Simple, reliable and cost-effective methods for detecting pathogens are a vital part of diagnostics inside and outside the clinic, in particular in the developing world. Paper based colorimetric techniques are a promising approach for biosensors and bioassays as they can be used at the point of sampling and require little equipment. This study reports on the development of a colorimetric cellulose bioassay that can detect pathogen DNA with covalently attached single-stranded DNA probes. Chemical activation of cellulose via tosylation and oxidation was investigated. The successful activation of cellulose was characterised by Fourier transform infrared spectroscopy, scanning electron microscopy and elemental analysis. Sulfhydryl and amine functionalised oligonucleotide probes complementary to a segment of IS6110 element in Mycobacterium tuberculosis genome were covalently immobilised on the cellulose strips for recognition of target nucleic acid. The detection of biotinylated target oligonucleotides was achieved with horseradish peroxidase (HRP) linked to streptavidin that binds biotin with high affinity. HRP catalysed the oxdidation of tetramethylbenzidine by hydrogen peroxide. The successful assay was confirmed by the appearance of blue coloured spots on cellulose strips incubated with biotinylated target oligonucleotides complementary to the surface attached probe. The study also showed that tosylated cellulose is more reliable for the detection of targets. Initial experiments have shown sensitivity upto 0.1 µM and considerable specificity. High probe immobilization efficiencies (>90%) have been observed. The assay was also effectively demonstrated with mycobacterial DNA. Additionally, the development of a label free assay based on a dual-probe approach was investigated, but did not yield conclusive results. The developed assay has the potential for use as a simple test for the detection of pathogen DNA in clinical samples since it requires minimal equipment and is cost effective. In addition, it also shows the potential use of tosylated cellulose as a prospective surface for attaching other types of biomolecules in an active conformation.

No description available.

Sclerotinia drop of lettuce, caused by the pathogenic fungi Sclerotinia minor and S. sclerotiorum, is a serious disease in most regions were this crop is grown. Conventional fungicides, such as Rovral (iprodione) and Endura (boscalid), are usually applied after lettuce is thinned and once more 2 to 3 weeks later. Two biological products, Contans (Coniothyrium minitans) and Serenade (Bacillus subtilis), are also available. In earlier field trials conducted from 2001 to 2004 in the presence of S. minor, the mean reduction in disease by Contans, Serenade and Endura was 36, 21 and 51%, respectively. The main objective of the current study was to determine the efficacy of the biological products Contans and Serenade, applied alone or in combination with each other or the conventional fungicide Endura, within a biologically intensive integrated pest management system for Sclerotinia drop on lettuce caused by S. minor. The study was conducted at the The University of Arizona, Yuma Valley Agricultural Center. Sclerotia of Sclerotinia minor were produced in the laboratory. Lettuce ‘Winterhaven’ was seeded and sclerotia were applied to the plots on Nov 14, 2005 and the final disease assessment at plant maturity was made Mar 13, 2006. There was a high degree of variability among the replicate plots for each treatment in this trial, as well as an overall low disease incidence, which made statistical comparisons of data difficult. The only treatment in this trial that significantly lowered disease incidence compared to nontreated plots was an initial application of Contans at seeding followed by Endura at thinning. This study was established in a field containing well-draining loam soil, which combined with the lack of any rainfall and the use of furrow irrigations in January and February, which kept the tops of beds dry, likely contributed to the excessive variability and low incidence of disease.

Powdery mildew on lettuce is caused by the fungus Golovinomyces cichoracearum (Erysiphe cichoracearum). The disease is favored by moderate to warm temperatures and relatively dry weather conditions. Several fungicides were evaluated for their ability to suppress development of powdery mildew on lettuce in 2006. Powdery mildew was first detected Jan 26 in this trial. The data in the accompanying table illustrate the degree of control obtained by applications of the various materials tested in this trial. Among treatments, the degree of powdery mildew suppression ranged from virtually complete to minimal; however, all treatments significantly reduced the severity of both mildew diseases compared to nontreated plants. This trial was initiated as a combined downy and powdery mildew trial; therefore, some of the products were placed in the test specifically for downy mildew. Due to low humidity levels and no rainfall during the trial, no downy mildew developed.

