Population structure and responses to disturbance of the basidiomycete Resinicium bicolor

Summary Resinicium bicolor (Alb. & Schw. ex Fr.) Parm. [=Odontia bicolor (Alb. & Schw. ex Fr.) Bres.] is an outcrossing resupinate basidiomycete associated with root and butt rots of trees, but is itself only very weakly pathogenic. The distribution of genets among every spruce stump in a 70-year-old 1250 m² spruce stand was analysed using somatic incompatibility testing. R. bicolor was present on 40% of 8-to 10-year-old stumps. Nineteen genets were found occupying 32 stumps; yielding probabilities of colonisation following establishment by basidiospores of 0.20–0.24 and by mycelial extension or dispersal of 0.16–0.20. The probability of colonisation decreased with increasing distance from a point of establishment. R. bicolor responded to both enrichment and destructive disturbances by the formation of an extensive cord system which enabled it to colonise discontinuously distributed resources and to overgrow fungi adjacent to it in a single resource unit, including Heterobasidion annosum.     

Structural,Biochemical, and Computational Characterization of the Glycoside Hydrolase Family 7 Cellobiohydrolase of the Tree-killing Fungus Heterobasidion irregulare

Root rot fungi of the Heterobasidion annosum complex are the most damaging pathogens in temperate forests, and the recently sequenced Heterobasidion irregulare genome revealed over 280 carbohydrate-active enzymes. Here, H. irregulare was grown on biomass, and the most abundant protein in the culture filtrate was identified as the only family 7 glycoside hydrolase in the genome, which consists of a single catalytic domain, lacking a linker and carbohydrate-binding module. The enzyme, HirCel7A, was characterized biochemically to determine the optimal conditions for activity. HirCel7A was crystallized and the structure, refined at 1.7 Å resolution, confirms that HirCel7A is a cellobiohydrolase rather than an endoglucanase, with a cellulose-binding tunnel that is more closed than Phanerochaete chrysosporium Cel7D and more open than Hypocrea jecorina Cel7A, suggesting intermediate enzyme properties. Molecular simulations were conducted to ascertain differences in enzyme-ligand interactions, ligand solvation, and loop flexibility between the family 7 glycoside hydrolase cellobiohydrolases from H. irregulare, H. jecorina, and P. chrysosporium. The structural comparisons and simulations suggest significant differences in enzyme-ligand interactions at the tunnel entrance in the −7 to −4 binding sites and suggest that a tyrosine residue at the tunnel entrance of HirCel7A may serve as an additional ligand-binding site. Additionally, the loops over the active site in H. jecorina Cel7A are more closed than loops in the other two enzymes, which has implications for the degree of processivity, endo-initiation, and substrate dissociation. Overall, this study highlights molecular level features important to understanding this biologically and industrially important family of glycoside hydrolases.     

Root inoculation with a forest soil streptomycete leads to locally and systemically increased resistance against phytopathogens in Norway spruce

Soil streptomycetes are commonly antagonistic against plant pathogens. However, interactions involving increased defense responses in the host plant, leading to suppression of plant disease development, have not yet been detailed. Here, the mechanisms were studied of disease suppression by Streptomyces sp. GB 4-2 against Heterobasidion root and butt rot in Norway spruce (Picea abies) seedlings. GB 4-2 promoted mycelial growth of the phytopathogenic fungus, germination rate of fungal spores, extension of germ tubes and early colonization of outer cortical layers of the plant root. Reduced colonization of the inner cortical cell layers was accompanied by the induction of cell wall appositions, and increased xylem formation in the vascular cylinder emerged after bacterium-fungus coinoculation. Bacterial treatment led to decreased water content in roots and needles and increased photosynthetic yield (F(v)/F(m)) and peroxidase activities in needles. The infection of needles by Botrytis cinerea was reduced by bacterial pretreatment. Complex interactions of GB 4-2 with Norway spruce and Heterobasidion abietinum were discovered. The bacterium promoted the growth of the phytopathogenic fungus but induced plant defense responses. Host responses indicate that GB 4-2 induces both local and systemic defense responses in Norway spruce.     

单孢融合性实验法鉴定采自四川北部的小孔异担子菌(英文)

在四川北部九寨沟和黄龙保护区的云杉、松树和铁杉上发现异担子菌 ,并从 6号标本中分离到 73个单孢菌株。在每个标本中随机选取 2个菌株分别与欧洲的原始多年异担子菌、小孔异担子菌和冷杉异担子菌的单孢菌株进行融合性交配。试验表明 ,这 6号标本都是小孔异担子菌。四川的菌株与欧洲的原始多年异担子菌交配不融合 ,而与欧洲的小孔异担子菌完全融合 ,并在交配后的菌落中形成锁状联合 ,且在交配的菌落中不产生拮抗线。虽然四川的菌株与欧洲的冷杉异担子菌有较高的融合性 ,但这些交配大部分为单项交配 ,即只在四川一侧的菌落中产生锁状联合 ,而且在交配的菌落中多数产生拮抗线。研究样品全部采自天然林 ,小孔异担子菌在四川经营林分中的致病性还有待进一步调查。     

Callus cultures and bark from Norway spruce clones show similar cellular features and relative resistance to fungal pathogens

