广西地不容内生真菌疣孢漆斑菌DBR-11的抑菌活性

为了研究广西地不容内生真菌的抑菌活性,从广西地不容块根中分离纯化出18株内生真菌,采用拮抗试验测定了其对10种植物病原真菌的抑制活性,发现疣孢漆斑菌DBR-11对多数供试菌有很好的抑制活性。进一步测定了DBR-11菌株发酵产物的抑菌活性,发现发酵产物的乙酸乙酯萃取物对病原菌的抑制活性较高,质量浓度为5 g/L时,对10种植物病原真菌的抑菌率均在90%以上,有效中浓度(EC50)为0.007 2~0.446 5 g/L,其中对烟草黑胫病菌的毒力最高,EC50值为0.007 2 g/L;发酵产物的乙酸乙酯萃取物质量浓度为2 g/L时,72 h对15种供试动物病原菌中的7种能完全抑制,最低抑制浓度(MIC)为0.062 5~0.25 g/L,其中对痢疾志贺氏菌的抑制活性最高,MIC值为0.062 5 g/L。DBR-11发酵产物乙酸乙酯萃取物有广谱的抗菌活性,具有潜在的应用价值。     

湖北武汉茶树内生真菌多样性及种群结构

为了解武汉地区茶树内生真菌的多样性,从鄂茶1号、鄂茶5号、福鼎大白茶的茎、叶共210个组织块中,分离获得166株内生真菌菌株。结果表明,茶树受内生真菌侵染程度较高,74.29%的组织有内生真菌存在。根据r DNA-ITS系统发育分析,从中分离鉴定内生真菌15属,27个分类单元,主要有链格孢(Alternaria)、葡萄座腔菌(Botryosphaeria)、刺盘孢(Colletotrichum)、间座壳属(Diaporthe)、镰孢霉(Fusarium)、赤霉(Gibberella)、小从壳菌(Glomerella)、球座菌(Guignardia)、新壳梭孢菌(Neofusicoccum)、拟盘多毛孢(Pestalotiopsis)、拟茎点霉(Phomopsis)、格孢腔菌(Pleosporales)、拟青霉(Paecilomyces)、腐霉(Pythium)、丝核菌(Rhizoctonia)和2种未知真菌。根据相对频率,葡萄座腔菌(Botryosphaeria dothidea)、盘长孢状刺盘孢(Colletotrichum gloeosporioides)、芒果球座菌(Guignardia mangiferae)等在茶树中比较常见。不同组织部位、不同品种内生真菌分布及组成存在差异,表现出较丰富的多样性。     

无瓣海桑果实内生真菌的分离鉴定及其抑菌活性

基于18S r DNA序列分析和Gen Bank同源性比较,研究采用平板分离法从红树无瓣海桑(Sonneratia apetala)果实中分离得到20株内生真菌,共18个属。分别对20株内生真菌的发酵液进行抑菌活性筛选,其中BGMRC1036T的代谢产物对荔枝炭疽病菌(Colletotrichum gloeosporioides)有较好的抑制作用,BGMRC1123T的代谢产物和BGMRC11095T的代谢产物对番木瓜炭疽病菌有较好的抑制作用,BGMRC1008T的代谢产物对香蕉黑星病菌(Cladosporium cucumerinum)表现出较好抗菌活性。     

不同配比香菇菌渣基质对鲜食黄瓜幼苗生长的影响

[目的]研究不同堆沤处理香菇菌渣复合基质对鲜食黄瓜幼苗生长的影响。[方法]采用A、B、C 3种堆沤方法对香菇菌渣进行处理,测定和分析发酵过程中菌渣温度和EC值变化,将处理后的菌渣与草炭、珍珠岩、蛭石组成复合基质,并以“鼎丰98F1”鲜食黄瓜为供试品种进行育苗试验。[结果]采用B方法进行堆沤处理温度高时间短,腐熟效果较好;复合基质T5和T6的理化性质基本都在理想基质范围值内;除在出苗率方面低于对照外,处理T5（腐熟菌渣∶珍珠岩∶蛭石＝1∶2∶1）在株高、根长和干鲜重等方面均明显高于对照（CK）,处理T6（腐熟菌渣∶珍珠岩∶蛭石＝2∶1∶1）在根冠比和壮苗指数值上均为最大,且明显优于对照（CK）。[结论]复合基质T5和T6可以作为鲜食黄瓜的无土育苗基质在生产中应用。     

The endophytic fungus Piriformospora indica protects wheat from fusarium crown rot disease in simulated UK autumn conditions

