Chlorosphaeropsis epidemica,spec. nova,eine neue Grünalge mit endophytischer Lebensweise

The new parasitic algaChlorosphaeropsis epidemica is a member of theChlorosarcinales and lives endophytically in the leaf-tissue ofPotamogeton species. It forms stacks of cells and appears in great numbers, especially in dying leaf-blades. Sexual reproduction is described for the first time in this genus. Apart from its endophytic way of life,Chl. epidemica is characterized by mononuclear cells with a parietal chloroplast of irregular thickness and with a few perforations, sometimes of complicated reticulate shape, and with 1–5 pyrenoids. The usually sphaerical gametes (respectively zoospores) are provided with two equal flagella, a parietal plastid, one pyrenoid and one stigma. In connection with the asexual reproduction the terms vegetative division and desmoschisis are critically discussed.

     

内生真菌和内生细菌接种对雷公藤生长和次生代谢产物积累的影响

为探究内生真菌与内生细菌对雷公藤(Tripterygium wilfordii)的生长和次生代谢产物积累的相互作用,用内生真菌NS33、NS6和内生细菌LG3、LY1单独或跨界联合接种雷公藤,对雷公藤的生长和雷公藤甲素、雷公藤红素合成进行了研究。结果表明,单独或混合培养的菌株具有分泌铁载体、吲哚乙酸(IAA)和溶磷能力,对种子萌发、芽伸长和根系活力有显著促进作用。接种菌株NS33、NS6、NS6-LG3和NS6-LY1均显著促进了雷公藤组培苗的生长。单独或联合接种菌株均能显著提高雷公藤组培苗雷公藤甲素和雷公藤红素的积累,其中NS33-LG3和LG3的作用最显著。菌株NS33与LG3能够协同促进IAA的分泌、小麦幼苗根系活力和雷公藤红素的积累;菌株NS6与LY1协同提高了雷公藤组培苗的高度、质量和雷公藤红素的积累。因此,内生真菌与内生细菌联合接种对雷公藤生长和次生代谢产物积累具有一定的协同效应,显示出实际应用潜力。     

Root-Associated Endophytic Bacterial Community Composition of Asparagus officinalis of Three Different Varieties

Asparagus (Asparagus officinalis L) is an economically important crop, rich in nutrients, and is also conducive to solving ecological and environmental problems. Plants may acquire benefits from root-associated endophytic bacteria. However, the composition of the endophytic bacterial community associated with the roots of asparagus is poorly elucidated. In this study, the nine root samples of asparagus from three different varieties including Asparagus officinalis var. Grande (GLD), A. officinalis var. Jinglvlu3 (JL3) and A. officinalis var. Jingzilu2 (JZL) were investigated by high-throughput sequencing technology of the 16S rDNA V5-V7 hypervariable region of endophytic bacteria. A total of 16 phyla, 29 classes, 90 orders, 171 families, and 312 genera were identified. Endophytic bacteria diversity and bacteria structure was different among the three varieties and was influenced by rhizosphere soil properties and varieties. In the GLD variety, the main phyla were Proteobacteria, Actinobacteria, and Firmicutes. The main phylum in JL3 and JZL varieties was Proteobacteria. The observations showed that GLD had the highest diversity of endophytes as indicated by the Shannon index (GLD > JZL > JL3). The order of the endophytes richness was GLD > JL3 > JZL. The PCA and PCoA analysis revealed the microbial communities were different between three different asparagus varieties, and the microbial composition of GLD and JZL was more similar. This report provides an important reference for the study of endophytic microorganisms of asparagus. Supplementary informationThe online version contains supplementary material available at (10.1007/s12088-021-00926-6) contains supplementary material, which is available to authorized users.     

