蛋白-O-岩藻糖基转移酶1基因CfPOFUT1在果生炭疽菌中的功能分析

【目的】蛋白-O-岩藻糖基转移酶1 (protein O-fucosyltransferase 1,POFUT1)是催化蛋白质O-岩藻糖基化的关键酶,在动物和人体内被证明调控一系列的生理病理过程,然而POFUT1基因在果生炭疽菌乃至真菌中还未见报道。本研究旨在克隆果生炭疽菌中CfPOFUT1基因,并分析其生物学功能。【方法】利用RT-PCR技术扩增CfPOFUT1的基因并进行生物信息学分析,构建了CfPOFUT1基因的沉默和过表达载体,通过PEG介导法将载体导入原生质体中获得CfPOFUT1基因的沉默和过表达突变体。测定了野生型菌株、CfPOFUT1沉默菌株和过表达菌株在PDA上的菌丝生长、分生孢子产生、萌发与附着胞形成、胁迫应答和致病力、杀菌剂敏感性等生物学表型。【结果】与野生型菌株相比,基因过表达突变体产孢量显著增加,致病力增强,对嘧菌酯敏感性降低,但对多菌灵和咪鲜胺敏感性增强。基因沉默突变体产孢量减少,细胞壁完整性、内质网应激敏感性提高,致病力减弱,对嘧菌酯敏感性提高,但对多菌灵和咪鲜胺敏感性降低。【结论】CfPOFUT1基因参与调控果生炭疽菌分生孢子产量,细胞壁完整性、内质网对应激和药剂敏感性,并对其致病性也具有一定的影响。     

Discovery of oxathiapiprolin,a new oomycete fungicide that targets an oxysterol binding protein

Oxathiapiprolin is the first member of a new class of piperidinyl thiazole isoxazoline fungicides with exceptional activity against plant diseases caused by oomycete pathogens. It acts via inhibition of a novel fungal target—an oxysterol binding protein—resulting in excellent preventative, curative and residual efficacy against key diseases of grapes, potatoes and vegetables. Oxathiapiprolin is being developed globally as DuPont™ Zorvec™ disease control with first registration and sales anticipated in 2015. The discovery, synthesis, optimization and biological efficacy are presented.     

Diversity in Phytochemical Composition and Medicinal Value of Murraya paniculata

Murraya paniculata is herbal medicinal plant which is traditionally being used for management of cardiovascular, intestinal and respiratory (air way) disorders. This evergreen plant of tropical regions is a member of Rutaceace family. The goal of this review is to analyze and report the biological activities and active phytochemicals reported from Murraya paniculata (M. paniculata) extracts and essential oil. The data was searched using different search engines and using specific key words including M. paniculata, herbal medicine, phytochemicals, extract, essential oil, pharmacological activities. M. paniculata has been found to have wide range of pharmacological activities, including antinociceptive, antianxiety, antioxidant, antidepressant, antibacterial, analgesic and anti-diabetic properties. A diverse range of phytochemicals, including phenols, coumarins, terpenoids, flavonoids, and alkaloids have been isolated from various portions of the plant and tested for a variety of biological activities. This review will provide more information and stimulate additional research to develop more effective and cost-efficient alternative medicine from this plant.     

Activating Blood Circulation,Anti-Inflammatory and Diuretic Effects of Leonurus japonicus Extract on a Rat Model of Trauma Blood Stasis and Its Phytochemical Profiling

Leonurus japonicus Houtt. has been traditionally used to treat many ailments. This study evaluated the activating blood circulation, anti-inflammatory, and diuretic effects of L. japonicus extract (LJ) and identified its phytochemicals. In this work, the phytochemicals in LJ were identified using liquid chromatography mass spectrometry. Rats were randomly assigned to three groups (n=8): Control group was treated with saline, while the Model group (saline) and LJ group (426 mg/kg) had induced traumatic injury. All rats were treated with once by daily oral gavage for one week. The biochemical indices and protein expression were measured. Herein, 79 constituents were identified in LJ, which were effective in elevating body weight, food consumption, water intake, and urinary excretion volume, as well as in ameliorating traumatic muscle tissues in model rats. In addition, LJ prominently decreased the contents of plasma viscosity, platelet aggregation rate, thrombin time, prothrombin time, activated partial thromboplastin time, fibrinogen, thromboxane B2 (TXB2), TXB2/6-keto-prostaglandin F1α (6-keto-PGF1α), urokinase-type plasminogen activator (u-PA), plasminogen activator inhibitor 1 (PAI-1), PAI-1/tissue-type PA (t-PA), and PAI-1/u-PA, while significantly increasing antithrombin III, 6-keto-PGF1α, and t-PA contents. Furthermore, LJ notably inhibited tumor necrosis factor alpha, interleukin 6 (IL-6), IL-8, angiotensin II, antidiuretic hormone, aldosterone, aquaporin 1 (AQP1), AQP2, and AQP3 levels, and markedly elevating IL-10 and natriuretic peptide levels. Finally, LJ markedly reduced the protein expression of AQP1, AQP2, and AQP3 compared to the model group. Collectively, LJ possessed prominent activating blood circulation, anti-inflammatory, and diuretic effects, thus supporting the clinical application of L. japonicus.     

