Rat adrenal uptake and metabolism of high density lipoprotein cholesteryl ester

Metabolism of high density lipoprotein (HDL) cholesteryl ester (CE) by cultured rat adrenal cells was studied. Addition of [3H]CE-HDL to cells pretreated with adrenocorticotrophin in lipoprotein poor media resulted in a time- and concentration-dependent accumulation of [3H]cholesteryl ester and production of [3H]cholesterol and [3H]corticosterone. HDL-CE metabolism could be described as the sum of a high affinity ([ HDL-cholesterol]1/2 max = 16 micrograms/ml) and low affinity ([ HDL-cholesterol]1/2 max greater than 70 micrograms/ml) process. [3H]Cholesterol was found both intracellularly and in the media. Accumulation of [3H]cholesteryl ester could not be attributed to uptake and re-esterification of unesterified cholesterol since addition of Sandoz 58-035, an inhibitor of acyl coenzyme A:cholesterol acyltransferase, did not prevent ester accumulation. Moreover, addition of chloroquine did not inhibit cholesteryl ester hydrolysis indicating that hydrolysis was not lysosomally mediated. Aminoglutethimide prevented conversion of [3H]CE-HDL to steroid hormones but did not inhibit [3H]cholesteryl ester uptake. Cellular accumulation of [3H] cholesteryl ester exceeded accumulation of 125I-apoproteins 5-fold at 1 h and 35-fold at 24 h indicating selective uptake of cholesteryl ester moiety. We conclude that rat adrenal cells possess a mechanism for selective uptake of HDL cholesteryl esters which provides substrate for steroidogenesis. These results constitute the first direct demonstration that cholesteryl esters in HDL can be used as steroidogenic substrate by the rat adrenal cortex

Evaluation of Grape Powdery Mildew Forecasting Programs Grape (Vitis vinifera 'Chardonnay') Powdery Mildew (Uncinula necator)

Three forecasting programs for scheduling fungicide applications were selected for comparison with the standard Oregon phenology based program. The California (Gubler-Thomas, UC-Davis) program used leaf wetness and temperature early in the year to predict ascospore infection periods and only temperature during the summer to predict conidial infection periods. The New York (Gadoury) program was based on rainfall and temperature. The German (Oi Diag) program incorporated relative humidity along with temperature and rainfall. Treatments were arranged in a randomized complete block design in a block of 'Chardonnay' planted in 1985 on a 7 x 10 ft spacing. Vines were trained to a bilateral cordon with spur pruning. Shoot thinning occurred 12-13 May to provide uniform cane density. Each treatment was replicated on 3 sets of 5 vines. Treatments were applied using a handgun sprayer at 300 psi at a rate of 200 gal water/A for applications between 1 May (budbreak) and 13 May (6" growth). Treatments were applied using a hooded boom sprayer at 300 psi at a rate of 200 gal water/A for all applications after 13 May. Approximately 3.5 gal of spray suspension was applied per 15 vines (150 gal water/A) between 1 May and 13 May, 4.5 gal between 21 May and 28 Jul, and 5 gal (200 gal water/A) for the rest of the applications. Treatments were applied as required by the guidelines for each program. However, additional conditions for stopping programs at or just after verasion were incorporated as requested by Oregon growers. The standard program and the water control were applied on 13 May (6" shoots), 21 May (12" shoots), 2 Jun (prebloom, EL growth stage 17), 16 Jun (90% bloom), 1 Jul, 15 Jul (bunch closure), 22 Jul, 5 Aug, and 12 Aug (verasion). No Botrytis control measures, including leaf removal, were applied to test vines. All programs used one of two fimgicides, Thiolux DF at 3 lb/100 gal water or Rally at 2 oz/100 gal water for each application. Trap plants of 'Cabernet Sauvignon' were placed next to nontreated vines for 24 hour periods within the same block of grapes from 8 May to 3 Jul. After 24 hours of exposure, plants were transferred to a greenhouse several miles away for incubation under conditions favorable for powdery mildew development

Development of a grower-conducted inoculum detection assay for management of grape powdery mildew

