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.     

遥感作物制图辅助核事故农业风险决策

【目的】 将作物时空分布数据应用于核事故农业风险决策支持系统,体现遥感作物制图在核事故农业风险决策中的重要性。【方法】 文章以大亚湾核电基地为研究案例,对其周边地区的作物轮作系统进行遥感制图;作物时空分布数据经后处理,上传至核事故农业风险决策支持系统,实现作物样本任务的自动生成,以及放射性核素浓度的时空分布模拟。【结果】 提出的遥感制图方法可以在耕地破碎、云雨繁密区识别作物轮作系统,快速、准确地提供大范围作物时空分布数据。经过处理的作物时空分布数据,能够方便地应用于决策支持系统,辅助完成特定或优先区作物样本任务点的自动生成,以及放射性核素浓度时空分布的模拟。【结论】 遥感作物制图与核事故农业风险决策支持系统相结合,可进一步提高采样的有效性,提升放射性核素空间和时间分布模拟与预测的准确性。从而帮助决策者制定核污染监测和评估策略、修复计划,科学指导农业生产的恢复。未来,有必要深入研究遥感作物制图在核事故农业风险决策中的应用,充分发挥遥感技术与数据的优势,规避核事故对农业生产带来的风险。     

Identification of <Emphasis Type="Italic">Ditylenchus</Emphasis> species associated with Fabaceae seeds based on a specific polymerase chain reaction of ribosomal DNA-ITS regions

A technique based on the use of specific primers for polymerase chain reaction (PCR) was developed for the identification of the stem and bulb nematode belonging to the Ditylenchus dipsaci species complex. The internal transcribed spacer region ITS1 and ITS2, the gene 5.8 S and part of genes 18 S and 26 S of twenty populations of the D. dipsaci species complex belonging to both D. dipsaci sensu stricto and Ditylenchus sp. B (corresponding to populations of giant individuals associated to Vicia faba) and three congeneric species were amplified with two universal ribosomal primers. PCR-amplified DNA samples were digested with five restriction enzymes in order to reveal some polymorphism allowing the identification of D. dipsaci populations associated with Fabaceae seeds. The polymorphism among species was confirmed by the sequencing of the PCR products. A primer (DdpS2) was designed in a region conserved in all populations of both D. dipsaci sensu stricto and D. sp. B studied in the present work. The other Anguinidae species (except a few species from Central Asia associated to Astereaceae and D. sp. G associated to Plantago maritima) differ in two to four nucleotides at the 3′ extremity of this region. This sequence portion coincides with a TspEI restriction site. In combination with a primer located in the ribosomal region, this first primer is a good candidate for identification by PCR of populations of the D. dipsaci species complex found in Fabaceae seeds. A second primer (DdpS1) was designed in a similar way and was specific to D. dipsaci sensu stricto. The utility of these two sets of primers is discussed against the background of quarantine regulation.