An inexpensive high-throughput method to extract high yields of good quality DNA from fungi

We developed an efficient method for high-throughput extraction of high-quality DNA from various fungi. In this method, fungal mycelia were cultured and harvested on the surfaces of membranes on media plates. We degraded cell walls using a lytic enzyme (Yatalase). Purification was performed on 96-well glass fibre filter plates. DNA was successfully extracted from various fungi provided (102 genus 132 species) at high yields and quality, and proved suitable for storage, polymerase chain reaction amplification and restriction enzyme digestion. The method described is rapid, inexpensive and automation friendly. This enables the simultaneous extraction of large numbers of samples, significantly improving the potential throughput in genomics, particularly in diagnostic and population studies.     

春兰菌根的显微结构及菌根真菌的分离

以盆栽春兰和野生春兰的根为材料,通过切片观察菌根真菌的分布特点和菌根的显微结构。同时采用组织分离方法从盆栽和野生春兰根分别分离得到6个和15个真菌菌株,分离菌株的出现频率分别为16．7％～50％和6．7％～73．3％,并由此判断CGP-3为盆栽春兰菌根的优势菌种,CGW-3为野生春兰菌根的优势菌种。经形态学观察将其初步分为6个分类单元,其中CGP-2和CGW-2分别具有拟日本曲霉（Aspergillus japonicus-like）和拟木霉属（Trichoderma sp.-like）的形态特征。此外,对兰科棺物与内生真菌间的专一件讲行了讨论。     

Phosphorus nutrition and mycorrhiza effects on grass leaf growth. P status- and size-mediated effects on growth zone kinematics

This study tested whether leaf elongation rate (LER, mm h(-1)) and its components--average relative elemental growth rate (REGRavg, mm mm(-1) h(-1)) and leaf growth zone length (L(LGZ), mm)--are related to phosphorus (P) concentration in the growth zone (P(LGZ) mg P g(-1) tissue water) of Lolium perenne L. cv. Condesa and whether such relationships are modified by the arbuscular mycorrhizal fungus (AMF) Glomus hoi. Mycorrhizal and non-mycorrhizal plants were grown at a range of P supply rates and analysed at either the same plant age or the same tiller size (defined by the length of the sheath of the youngest fully expanded leaf). Both improved P supply (up to 95%) and AMF (up to 21%) strongly increased LER. In tillers of even-aged plants, this was due to increased REGRavg and L(LGZ). In even-sized tillers, it was exclusively due to increased REGRavg. REGRavg was strictly related to P(LGZ) (r2 = 0.95) and independent of tiller size. Conversely, L(LGZ) strictly depended on tiller size (r2 = 0.88) and not on P(LGZ). Hence, P status affected leaf growth directly only through effects on relative tissue expansion rates. Symbiosis with AMF did not modify these relationships. Thus, no evidence for P status-independent effects of AMF on LER was found.     

不同干旱胁迫条件下丛枝菌根真菌对 木棉叶绿素荧光参数的影响

为了探明丛枝菌根真菌(AMF)与木棉(Bombax ceiba Linn.)抗旱性的关系,采用盆栽法对不同干旱胁迫条件下接菌组(1 g土壤含100个AMF孢子)和未接菌组木棉叶绿素荧光参数进行了比较分析.结果表明:中度干旱胁迫(土壤相对含水量45%~50%)和重度干旱胁迫(土壤相对含水量30%~35%)条件下接菌组的初始荧光(Fo)均低于未接菌组;与对照(土壤相对含水量65%~70%)相比,重度干旱胁迫5、15和35 d,接菌组的Fo值分别升高了10.39%、20.93%和14.14%,未接菌组的Fo值分别升高了26.95%、48.75%和71.35%.接菌组的PSⅡ最大光化学效率(Fv/Fm)和PSⅡ潜在活性(Fv/Fo)在中度和重度干旱胁迫5、15和35 d均高于未接菌组.与对照相比,重度干旱胁迫5、15和35 d,接菌组的Fv/Fm值分别降低了9.09%、6.85%和22.06%,未接菌组的Fv/Fm值分别降低了10.67%、16.21%和52.78%;接菌组的Fv/Fo值分别降低了28.14%、22.43%和46.01%,未接菌组的Fv/Fo值分别降低了28.38%、48.20%和75.96%.总体来看,在中度和重度干旱胁迫条件下,接菌组的PSⅡ实际光量子产量〔Y(Ⅱ)〕、PSⅡ非调节性能量耗散的量子产量〔Y(NO)〕、光化学淬灭系数(qP)和光合电子传递速率(ETR)高于对照,而PSⅡ调节性能量耗散的量子产量〔Y(NPQ)〕和非光化学淬灭系数(qN)低于对照;未接菌组的Y(Ⅱ)、qP、qN和ETR值低于对照,而Y(NO)和Y(NPQ)值则高于对照.研究结果显示:AMF能够减轻干旱尤其是重度干旱胁迫条件下木棉受伤害和光抑制的程度,并提高其叶片PSⅡ反应中心的活性,从而增强木棉在干旱胁迫条件下的生存能力.     

