Phytotoxicity on cotton ex-plants of an 18.5 kDa protein from culture filtrates of Verticillium dahliae

A phytotoxic protein that evokes the typical symptoms of Verticillium wilt disease in seedlings of Gossypium hirsutum L. (Upland cotton) was isolated from culture filtrates of Verticillium dahliae. The protein was purified by ammonium sulfate precipitation, Sephadex-G100 fractionation, and native PAGE. The 18.5 kDa protein, designated VD18.5, appears to be a single subunit protein with an isoelectric point between 3 and 5. VD18.5 induces symptoms of leaf dehydration, chlorosis, necrosis and stem discoloration in seedlings of the disease susceptible cotton cultivar Siokra 1–4. The LD₅₀ of VD18.5 on protoplasts of Siokra 1–4 was 18 μg mL⁻¹. VD18.5 had no noticeable effect on Pima S-7, which is a disease resistant cultivar. Phytotoxic activity was partially destroyed at high temperature and was abolished by digestion with proteinase K. Mass spectrometry fingerprinting and protein sequence data from VD18.5 yielded no significant matches when submitted to the Mascot search engine and NCBI non-redundant protein databases, respectively. These results suggest that VD18.5 is a novel protein that may be involved in the development of some of the symptoms associated with Verticillium wilt disease in the cotton plant.     

Soil conditions and land use intensification effects on soil microbial communities across a range of European field sites

Intensive land use practices necessary for providing food and raw materials are known to have a deleterious effect on soil. However, the effects that such practices have on soil microbes are less well understood. To investigate the effects of land use intensification on soil microbial communities we used a combined T-RFLP and pyrosequencing approach to study bacteria, archaea and fungi in spring and autumn at five long term observatories (LTOs) in Europe; each with a particular land use type and contrasting levels of intensification (low and high). Generally, due to large gradients in soil variables, both molecular methods revealed that soil microbial communities were structured according to differences in soil conditions between the LTOs, more so than land use intensity. Moreover, variance partitioning analysis also showed that soil properties better explained the differences in microbial communities than land use intensity effects. Predictable responses in dominant bacterial, archaeal and fungal taxa to edaphic conditions (e.g. soil pH and resource availability) were apparent between the LTOs. Some effects of land use intensification at individual field sites were observed. However, these effects were manifest when land use change affected soil conditions. Uniquely, this study details the responses of different microbial groups to soil type and land use intensification, and their relative importance across a range of European field sites. These findings reinforce our understanding of drivers impacting soil microbial community structure at both field and larger geographic scales.     

Response of the soil fungal community to multi-factor environmental changes in a temperate forest

Both environmental and climatic changes are known to influence soil microbial biomes in terrestrial ecosystems. However, there are limited data defining the interactive effects of multi-factor environmental disturbances, including N-deposition, precipitation, and air temperature, on soil fungal communities in temperate forests. A 3-year outdoor pot experiment was conducted to examine the temporal shifts of soil fungal communities in a temperate forest following N-addition, precipitation and air temperature changes. The shifts in the structure and composition of soil fungal communities were characterized by denaturing gradient gel electrophoresis and DNA sequencing. N-addition regimen induced significant alterations in the composition of soil fungal communities, and this effect was different at both higher and lower altitudes. The response of the soil fungal community to N-addition was much stronger in precipitation-reduced soils compared to soils experiencing enhanced precipitation. The combined treatment of N-addition and reduced precipitation caused more pronounced changes in the lower altitude versus those in the higher one. Certain fungal species in the subphylum Pezizomycotina and Saccharomycotina distinctively responded to N fertilization and soil water control at both altitudes. Redundancy discrimination analysis showed that changes in environmental factors and soil physicochemical properties explained 43.7% of the total variability in the soil fungal community at this forest ecosystem. Variations in the soil fungal community were significantly related to the altitude, soil temperature, total soil N content (TN) and pH value (P < 0.05). We present evidence for the interactive effects of N-addition, water manipulation and air temperature to reshape soil fungal communities in the temperate forest. Our data could provide new insights into predicting the response of soil micro-ecosystem to climatic changes.     

4种微生物菌剂对多年连作甜瓜土壤真菌群落的影响

为了探究微生物菌剂对土壤真菌群落及成员的影响,利用4种微生物菌剂(A.谷乐丰聚谷氨酸微生物菌剂,B.启高系列生物菌剂,C.施特葆系列菌剂,D.新型菌剂NBmelon)对土壤进行处理,样品经DNA抽提、PCR扩增和真菌ITS测序,进行分析制图.结果显示,处理B和A的Shannon指数、Simpson指数高于CK,处理D、...     

