Ergovaline and peramine production in endophyte-infected tall fescue: Independent regulation and effects of plant and endophyte genotype

Peramine and ergovaline have ecological and economical significance because they possess insect and/or mammalian toxicity properties. The relationship between these endophytically derived alkaloids in tall fescue (Festuca arundinacea Schreb.) is unknown. Seasonal and plant tissue effects on the concentration of peramine and ergovaline was investigated in field and greenhouse experiments. The relationship between the alkaloids and the regulatory effects of endophyte and plant on their content were investigated among progeny of reciprocal crosses between high- and low-ergovaline and peramine plant-endophyte combinations. Variation in peramine concentration ranged from 750 to 1742 µg/kg in greenhouse-grown plants, and there was no seasonal trend in peramine content. There was a correlation (r=0.69) between peramine and ergovaline content among leaf tissues of field-grown plants, but there was no correlation between the alkaloids in the culm (r=0.20) or panicle (r=0.17) tissues. Mean leaf ergovaline concentration of progeny derived from the low-ergovaline parent (163 µg/kg) was less than the midparent value (228 µg/kg), but mean of progeny from the high-ergovaline parent was not different from the mid-parent value. Ranges within each progeny set were approximately double their mean. Mean leaf peramine concentrations of the progeny sets were similar to the mid-parent value (3354 µg/kg) but ranges were from 1716 to 8753 µg/kg. There was no correlation between leaf peramine and ergovaline (r=0.01). These data suggest that endophytically produced alkaloids are independently regulated and are controlled by both plant and endophyte genotype.     

头孢菌素C发酵过程中植物油作用机理的研究

由于葡萄糖效应的存在,目前在头孢菌素C的发酵过程中一般采用植物油为主要碳源。国内外的研究成果都证明添加植物油对头孢菌素C的产量有明显的促进作用。故其作用机理的研究显得格外重要。本文通过分析前人有关顶头孢霉菌C培养过程中植物油作用机理的研究成果,从不同方面综述了植物油对头孢菌素C合成的作用机理,并对其未来的研究方向进行了探讨。     

双对苯醌的抗氧化活性分析及其固体发酵条件优化

为探索生物活性未知的双对苯醌(2,7-dihydroxy-3,6,9-trimethyl-9H-xanthene-1,4,5,8-tetraone,DTXT)的抗氧化活性,并提高其发酵产量,考察DTXT的还原力以及对超氧阴离子自由基、羟自由基、1,1-二苯基-2-三硝基苯肼(1,1-diphenyl-2-picrylhydrazyl,DPPH)自由基的清除效果,在单因素试验基础上,采用响应面法优化了DTXT产生菌瓶生顶孢霉(Acremonium cavaraeanum)CA022菌株的固体发酵培养基。结果表明：在200 μg/mL质量浓度下,DTXT的还原力与芦丁差异不显著,高于VE和2,6-二叔丁基-4甲基苯酚,对超氧阴离子自由基清除率达到67.00%,对羟自由基清除率达到78.83%,对DPPH自由基清除率达到76.53%。通过响应面试验,得到最佳培养基配方为葡萄糖0.773%、硝酸钠0.185%、H₃BO₃ 0.032%、VB₁ 100 μg/100 g,在此条件下实际获得的DTXT产量为4 150.8 mg/kg...     

Isolation of feeding deterrents against argentine stem weevil from ryegrass infected with the endophyteAcremonium loliae

Infection of ryegrass (Lolium perenne) by an endophytic fungus (Acremonium loliae) confers resistance against the Argentine stem weevil (Listronotus bonariensis). Extracts from ryegrass clones, infected and uninfected withA. loliae, were compared in a feeding choice bioassay, and several fractions were identified which affected stem weevil feeding behavior. One stem weevil feeding deterrent, peramine C₁₂H₁₇N₅O, has been isolated from infected ryegrass and partially characterized as a basic indole derivative. Extracts from culturedA. loliae had no effect on stem weevil feeding behavior nor was peramine detected in the fungal cultures examined. Peramine and the other active substances are hydrophilic in contrast to the lipophilic properties reported for the neurotoxic lolitrems also isolated from ryegrass infected withA. loliae and associated with causing ryegrass staggers disorder in livestock. It is suggested that ryegrass staggers and stem weevil feeding deterrency may arise by different biochemical mechanisms.     

海洋枝顶孢霉BH0531发酵液杀线虫组分的分离及活性测定

实验采用生物活性跟踪法,通过超滤、Sephadex凝胶层析、离子交换树脂、硅胶板薄层层析,以及特异性鉴别染色等方法对海洋枝顶孢霉BH0531发酵液的杀线虫活性组分进行分级分离和初步的性质分析。结果表明,Sephadex凝胶柱层析（G-25,G-10）可有效分离获得杀线虫活性组分,3 d的线虫校正死亡率达98.33%;以正丁醇:水:无水乙醇=1∶1∶1(V/V)为展开剂的薄层层析,可将活性组分有效分离,Rf = 0.31,斑点洗脱物的线虫校正死亡率达到94.00%;定性染色结果显示,茚三酮反应阳性,证实有α-氨基存在,茚三酮吡啶呈红色,显现出寡肽性质;双甲酮染色呈亮黄色,表明具有酮糖性质。结果表明,BH0531杀线虫活性代谢物应为含有酮糖基团的短肽类化合物。     

