PCV2 Rep蛋白N-糖基化位点突变对病毒复制的影响

N-糖基化对病毒的感染与增殖均具有重要作用,与PCV2复制相关的Rep蛋白含有三个N-糖基化位点。为了分析PCV2Rep蛋白N-糖基化位点突变对病毒复制的影响,本试验构建了三个双拷贝突变体感染性克隆2M23、2M256、2M286,并成功拯救病毒。通过间接免疫荧光检测病毒的拯救效果,TCID50测定病毒的感染力,荧光定量PCR检测细胞病毒的载量。结果显示,PCV2Rep蛋白的23~25aa、256~258aa N-糖基化位点突变后降低病毒的复制能力,而286~288aa突变后增强病毒的复制能力,为进一步阐明PCV2的复制及致病机制提供参考。     

Polymorphism of 14α-demethylase Gene (CYP51) in the Cereal Eyespot Fungi Tapesia acuformis and Tapesia yallundae

Cereal eyespot fungi Tapesia acuformis and Tapesia yallundae are closely related species which show different behaviours upon treatment with sterol 14-demethylase inhibitors (DMIs). T. acuformis is naturally resistant to DMIs belonging to the triazole family and susceptible to the imidazole ones, whilst T. yallundae is sensitive to both inhibitors. Cloning of the target enzyme gene, CYP51, from the two species revealed an important polymorphism between them. Further sequencing of CYP51 from sixteen T. acuformis and eleven T. yallundae strains with different phenotypes with regards to resistance to DMIs confirmed that at least eleven variations are species related. Among them, a conserved phenylalanine residue at position 180, found both in T. yallundae and in all known CYP51 proteins from filamentous fungi and yeast, was replaced in T. acuformis by a leucine. Therefore, a leucine at 180 could be possibly involved in natural resistance of T. acuformis to triazoles. Other mutations were observed in some resistant strains, sometimes simultaneously, but in contrast to what was reported for other filamentous fungi, where a mutation at the 136 position of the CYP51 gene product seemed to correlate with resistance to DMIs, we did not find a clear relationship between a given mutation and a particular phenotype. This result suggests that resistance to DMIs could have a polygenic nature in Tapesia. We took advantage of species-related variations to develop a PCR-based assay allowing rapid and easy discrimination between field strains of the two species.     

A Greenhouse Test for Screening Sugar Beet (Beta Vulgaris) for Resistance to Rhizoctonia Solani

Rhizoctonia solani Kühn is a serious plant pathogenic fungus, causing various types of damage to sugar beet (Beta vulgaris L.). In Europe, the disease is spreading and becoming a threat for the growing of this crop. Plant resistance seems to be the most practical and economical way to control the disease. Experiments were carried out to optimise a greenhouse procedure to screen plants of sugar beet for resistance to R. solani. In the first experiment, two susceptible accessions were evaluated for root and leaf symptoms, after being grown in seven different soil mixtures and inoculated with R. solani. The fungus infected all plants. It was concluded that leaf symptoms were not reliable for the rating of disease severity. Statistically significant differences between the soil mixtures were observed, and there were no significant differences between the two accessions. The two soil mixtures, showing the most severe disease symptoms, were selected for a second experiment, including both resistant and susceptible accessions. As in the first experiment, root symptoms were recorded using a 1–7 scale, and a significant expression of resistance was observed. The average severity of the disease in the greenhouse experiment generally was comparable with the infection in field experiments, and the ranking of the accessions was the same in the two types of experiments. It was concluded that evaluation procedures in the greenhouse could be used as a rapid assay to screen sugar beet plants for resistance to R. solani.     

南方水稻区域试验品种对水稻白叶枯病抗性调查

1983-2007年,对南方稻区水稻区域试验品种、组合的白叶枯病抗性进行了研究。结果表明,在1 805个参试品种、组合中,78个表现抗白叶枯病,抗病品种频率为4.3%;其中,常规水稻74个,占抗病品种的94.9%。在1 219个籼型杂交水稻中,抗病的组合3个,中抗的76个,抗病和中抗组合频率分别为0.2%和6.2%。不同类型和不同生态型水稻的抗病品种频率差异显著,晚粳>早籼>中籼>晚籼;常规水稻>杂交水稻。籼型杂交水稻中抗病组合频率为:杂交晚稻>杂交早稻>杂交中稻。1990-2004年,17个抗或中抗白叶枯病的优良水稻新品种、组合通过国家审定。     