产紫杉醇内生真菌枝状枝孢霉MD2的发酵条件优化

[目的]通过优化内生真菌枝状枝孢霉MD2的发酵条件,提高10-去乙酰巴卡亭Ⅲ (10-DAB)和紫杉醇(Taxol)的产量.[方法]采用单因素试验分析不同的培养基初始pH值、培养温度、摇床转速和培养时间对10-DAB和紫杉醇产量的影响,优化枝状枝孢霉MD2的培养条件;以YES为基本培养基,采用单因素试验和正交试验分析添加苯甲酸钠、苯丙氨酸、丝氨酸和甘氨酸4种前体物对10-DAB和紫杉醇产量的影响,优化枝状枝孢霉MD2的培养基组分.[结果]优化后发酵条件为:在初始pH为5.0的300 mL YES培养基中,添加15 mg/L苯甲酸钠、25 mg/L苯丙氨酸、5 mg/L丝氨酸、15 mg/L甘氨酸,接种1 mL枝状枝孢霉MD2的孢子悬液(107-10s个孢子/mL),28.0℃、220 r/min发酵培养12d.在此条件下,枝状枝孢霉MD2的生物量、10-DAB和紫杉醇的产量分别为15.5 g/L、471.5 μg/L和569.5 μg/L,与初始发酵条件相比,分别提高了1.3、3.6和3.4倍.[结论]首次获得了枝状枝孢霉MD2生产10-DAB和紫杉醇的较适摇瓶发酵条件,可为进一步放大发酵培养提供参考.     

对叶榕榕果内生细菌分离鉴定及抑菌活性的研究

【目的】为植物-内生细菌的生态关系研究, 以及植物内生细菌资源的利用提供一定的依据。【方法】采用微生物学传统分离培养的方法从对叶榕果实中分离到内生细菌54株, 通过限制性酶切分析(ARDRA)共有16个操作分类单元(Operational taxonomic units, OTUs), 对16株代表菌株16S rDNA序列系统发育分析。【结果】16个菌株都找到了与其相似性最高的菌株, 相似性达到95%?100%。其中6株为芽孢杆菌Bacillus属, 为对叶榕果实内生细菌优势菌属; 3株为Staphylococcus属, 2株为Pseudomonas属, 1株为Serrata属, 1株为非培养细菌的同源菌, 1株为Kocuria属, 1株为Delftia属, 还有1株为Acinetobacter属。【结论】这16株内生菌在系统发育树中明显聚为两大支; 在参与抑菌试验的14株内生菌中, 有13株对受试菌有不同程度的拮抗作用。尤其是其中的芽孢菌属的Swx15和不动杆菌属的Swx25菌株, 抑菌作用较强, 且有较广的抑菌谱性。     

A comparison of fungal endophytic community diversity in tree leaves of rural and urban temperate forests of Kanto district,eastern Japan

To clarify the effects of forest fragmentation and a change in tree species composition following urbanization on endophytic fungal communities, we isolated fungal endophytes from the foliage of nine tree species in suburban (Kashiwa City, Chiba) and rural (Mt. Wagakuni, Ibaraki; Mt. Takao, Tokyo) forests and compared the fungal communities between sites and host tree species. Host specificity was evaluated using the index of host specificity (Si), and the number of isolated species, total isolation frequency, and the diversity index were calculated. From just one to several host-specific species were recognized in all host tree species at all sites. The total isolation frequency of all fungal species on Quercus myrsinaefolia, Quercus serrata, and Chamaecyparis obtusa and the total isolation frequency of host-specific species on Q. myrsinaefolia, Q. serrata, and Eurya japonica were significantly lower in Kashiwa than in the rural forests. The similarity indices (nonmetric multidimensional scaling (NMS) and C_MH) of endophytic communities among different tree species were higher in Kashiwa, as many tree species shared the same fungal species in the suburban forest. Endophytic fungi with a broad host range were grouped into four clusters suggesting their preference for conifer/broadleaves and evergreen/deciduous trees. Forest fragmentation and isolation by urbanization have been shown to cause the decline of host-specific fungal species and a decrease in β diversity of endophytic communities, i.e., endophytic communities associated with tree leaves in suburban forests were found to be depauperate.     

Effects of simultaneous infections of endophytic fungi and arbuscular mycorrhizal fungi on the growth of their shared host grass Achnatherum sibiricum under varying N and P supply

Grasses can be infected by endophytic fungi and arbuscular mycorrhizal (AM) fungi simultaneously. Here, we investigated the interactions of a native grass, Achnatherum sibiricum, with both endophytic and AM fungi (Glomus mosseae, GM and Glomus etunicatum, GE) at different nitrogen (N) and phosphorus (P) levels. The results showed that endophyte infection significantly suppressed the colonization rates and spore density of GE, but had no effect on those of GM. Endophyte infection increased shoot biomass regardless of the nutrient conditions. The effects of AM fungi on host growth were dependent on mycorrhizal species. There was no significant interaction between endophytic fungi and GE on host growth; however, a significant interaction between endophytic fungi and GE occurred in total phenolic concentrations and P uptake. As for GM, a significant interaction among endophytic fungi, AM fungi and nutrient availability occurred in shoot growth. Under sufficient N and P conditions, endophyte infection alleviated the detrimental effects of GM colonization on host growth.     

