Unravelling Diaporthe species associated with Camellia

The genus Diaporthe (syn. Phomopsis) comprises important pathogens, endophytes or saprobes with diverse host associations and worldwide distribution. Phomopsis theae is the first and hitherto the only recorded Diaporthe species on Camellia in China. The aim of this study was to investigate the Diaporthe species associated with symptomatic and asymptomatic tissues of Camellia spp. from several provinces in China. Eighty-three strains were isolated in the present study. Based on the multi-locus (ITS, HIS, TEF1, TUB) phylogenetic analyses and phenotypic characters, four novel species (D. apiculata Y.H. Gao & L. Cai, D. compacta Y.H. Gao & L. Cai, D. oraccinii Y.H. Gao & L. Cai, D. penetriteum Y.H. Gao & L. Cai), and three known species (D. discoidispora, D. hongkongensis, D. ueckerae) were identified. Five strains were assigned to D. amygdali species complex and 17 strains to D. eres species complex respectively, but they could not be further identified to species level using current multi-locus phylogenetic analysis and morphological characters. Of the identified species, D. compacta and D. discoidispora are only known as endophytes. Diaporthe hongkongensis is the dominant species on Camellia, accounting for 53.3% of the frequency of occurrence. Diaporthe lithocarpus is synonymized with D. hongkongensis; D. miriciae is synonymized with D. ueckerae. Our study revealed a high diversity of undescribed Diaporthe species on Camellia.     

Antimycobacterial pimarane diterpenes from the Fungus Diaporthe sp

Two new pimarane diterpenes, diaportheins A (1) and B (2), were isolated from a culture broth of the fungus Diaporthe sp. BCC 6140. Diaporthein B (2) strongly inhibited the growth of Mycobacterium tuberculosis with the MIC value of 3.1 microg/mL, while diaporthein A (1) showed only mild activity (MIC value of 200 microg/mL).     

Reaction of Diaporthe longicolla to a strain of Sarocladium kiliense

Phomopsis seed decay of soybean [Glycine max (L.) Merr.] is a seedborne fungal disease caused by Diaporthe longicolla that causes yield losses and reduced seed quality. Biocontrol of this pathogen by a strain of Sarocladium kiliense isolated from a culture of D. longicolla was investigated in vitro and in vivo. A zone of inhibition formed between the two fungi in vitro, but was poorly sustained, and inhibition of conidial germination of D. longicolla by a culture filtrate of S. kiliense was not significant, though D. longicolla mycelial growth was inhibited by a 25% culture filtrate. Co-inoculation of both fungi failed to reduce seed rot or increase seed germination in greenhouse and growth chamber experiments, respectively. Co-inoculation of both fungi also failed to reduce pycnidial development in colonies of D. longicolla growing on leaf pieces, but soaking of the leaf pieces with S. kiliense conidia for one or three days prior to inoculation with D. longicolla eliminated pycnidial development completely. Although it was not possible to reproduce the parasitism of S. kiliense on D. longicolla under the experimental and environmental conditions used, the potential to use S. kiliense as a protectant biocontrol for soybean fungal pathogens should be further investigated.     

Alkaloids from the mangrove endophytic fungus Diaporthe phaseolorum SKS019

Six new alkaloids including four new chromeno[3,2-c]pyridines, diaporphasines A-D (1–4), and two new isoindolinones, meyeroguillines C and D (6–7), as well as three known compounds meyeroguilline A (5), 5-deoxybostrycoidin (8), and fusaristatin A (9), were isolated from an endophytic fungus Diaporthe phaseolorum SKS019. Their structures were determined by analysis of 1D and 2D NMR and mass spectroscopic data. Compounds 1–9 are alkaloid components reported for the first time from the Diaporthe sp., and diaporphasines A-D (1–4) are the third examples of alkaloids possessing the unique chromeno[3,2-c]pyridine nucleus. All isolated compounds 1–9 were evaluated for their cytotoxic activity in vitro using MDA-MB-435, HepG2, MCF10A, HCT116, and NCI-H460 human cell lines. Compound 8 exhibited cytotoxicity against MDA-MB-435 and NCI-H460 human cancer cell lines with IC₅₀ values of 5.32 and 6.57 μM, respectively, and compound 9 showed growth-inhibitory activity against MDA-MB-435 human cancer cell line with IC₅₀ value of 8.15 μM.     

