AM真菌与镉互作影响桑生长和无机元素吸收转运

为评价Cd胁迫下接种AM真菌(AMF)对减轻桑树Cd毒害及Cd迁移规律的影响,采用了分室培养法开展试验。在菌根室种植无菌桑苗并接种AMF(Gigaspora rosea),菌丝室设置兼菌丝收集器功能的Cd陷阱。陷阱中的Cd处理浓度为0、5、20、40mg/kg,菌根室每室种植3株桑苗,6个重复,对照不接种AMF。培养60d后检测桑苗的菌根侵染率、不同部位的生长量、AMF菌丝生物量、4种矿质营养元素组分及Cd在AMF菌丝体和桑株中的定位及转运情况。结果表明:在0–40mg/kg的Cd处理下,不接种AMF处理的对照桑,其菌根侵染率、AMF菌丝生物量以及镉含量都为0。接种AMF处理中,当Cd<5mg/kg时,桑苗菌根侵染率、AMF菌丝生物量、桑苗鲜生物量、枝叶中N、K、Ca、Mg以及根系N、K、Ca矿质元素的积累都有促进作用,在5mg/kg时促生效果最佳;当Cd>5mg/kg时,随着Cd浓度的升高,桑苗菌根侵染率、AMF菌丝生物量、桑苗鲜生物量、枝叶中N、K、Ca、Mg以及根系N、K、Ca矿质元素的积累都受到渐强的抑制;陷阱中Cd浓度高低与根系中Mg元素的吸收量之间显著负相关;接种AMF可以显著提高桑株生物量和矿质营养的积累;Cd主要集中在桑苗的根部,Cd处理浓度与AMF菌丝体中以及桑株中的Cd含量均显著正相关;Cd在菌根桑苗中的迁移率(<9%)和不同部位(根-茎、根-叶)的转移系数(<0.1)都极其低。结果证明:Cd胁迫对桑苗的菌根侵染率、营养生长、AMF菌丝生长和矿质吸收(除Mg外)都存在低促高抑的剂量效应;AMF对Cd胁迫有一定的抗性,接种AMF显著促进植物的生长以及矿质营养元素的吸收和转运;重金属Cd在菌根桑中的分布、迁移是不均匀的,具有一定的独特性。     

金沙江支流普渡河、小江干热河谷的丛枝菌根

调查具有不同植物群落的金沙江支流普渡河、小江干热河谷中91种常见植物的丛枝菌根真菌的侵染率及孢子密度。普渡河样地调查了56种植物,其中54种(96%)植物能形成典型的丛枝菌根,其平均孢子密度为1423±175/100g土;小江样地35种植物中有34种(97%)植物能形成典型的丛枝菌根,其平均孢子密度为601±103/100g土。单因素方差分析表明两个样地植物的AMF总感染率差异不显著,但其根际土壤中AMF孢子密度却存在显著差异,小江样地的AMF孢子密度明显低于普渡河样地。相关性分析表明,干热河谷植物的AMF感染率与其根际土壤中的AMF孢子密度之间不存在相关性。此外,调查还发现91种植物中,有61种植物(67%)在形成AM的同时,也被黑色有隔内生菌感染。     

丛枝菌根共生体中碳、氮代谢及其相互关系

丛枝菌根共生体(arbuscular mycorrhiza, AM)是丛枝菌根真菌(arbuscular mycorrhizal fungi, AMF)与宿主植物之间形成的互惠共生形式.共生体中的碳、氮交换和代谢影响着宿主植物和共生真菌之间的营养平衡和资源重新分配,在物质和能量循环中发挥着重要作用.宿主植物光合固定的碳输送到真菌内,并且分解和释放真菌所需的生命物质和能量,包括促进孢子萌发、菌丝生长和提高氮等营养元素的吸收;而菌根真菌利用宿主植物提供的碳骨架和能量,发生氮的转化和运输,最终传递给宿主植物供其利用.本文综述了丛枝菌根共生体中碳、氮传输和代谢的主要模式,碳、氮的交互影响和调控机制,以促进丛枝菌根在可持续农业和生态系统中的应用.     