The root endophytic fungus Piriformospora indica (Sebacinacea) forms mutualistic symbioses with a broad range of host plants, increasing their biomass production and resistance to fungal pathogens. This study evaluated the effect of P. indica on fusarium crown rot disease of wheat, under in vitro and glasshouse conditions. Interaction of P. indica and Fusarium isolates under axenic culture conditions indicated no direct antagonistic activity of P. indica against Fusarium isolates. Seedlings of wheat were inoculated with P. indica and pathogenic Fusarium culmorum or F. graminearum and grown in sterilized soil‐free medium or in a non‐sterilized mix of soil and sand. Fusarium alone reduced emergence and led to visible browning and reduced root growth. Roots of seedlings in pots inoculated with both Fusarium isolates and P. indica were free of visible symptoms; seed emergence and root biomass were equivalent to the uninoculated. DNA was quantified by real‐time polymerase chain reaction (qPCR). The ratio of FusariumDNA to wheat DNA rose rapidly in the plants inoculated with Fusarium alone; isolates and species were not significantly different. Piriformospora indica inoculation reduced the ratio of Fusarium to host DNA in the root systems. The reduction increased with time. The ratio of P. indica to wheat DNA initially rose but then declined in root systems without Fusarium. With Fusarium, the ratio rose throughout the experiment. The absolute amount of FusariumDNA in root systems increased in the absence of P. indica but was static in plants co‐inoculated with P. indica.     

内生细菌EBS05对烟草诱导抗性的信号转导途径研究

 樟树内生枯草芽胞杆菌EBS05是一株对多种植物病原菌具有较强拮抗活性,并能诱导烟草系统抗性的生防菌株。本文以缺失Surfactin A合成相关基因的突变菌株EBS05^T为材料,研究了内生细菌EBS05对烟草诱导系统抗性的激发子及其信号转导途径。结果表明,菌株EBS05产生的Surfactin A是诱导烟草对TMV系统抗性的有效激发子;Surfactin A诱导处理后,SA信号转导途径下游的关键调节基因NPR1首先被激活,并持续超量表达,进而触发PR1b和PR1a基因持续超量表达,表明Surfactin A诱导烟草对TMV的系统抗性是通过激活SA信号转导途径实现的。同时,Surfactin A诱导处理后24~72 h,JA/ET信号转导途径调节基因PDF1.2被激活,且超量表达,表明在Surfactin A诱导烟草对TMV系统抗性的信号转导过程中,可能存在SA信号途径和JA/ET信号途径的交叉协同作用。     

Suppression of Meloidogyne incognita by the endophytic fungus Acremonium implicatum from tomato root galls

Specific endophytes with biocontrol potential might occur in diseased plant tissues. We isolated an endophytic fungus from tomato root galls infected with Meloidogyne incognita and identified it as Acremonium implicatum based on morphology and internal transcribed spacer sequences. Its biocontrol potential was tested in vitro and in pot and field experiments. In the in vitro test, 96.0% of second-stage juveniles of M. incognita were killed by a culture filtrate of A. implicatum after 48 h. The fungus also suppressed egg hatching, with only 36.3% of treated eggs hatching compared with 87.3% of control eggs. Pot experiments showed that A. implicatum inhibited the formation of root galls, with 40.6 galls per treated plant compared with 121.6 on control plants. A. implicatum reduced the nematode population in soil, with 151.1 nematodes per 100 g treated soil and 375.1 in control soil. Field experiments demonstrated that the root gall index of treated plants (25) was markedly lower than that of control plants (96). In conclusion, A. implicatum has excellent potential for the biocontrol of M. incognita.     