In field experiments, clones of Norway spruce [Picea abies (L.) Karst.] showed different degrees of resistance against pathogenic fungi inoculated into the bark that correlate with differences in polyphenolic parenchyma (PP) cells of the bark. Cells of spruce callus cultures, particularly towards the callus surface, resemble PP cells and this study looks at changes in callus cells during infection and the relative resistance of cultures from clones of low (weak) or high (strong) resistance to fungal infection. Callus cultures, initiated from trees with different resistance, were co-inoculated with Ceratocystis polonica (Siem.) C. Moreau and Heterobasidion annosum (Fr.) Bref. Callus cells from strong clones resemble PP cells of bark tissue from strong clones, having more polyphenolic bodies, while callus cells from weak clones are more similar to PP cells from those clones, which have less extensive phenolic bodies. Callus cultures from trees with weak resistance were more quickly overgrown by both species of pathogenic fungi than cultures from trees with strong resistance. Callus cells of infected cultures showed changes similar to activated PP cells of bark, including enhanced accumulation of polyphenolics. Phenolic bodies were more numerous and more extensive (larger and denser) in callus cells of strong versus weak clones under all conditions. Thus, callus cells may perform similar functions in defense as PP cells in the bark. Callus from trees of varying resistance seem to reflect the relative resistance of the trees from which they are derived, and this study indicates that some mechanisms of resistance can be studied using callus from trees of different resistance.     

Trikaryon formation and nuclear selection in pairings between heterokaryons and homokaryons of the root rot pathogen Heterobasidion parviporum

Pairings between heterokaryons and homokaryons of Agaricomycete fungi (he-ho pairings) can lead to either heterokaryotization of the homokaryon or displacement of the homokaryotic nucleus through migration of nuclei from the heterokaryon into the homokaryon. In species of Agaricomycetes with multinucleate cells (>2 nuclei per cell), he-ho pairings could result in the stable or transient formation of a hypha with three genetically different nuclei (trikaryons). In this study, he-ho pairings were conducted using the multinucleate Agaricomycete Heterobasidion parviporum to determine whether trikaryons can be formed in the laboratory and whether nuclear genotype affects migration and heterokaryon formation. Nuclei were tracked by genotyping the heterokaryotic mycelium using nucleus-specific microsatellite markers. The data indicated that certain nuclear combinations were favored, and that nuclei from some strains had a higher rate of migration. A high percentage of trikaryons (19 %) displaying three microsatellite alleles per locus were identified among subcultures of the he-ho pairings. Using hyphal tip and conidial isolation, we verified that nuclei of three different mating types can inhabit the same mycelium, and one of the trikaryotic strains was judged to be semi-stable over multiple sub-culturing steps, with some hyphal tips that retained three alleles and others that reduced to two alleles per locus. These results demonstrate that nuclear competition and selection are possible outcomes of heterokaryon-homokaryon interactions in H. parviporum and confirm that ratios of component nuclei in heterokaryons are not strictly 1:1. The high rate of trikaryon formation in this study suggests that fungi with multinucleate cells may have the potential for greater genetic diversity and recombination relative to dikaryotic fungi.     

针叶树病原菌异担子菌属的物种多样性及相关检疫建议

异担子菌属Heterobasidion种类是北半球针叶树最严重的森林病原菌,在世界范围内能侵染27种针叶树,对欧洲和北美洲的经营林已造成重大损失。基于传统形态学研究,认为异担子菌属有2个种,即多年异担子菌H. annosum和岛生异担子菌H. insulare,然而单孢交配实验研究证明2个种为复合种。分子系统发育分析研究表明：异担子菌属包括15个种,其中5个种为森林病原菌[冷杉异担子菌H. abietinum、多年异担子菌（狭义）H. annosum s. str.、变孔异担子菌H. irregulare、西方异担子菌H. occidentale和小孔异担子菌H. parviporum];10个种为腐生菌（淀粉孢异担子菌H. amyloideum、南洋杉异担子菌H. araucariae、阿曼德异担子菌H. armandii、南方异担子菌H. australe、岛生异担子菌H. insulare、林芝异担子菌H. linzhiense、东方异担子菌H. orientale、拟岛生异担子菌H. subinsulare、拟小孔异担子菌H. subparviporum和西藏异担子菌H. tibeticum）。多年异担子菌（狭义）H. annosum s. str.、小孔异担子菌H. parviporum和冷杉异担子菌H. abietinum分布于欧洲,分别是松属、云杉属和冷杉属林木的病原菌。变孔异担子菌H. irregulare和西方异担子菌H. occidentale分布于北美洲,前者侵染松属和柏属树木,后者侵染冷杉属、铁杉属、黄杉属和巨杉属树木。虽然广义的多年异担子菌H. annosum sensu lato曾在我国报道,但基于目前研究结果表明该种为拟小孔异担子菌H. subparviporum。目前世界上最具侵染力的5种病原菌即狭义的多年异担子菌H. annosum s. str.、小孔异担子菌H. parviporum、冷杉异担子菌H. abietinum、变孔异担子菌H. irregulare和西方异担子菌H. occidentale还未在我国发现,也未列为对外检疫对象,因此建议将其列为进境植物检疫性有害生物。RNA聚合酶II大亚基序列（RPB1）在异担子菌属种类鉴定中敏感性和特异性最高,研究证明该分子方法可应用于鉴别不同异担子菌种类的有效基因标记,在海关部门进行原木和木质产品的检疫中可运用该分子标记进行林木病原异担子菌的检测。