Effect of fungal metabolite peramine and analogs on feeding and development of argentine stem weevil (Listronotus bonariensis)

Peramine, a pyrrolopyrazine alkaloid produced by the fungal endophyte of perennial ryegrassAcremonium lolii, deterred the feeding of both adults and larvae of the graminacious herbivore, the Argentine stem weevil (Listronotus bonariensis), at 0.1 g/g and 10 g/g, respectively. In a no-choice test fewer stem weevil larvae fed and developed on diet containing as little as 2 g/g peramine. The proportion of larvae which did not develop beyond the first instar was higher on diet containing peramine and appeared to be due to a higher proportion of larvae which did not feed. For larvae which fed on the peramine-containing diet, feeding scores and times to pupation were not significantly different from those of controls. A number of simple peramine analogues showed feeding-deterrent activity against adult weevils, indicating the importance of the pyrrolopyrazine ring system of peramine in determining feeding-deterrent activity.     

海洋枝顶孢霉BH0531发酵液对根结线虫卵及二龄幼虫的影响

实验以根结线虫的卵、卵囊和二龄幼虫（J2）为研究对象,对海洋枝顶孢霉（Acremonium sp.）BH0531发酵液处理根结线虫不同发育阶段的杀线作用进行了效果评价,并以J2幼虫为靶标,与市面上的灭线磷及阿维菌素的药效进行了初步对比。结果显示,在实验条件下,海洋枝顶孢霉BH0531发酵液对根结线虫卵囊和分散卵的孵化以及J2幼虫的活力均具有明显抑制作用,其对卵囊孵化的抑制作用达40.18%;对分散卵孵化的抑制作用可达84.28%;而对J2幼虫的杀线作用最为显著,校正死亡率高达97.41%,高于实验条件下灭线磷及阿维菌素的测定值,在根结线虫的生物防控方面显现出潜在的研究价值。     

地生枝顶孢RCEF0260多糖提取工艺的研究

本文对地生枝顶孢RCEF0260菌株液体深层发酵液中胞内、胞外多糖的提取工艺进行了研究。结果发现采用机械法处理RCEF0260菌丝体,多糖得率从21.5g/kg干菌体提高到27.0g/kg干菌体,利用正交试验结果得到胞内多糖的提取工艺参数温度60℃,加水量2.5mL.g-1,浸提时间为1.5h,次数2次;RCEF0260胞外多糖的提取工艺参数乙醇终浓度为65%,发酵液浓缩到1/2,浓缩的温度为40℃。     

海洋真菌BH0531对黄瓜促生长作用的研究

通过室内盆栽试验研究了海洋枝顶孢霉（Acremonium sp.）BH0531次生代谢产物对黄瓜根结线虫病的防治效果及对黄瓜植株生长的促进作用。实验以根结线虫（Meloidogyne incognita）侵染后的黄瓜为材料,在土壤中加入质量浓度为5 mg/mL、10 mg/mL、20 mg/mL、40 mg/mL菌株BH0531发酵液40 mL,用等体积清水作为对照,测定黄瓜的株高、茎粗、叶片的可溶性蛋白及可溶性糖含量、叶绿素含量和根系活力。结果表明,在试验浓度范围内施入发酵液,对根结线虫病的防效最高达72.7%,同时施入发酵液显著增加叶片可溶性糖和叶绿素含量,提高黄瓜的根系活力。因此,海洋枝顶孢霉BH0531的代谢物不仅对黄瓜根结线虫病有显著的控制效果,而且对黄瓜生长有一定的促进作用。     

ParasiticMeloidogyne and mutualisticAcremonium increase chitinase in tall fescue

Tall fescue (Festuca arundinacea Schreb.) is a C-3 perennial grass noted for its persistence in harsh environments. Tall fescue persistence is enhanced byAcremonium coenophialum, a mutualistic fungal endophyte that increases resistance to drought, pathogens, and insects. This research was conducted to identify and elicit biochemical mechanism(s) that could account for tall fescue persistence. In initial studies, two cultivars known to differ in persistence were analyzed for chitinase, an antifungal hydrolase associated with disease resistance in other plants.Acremonium-infected Kentucky 31 (KY31), a persistent cultivar, and Johnstone, a nonpersistent cultivar, were inoculated with the parasitic nematode,Meloidogyne marylandi, grown for 50 days, and analyzed at 10-day intervals. Chitinase fluctuated throughout the 50-day period of seedling development, and activity was highest in the persistentAcremonium-infected KY31. In addition, chitinase was elicited by parasiticM. marylandi and expressed systemically. Subsequent studies were conducted to determine whether or not mutualisticAcremonium could increase chitinase activity. Genetically identical KY31, with and withoutAcremonium, were grown for 25 days and analyzed for chitinase at 5-day intervals. After 20 days,Acremonium-infected KY31 expressed more chitinase thanAcremonium-free KY31. We concluded that chitinase is related to tall fescue persistence; it was highest in the most persistent cultivar, increased under pathogen attack, and increased in the presence ofAcremonium, a symbiont known to enhance disease resistance.Contribution from University of Missouri Agricultural Experiment Station, Journal Series No. 11,472.