Sclerotinia drop on lettuce is caused by two soil-borne fungi, Sclerotinia minor and S. sclerotiorum. This disease is favored by moist soil and moderate temperatures. Some registered products as well as new chemistries in development were compared for their ability to suppress Sclerotinia drop on lettuce during the winter vegetable growing season in 2006-2007. Sclerotia of each pathogen were incorporated into plots after lettuce thinning and just before the first application of test compounds. In plots infested with S. minor, Sclerotinia drop was reduced most effectively by some of the treatments containing boscalid, (Endura), fluazinam (Omega), Coniothyrium minitans (Contans), potassium silicate (Silmatrix), and polyoxin D (Endorse), where disease reduction compared to untreated plants ranged from 34 to 53%.. In plots infested with S. sclerotiorum, disease was reduced most effectively by application of iprodione (Rovral) and Omega followed by cyprodinil+fludioxonil (Switch), where Sclerotinia drop reduction compared to untreated plants ranged from 40 to 52%. Several other treatments provided significant but lessor degrees of disease control in the presence of S. minor and S. sclerotiorum. Two applications of the biopesticide Contans significantly reduced disease in plots infested with either S. minor or S. sclerotiorum. There was no statistical difference in disease control for either pathogen between application of Endura at 50 or 100 gal/acre. Two applications of Endura resulted in significantly higher disease control compared to one application of this product for plots infested with S. sclerotiorum; however, there was no difference in plots containing S. minor. For a valid comparison of products for control of Sclerotinia drop of lettuce, it is important to compare the results obtained from more than one field study. The reader is urged to review previous studies in addition to this report to get an accurate picture of the relative efficacy of tested compounds for control of Sclerotinia drop. Fungicides are not the only tools available to growers to manage Sclerotinia drop. Cultural methods, such as soil solarization or soil flooding in the summer, as well as crop rotation, can greatly reduce the number of viable sclerotia in an infested field. Use of these cultural methods alone or in combination with fungicide treatments can result in dramatic reductions in the incidence of Sclerotinia drop of lettuce.

Powdery mildew, caused by the fungus Golovinomyces cichoracearum (formerly known as Erysiphe cichoracearum), can develop rapidly in spring lettuce during March and April in western Arizona, as the crop nears maturity, when moderate to warm temperatures and dry environmental conditions prevail. The first signs of disease can occur as early as December or January. Successful chemical control of powdery mildew requires the presence of an effective fungicide on plants before disease onset, followed by successive applications of materials to maintain disease control until harvest. A field trial was conducted in 2007 to test and compare the efficacy of some registered as well as new fungicides, applied alone or in a rotational treatment program, for management of powdery mildew. Foliar applications of treatments were made Jan 24, Feb 1, Feb 16 and Feb 28, 2007. Among treatments, the degree of powdery mildew control ranged from virtually complete to minimal; however, all treatments significantly reduced disease severity compared to untreated plants. Treatments that reduced the severity of powdery mildew more than 90% compared to untreated plants included Procure alternated with Quintec, Quintec, Microthiol Disperss, and Procure alternated with Microthiol Disperss. Yield loss due to rejected lettuce heads usually would begin to occur when the powdery mildew disease rating exceeds 2.0. Fungicide treatments that kept disease severity below this level (in addition to the treatments already listed) include V-10118, Procure alternated with Quadris, Procure, Switch, Forum + Cabrio, Endorse, Maneb + Reason, and Endura. This trial was initiated as a combined downy and powdery mildew trial; therefore, some of the products were placed in the test specifically for downy mildew. Due to low humidity levels and no rainfall during the trial, no downy mildew developed; however, some of these downy mildew fungicides, such as Forum, Maneb, and Reason, significantly suppressed powdery mildew. Phytotoxicity symptoms were not noted on lettuce for any of the materials tested.

Phytophthora blight of peppers (Capsicum annuum) is caused by the oomycete pathogen Phytophthora capsici. In Arizona, the root and crown rot phase of the disease initially can appear on plants early in the growing season in areas of the field where soil remains saturated with water after an irrigation or rainfall event. Disease severity can increase dramatically due to summer rains during July and August in the southeastern Arizona production area. Fungicides are an important component of a Phytophthora disease management system, when used in combination with other management practices such as crop rotation, raised beds, and water management. The efficacy of the systemic fungicide mefenoxam (Ridomil Gold) for control of Phytophthora blight on pepper has been documented; however, in many pepper production regions, populations of the pathogen insensitive to this fungicide have developed. Other chemistries, including dimethomorph (Acrobat) as well as some new fungicides in development, have activity on some species of Phytophthora and associated diseases on crops other than pepper. The objective of the following study was to evaluate these additional chemistries for efficacy in suppressing development of root and crown rot on pepper plants grown in soil naturally infested with Phytophthora capsici in a greenhouse environment. The mean duration of survival for Aristotle bell pepper plants in untreated soil infested with P. capsici was 29 days. On the other hand, a significant increase in pepper plant survival was achieved when soil was treated with Reason (fenamidone) + Previcur Flex (propamocarb), SA-110201, Ranman (cyazofamid), Omega (fluazinam), Ridomil Gold (mefenoxam), V-10161(fluopicolide), Forum (dimethomorph), NOA-446510 (mandipropamid), IR-6141 (kiralaxyl), and Maestro (captan). The data from this study suggest that several fungicides currently not registered for use on peppers may be effective components of a management program for Phytophthora crown and root rot. The data is promising; however, additional studies in field soil naturally infested with P. capsici are needed to confirm these preliminary findings as well as to determine the optimal application rate and timing for each new chemistry.

