Expression and homology modelling of sterol 14α‐demethylase of Magnaporthe grisea and its interaction with azoles

BACKGROUND: Magnaporthe grisea (Hebert) ME Barr infection is one of the most serious diseases for cultivated rice in the world. Sterol 14α‐demethylase (CYP51) is an important drug target for microbial pathogenic infections. To exploit specific and effective fungicides for M. grisea better, the authors have analysed the characteristics of interaction between sterol 14α‐demethylase from M. grisea (MGCYP51) and azoles. MGCYP51 with truncation of N‐terminal residues was cloned and expressed in E. coli, difference binding spectra of MGCYP51 induced by addition of four commercial azoles were determined and molecular modelling of MGCYP51 based on the crystal structure of Mycobacterium tuberculosis Lehmann & Newman and docking with the azoles were performed. RESULTS: The affinity of the azoles for MGCYP51 was positively correlated with their hydrophobicity. Amino acid residues Tyr112, Phe120, Phe220, His308 and Phe497 of MGCYP51, forming a large hydrophobic cavity, are the key residues interacting with azole fungicides. Furthermore, Phe220 and Phe497 are fungus and species specific respectively. CONCLUSION: The results suggest that the more potent azole fungicides for MGCYP51 should possess more hydrophobic groups interacting with residues Phe220 and Phe497. Copyright © 2009 Society of Chemical Industry     

Does agricultural use of azole fungicides contribute to resistance in the human pathogen Aspergillus fumigatus?

Azole resistance in human fungal pathogens has increased over the past twenty years, especially in immunocompromised patients. Similarities between medical and agricultural azoles, and extensive azole (14α‐demethylase inhibitor, DMI) use in crop protection, prompted speculation that resistance in patients with aspergillosis originated in the environment. Aspergillus species, and especially Aspergillus fumigatus, are the largest cause of patient deaths from fungi. Azole levels in soils following crop spraying, and differences in sensitivity between medical and agricultural azoles (DMIs), indicate weaker selection in cropping systems than in patients receiving azole therapy. Most fungi have just one CYP51 paralogue (isozyme CYP51B), but in Aspergillus sp. mutations conferring azole resistance are largely confined to a second paralogue, CYP51A. Binding within the active centre is similar for medical and agricultural azoles but differences elsewhere between the two paralogues may ensure selection depends on the DMI used on crops. Two imidazoles, imazalil and prochloraz, have been widely used since the early 1970s, yet unlike triazoles they have not been linked to resistance in patients. Evidence that DMIs are the origin, or increase the frequency, of azole resistance in human fungal pathogens is lacking. Limiting DMI use would have serious impacts on disease control in many crops, and remove key tools in anti‐resistance strategies. © 2017 Society of Chemical Industry     

大果木姜子浸提液对艾纳香幼苗的化感作用研究

为了在贵州大面积推广种植艾纳香与大果木姜子,通过盆栽方式,研究大果木姜子浸提液对艾纳香化感作用,测定艾纳香的叶片相对含水量、生物量尧游离脯氨酸含量、可溶性蛋白质含量、可溶性糖含量、可溶性淀粉含量、根系活力、MDA、POD、SOD、CAT 等活性变化,得出敏感指数与综合效应等指标。结果表明：随着大果木姜子浸提液浓度的增加,艾纳香抗氧化系统的3种酶活、丙二醛含量、渗透调节、有机物质和根系活力都有一定程度改变,对植物的相对含水量和生物量造成影响,各物质在处理的过程中表现出的化感作用为抑制,且随浓度增大抑制作用增强。因此在对艾纳香和大果木姜子的间作过程中要及时清理大果木姜子的凋落物以减弱抑制作用。     

