酯酶同工酶在食用菌菌种选育中的应用研究

采用聚丙烯酰胺凝胶电泳 ,分析了不同培养时间和不同培养基种类对平菇、杏鲍菇、白灵菇酯酶同工酶的影响 ,结果表明在不同培养时间和不同培养基条件下 ,酯酶同工酶酶谱的酶带呈现不同的多态性 ,由此提示同工酶在食用菌菌种选育的应用上应固定最佳培养时间及最适培养基种类 ,并在所有操作条件一致的情况下 ,才能得到稳定准确的分析结果     

迷迭香精油对几种植物病原菌的抑菌活性研究

采用生长速率法和抑菌圈法测定了迷迭香精油对甜瓜蔓枯病菌、西瓜炭疽病菌、甜瓜灰霉病菌、甜瓜枯萎病菌、梨树腐烂病菌、柑橘黑腐病菌、油菜菌核病菌、蚕豆轮纹病菌和桃细菌性穿孔病菌的抑菌活性.结果表明:迷迭香精油对8种病原真菌都具有很好的抑菌活性,其EC50分别为843.9914、1 084.2372、483.9457、1 735.5866、1 011.7869、901.2150、356.8803、1 197.3657 μL/L;对桃细菌性穿孔病菌也具有一定的抑菌活性,浓度为100 mL/L时,抑菌圈直径为1.00 cm,最低抑菌浓度为1 000 μL/L.     

The use of Verticillium dahliae and Diplodia scrobiculata to induce resistance in Pinus halepensis against Diplodia pinea infection

The pathogenic fungi Verticillium alboatrum and Diplodia scrobiculata were assayed for biological control of Diplodia pinea on Aleppo pine (Pinus halepensis) in Catalonia (north-eastern Spain). Young shoots were pre-treated with inoculations of either V. dahliae or D. scrobiculata, by placing colonized agar plugs on wounds made by removing a single needle fascicle. An inoculation with D. pinea was performed 15 days later. Two months after the shoot inoculations, the canker length on the stems was measured and the percentage of shoot dieback calculated. Verticillium dahliae and D. scrobiculata were found to significantly reduce the canker length of D. pinea (P < 0.05) when compared with positive controls. Diplodia pinea was slightly more sensitive to V. dahliae than to D. scrobiculata, but no significant differences (P > 0.05) were observed in the mean canker length between the two treatments. Trees pre-inoculated with V. dahliae resulted in 31.12% shoot dieback, while those pre-inoculated with D. scrobiculata resulted in 32.18% shoot dieback, compared with positive controls (42.85%).     

分子生物学技术在真菌种性检测中的应用

本文介绍了当前应用于真菌分类学中的六种分子生物学技术（G C值、DNA互补,RNA技术,RFLP、RAPD五种核酸技术和一种蛋白质技术——同工酶技术）的原理、操作。适用范围及其应用状况,为食、药用真菌研究者和生产者提供参考。     

Environmental stress response limits microbial necromass contributions to soil organic carbon

The majority of dead organic material enters the soil carbon pool following initial incorporation into microbial biomass. The decomposition of microbial necromass carbon (C) is, therefore, an important process governing the balance between terrestrial and atmospheric C pools. We tested how abiotic stress (drought), biotic interactions (invertebrate grazing) and physical disturbance influence the biochemistry (C:N ratio and calcium oxalate production) of living fungal cells, and the subsequent stabilization of fungal-derived C after senescence. We traced the fate of ¹³C-labeled necromass from ‘stressed’ and ‘unstressed’ fungi into living soil microbes, dissolved organic carbon (DOC), total soil carbon and respired CO₂. All stressors stimulated the production of calcium oxalate crystals and enhanced the C:N ratios of living fungal mycelia, leading to the formation of ‘recalcitrant’ necromass. Although we were unable to detect consistent effects of stress on the mineralization rates of fungal necromass, a greater proportion of the non-stressed (labile) fungal necromass C was stabilised in soil. Our finding is consistent with the emerging understanding that recalcitrant material is entirely decomposed within soil, but incorporated less efficiently into living microbial biomass and, ultimately, into stable SOC.     