Sclerotinia drop on lettuce is caused by two soil-borne fungi, Sclerotinia minor and S. sclerotiorum. Moist soil and moderate temperatures favor this disease. Some registered products as well as new chemistries in development were compared for their ability to suppress Sclerotinia drop on lettuce during the winter vegetable growing season in 2005-2006. Sclerotia of each pathogen were incorporated into plots after lettuce thinning and just before the first application of test compounds. In plots infested with S. minor, a significant reduction in disease compared to untreated plots was achieved with Omega, Rovral, Endorse, Endura, and Switch. For plots containing S. sclerotiorum, disease was significantly reduced by Contans, Rovral, Omega and Endura. For a valid comparison of products for control of Sclerotinia drop of lettuce, it is important to compare the results obtained from more than one field study. The reader is urged to review previous studies in addition to this report to get an accurate picture of the relative efficacy of tested compounds for control of Sclerotinia drop. Fungicides are not the only tools available to growers to manage Sclerotinia drop. Cultural methods, such as soil solarization or soil flooding in the summer, as well as crop rotation, can greatly reduce the number of viable sclerotia in an infested field. Use of these cultural methods alone or in combination with fungicide treatments can result in dramatic reductions in the incidence of Sclerotinia drop of lettuce.

Powdery mildew occurs annually on melons in Arizona. Podosphaera xanthii (Sphaerotheca fuliginea) is the plant pathogenic fungus that causes powdery mildew on cucurbits, such as cantaloupe, honeydew, watermelon, cucumber and squash. Development of powdery mildew on melons is favored by moderate temperatures and relative humidity, succulent plant growth and reduced light intensity brought about by a dense plant canopy. Existing products as well as materials under development were evaluated and compared for efficacy in management of powdery mildew on cantaloupe in a field trial conducted during the spring of 2006 at the Yuma Valley Agricultural Center. A high level of disease had developed when disease severity data was recorded at crop maturity (Jun 15). Among treatments, the degree of powdery mildew control ranged from 0 to 100%. Among tested products, several are registered for use in Arizona for control of powdery mildew on melons. Using a mixture of products or rotating among efficacious fungicides with different modes of action is important to minimize the development of insensitivity in the pathogen population to one or more of these active ingredients.

Phytophthora blight of peppers (Capsicum annuum) is caused by the oomycete pathogen Phytophthora capsici. In Arizona, the root and crown rot phase of the disease initially can appear on plants early in the growing season in areas of the field where soil remains saturated with water after an irrigation or rainfall event. Disease severity can increase dramatically due to summer rains during July and August in the southeastern Arizona production area. The efficacy of the systemic fungicide mefenoxam (Ridomil Gold)) for control of Phytophthora blight on pepper has been documented; however, in many pepper production regions, populations of the pathogen insensitive to this fungicide have developed. Other chemistries, including dimethomorph (Acrobat) as well as some new fungicides in development, have activity on some species of Phytophthora and associated diseases on crops other than pepper. The objective of the following study was to evaluate additional chemistries for efficacy in suppressing development of root and crown rot on pepper plants grown in soil naturally infested with Phytophthora capsici. In the first trial, nontreated pepper plants were all dead after an average elapsed time of 5 days in soil infested with P. capsici. In the same trial, no plants died after 66 days when the soil was treated with Ranman (cyazofamid), V-10161 (fluopicolide), and Reason (fenamidone) + Previcur Flex (propamocarb). Additionally, only one out of five pepper plants died when treated with Omega (fluazinam), NOA-446510 (mandipropamid) and AgriFos (mono- and di-potassium salts of phosphorous acid). For all of these treatments, the duration of plant survival and fresh weight of plant shoots and roots did not differ significantly from plants grown in sterilized soil. Similar results were obtained in the second trial. The results from these trials suggest that several fungicides currently not registered for use on peppers may be effective components of a management program for Phytophthora root and crown rot. The data is promising; however, additional studies in field soil naturally infested with P. capsici are needed to confirm the preliminary findings of these initial experiments.