Variation of Fungicide Resistance in Czech Populations of Pseudoperonospora cubensis

During the growing seasons between the years 2001 and 2004, 98 isolates of Pseudoperonospora cubensis from nine regions of Czech Republic were collected and screened for tolerance/resistance to the three frequently used fungicides (propamocarb, fosetyl‐Al, metalaxyl). Fungicides were tested in five different concentrations, using a floating disc bioassay. Fungicide effectiveness varied considerably. Propamocarb appeared most effective and all the isolates collected in the years 2001–2003 were found sensitive to all tested concentrations [607–9712 μg active ingredient (a.i.)/ml]. In 2004, some strains with increased resistance to propamocarb were detected. These strains were characterized by tolerance at the lowest concentrations (607 μg a.i./ml, eventually on 1214 μg a.i./ml); however, they were controlled by 2428 μg a.i./ml. Fosetyl‐Al was effective at the recommended concentration of 1600 μg a.i./ml against all isolates. However, the occurrence of isolates (collected in 2001) which sporulated at low concentrations (400 and 800 μg a.i./ml) indicated that the selection for tolerance occurs in the pathogen population. Nevertheless, this phenomenon was not confirmed with the P. cubensis isolates collected between the years 2002 and 2004. Metalaxyl was found ineffective, because 97% of the isolates showed the resistance to the recommended concentration (200 μg a.i./ml), and the other 3% of isolates expressed tolerant response. The majority of the isolates showed profuse and/or limited sporulation at higher concentrations (400 and 800 μg a.i./ml). A substantial shift to highly metalaxyl resistant strains was evident in the Czech P. cubensis populations during 2001–2004.     

Assessment of Phytochemical Analysis,Nutritional Composition and Antimicrobial Activity of<i> Moringa oleifera</i>

Moringa oleifera is a miracle plant rich in nutrients, antioxidants, and antibiotic properties. Present study was designed to evaluate various biochemical attributes of leaves and flowers of M. oleifera. Plant parts (leaves, flowers) of M. oleifera, collected from different roadsides of Multan district, Punjab, Pakistan, were used as experimental material. Result indicates that alkaloids, saponin, carbohydrates, fats, and protein had a high value in the aqueous extract of both leaves and flowers of M. oleifera. Whereas phenol content was high in methanolic leaves extract and the phenol contents were high in aqueous extract of flowers. The extract yield of M. oleifera leaves and flowers both showed a higher percentage in aqueous extract (57.5%), followed by methanol extract and lowest in ethyl acetate extract. Flavonoids contents were higher in ethyl acetate extract of leaves (33.67%) and aqueous extract of flowers (53.71%). While crude fiber was high in methanolic extract of leaves (12.40%) and in flowers crude fiber was high in ethyl acetate extract (15.86%). The moisture contents were higher in leaves (8.87%) than flowers (7.3%) and similarly, ash percentage in flowers (52.60%) than leaves (41.84%). Ethyl acetate extracts of M. oleifera leaves show antibacterial activity against Pseudomonas aeruginosa while methanolic extract of M. oleifera flowers shows antibacterial activity against Xanthomonas sp. Maximum growth inhibits show in all extracts of leaves against Aspergillus flavus, F. oxysporum, and P. glabrum except for the concentrated aqueous extract of leaves. While in flowers maximum growth inhibits all extracts against P. glabrum, A. niger, and A. flavus except the diluted ethyl acetate extract. Phytochemicals present in different parts of moringa have significant edible and commercial potential. Moringa extracts exhibited significant antimicrobial activity, therefore have applications in pharmaceuticals.