Characterizing the variation in aggressiveness and sporulation of the NA1 and EU1 lineages of Phytophthora ramorum in Oregon

Phytophthora ramorum, the cause of sudden oak death, is an invasive pathogen present in parts of coastal California and south-western Oregon forests. The majority of these forest infestations have been caused by the NA1 clonal lineage. In 2015, the EU1 lineage of P. ramorum was isolated from a tanoak (Notholithocarpus densiflorus) tree located in a mixed-conifer forest of Curry County, Oregon. In order to evaluate the threat to Oregon forests of the EU1 lineage relative to the established NA1 lineage, a series of experiments was conducted comparing aggressiveness and sporulation of NA1 and EU1 isolates on logs and seedlings in the growth chamber and forest. There was no significant difference in lesion size on logs inoculated with NA1 and EU1 isolates for any of the tree species tested. Across all seedling experiments differences among isolates within lineage, in terms of both aggressiveness and sporulation, were more commonly observed than differences among lineages. Site to site variation in tanoak sporulation, as measured by rain bucket baiting, appears to be correlated with the number of P. ramorum-positive seedlings detected at each site.     

利用表型芯片技术筛选利于枯草芽胞杆菌Bacillus subtilis BAB-1生长与芽胞形成的营养物质

为获得枯草芽胞杆菌Bacillus subtilis BAB-1菌株的芽胞高效率生产技术,采用Biolog微生物细胞表型芯片(phenotype microarray,PM)技术分析了适合该菌株细胞生长的营养物质、渗透压和pH等生长环境条件;通过单因素试验分析了5种碳源及5种氮源对该菌株生长及芽胞形成的影响。结果表明,BAB-1菌株能够利用114种碳源、228种氮源、29种磷源及29种硫源作为其营养物质,其中52种碳源、66种氮源、3种硫源及1种磷源能够明显促进该菌株的细胞生长;BAB-1菌株对渗透压的忍受能力较强,在1%～10%氯化钠、3%～6%氯化钾、2%～5%硫酸钠、5%～20%乙二醇、1%～6%甲酸钠、2%～7%尿素、1%～12%乳酸钠、20～200 mmol/L磷酸钠、10～100 mmol/L硝酸钠、10～100 mmol/L硫酸铵中均能较好的生长;BAB-1菌株对pH的耐受范围较广,在4.5≤pH≤10.0条件下均能够生长。葡萄糖与氯化铵分别是最适宜BAB-1菌株生长的碳源与氮源,在不同培养时间其菌体总量均最高,48 h时该菌株的菌体总量分别达到6.90×10~8CFU/mL与6.87×10~8CFU/mL;培养12 h时,D-木糖、D-核糖、L-阿拉伯糖及熊果苷能够显著提高BAB-1菌株的芽胞形成率,其中L-阿拉伯糖与熊果苷的芽胞形成率较高,分别为61.94%与66.92%;培养12～36 h,L-脯氨酸、L-谷氨酰胺及L-天冬酰胺能够显著提高BAB-1菌株的芽胞形成率,芽胞形成率介于27.88%～82.37%之间。表明表型芯片技术可以应用于枯草芽胞杆菌生长及芽胞形成营养物质的高通量分析及培养工艺的优化中。     

Dieback and mortality of Nothofagus in Britain: ecology,pathogenicity and sporulation potential of the causal agent Phytophthora pseudosyringae

Since 2009 extensive dieback and mortality of Nothofagus obliqua, associated with bleeding cankers on stems and branches, has been observed in the UK. The causal agent was identified as Phytophthora pseudosyringae, based on morphological and analysis of the internal transcribed spacer (ITS) sequences. Between 2011 and 2013, a survey assessed the frequency and nature of these P. pseudosyringae infections. Mature trees of Nothofagus with stem lesions caused by P. pseudosyringae were found across England, Scotland and Wales. Additional symptoms such as twig blight and leaf necrosis indicated that aerial infection was occurring. Besides N. obliqua, other hosts regularly encountered included Nothofagus alpina, Fagus sylvatica and Vaccinium myrtillus. In pathogenicity tests involving inoculation of logs, P. pseudosyringae was shown to be an aggressive bark pathogen of N. obliqua and F. sylvatica, but significantly less aggressive on N. alpina. Foliage susceptibility and sporulation tests showed marked differences between the six host species tested. Leaves of N. obliqua and V. myrtillus were highly susceptible. Leaves of N. alpina were moderately susceptible, those of Rhododendron ponticum slightly susceptible and those of F. sylvatica not susceptible at all. High levels of sporulation were observed only on inoculated N. obliqua and V. myrtillus leaves. This suggests that P. pseudosyringae may sporulate heavily on N. obliqua foliage in the field and that this inoculum initiates the aerial lesions observed on the shoots, branches and stems. The results also suggest that P. pseudosyringae has the potential to pose a serious threat to N. obliqua and other Nothofagus species in their Southern Hemisphere native ranges.     

