EPSPS gene amplification in glyphosate-resistant Italian ryegrass (Lolium perenne ssp. multiflorum) from Arkansas

BACKGROUND: Resistance to glyphosate in weed species is a major challenge for the sustainability of glyphosate use in crop and non‐crop systems. A glyphosate‐resistant Italian ryegrass population has been identified in Arkansas. This research was conducted to elucidate its resistance mechanism. RESULTS: The investigation was conducted on resistant and susceptible plants from a population in Desha County, Arkansas (Des03). The amounts of glyphosate that caused 50% overall visual injury were 7 to 13 times greater than those for susceptible plants from the same population. The EPSPS gene did not contain any point mutation that has previously been associated with resistance to glyphosate, nor were there any other mutations on the EPSPS gene unique to the Des03 resistant plants. The resistant plants had 6‐fold higher basal EPSPS enzyme activities than the susceptible plants, but their I₅₀ values in response to glyphosate were similar. The resistant plants contained up to 25 more copies of EPSPS gene than the susceptible plants. The level of resistance to glyphosate correlated with increases in EPSPS enzyme activity and EPSPS copy number. CONCLUSION: Increased EPSPS gene amplification and EPSPS enzyme activity confer resistance to glyphosate in the Des03 population. This is the first report of EPSPS gene amplification in glyphosate‐resistant Italian ryegrass. Other resistance mechanism(s) may also be involved. Copyright © 2012 Society of Chemical Industry     

Simple species identification of Trichinella isolates by amplification and sequencing of the 5S ribosomal DNA intergenic spacer region

We developed a PCR-based assay using a single primer pair to amplify the 5S ribosomal DNA intergenic spacer region to identify Trichinella isolates. In our method, amplified products are directly sequenced on both strands and compared to GenBank sequences. Using this method, we were able to identify Trichinella spiralis, T. britovi and T. nativa. This method permits rapid species identification of Trichinella isolates; however, further evaluation is required before recommending this approach for routine use.     

Cultivation and expansion of canine Schwann cells using reexplantation

Despite the numerous available possibilities for the surgical treatment of peripheral nerve lesions found in the dog, the success of these treatments is often unsatisfactory. It has been proven that Schwann cells (SC) have a positive influence on the regeneration of nerve stumps. Implanting a guidance channel seeded with autologous SC at the lesion site could be a new therapeutic approach. The aim of this research was to investigate the in vitro cultivation and expansion of canine SC as the main requirement for the treatment referred to above. Biopsies were carried out on 17 nerve samples originating from dogs of different breed, age, gender and condition. The reexplantation method was employed, followed by dissociation using hyaluronidase, collagenase and trypsin and further expansion. The samples were divided into six groups which were treated with a varying combination of mitogens (forskolin, bovine PEX, choleratoxin, heregulin). To obtain the quantities of SC, the specimens were immunostained by a p75-antibody. By employing a growing number of agents it was possible to obtain an increase in both the quantity of cells and purity of cultures. A maximum of 16x10(5) cells per millilitre of suspension was achieved. The largest SC purity measured 27.1%. The maximum SC quantity achieved was 43.3x10(4) SC per millilitre.     

Matrix metalloproteinase inhibition in corneal ulceration.

The primary objective of current treatment strategies for infectious keratitis is to sterilize the ulcer as rapidly as possible with topically administered antibiotics. Ulcerative processes can proceed in some cases, despite the absence of microbes, as a result of remaining corneal and tear film MMPs. Combining antibiotic therapy with MMP inhibitors can speed corneal healing, because MMPs play an important role in corneal ulceration and stromal liquefaction. MMPs from the rabbit, horse, and human being are inhibited by metal-binding agents EDTA, NAC, and doxycycline as well as by the serum antiprotease alpha2-macroglobulin. It is not yet certain which proteinase inhibitor has the most favorable therapeutic index for clinical use, although we prefer serum because of its effects on multiple types of proteinases. The MMP inhibitors do have significant therapeutic promise in the treatment of corneal ulceration.     

Generation of reactive oxygen species by a novel anthraquinone derivative in the cyanobacterium Planktothrix perornata (Skuja)

A water-soluble anthraquinone derivative (2-[methylamino-N-(1′-methylethyl)]-9,10-anthraquinone monophosphate), previously found to be selectively toxic towards Planktothrix perornata at submicromolar concentrations, was studied to determine its toxic mode of action towards this cyanobacterium. Chlorophyll fluorescence was monitored as an indicator of photosynthetic efficiency, and measurement of reactive oxygen species (ROS) was performed using the ROS-sensitive probe 2′,7′-dichlorodihydrofluorescein diacetate. The effects of the herbicide paraquat (a ROS generator) as well as ascorbate and α-tocopherol (ROS scavengers) on ROS formation by P. perornata were studied. Also, the effects of different concentrations of ascorbate, α-tocopherol, and the herbicide diuron on reducing the toxicity of the water-soluble anthraquinone derivative towards P. perornata were determined. Our results indicate that the water-soluble anthraquinone derivative does not inhibit photosynthetic electron transport directly, but does generate ROS at levels that may cause toxicity towards P. perornata.     

磷改良剂对中度和高度含碳酸盐的菜园土壤中Cd和Pb环境有效性和植物有效性的影响

The behaviour of metals mainly depends on soil p H, carbonate contents and contamination level, which should be considered for the management of contaminated soils. In this study, kitchen garden topsoils(0–25 cm) were sampled from the area around three smelters in France, with different Cd and Pb contamination levels. Effect of a phosphate amendment(a mixture of diammonium phosphate and hydroxyapatite) on the environmental availability and phytoavailability of Cd and Pb was evaluated by different chemical extractions and cultivating lettuce(Lactuca sativa L.), respectively. Changes in the distribution of Cd and Pb were found in most contaminated soils after phosphate amendment. An increase of Cd and Pb in the residual phase was highlighted in almost all carbonated contaminated soils, whereas a decrease of Pb in the exchangeable, water and acid-soluble phase was observed in most contaminated soils with the lowest carbonate contents. The concentrations of extractable Cd and Pb using calcium chloride and acetic and citric acids generally decreased after the soil amendment. Lettuces grown on amended soils were acceptable for human consumption as regard to Pb concentration. In contrast, some lettuces were unacceptable for human consumption, since the concentrations of Cd in the leaves were higher than the European legislation limit. Surprisingly, in carbonated soils with very low concentration of Cd, the Cd concentrations in lettuce reached up to the European legislation limit, making the lettuce unacceptable for human consumption.Our study highlighted the fact that the total metal concentration in soils does not always allow to predict the metal accumulation in the edible parts of vegetables in order to make a judgement about their acceptability or unacceptability for human consumption.     

Activity of quinones on colletotrichum species

The antifungal activity of 1,4-naphthoquinones, 1,2-naphthoquinones, 1,4-benzoquinones, and anthraquinones from our natural products collection was tested by direct bioautography to identify natural products with potential use in agricultural fungal pathogen control. Quinones demonstrated good to moderate antifungal activity against Colletotrichum spp. Colletotrichum fragariae was the most sensitive species to quinone-based chemistry, Colletotrichum gloeosporioides had intermediate sensitivity, while Colletotrichum acutatum was the species least sensitive to these compounds.