Biomineralization of gold by Mucor plumbeus: The progress in understanding the mechanism of nanoparticles’ formation

This contribution describes the deposition of gold nanoparticles by microbial reduction of Au(III) ions using the mycelium of Mucor plumbeus. Biosorption as the major mechanism of Au(III) ions binding by the fungal cells and the reduction of them to the form of Au(0) on/in the cell wall, followed by the transportation of the synthesized gold nanoparticles to the cytoplasm, is postulated. The probable mechanism behind the reduction of Au(III) ions is discussed, leading to the conclusion that this process is nonenzymatic one. Chitosan of the fungal cell wall is most likely to be the major molecule involved in biomineralization of gold by the mycelium of M. plumbeus. Separation of gold nanoparticles from the cells has been carried out by the ultrasonic disintegration and the obtained nanostructures were characterized by UV‐vis spectroscopy and transmission electron micrograph analysis. © 2017 American Institute of Chemical Engineers Biotechnol. Prog., 33:1381–1392, 2017     

Arbuscular mycorrhizal fungi associated with Populus-Salix stands in a semiarid riparian ecosystem

This study examined the activity, species richness, and species composition of the arbuscular mycorrhizal fungal (AMF) community of Populus-Salix stands on the Verde River (Arizona, USA), quantified patterns of AMF richness and colonization along complex floodplain gradients, and identified environmental variables responsible for structuring the AMF community. Samples from 61 Populus-Salix stands were analyzed for AMF and herbaceous composition, AMF colonization, gravimetric soil moisture, soil texture, per cent organic matter, pH, and concentrations of nitrate, bicarbonate phosphorus and exchangeable potassium. AMF species richness declined with stand age and distance from and elevation above the channel and was positively related to perennial species cover and richness and gravimetric soil moisture. Distance from and elevation above the active channel, forest age, annual species cover, perennial species richness, and exchangeable potassium concentration all played a role in structuring the AMF community in this riparian area. Most AMF species were found across a wide range of soil conditions, but a subset of species tended to occur more often in hydric areas. This group of riparian affiliate AMF species includes several not previously encountered in the surrounding Sonoran desert.     

Mycorrhizal suppression and phosphorus addition influence the stability of plant community composition and function in a temperate steppe

Nutrient enrichment can reduce ecosystem stability, typically measured as temporal stability of a single function, e.g. plant productivity. Moreover, nutrient enrichment can alter plant–soil interactions (e.g. mycorrhizal symbiosis) that determine plant community composition and productivity. Thus, it is likely that nutrient enrichment and interactions between plants and their soil communities co-determine the stability in plant community composition and productivity. Yet our understanding as to how nutrient enrichment affects multiple facets of ecosystem stability, such as functional and compositional stability, and the role of above–belowground interactions are still lacking. We tested how mycorrhizal suppression and phosphorus (P) addition influenced multiple facets of ecosystem stability in a three-year field study in a temperate steppe. Here we focused on the functional and compositional stability of plant community; functional stability is the temporal community variance in primary productivity; compositional stability is represented by compositional resistance, turnover, species extinction and invasion. Community variance was partitioned into population variance defined as community productivity weighted average of the species temporal variance in performance, and species synchrony defined as the degree of temporal positive covariation among species. Compared to treatments with mycorrhizal suppression, the intact AM fungal communities reduced community variance in primary productivity by reducing species synchrony at high levels of P addition. Species synchrony and population variance were linearly associated with community variance with the intact AM fungal communities, while these relationships were decoupled or weakened by mycorrhizal suppression. The intact AM fungal communities promoted the compositional resistance of plant communities by reducing compositional turnover, but this effect was suppressed by P addition. P addition increased the number of species extinctions and thus promoted compositional turnover. Our study shows P addition and AM fungal communities can jointly and independently modify the various components of ecosystem stability in terms of plant community productivity and composition.     

Identification of specificity-defining amino acids of the wheat immune receptor Pm2 and powdery mildew effector AvrPm2

Plant nucleotide-binding leucine-rich repeat receptors (NLRs) act as intracellular sensors for pathogen-derived effector proteins and trigger an immune response, frequently resulting in the hypersensitive cell death response (HR) of the infected host cell. The wheat (Triticum aestivum) NLR Pm2 confers resistance against the fungal pathogen Blumeria graminis f. sp. tritici (Bgt) if the isolate contains the specific RNase-like effector AvrPm2. We identified and isolated seven new Pm2 alleles (Pm2e–i) in the wheat D-genome ancestor Aegilops tauschii and two new natural AvrPm2 haplotypes from Bgt. Upon transient co-expression in Nicotiana benthamiana, we observed a variant-specific HR of the Pm2 variants Pm2a and Pm2i towards AvrPm2 or its homolog from the AvrPm2 effector family, BgtE-5843, respectively. Through the introduction of naturally occurring non-synonymous single nucleotide polymorphisms and structure-guided mutations, we identified single amino acids in both the wheat NLR Pm2 and the fungal effector proteins AvrPm2 and BgtE-5843 responsible for the variant-specific HR of the Pm2 variants. Exchanging these amino acids led to a modified HR of the Pm2–AvrPm2 interaction and allowed the identification of the effector head epitope, a 20-amino-acid long unit of AvrPm2 involved in the HR. Swapping of the AvrPm2 head epitope to the non-HR-triggering AvrPm2 family member BgtE-5846 led to gain of a HR by Pm2a. Our study presents a molecular approach to identify crucial effector surface structures involved in the HR and demonstrates that natural and induced diversity in an immune receptor and its corresponding effectors can provide the basis for understanding and modifying NLR–effector specificity.     

丛枝菌根真菌伴生细菌的研究进展

在丛枝菌根真菌（Arbuscular mycorrhizal fungi,AMF）的孢子、菌丝的表面或内部栖息着细菌,称之为AMF伴生细菌。AMF伴生细菌种类多样、分布广泛,生态位点包括孢子壁的表面或内部、细胞质、菌丝、孢子果等。其可能的生物学意义包括影响AMF孢子萌发、菌丝生长、菌根形成等过程。由于伴生细菌与AMF联系紧密,其对AMF和土壤微生物生态学具有重要的意义。国际上在该领域的研究已有30多年的历史,就其研究进展进行综述。