一种观察放线菌气生菌丝的简单方法

放线菌是由纤细的菌丝所形成的单细胞菌丝体。菌丝体又包括二个部分,潜入在培养基中的为营养菌丝(亦称为基内菌丝);生长在培养基表面的为气生菌丝,气生菌丝分化出形成孢子的菌丝称作孢子丝。孢子丝的形状可呈螺旋形、波浪弯曲形或分枝状等。孢子丝的形状常常作为分类的重要依据。     

暖季型草坪草AMF 侵染与孢子密度季节动态研究

以广州地区常见暖季型草坪草(地毯草Axonopus compressus、狗牙根Cynodon dactylon、结缕草Zoysia japonica)为研究对象, 通过测定植物根系AMF 侵染率、根际土壤AMF 孢子密度以及土壤理化性质, 探讨AMF 侵染率和孢子密度的季节动态变化特征, 以及与土壤理化性质之间的关系。结果表明: (1)三种草坪草的根系AMF 侵染以菌丝侵染为主, 总体表现为夏季最高, 秋季和冬季次之, 春季最低, 囊泡侵染率较低, 很少丛枝侵染; (2)季节对根际土壤AMF 孢子密度有一定的影响, 草坪草根际土壤孢子密度最大值多出现在夏季或冬季, 而春季和秋季则较低; (3)对于土壤因子, AMF 菌丝侵染率与土壤有机质含量成极显著正相关, 而囊泡侵染率与电导率成显著正相关, 与全氮和全磷成极显著负相关; 对于气候因子, AMF 总侵染率和菌丝侵染率均与月均气温和月均降水成极显著正相关。     

丛枝菌根真菌状巨孢囊霉孢子伴生细菌的绿色荧光蛋白标记及定殖分析

摘要：【目的】分析丛枝菌根（Arbuscualr Mycorrhizal, AM）真菌珠状巨孢囊霉(Gigaspora margarita) MAFF 520054孢子伴生细菌的定殖情况,明确其生态位点,以及为进一步分析其种群生态或功能提供信息。【方法】以载体pNF8（gfp-mut1）对6株珠状巨孢囊霉MAFF 520054 孢子伴生细菌进行绿色荧光蛋白（GFP）标记,并通过荧光显微镜和平板计数的方法研究标记菌株对真菌宿主的定殖位点和不同条件下的定殖数量动态。【结果】对粘状芽孢杆菌（Peanibacillus spp.）M060106-1和M061122-6、芽孢杆菌（Bacillus sp.）M061122-10和短小芽孢杆菌（Brevibacillus sp.）M061122-12成功进行了GFP标记,其均具有较好的质粒稳定性,且与出发株的基本性状一致,适合短期内进行环境定殖研究。所有菌株均能定殖珠状巨孢囊霉MAFF 520054孢子壁,而M061122-6和M061122-12还能够定殖其菌丝;不同pH值条件下,各菌株定殖珠状巨孢囊霉MAFF 520054孢子的数量动态均为先上升后下降,pH值对各菌株的定殖数量有不同的影响;各GFP菌株对低活力的珠状巨孢囊霉孢子定殖数量高于高活力的孢子,且对高活力孢子的定殖数量动态不同。【结论】分离的珠状巨孢囊霉孢子伴生细菌能够重新定殖其孢子,菌株的定殖能力受其特性及外界因子的影响,为进一步分析AM真菌伴生细菌的种群生态及功能提供了信息。     

丛枝菌根真菌系统分类及群落研究技术进展

丛枝菌根真菌（AMF）是自然生态系统重要的组成部分,能与植物根系形成互惠共生体.传统的AMF分类主要依赖于对土壤无性孢子的形态鉴定,具有一定的局限性.近年来基于核酸分析的分子鉴定技术使AMF的分类更具科学性和准确性,补充和完善了基于孢子形态鉴定所建立的分类系统.AMF群落研究依赖于AMF的分类鉴定,主要包括孢子形态鉴定和分子生物学分析两类研究法.本文综述了AMF的分类系统和群落研究方法,着重介绍了近年来应用较多的AMF群落研究的分子生物学技术.作者认为,采取形态与分子相结合的办法将有助于推动AMF群落研究和AMF自然分类系统的建立和完善.     

银杏根际丛枝菌根真菌生长与根系黄酮含量的相关性研究

试验通过调查银杏根际丛枝菌根真菌的菌丝体长度、孢子密度及根系菌根侵染率,并测定银杏根系黄酮含量的季节性变化,研究二者的变化规律,分析相关关系。试验结果表明,年周期内银杏根际丛枝菌根真菌菌丝体长度、孢子密度与根系黄酮类化合物含量呈规律性变化:1月~3月根际丛枝菌根真菌菌丝长度有限,孢子密度、根系菌根侵染率与根系黄酮类化合物含量都最低;3月上中旬皆迅速增加,到9月达到全年最高峰;11月后,丛枝菌根真菌菌丝长度、孢子密度及根系菌根侵染率与根系黄酮类化合物含量均有所下降。通过数学模型分析,银杏根际丛枝菌根真菌菌丝长度、孢子密度及根系菌根侵染率与根系黄酮类化合物含量表现显著的正相关。     