Fungal and bacterial N2O production regulated by soil amendments of simple and complex substrates

Fungal N₂O production results from a respiratory denitrification that reduces NO₃⁻/NO₂⁻ in response to the oxidation of an electron donor, often organic C. Despite similar heterotrophic nature, fungal denitrifiers may differ from bacterial ones in exploiting diverse resources. We hypothesized that complex C compounds and substances could favor the growth of fungi over bacteria, and thereby leading to fungal dominance for soil N₂O emissions. Effects of substrate quality on fungal and bacterial N₂O production were, therefore, examined in a 44-d incubation after soils were amended with four different substrates, i.e., glucose, cellulose, winter pea, and switchgrass at 2 mg C g⁻¹ soil. During periodic measurements of soil N₂O fluxes at 80% soil water-filled pore space and with the supply of KNO₃, substrate treatments were further subjected to four antibiotic treatments, i.e., no antibiotics or soil addition of streptomycin, cycloheximide or both so that fungal and bacterial N₂O production could be separated. Up to d 8 when antibiotic inhibition on substrate-induced microbial activity and/or growth was still detectable, bacterial N₂O production was generally greater in glucose- than in cellulose-amended soils and also in winter pea- than in switchgrass-amended soils. In contrast, fungal N₂O production was more enhanced in soils amended with cellulose than with glucose. Therefore, fungal-to-bacterial contribution ratios were greater in complex than in simple C substrates. These ratios were positively correlated with fungal-to-bacterial activity ratios, i.e., CO₂ production ratios, suggesting that substrate-associated fungal or bacterial preferential activity and/or growth might be the cause. Considering substrate depletion over time and thereby becoming limited for microbial N₂O production, measurements of soil N₂O fluxes were also carried out with additional supply of glucose, irrespective of different substrate treatments. This measurement condition might lead to potentially high rates of fungal and bacterial N₂O production. As expected, bacterial N₂O production was greater with added glucose than with added cellulose on d 4 and d 8. However, this pattern was broken on d 28, with bacterial N₂O production lower with added glucose than with added cellulose. In contrast, plant residue impacts on soil N₂O fluxes were consistent over 44-d, with greater bacterial contribution, lower fungal contribution, and thus lower fungal-to-bacterial contribution ratios in winter pea- than in switchgrass-amended soils. Real-time PCR analysis also demonstrated that the ratios of 16S rDNA to ITS and the copy numbers of bacterial denitrifying genes were greater in winter pea- than in switchgrass-amended soils. Despite some inconsistency found on the impacts of cellulose versus glucose on fungal and bacterial leading roles for N₂O production, the results generally supported the working hypothesis that complex substrates promoted fungal dominance for soil N₂O emissions.     

印度梨形孢真菌促进芝麻生长并提高芝麻抗旱性

将印度梨形孢（Piriformospora indica,Pi）真菌接种芝麻幼苗根部,分别在大田及温室条件下研究供试真菌对芝麻生长和抗旱性的效应。大田试验主要调查产量性状,温室试验在芝麻初花期进行15d的持续干旱处理,测定株高等形态性状、过氧化氢酶活性等生理指标。结果表明,接种及未接种Pi真菌的芝麻植株在干旱处理前后叶长、叶宽差异达到极显著水平,株高、全展叶片数、基部茎粗、叶绿素含量、根颈粗差异达到显著水平;在人工干旱胁迫条件下,接种Pi真菌后芝麻植株能维持较高的过氧化氢酶（CAT）活性和较高含量的脯氨酸（Pro）,降低丙二醛（MDA）含量,从而减轻干旱胁迫伤害。在千粒重及单株籽粒重两个产量性状上,大田和温室接种的芝麻与未接种的差异均达到显著水平。接种Pi真菌不仅能促进芝麻的生长,而且在干旱条件下,使得芝麻表现出较强的抗旱性,显著提高芝麻产量。     

