextrose agar (PDA) acidified with 1 ml of 85% lactic acid per liter as well as onto one-quarter-strength PDA. A fungus morphologically consistent with Plectosporium tabacinum (van Beyma) M.E. Palm, W. Gams, & H.I. Nirenberg (synonyms M. tabacinum (von Arx, 1984) and Fusarium tabacinum (Gams & Gerlagh, 1968) (3) was cultured from the basil stems and identified as P. tabacinum by R. Wick. Cultures sent to J. McKemy and J. Bischof (USDA/APHIS/PPQ) and W. Elmer (Connecticut Agricultural Experiment Station) also were identified as P. tabacinum. Amplification of the 323-bp internal transcribed spacer (ITS) region (ITS1, 5.8S rRNA gene, ITS2) and subsequent BLAST alignments of the resulting sequence indicated a 98% match for Plectosphaerella cucumerina (anamorph P. tabacinum) (GenBank Accession No. U17399; MIDI Inc., Newark, DE). Inoculations were performed on basil plants grown in peat-based soilless medium in a greenhouse for 6 weeks. Immediately before inoculation, the roots were washed with tap water to remove the peat-based medium. A single basil plant was placed in each of eight, 125-ml Erlenmeyer flasks. Four flasks were filled with 100 ml of deionized water as negative controls and four were filled with a 1 × 106 CFU/ml water suspension of P. tabacinum so that the liquid reached the crown of the basil plant. Basil plants in the Erlenmeyer flasks were incubated on a laboratory bench at 23°C. After 24 h, the solutions in all flasks were discarded and each flask and root system was rinsed three times with deionized water. The plants were then incubated in deionized water on the laboratory bench for four to five additional days. Within 4 days, dark brown-to-black stem lesions similar to those observed originally on basil plants in the hydroponic production greenhouse developed on the plants at the water interface and extended up the stem. Lesions extended a mean of 22 mm above the water level on inoculated plants. Control plants remained symptomless. P. tabacinum was recovered from symptomatic tissue of inoculated plants to complete Koch's postulates. The experiment was repeated several times with similar results. Further evidence of pathogenicity was obtained by stem inoculation of basil plants growing in a soilless medium. These data indicate that P. tabacinum was the causal agent of the symptoms observed on the hydroponic basil. To our knowledge, this is the first report of P. tabacinum causing 'black leg' and reduced growth on basil in the United States and the first report in the world of P. tabacinum on hydroponic basil.

Research in the Agricultural Research Service/USDA: Introduction and Early History

a period of intensive skirmishing and skillful diplomacy had finally resulted in the appropriation (6). Dr. Atwater, already the Director of the Connecticut Agricultural Experiment Station, was appointed as Chief of Nutrition Investigations in the USDA’s Office of Experiment Stations. In the 10 y that Atwater headed the program, he coordinated or conducted work in 4 areas: types and amounts of foods consumed by different groups; chemical composition of foods; effects of cooking and food processing on nutritional quality; and amounts and types of nutrients people need to function at their best. These accomplishments led to his accoladeasthe‘‘FatherofNutritionScience’’intheUnitedStates. The human nutrition investigations were moved to Washington, DC, when Atwater retired in 1906 and the laboratory was put into the Office of Home Economics. The scientifically distinguished record proceeded in synchrony through the succeeding decades with discoveries of the vitamins and essential mineral elements. The energy metabolism work at the USDA was the lead in the nutrition field, but important work on human requirements for essential fatty acids, vitamin G (which became riboflavin), vitamin A requirements of adults, and human utilization of calcium from spinach also was accomplished.

The Determination of the Basic Amino Acids of Proteins: the Work of Hubert Vickery

The Determination of the Basic Amino Acids of Proteins: the Work of Hubert Vickery

Strategies for reducing the risk of Lyme borreliosis in North America

The incidence of Lyme borreliosis continues to increase in the United States. In 1991, when Lyme borreliosis first became a nationally reportable disease to the Centers for Disease Control and Prevention (CDC), a total of 9470 cases were reported; in contrast, by 2002 a total of 23,763 cases were reported, >2.5x the total in 1991. Area-wide acaricides can be highly effective in killing nymphal Ixodes scapularis, with >95% of nymphs killed in studies using cyfluthrin, deltamethrin, or carbaryl. The majority of residents living in households within the area hyperendemic for Lyme borreliosis will not, however, consider the use of area-wide acaricides. A survey of communities in 4 states (Connecticut, Massachusetts, New Jersey, New York) demonstrated that <25% of the populace have used area-wide acaricides on their own property. In searching for alternative methods of reducing Lyme borreliosis risk, host-targeted methods have been proven to be effective. Newly developed methods include the use of acaricides applied to deer feeder stations. This method is called the 4-poster method and has been shown in trials to reduce populations of nymphal I. scapularis by ⩾69%. In addition, rodent-targeted bait boxes containing fipronil have been shown to eliminate ticks on mice and negatively impact the population of questing I. scapularis and reduce the proportion of these ticks infected with Borrelia burgdorferi sensu stricto. Host eradication can also be utilized. On Monhegan Island, Maine, white-tailed deer were totally eradicated from the island from 1999 to 2000. By 2004, no immature I. scapularis could be found on rodents on Monhegan Island. Landscape management practices can also be utilized to reduce the risk of Lyme borreliosis as can personal protection procedures including regular tick checks. These practices have been nicely summarized in a new Tick Management Handbook produced by Dr. Kirby C. Stafford III with the Connecticut Agricultural Experiment Station. Although there is no magic bullet available to completely eliminate the risk of Lyme borreliosis from large geographic areas, the use of Integrated Pest Management (IPM) practices holds the prospect for reducing and managing Lyme borreliosis risk in the future.

GROWTH AND COMPOSITION OF SALAD GREENS AS AFFECTED BY ORGANIC COMPARED TO NITRATE FERTILIZER AND BY ENVIRONMENT IN HIGH TUNNELS1*

Form of fertilizer affects composition of salad greens. This response may vary with growth rate or environment during growth. The interaction of environment and form of fertilizer was examined for salad greens grown as successive plantings in unheated high tunnels. Eight species, including eleven plantings of lettuce and spinach, were grown with either a nitrate-based complete soluble fertilizer in perlite or with an organic fertilizer, namely leaf compost: perlite 1 : 1 v/v amended with cottonseed meal. For each species, relative growth rate and specific leaf area varied with time of year. Fertilizer had little effect on growth rate, but specific leaf area was generally greater for plants with organic fertilizer. For many species, leaf concentrations of nitrogen (N), phosphorus (P), potassium (K), and calcium (Ca) were raised by 10 to 20% with organic, compared to nitrate-based, fertilizer. How-ever, some species responded differently. For instance, organic fertilizer lowered Ca in spinach and nitrate and K in kale. The nutrient concentrations varied with time of year, but each nutrient had a different pattern. The seasonal variation was similar in magnitude to that due to fertilizer. Interaction of effects of season and fertilizer were significant only for N and P in lettuce. Salad greens grown in high tunnels had low concentrations of nitrate, and nitrate varied less with fertilizer or season than did total or reduced N. *Approved by the director of the Connecticut Agricultural Experiment Station.