柑橘木虱黄龙病菌携带量与其内生菌群相关性

【目的】分析柑橘木虱（Diaphorina citri）体内可能与黄龙病菌（Candidatus Liberibacter asiaticus）互作的内生细菌,为黄龙病菌的人工培养及其病害防控奠定基础。【方法】首先通过传统分离培养方法比较不同地理来源带黄龙病菌（带菌）和不带菌黄龙病菌（不带菌）的木虱中可培养内生细菌的差异。其次将带菌状况不同的木虱分别分为头、胸、腹3部分,经PCR扩增其16S rDNA的V6-V8区,用变性梯度凝胶电泳（PCR-DGGE）方法,比较带菌状况不同的木虱内生细菌的差异和木虱不同部位内生细菌的差异。选择3种差异的内生细菌：Bacillus sp.、Salmonella sp.、Enterobacter sp.,在8份带菌状况不同的木虱样品中通过q-PCR分别对其进行实时荧光定量分析,再以总细菌量为校正计算包括黄龙病菌在内的4种细菌的相对含量,数据经LSD检验,以各种细菌相对含量的-lg值作图,先比较同一样品中3种细菌分别与黄龙病菌的相对含量关系,再比较同种细菌在不同样品中的特性,分析3种内生细菌和黄龙病菌的互作关系。【结果】不带菌木虱中可培养内生菌菌落丰富度和菌落形成单位均大于带菌木虱中。在不带菌木虱中共获得14株形态不同的菌株,分属于芽孢杆菌属（Bacillus,3株）、欧文氏菌属（Erwinia,1株）、克雷伯氏杆菌属（Klebsiella,1株）、葡萄球菌属（Staphylococcus,2株）、节杆菌属（Arthrobacter,1株）、泛菌属（Pantoea,2株）、果胶杆菌属（Pectobacterium,1株）、沙门氏菌属等（Salmonella,1株）、链霉菌属（Streptomyces,1株）、Massilia brevitalea（1株）等10个细菌属。在带菌木虱中分得的4株细菌在不带菌木虱中均分离到,分属于克雷伯氏菌属、芽孢杆菌属、果胶杆菌属。其中Dc-11（嗜气芽孢杆菌属）在带菌木虱和不带菌木虱中分离频率均达到100%,表明其为木虱体内常驻细菌;对木虱不同部位（头、胸、腹）内生细菌16S rDNA的PCR-DGGE图谱显示,带菌与不带菌木虱中细菌种群差别明显,带菌状况相同的木虱不同部位之间差别不明显。其中优势条带10 （Wolbachia sp.）、12（Wolbachia pipientis）、13（Syncytium endosymbiont of Diaphorina citri）、14（Uncultured bacterium）、19（Serratia marcescens）、21和22（均为嗜麦芽寡养单胞菌Stenotrophomonas maltophilia）在木虱体内稳定存在,带菌木虱腹部特有优势内生细菌为Enterobacter sp.,木虱中同样也存在次级内生菌Wolbachia。q-PCR的结果验证了所选的3种细菌在前期传统分离培养和16S rDNA-PCR-DGGE中结果的可靠性,同时表明肠杆菌属在8份样品中与黄龙病菌呈正相关关系。【结论】黄龙病菌进入木虱体内会改变木虱内生细菌菌群种类和结构;嗜气芽孢杆菌（B. aerophilus）在柑橘木虱体内稳定存在,为木虱体内常驻菌群;Enterobacter sp.与黄龙病菌带菌量呈正相关,推测其可能与黄龙病菌互作。     

Nitrifier dominance of Arctic soil nitrous oxide emissions arises due to fungal competition with denitrifiers for nitrate

Arctic soils emit nitrous oxide, which is a potent greenhouse gas and also represents an important loss of nitrogen to oligotrophic Arctic ecosystems. However, little is known about the temperature sensitivity of nitrous oxide release in Arctic soils or the organisms mainly responsible for it. We investigated controls on nitrous oxide emissions in an Arctic soil across a typical temperature range (between 4 and 13 °C) on Truelove Lowland, Devon Island, Canada (75°40′N 84°35′W) at two different moisture contents. When fertilized with ammonia or nitrate, nitrous oxide emissions and temperature dependence of nitrous oxide emissions were insensitive to soil moisture content but linked to nitrification rates. Stable isotope analysis revealed that nitrous oxide was predominantly released by nitrifiers. However, nitrous oxide emissions were not linked to nitrifier prevalence with an insignificant (P < 0.219) increase in amoA genes and a (P < 0.01) decrease in archaeal nitrifiers. In contrast, denitrifier nosZ prevalence was 10,000 times greater than that of nitrifiers and was related to nitrous oxide emission potential when soils were fertilized with nitrate. Manipulating water-filled pore space should have changed the pattern of N₂O emissions. We used selective inhibitors to further explore why denitrification did not occur under field conditions when we manipulated water-filled pore space or when we used ¹⁵N analysis. When fungi were inhibited in the soil, nitrous oxide emissions from denitrifiers increased with no change in nitrous oxide released by nitrifiers. When fungi were active in the soil, there was little available nitrate but when fungi were inhibited, available soil nitrate increased over the incubation period. The dominance of nitrifiers in nitrous oxide emissions from Arctic soils under field conditions is linked to the competition for nitrate between fungi and denitrifiers.     

Compartmentalization of the soil animal food web as indicated by dual analysis of stable isotope ratios (N/N and C/C)

The soil animal food web has become a focus of recent ecological research but trophic relationships still remain enigmatic for many taxa. Analysis of stable isotope ratios of N and C provides a powerful tool for disentangling food web structure. In this study, animals, roots, soil and litter material from a temperate deciduous forest were analysed. The combined measurement of δ¹⁵N and δ¹³C provided insights into the compartmentalization of the soil animal food web. Leaf litter feeders were separated from animals relying mainly on recent belowground carbon resources and from animals feeding on older carbon. The trophic pathway of leaf litter-feeding species appears to be a dead end, presumably because leaf litter feeders (mainly diplopods and oribatid mites) are unavailable to predators due to large size and/or strong sclerotization. Endogeic earthworms that rely on older carbon also appear to exist in predator-free space. The data suggest that the largest trophic compartment constitutes of ectomycorrhizal feeders and their predators. Additionally, there is a smaller trophic compartment consisting of predators likely feeding on enchytraeids and potentially nematodes.