不同程度重金属污染对稻田土壤真菌群落结构的影响

为了研究土壤真菌群落结构在不同程度重金属污染中的变化,本文用Illumina Hi Seq高通量测序技术分析了苏南地区某金属冶炼厂和加工产业区的重金属污染水稻土的真菌群落结构,发现不同程度重金属污染对水稻季土壤真菌丰度和群落结构均有显著影响。经过真菌主成分分析发现,PC1影响因素对样品处理差异的贡献率是35.96%,PC2影响因素对样品处理差异的贡献率是21.48%;通过真菌冗余度分析发现,重金属Pb和Cu污染对土壤真菌群落结构的影响显著;通过对真菌属水平的相对丰度分析表明,重金属污染会显著降低敏感真菌的丰度,如被孢霉属相对丰度最高降低了87.50%、木霉属最高降低了99.46%、离壳菌属和菇属最高降低了100.00%,同时耐性真菌的相对丰度会提高,如类球囊霉属的相对丰度最高增加了98倍、四枝孢霉属最高增加了56倍、根囊壶菌属最高增加了2.62倍。综上所述,不同程度重金属污染对稻田土壤真菌群落结构有显著影响,且随着污染程度的增加,抗逆真菌相对数量和种类显著增加,敏感真菌的相对数量急剧减少,真菌群落结构随着重金属污染程度增加进一步分化。     

Fungal biomass in pastures increases with age and reduced N input

Previous studies have shown that soil fungal biomass increases towards more natural, mature systems. Shifts to a fungal-based soil food web have previously been observed with abandonment of agricultural fields and extensification of agriculture. In a previous field experiment we found increased fungal biomass with reduced N fertilisation. Here, we explore relationships between fungi, bacteria, N input and grassland age on real dairy farms in the Netherlands. We hypothesised that also in pastures that are still in production there is a negative relationship between fungal biomass and fertilisation, and that fungal biomass increases with grassland age in pastures that are still in production. We expected the fungal/bacterial biomass ratio to show the same responses, as this ratio has often been used as an indicator for management changes. We sampled 48 pastures from eight organic dairy farms. Sites differed in age and fertilisation rate. We determined fungal and bacterial biomass, as well as ergosterol (a fungal biomarker). Fungal and bacterial biomass and ergosterol, showed a negative relationship with N application rate, and correlated positively with organic matter percentage. In old pastures, fungal biomass and ergosterol were higher than in younger pastures. Because bacterial biomass responded in the same way as fungal biomass, the F/B ratio remained constant, and can therefore—in our data set—not be used as an indicator for changing management. We conclude that the changes in fungal and bacterial biomass were driven by changes in organic matter quality and quantity. The negative relationship we found between N application rate and fungal biomass adds to earlier work and confirms the presence of this relationship in pastures with relatively small differences in management intensities. Earlier studies on shifts in fungal biomass focused on ex-agricultural fields or restoration projects. Here we show that fungal biomass is also higher in older agricultural pastures.     

高砷硫铁矿周边农田土壤砷分布及真菌群落结构特征研究

本文主要以高砷硫铁矿区周边农田中的真菌群落为研究对象,研究利用Miseq高通量测序技术系统的分析了矿区周边不同区域土壤中真菌的群落特征,对硫铁矿区周边土壤真菌群落的适应机制进行了探究分析。结果表明,(1)砷、铁复合污染土壤可促使土壤真菌丰富度降低,而对土壤真菌多样的影响则表现为先促进后抑制;(2)砷、铁复合污染抑制土壤中Ascomycota(子囊菌门)代谢,促进Basidiomycota(担子菌门)代谢,而对Zygomycota(接合菌门)则表现为低浓度促进,高浓度抑制,对Fusarium(镰刀菌属)、Pseudallescheria(假霉样真菌属)、Cryptococcus(隐球菌属)也呈现“低浓度促进、高浓度抑制”作用;(3)砷、铁的复合物污染使土壤中真菌群落结构产生了明显差异,环境因子对土壤环境真菌群落结构差异影响大小依次为有效砷、总砷、总铁。