夏块菌与青刚栎形成外生菌根形态变化的研究

夏块菌（Tuber aestivum）是一种具有较高经济价值的菌根食用菌。对夏块菌与青刚栎（Cycloba-lanopsis glauca）在形成菌根过程中不同阶段的菌根形态变化进行了研究,结果显示：用夏块菌孢子液接种青刚栎苗后,第14天起开始形成淡乳色的外生菌根,外延菌丝刚毛状;第一至第二个月可形成黄褐色、褐色外生菌根,外延菌丝刚毛或羊毛状。外生菌根为单根,长1～4mm,直径150～250μm。菌套厚12～20μm,平坦或自菌根延伸出刚毛状菌丝,外延菌丝束黄绿色;哈替氏网菌丝直径1～1.5μm。菌根老化后变暗褐或萎缩。外延菌丝束呈黄绿色是夏块菌菌根区别于其它块菌菌根最重要的形态特征。     

宿主植物栽培密度对AM真菌生长发育的影响

温室盆栽条件下宿主植物高粱(SorghumvulgarePers.)的栽培密度对丛枝菌根(Arbuscularmycorrhizae,AM)真菌Glomusmosseae(Nicol.&Gerd.)Gerdemann&Trappe生长发育的影响试验结果表明:60株/盆密度处理的根外菌丝量及孢子数均高于其它处理。在一定栽培密度下(20~60株/盆),植株根系可溶性糖浓度与根外菌丝量呈显著负相关,与菌根侵染率呈显著正相关。植株根中磷浓度与根外菌丝量、根外菌丝量与孢子数均呈显著正相关。植株根中磷浓度与菌根侵染率呈显著负相关。结果说明:适当密植虽对植株生长有一定影响,但却促进了真菌的生长,此时菌根共生体有可能由互惠共生开始向偏利共生或弱寄生转化。密植作为一种调控手段,在菌剂生产中能获得较大数量的侵染根段、菌丝及孢子等繁殖体。     

Arbuscular mycorrhizal fungi stimulate organic phosphate mobilization associated with changing bacterial community structure under field conditions

The extraradical hyphae of arbuscular mycorrhizal fungi (AMF) harbour and interact with a microbial community performing multiple functions. However, how the AMF‐microbiome interaction influences the phosphorus (P) acquisition efficiency of the mycorrhizal pathway is unclear. Here we investigated whether AMF and their hyphal microbiome play a role in promoting organic phosphorus (P) mineralizing under field conditions. We developed an AMF hyphae in‐growth core system for the field using PVC tubes sealed with membrane with different size of pores (30 or 0.45 μm) to allow or deny AMF hyphae access to a patch of organic P in root‐free soil. AMF and their hyphae associated microbiome played a role in enhancing soil organic P mineralization in situ in the field, which was shown to be a function of the change in bacteria community on the hyphae surface. The bacterial communities attached to the AMF hyphae surface were significantly different from those in the bulk soil. Importantly, AMF hyphae recruited bacteria that produced alkaline phosphatase and provided a function that was absent from the hyphae. These results demonstrate the importance of understanding trophic interactions to be able to gain insight into the functional controls of nutrient cycles in the rhizosphere.     

缩球囊霉孢子萌发及细胞核在孢子和芽管中分布的研究*

本文研究了从本校分离的丛枝菌根真菌缩球囊霉Glomus constrictum的孢子萌发和萌发孢子的细胞核DAPI(46-diamidino-2-phenylindol)染色。结果显示,G. constrictum孢子直径为179.5-198.7μm ,顶生于产孢菌丝上。经表面消毒处理后,孢子在水琼脂平板上7天开始萌发。DAPI染色后,经稀释荧光计数,单个孢子细胞核数目约为5300,在孢子中无序分布, 细胞核直径约为9.9-11.2μm 。孢子萌发过程中,细胞核总数无明显变化,只是部分细胞核从孢子流向了萌发伸长的菌丝。     

Isolation and identification of soil bacteria growing at the expense of arbuscular mycorrhizal fungi