不同向日葵品种对黄萎病的抗性*

采用浸根接种法鉴定不同向日葵种质资源对黄萎病的抗性差异。结果表明：40份供试向日葵品种依据抗性划分标准共分为3类,其中中抗品种有5份如：‘118’、‘新葵杂5号’等;低抗品种有8份如‘767’、‘KJ003’等;其余27份供试材料均呈现感病;没有鉴定出免疫和高抗的向日葵品种。     

三峡库区野生红三叶苗期抗旱性灰色关联分析

以三峡库区采集的9份野生红三叶和3份国内外品种为材料,在干旱胁迫下,测定其地上生物量、地下生物量和脯氨酸含量等10项指标,进行灰色关联度分析。结果表明,筛选出地下生物量、脯氨酸含量和叶绿素a含量3项可对红三叶抗旱性进行评价的指标;根据红三叶抗旱性评估模型综合评估值(φ_K)得出材料的抗旱性排序为:瑞德(Red)>CQ5>CQ9>多丽(Dory)>CQ2>CQ4>CQ7>CQ3>CQ1>巫溪>CQ8>CQ6。     

4种高原乡土禾草的抗旱生理比较

采用盆栽法对梭罗草（Roegneria thoroldiana）、草地早熟禾（Poa pratensis）、垂穗披碱草（Elymus nutans）和中华羊茅（Festuca sinensis）4种高原乡土禾本科牧草幼苗的抗旱性进行了比较。结果表明,一定程度的干旱胁迫后,4种禾草的游离脯氨酸含量、可溶性蛋白含量、可溶性糖含量、过氧化氢酶活性、过氧化物酶活性等均有所增加,且叶绿素分解、细胞膜透性增大、电解质外渗率增加,牧草组织细胞受到伤害。不同牧草在干旱胁迫下生理生化指标的变化不同,综合各项指标,运用隶属函数分析表明,4种禾草抗旱性强弱的排序为梭罗草>草地早熟禾>垂穗披碱草>中华羊茅。     

Resistance to a triple-combination anthelmintic in Trichostrongylus spp. on a commercial sheep farm in New Zealand

Abstract

AIM: To evaluate resistance to anthelmintics containing abamectin, levamisole, and oxfendazole (AB-LEV-OX), derquantal and abamectin (DEQ-AB), moxidectin, and monepantel in naturally acquired gastrointestinal nematodes present on a sheep farm.

METHODS: Faecal nematode egg count reduction tests (FECRT) were carried out on lambs that were approximately 7 months-old and infected with naturally acquired nematodes. Lambs were randomly allocated to one of five groups (n=15 per group): treatment with 2?mg/kg derquantel and 0.2?mg/kg abamectin; 0.2?mg/kg abamectin, 8?mg/kg levamisole HCl and 4.5?mg/kg oxfendazole; 2.5?mg/kg monepantel; 0.2?mg/kg moxidectin, or no treatment. Post-treatment samples were collected 12 days later. Abomasa and small intestines were collected from two slaughtered lambs from each of the DEQ-AB, AB-LEV-OX, moxidectin and control groups 15 days after treatment, for nematode counting.

RESULTS: The FECRT demonstrated that efficacy was 90.3 (95% CI=84.2–94.1)% for AB-LEV-OX, 54.5 (95% CI=28.4–71.1)% for moxidectin, 99.2 (95% CI=97.4–99.8)% for DEQ-AB and 100% for monepantel, across all genera. For Trichostrongylus spp. efficacy was 85.5% for AB-LEV-OX and 46.7% for moxidectin. Haemonchus spp. were fully susceptible to all treatments. Post-treatment nematode counts indicated that the resistant Trichostrongylus spp. were from the small intestine.

CONCLUSIONS: Anthelmintic resistance to both AB-LEV-OX and moxidectin was present in the Trichostrongylus genus on a commercial sheep farm. Monepantel and DEQ-AB were both effective against Trichostrongylus spp. based on FECRT results.

CLINICAL RELEVANCE: This finding of resistance to an AB-LEV-OX triple-combination anthelmintic in the Trichostrongylus genus in sheep in New Zealand further limits anthelmintic treatment options available, and calls into question whether this combination is suitable for use as a quarantine treatment.