Fungal endophyte Phomopsis liquidambari B3 enriches the diversity of nodular culturable endophytic bacteria associated with continuous cropping of peanut

Previously, the application of the fungal endophyte Phomopsis liquidambari B3 exhibited notable effect in alleviating soil obstacles caused by continuous cropping of peanut. It was supposed that P. liquidambari B3 could enhance the efficiency of nodulation and nitrogen fixation by enriching the diversity of nodular endophytic bacteria under continous monocropping system. To verify this hypothesis, a pot experiment was conducted under natural conditions. Four treatments were set as follow: discontinuous cropping soil (NS), continuous cropping soil (CK), continuous cropping soil inoculate with actived P. liquidambari B3 (CSB3), and continuous cropping soil inoculate with sterilized P. liquidambari B3 (CSIB3). As a result, a total of 120 isolates were obtained. Our study clearly declared that the addition of fungal endophyte effectively enriched the genetic diversity and the community composition in CSB3. In contrast, a low genetic diversity level and simplex community structure were exhibited in CK and CSIB3. Meanwhile, the positive percentage rate and corresponding community composition of the plant growth-promoting isolates in NS and CSB3 were larger and more abundant than the other. This is the first time to describe the effects of fungal endophytes on the diversity of the nodular culturable endophytic bacteria associated with continuous cropping of peanut.     

再植枸杞根际真菌群落对长期连作的响应研究

受枸杞自身种植特点和道地产区土地资源限制,连作障碍已成为制约宁夏枸杞产业可持续发展的主要因素之一,导致严重的经济损失和生态问题。前期研究表明,连作能够显著影响再植枸杞根际土壤细菌的群落结构和多样性,但就连作对真菌群落结构和功能的影响目前仍不清楚。本文利用实时荧光定量PCR和高通量测序技术,研究连作对再植枸杞根际真菌群落丰度及多样性的影响。实时荧光定量PCR分析表明,与对照样地相比,连作显著促进再植枸杞根际及非根际土壤中细菌和真菌的丰度。但连作对真菌的促进作用明显高于细菌,导致细菌/真菌比例失衡,使再植枸杞根际及非根际土壤微生物环境偏向于真菌型。对测序结果的分析发现,所测定样地中枸杞根际及非根际土壤中的优势真菌门分别为子囊菌门、担子菌门、接合菌门、壶菌门及球囊菌门,其中连作地再植枸杞根际子囊菌门的相对丰度较对照样地显著降低,而接合菌门的相对丰度则显著增加(P0.05)。FUNGuild真菌功能预测也证实连作显著抑制再植枸杞根际土壤丛枝菌根真菌的相对丰度(P0.05)。基于距离的冗余分析(db-RDA)结果表明,土壤pH、电导率、硝态氮和有效磷含量是影响枸杞根际土壤真菌群落变化的主要因子(P0.05),而土壤硝态氮和有效磷含量则是解释非根际土壤真菌群落变化的主要因子(P0.05)。这些结果说明长期施用化肥可能是改变连作地再植枸杞根际土壤真菌群落结构及枸杞-真菌互作关系的主要因素之一。这一研究结果为理解枸杞连作障碍的形成机制提供理论基础。     

Ergosterol and microbial biomass relationship in soil

Ergosterol and microbial biomass C were measured in 26 arable, 16 grassland and 30 forest soils. The ergosterol content ranged from 0.75 to 12.94 g g^-1 soil. The geometric mean ergosterol content of grassland and forest soils was around 5.5 g g^-1, that of the arable soils 2.14 g g^-1. The ergosterol was significantly correlated with biomass C in the entire group of soils, but not in the subgroups of grassland and forest soils. The geometric mean of the ergosterol: microbial biomass C ratio was 6.0 mg g^-1, increasing in the order grassland (5.1), arable land (5.4) and woodland (7.2). The ergosterol:microbial biomass C ratio had a strong negative relationship with the decreasing cation exchange capacity and soil pH, indicating that the fungal part of the total microbial biomass in soils increased when the buffer capacity decreased. The average ergosterol concentration calculated from literature data was 5.1 mg g^-1 fungal dry weight. Assuming that fungi contain 46% C, the conversion factor from micrograms ergosterol to micrograms fungal biomass C is 90. For soil samples, neither saponification of the extract nor the more effective direct saponification during extraction seems to be really necessary.     

Relative abundance and activity of melanized hyphae in different soil ecosystems

Here we report on the frequency of melanized fungal hyphae in 323 soils, covering different land use types. The proportion of total hyphae that was melanized averaged 61%. Arable fields with loamy sand, heathlands and city parks on sandy soils had the highest percentage of melanized hyphae. In addition to the frequency determinations, a microcosm study was performed on the role of melanized hyphae in two different ecosystems: an ex-arable field and a forest. Melanized hyphae appeared to be part of the active hyphae in the forest soil but not in the ex-arable soil. In conclusion, our results indicate that (1) melanized hyphae represent a large proportion of the total fungal biomass in soils and that (2) their function might differ between ecosystems.