Soil-microorganism symbioses are of fundamental importance for plant adaptation to the environment. Research in microbial ecology has revealed that some soil bacteria are associated with arbuscular mycorrhizal fungi (AMF). However, these interactions may be much more complex than originally thought. To assess the type of bacteria associated with AMF, we initially isolated spores of Glomus irregulare from an Agrostis stolonifera rhizosphere. The spores were washed with sterile water and plated onto G. irregulare mycelium growing in vitro in a root-free compartment of bicompartmented Petri dishes. We hypothesized that this system should select for bacteria closely associated with the fungus because the only nutrients available to the bacteria were those derived from the hyphae. Twenty-nine bacterial colonies growing on the AMF hyphae were subcultured and identified using 16S rRNA gene sequences. All bacterial isolates showed high sequence identity to Bacillus cereus, Bacillus megaterium, Bacillus simplex, Kocuria rhizophila, Microbacterium ginsengisoli, Sphingomonas sp. and Variovorax paradoxus. We also assessed bacterial diversity on the surface of spores by PCR-denaturating gradient gel electrophoresis. Finally, we used live cellular imaging to show that the bacteria isolated can grow on the surface of hyphae with different growing patterns in contrast to Escherichia coli as a control.     

钦州湾红树林丛枝菌根初步研究

海漆 (Excoecaria agallocha)、桐花树 (Aegiceras corniculatum)、秋茄 (Kandlia candel)、白骨壤 (Avi-cennia marina)是钦州湾分布广、生长量大的 4种红树植物 ,该项研究初步调查了丛枝菌根真菌在这四种植物根系的定居情况。在这四种植物的根际土壤均分离到丛枝菌根真菌孢子 ,其孢子密度以海漆根际的最高 ,其次为桐花树、秋茄、白骨壤。同时 ,海漆根系的侵染率和物种丰富度也最高 ,但在所取的白骨壤根样中没有观察到丛枝菌根的侵染。菌根侵染主要是以根内菌丝、胞内菌丝膨大扭曲、泡囊等形态出现。海水和土壤质地是影响菌根侵染率的主要因素。在所采土样中仅发现球囊霉属 (Glomus)、无梗囊霉属 (Acaulospora)丛枝菌根真菌 ,计有 1 7种和多个未确定种。     

N-P fertilization did not reduce AMF abundance or diversity but alter AMF composition in an alpine grassland infested by a root hemiparasitic plant

Fertilization has been shown to have suppressive effects on arbuscular mycorrhizal fungi (AMF) and root hemiparasites separately in numerous investigations, but its effects on AMF in the presence of root hemiparasites remain untested. In view of the contrasting nutritional effects of AMF and root hemiparasites on host plants, we tested the hypothesis that fertilization may not show strong suppressive effects on AMF when a plant community was infested by abundant hemiparasitic plants. Plants and soil samples were collected from experimental field plots in Bayanbulak Grassland, where N and P fertilizers had been applied for three continuous years for control against a spreading root hemiparasite, Pedicularis kansuensis. Shoot and root biomass of each plant functional group were determined. Root AMF colonization levels, soil spore abundance, and extraradical hyphae length density were measured for three soil depths (0-10 cm, 10-20 cm, 20-30 cm). Partial 18S rRNA gene sequencing was used to detect AMF diversity and community composition. In addition, we analyzed the relationship between relative abundance of different AMF genera and environmental factors using Spearman's correlation method. In contrast to suppressive effects reported by many previous studies, fertilization showed no significant effects on AMF root colonization or AMF species diversity in the soil. Instead, a marked increase in soil spore abundance and extraradical hyphae length density were observed. However, fertilization altered relative abundance and AMF composition in the soil. Our results support the hypothesis that fertilization does not significantly influence the abundance and diversity of AMF in a plant community infested by P. kansuensis.     

Effect of physical, chemical and environmental characteristics on arbuscular mycorrhizal fungi in Brachiaria decumbens (Stapf) pastures

Aim:  To evaluate the effects of soil physical and chemical factors (pH, conductivity, humidity, available phosphorus and organic matter) and environmental factors (temperature, relative air humidity, altitude and atmospheric pressure) on arbuscular mycorrhizal fungi (AMF)– Brachiaria decumbens grass relationship. Furthermore to establish patterns of microbiological responses that allow to differentiate the study sites in two relief types.
Methods and Results:  Mycorrhizal characteristics (spore density, external hyphae and root colonizations by hyphae, vesicles and arbuscules), physical and chemical factors in soil and environmental factors were measured.
Conclusions:  The effect of physical, chemical and environmental factors on microbiological variables was related to the type of relief 'valley and hilly terrain'; the AMF behaviour was affected only over narrower ranges of evaluated variables. Similarly, the colonization of B. decumbens roots by AMF hyphae, vesicles and the mycorrhizal spore density follow different patterns according to the relief type.
Significance and Impact of the Study:  The type of relief is one of the factors to be taken into consideration to evaluate the AMF inoculum and root colonization of these pastures, because of the influence of slope – as physical property of soil – on AMF.     