Management of grape powdery mildew (Erysiphe necator) and other polycyclic diseases often relies on calendar-based pesticide application schedules that assume the presence of inoculum. An inexpensive, loop-mediated isothermal amplification (LAMP) assay was designed to quickly detect airborne inoculum of E. necator to determine when to initiate a fungicide application programme. Field efficacy was tested in 2010 and 2011 in several commercial and research vineyards in the Willamette Valley of Oregon from pre-bud break to v eraison. In each vineyard, three impaction spore traps were placed adjacent to the trunk. One trap was maintained and used by the grower to conduct the LAMP assay (G-LAMP) on-site and the other two traps were used for laboratory-conducted LAMP (L-LAMP) and quantitative PCR assay (qPCR). Using the qPCR as a gold standard, L-LAMP was comparable with qPCR in both years, and G-LAMP was comparable to qPCR in 2011. Latent class analysis indicated that qPCR had a true positive proportion of 98% in 2010 and 89% in 2011 and true negative proportion of 96% in 2010 and 64% in 2011. An average of 3 3 fewer fungicide applications were used when they were initiated based on spore detection relative to the grower standard practice. There were no significant differences in berry or leaf incidence between plots with fungicides initiated at detection or grower standard practice plots, suggesting that growers using LAMP to initiate fungicide applications can use fewer fungicide applications to manage powdery mildew compared to standard practices

The Infection and Impact of Azorhizobium Caulinodans ORS571 on Wheat (Triticum Aestivum L.)

Based on our previous study, cereal crop wheat (Triticum aestivum L.) could be infected by rhizobia Azorhizobium caulinodans ORS571, and form para-nodules with the induction of 2.4-dichlorophenoxyacetic acid, a common plant growth regulator. To enhance this infection and the potential agricultural application, we compared six different infection methods (Direct seed dip; Seed germination dip; Pruned-root dip; Foliar spray; Circum-soil dip; Seed dip and circum-soil dip) for achieving the high efficient infection of A. caulinodans into wheat plants by employing a green fluorescent protein (gfp)-labeled Azorhizobium caulinodans strain ORS571. With proper methods, copious rhizobia could enter the interior and promote the growth of wheat to the hilt. Circum-soil dip was proved to be the most efficient method, seed germination dip and pruned-root dip is the last recommended to infect wheat, seed germination dip and seed dip and circum-soil dip showed better effects on plant growth, pruned-root dip did not show too much effect on plant growth. This study laid the foundation for understanding the interaction between rhizobia and cereal crops and the growth-promoting function of rhizobia

Development of a quantitative loop-mediated isothermal amplification assay for the field detection of Erysiphe necator

Plant pathogen detection systems have been useful tools to monitor inoculum presence and initiate management schedules. More recently, a loop-mediated isothermal amplification (LAMP) assay was successfully designed for field use in the grape powdery mildew pathosystem; however, false negatives or false positives were prevalent in grower-conducted assays due to the difficulty in perceiving the magnesium pyrophosphate precipitate at low DNA concentrations. A quantitative LAMP (qLAMP) assay using a fluorescence resonance energy transfer-based probe was assessed by grape growers in the Willamette Valley of Oregon. Custom impaction spore samplers were placed at a research vineyard and six commercial vineyard locations, and were tested bi-weekly by the lab and by growers. Grower-conducted qLAMP assays used a beta-version of the Smart-DART handheld LAMP reaction devices (Diagenetix, Inc., Honolulu, HI, USA), connected to Android 4.4 enabled, Bluetooth-capable Nexus 7 tablets for output. Quantification by a quantitative PCR assay was assumed correct to compare the lab and grower qLAMP assay quantification. Growers were able to conduct and interpret qLAMP results; however, the Erysiphe necator inoculum quantification was unreliable using the beta-Smart-DART devices. The qLAMP assay developed was sensitive to one spore in early testing of the assay, but decreased to >20 spores by the end of the trial. The qLAMP assay is not likely a suitable management tool for grape powdery mildew due to losses in sensitivity and decreasing costs and portability for other, more reliable molecular tools