导热油炉电热管阵列故障自诊断及容障运行的实现

介绍了导热油炉加热管阵列故障在检修工作中故障点判定的繁琐、耗时及对生产、安全的不利影响,提出用单片机嵌入其控制系统中作在线自动诊断及容障运行的实现方法。     

Effects of CENP-W down-regulation on human glioma U87 cells

AIM: To study the effect of centromere protein W (CENP-W) down-regulation on human glioma U87 cells.METHODS: Small interfering RNA (siRNA) was used to inhibit the expression of CENP-W in the U87 cells. The interference effect of siRNA was evaluated by RT-qPCR and Western blot. The proliferation of the cells was analyzed by MTT assay, BrdU staining and colony formation experiment. Transwell chamber assay was used to detect the invasion ability of the cells. The cell migration ability was measured by a scratch test. The changes of the cell cycle distribution and apoptosis were analyzed by flow cytometry.RESULTS: The results of MTT assay, colony formation experiment and BrdU staining showed that the cell proliferation and colony formation abilities in experimental group were significantly lower than those in control group and negative control group. The results of Transwell and scratch experiments showed that the migration and invasion abilities in experimental group were weaker than those in blank control group and negative control group. The results of flow cytometry analysis showed that the cell cycle distribution in experimental group was arrested in G₀/G₁ phase. The percentage of apoptotic cells in experimental group was higher than that in control group (P<0.05).CONCLUSION: Down-regulation of CENP-W expression inhibits the proliferation, migration and invasion of human glioma cells and promotes the apoptosis of the cells, suggesting that CENP-W may be a potential target of gene therapy for human glioma.     

五加的扦插繁殖研究

采用3因子2水平正交试验设计，探讨了不同扦插基质和IBA处理对五加老枝和嫩枝扦插繁殖的影响，结果表明：二年生或三年生插条优于当年生插条；河沙和红土的混合基质于珍珠岩和腐殖土的混合基质；用1000mg/LIBA蘸插条10s(秒）能提高五加插条的生根率和发梢率。     

Emergence and early evolution of fungicide resistance in North American populations of Zymoseptoria tritici

Although fungicide resistance in crop pathogens is a global threat to food production, surprisingly little is known about the evolutionary processes associated with the emergence and spread of fungicide resistance. Early stages in the evolution of fungicide resistance were evaluated using the wheat pathogen Zymoseptoria tritici, taking advantage of an isolate collection spanning 20 years in Oregon, USA, and including two sites with differing intensity of fungicide use. Sequences of the mitochondrial cytb protein conferring single‐mutation resistance to QoI fungicides and the nuclear CYP51 gene implicated in multiple‐mutation resistance to azole fungicides were analysed. Mutations associated with resistance to both fungicides were absent in the 1992 isolates, but frequent in the 2012 collection, with higher frequencies of resistance alleles found at the field site with more intensive fungicide use. Results suggest that the QoI resistance evolved independently in several lineages, and resulted in significant mitochondrial genome bottlenecks. In contrast, the CYP51 gene showed signatures of diversifying selection and intragenic recombination among three phylogenetic clades. The findings support a recent emergence of resistance to the two fungicide classes in Oregon, facilitated by selection for mutations in the associated resistance genes.     

影响降解菌W2修复异噁草酮污染土壤的三种因子的优化

［目的］ 确定降解菌W2对土壤中异噁草酮的最优生物修复条件。［方法］ 采用3因素5水平正交旋转组合设计,室外盆栽生物测定方法,研究降解菌W2接种量、土壤含水量和肥料添加量3种田间可控因子对降解菌W2修复异噁草酮污染土壤效果的影响。［结果］ 确定修复条件的优化数学回归模型为:y=62.363 9+5.872 8×C1-4.494 1×C2C3-1.262 1×C21-4.076 7×C22,不同因子对土壤修复影响大小顺序依次为土壤含水量、肥料添加量、降解菌W2接种量。［结论］ 降解菌W2对土壤中异噁草酮的最优生物修复条件为：降解菌W2接种量8.19～11.81 mL/kg（A650=0.4）,土壤含水量18.6%～20.84%,肥料添加量1.83～2.52 g/kg。在此范围内降解菌W2对异噁草酮有效成分浓度为500 μg/kg的风干土壤30 d后的降解率可达60%以上,可接近该修复天数的理论极值65.56%。