Bacteria associated with spores of the arbuscular mycorrhizal fungi Glomus geosporum and Glomus constrictum

Spores of the arbuscular mycorrhizal fungi (AMF) Glomus geosporum and Glomus constrictum were harvested from single-spore-derived pot cultures with either Plantago lanceolata or Hieracium pilosella as host plants. PCR-denaturing gradient gel electrophoresis analysis revealed that the bacterial communities associated with the spores depended more on AMF than host plant identity. The composition of the bacterial populations linked to the spores could be predominantly influenced by a specific spore wall composition or AMF exudate rather than by specific root exudates. The majority of the bacterial sequences that were common to both G. geosporum and G. constrictum spores were affiliated with taxonomic groups known to degrade biopolymers (Cellvibrio, Chondromyces, Flexibacter, Lysobacter, and Pseudomonas). Scanning electron microscopy of G. geosporum spores revealed that these bacteria are possibly feeding on the outer hyaline spore layer. The process of maturation and eventual germination of AMF spores might then benefit from the activity of the surface microorganisms degrading the outer hyaline wall layer.     

Effect of long-term tillage and mineral phosphorus fertilization on arbuscular mycorrhizal fungi in a humid continental zone of Eastern Canada

Aims

Evidence shows that tillage modifies soil properties, especially phosphorus (P) dynamics. Our objective was to disentangle long-term effects of P-fertilization and tillage on arbuscular mycorrhizal fungal (AMF) proliferation and community structure.

Methods

Changes in the community structure of AMF and in the density of their hyphae and spores induced by moldboard plow (MP) or no till (NT), and fertilization with 0, 17.5, or 35 kg?P?ha^?1 were sought in the 0–15 cm and 15–30 cm soil layers after soybean harvest, at a long-term (17 years) experimental site in a humid continental zone of eastern Canada. The relationships among AMF, soil and plant attributes were examined.

Results

The 0–15 cm and 15–30 cm soil layers had different properties under NT, but were similar under MP, after 17 years, and MP increased soil available P levels. Phosphorus fertilization increased P levels in soil and in soybean. Treatment effects on AMF spore and hyphal density at 0–15 cm were greater than that at 15–30 cm, whereas effects on AMF community structure did not change with soil depths. At 0–15 cm, P-fertilization increased AMF spore density and reduced AMF hyphal density, and MP reduced AMF spore density. A total of eight AMF phylotypes were detected. Phosphorus fertilization reduced AMF phylotype richness and Shannon diversity index. Soil P availability increased under MP and hence the influence of P-fertilization treatments on the frequency of AMF phylotype detection varied with tillage system; it declined with P-fertilization under MP, but increased under NT.

Conclusions

Phosphorus fertilization shifts resource partitioning in AMF propagules rather than in their hyphae, and degrades the genetic diversity of AMF in soil; tillage increases soil P availability and hence aggravates the impact of P-fertilization.     

Rhizosphere effect of nanoscale zero-valent iron on mycorrhiza-dependent maize assimilation

Rhizosphere effect of nanoscale zero-valent iron (nZVI) is crucial but little reported. Maize seeds were dressed with four nZVI concentrations (0, 1.0, 1.5, 2 g kg⁻¹) and inoculated with arbuscular mycorrhizal fungus (AMF) (Funneliformis mosseae). The SEM images illuminated that excessive nZVI particles (2 g kg⁻¹) were agglomerated on the surface of hyphae and spore, causing severe deformation and inactivation of AMF symbionts and thereafter inhibiting water uptake in maize seedlings. This restrained the scavenging effects of enzymatic (superoxide dismutase, peroxidase) and non-enzymatic compounds (proline & malondialdehyde) on ROS, and leaf photoreduction activity and gas exchange ability (p < 0.05). Interestingly, the inoculation with AMF effectively alleviated above negative effects. In contrast, appropriate dose of nZVI, that is, ≤1.5 g kg⁻¹, can be evenly distributed on the hyphae surface and form the ordered symbionts with AMF. This help massively to enhance hyphae growth and water and nutrient uptake. The enhanced mycorrhizal infection turned to promote rhizosphere symbiont activity and leaf Rubisco and Rubisco activase activity. Light compensation point was massively lowered, which increased photosynthetic carbon supply for AMF symbionts. Particularly, such priming effects were evidently enhanced by drought stress. Our findings provided a novel insight into functional role of nZVI in agriculture and AMF-led green production.