Effect of temperature on infection and development of powdery mildew on cucumber

Podosphaera xanthii and Golovinomyces orontii are the causal agents of cucurbit powdery mildew. The effect of temperature on conidial germination, infection and sporulation was studied under controlled conditions. Conidia were inoculated on cucumber leaf discs, and incubated at six constant temperatures (from 10 to 35 °C in 5 °C steps) for 3 to 72 h to evaluate conidial germination and infection, and for 6–15 days to evaluate sporulation intensity. Germination took place at all tested temperatures, but was close to zero at 35 °C. The longest germ tubes measured in this experiment were 141.74 μm for the secondary germ tube of P. xanthii at 20 °C after 48 h of incubation, and 67.92 μm for G. orontii for the primary germ tube at 20 °C after 48 h of incubation. The optimal temperatures for conidial germination, infection and sporulation were 24.4, 25.7 and 22.3 °C, respectively, for P. xanthii, and 17.9, 17.3 and 14.9 °C, respectively, for G. orontii. Equations were developed to describe conidial germination with a coefficient of determination (R²) of 0.85 and 0.90 for P. xanthii and G. orontii, respectively. Infection equations resulted in R² of 0.94 and 0.93 for P. xanthii and G. orontii, respectively; and for sporulation, R² of 0.75 and 0.76 for P. xanthii and G. orontii respectively, as a function of temperature. These results can be used to develop models for the risk of cucurbit powdery mildew under field conditions.     

Effect of Essential Oils from Citrus Varieties on in vitro Growth and Sporulation of Phaeoramularia angolensis Causing Citrus Leaf and Fruit Spot Disease

Citrus leaf and fruit spot disease caused by Phaeoramularia angolensis is a serious production constraint in tropical Africa. In previous studies, essential oils extracted from fruit peels of two tolerant varieties exhibited a strong antifungal activity in vitro against P. angolensis as compared to oils from susceptible ones. In order to investigate if the susceptibility of citrus varieties is associated with the antifungal activity of their essential oils, some 22 varieties of different susceptibility levels (tolerant, susceptible and highly susceptible) and belonging to different botanical groups were studied. Oils extracted from fruit peels were evaluated for their activity against radial growth and sporulation using the poisoned food technique. The optimal doses for growth inhibition and conidial reduction were 2500 and 1000 ppm, respectively. At these doses, radial growth and sporulation exceeded the untreated control respectively for four and nine varieties suggesting that oils from these varieties promote fungal development. In general, oils from the tolerant group were most effective in reducing radial growth irrespective of dose. The highly susceptible group ranked first in reducing sporulation at dose 1000 ppm (45.93%) while at higher doses of about 2000–2500 ppm, oils from the tolerant varieties could reduce sporulation up to 100%. The marked dose effect in reducing sporulation suggests that there may be different compounds acting with changing dose. Botanically, oils from pummelo (Citrus maxima, tolerant group), were best in reducing radial growth (>87% inhibition) while those from grapefruits (C. paradisi, highly susceptible group) were most effective in reducing sporulation (>64% reduction).     

食线虫性真菌Duddingtonia flagrans F088的生长和产孢特性

【目的】研究食线虫性真菌Duddingtonia flagrans生长特性和不同营养要素对产孢量的影响,为其规模化发酵工艺的确定提供参考。【方法】以D.flagrans F088分离株为研究对象,研究该菌培养过程中pH值、厚垣孢子产量、菌丝干质量等指标的变化;通过单因素试验及正交试验,研究几种常见碳源、氮源、无机盐、维生素和碳氮比对厚垣孢子产量和菌丝平均生长速率的影响。【结果】供试真菌在培养2 d后进入对数生长期,4 d时达到高峰(OD_(600)值为7.46,菌丝干质量为12.07 g/L),而pH快速下降至4.45;3 d开始产生厚垣孢子,4 d时呈爆发式增长。OD_(600)与菌丝干质量、厚垣孢子产量呈极显著正相关,与pH呈极显著负相关。单因素试验结果表明,产生厚垣孢子的最佳碳源为白糖,氮源为大豆蛋白胨,碳氮比为40∶1,维生素为V_(B_1)、无机盐为MgSO_4;菌丝生长较好的碳源为红糖,氮源为大豆蛋白胨,与对照相比,无机盐和维生素对菌丝生长无显著促进作用。4因素3水平正交试验结果显示,D.flagrans F088产生厚垣孢子和菌丝生长的最佳配方组合为:30 g/L白糖、4 g/L大豆蛋白胨、1 g/L MgSO_4和30 mg/L V_(B_1)。【结论】初步明确了D.flagrans F088培养过程中pH、厚垣孢子产量、菌丝干质量等的变化趋势,确定了该菌菌丝平均生长速率和厚垣孢子产量达到最佳时的营养要素和配方组成。     

黄曲条跳甲高致病力绿僵菌的筛选及培养特性研究

本研究测定了8株绿僵菌菌株对黄曲条跳甲成虫的致病力,旨在筛选出感染黄曲条跳甲的高致病力菌株,为生物防治提供新的资源。筛选获得1株对黄曲条跳甲有较高致病力的菌株Ma6;室内毒力测定结果显示,在25℃下,菌株Ma6孢子悬浮液浓度8.0×108个·mL~(-1)处理黄曲条跳甲成虫,其LT_(50)为4.09d;致病力复测试验结果表明,7d累计死亡率达到100%,对黄曲条跳甲成虫表现较强的致病力。菌株Ma6的生物学特性研究表明,其最适产孢培养基为PDA培养基,最适产孢温度为25℃,最适产孢pH为pH=7,光照条件为先黑暗6d再光照8d有利于其产孢。     

金丝桃素抗鸡球虫效果观察

为探明金丝桃素是否具有抗鸡球虫的作用,特进行了金丝桃素体外抗球虫孢子化试验及体内抗球虫试验.体外抗球虫孢子化试验结果表明,金丝桃素浓度达到0.03％即有明显的抑制球虫孢子化的作用;体内抗球虫试验也表明金丝桃素具有抗鸡球虫的活性,服用金丝桃素6、8、10、12 mg/只,其抗球虫指数分别为66.1、100.3、126.0、131.3.研究结果为拓展药物资源应用的新领域、研发抗球虫新药提供了良好基础.