Antifungal Monoterpene Derivatives from the Plant Endophytic Fungus Pestalotiopsis foedan

A new monoterpene lactone, (1R,4R,5R,8S)‐8‐hydroxy‐4,8‐dimethyl‐2‐oxabicyclo[3.3.1]nonan‐3‐one ( 1 ), along with one related known compound, (2R)‐2‐[(1R)‐4‐methylcyclohex‐3‐en‐1‐yl]propanoic acid ( 2 ), were isolated from the liquid culture of the plant endophytic fungus Pestalotiopsis foedan obtained from the branch of Bruguiera sexangula. The structure and absolute configuration of 1 were determined on the basis of extensive analysis of NMR spectra combined with computational methods via calculation of the optical rotation (OR) and ¹³C‐NMR. Both compounds exhibited strong antifungal activities against Botrytis cinerea and Phytophthora nicotianae with MIC values of 3.1 and 6.3 μg/ml, respectively, which are comparable to those of the known antifungal drug ketoconazole. Compound 2 also showed modest antifungal activity against Candida albicans with a MIC value of 50 μg/ml.     

Bioprospecting Chemical Diversity and Bioactivity in a Marine Derived Aspergillus terreus

A comparative metabolomic study of a marine derived fungus (Aspergillus terreus) grown under various culture conditions is presented. The fungus was grown in eleven different culture conditions using solid agar, broth cultures, or grain based media (OSMAC). Multivariate analysis of LC/MS data from the organic extracts revealed drastic differences in the metabolic profiles and guided our subsequent isolation efforts. The compound 7‐desmethylcitreoviridin was isolated and identified, and is fully described for the first time. In addition, 16 known fungal metabolites were also isolated and identified. All compounds were elucidated by detailed spectroscopic analysis and tested for antibacterial activities against five human pathogens and tested for cytotoxicity. This study demonstrates that LC/MS based multivariate analysis provides a simple yet powerful tool to analyze the metabolome of a single fungal strain grown under various conditions. This approach allows environmentally‐induced changes in metabolite expression to be rapidly visualized, and uses these differences to guide the discovery of new bioactive molecules.     

不同生境下五唇兰根部可培养内生细菌多样性研究

兰科植物内生细菌与菌根真菌的协作对宿主植物的生长、抗病、抗逆及植物修复环境能力等具有重要意义,揭示其内生细菌多样性及与生境之间的关系有助于阐明兰科植物的适应与进化机制。本研究基于16SrDNA序列分析探讨了不同生境下东南亚特有种五唇兰根部可培养内生细菌多样性及其空间异质性。结果表明:从不同生境下五唇兰根部共分离出内生细菌59株,其中从土生型五唇兰根部分离出内生细菌45株(76.27%),从石生型五唇兰根部分离出内生细菌14株(23.73%);基于内生细菌16SrDNA序列同源性分析及构建的系统发育树显示,五唇兰根部内生细菌分属于7属,即芽孢杆菌属(Bacillus)、伯克氏菌属(Burkholderia)、草酸菌属(Pandoraea)、土壤杆菌属(Agrobacterium)、类芽孢杆菌属(Paenibacillus)、泛菌属(Pantoea)、欧文氏菌属(Erwinia),其中优势属为芽孢杆菌属,次优势属为泛菌属和伯克氏菌属;多样性分析显示,土生型五唇兰根部内生细菌群落的Shannon多样性指数大于石生型五唇兰,不同生境下五唇兰根部内生细菌群落结构差异极显著(P0.01)。土生型五唇兰根部内生细菌群落优势属为芽孢杆菌属和泛菌属,石生型五唇兰根部内生细菌群落优势属为芽孢杆菌属和伯克氏菌属。