进境美国大豆中值得关注的真菌Diaporthe novem

采用常规平板分离法,从一批进境的美国大豆样品中获得1株可疑的间座壳属菌株MDD57.经形态学观察发现,该菌株在PDA培养基上产生分生孢子器,且同时产生大量α型和β型分生孢子,未见有性阶段.经ITS和tef1α基因扩增、核酸序列比对分析,发现该菌株同GenBank中2株Diaporthe novem菌株的基因序列同源性达...     

猕猴桃间座壳菌Diaporthe actinidiae的绿色荧光蛋白基因标记及侵染过程

果实软腐病是猕猴桃贮藏期间最严重的真菌病害,猕猴桃间座壳Diaporthe actinidiae是该病检出率最高且致病力最强的病原菌.该病菌从花前期开始侵染,至果实贮藏期才表现症状,侵染至发病周期较长,可借助荧光标记对其侵染过程进行研究.本研究采用PEG介导原生质体转化的技术,运用携带GFP及潮霉素抗性基因的双元载体p...     

Cytotoxic and antimicrobial metabolites from marine lignicolous fungi, Diaporthe sp

A new compound (1), named diaporthelactone, together with two known compounds (2 and 3) were isolated from the culture of Diaporthe sp., a marine fungus growing in the submerged rotten leaves of Kandelia candel in the mangrove nature conservation areas of Fugong, Fujian Province of China. The new compound was elucidated to be 1,3-dihydro-4-methoxy-7-methyl-3-oxo-5-isobenzofuran-carboxyaldehyde (1), which showed cytotoxic activity against KB and Raji cell lines (IC50 6.25 and 5.51 microg mL(-1), respectively). Two known compounds, 7-methoxy-4,6-dimethyl-3H-isobenzofuran-1-one (2) and mycoepoxydiene (3), were also demonstrated to exhibit cytotoxic activities for the first time. All three compounds were assessed for antimicrobial activity.     

贵州梨芽枯间座壳菌属Diaporthe的种类

为明确引起贵州梨芽枯病的间座壳菌种类,本研究进行了病害调查、病芽采集和病菌分离及纯化,根据其菌落特征与ITS序列鉴定,共获得94个间座壳属Diaporthe菌株,从中选取40个代表菌株进行多基因(ITS、TEF、CAL、HIS和TUB)系统发育分析和形态学观察,结果显示它们分属于间座壳属的8个种,包括核桃间座壳菌D. biguttulata、榆树间座壳菌D. eres、椭圆间座壳菌D. ellipsoidea、梭状间座壳菌D. fusicola、香港间座壳菌D. hongkongensis、塞纳间座壳菌D. sennae、大豆间座壳菌D. sojae和蜜柑间座壳菌D. unshiuensis,其中D. ellipsoidea、D. biguttulata和D. sennae为侵染梨的首次报道。玉露香梨离体枝条接种结果显示,8种间座壳的代表菌株均可产生与田间相同的症状,分离比最高的D. eres致病力较强,表明它们均为梨芽枯病的病原菌,其中D. eres为病原优势种。本研究首次报道了引起梨芽枯的间座壳菌种类组成,为该病的发生规律及防控技术研究提供了理论依据。     

中国云南省间座壳属两个新种

我国间座壳属真菌种类繁多,但对其分类研究滞后。通过对云南省间座壳属标本采集,基于形态学和核糖体内部转录间隔区、β微管蛋白基因、翻译延伸因子基因、钙调蛋白基因和组蛋白基因的DNA序列数据比较进行物种鉴定,描述了分别采集自八角黄皮和可可患病叶片上的间座壳属2个新种——八角黄皮间座壳和可可间座壳,并讨论了其与间座壳属物种的形态和系统发育关系。     

Cultural Growth Patterns and Incompatibility Reactions in Diaporthe and Phomopsis Populations

Comparative studies were carried out with cultures isolated from Phomopsis condiomata and/or Diaporthe ascomata found on naturally-infected living stems or on overwintered stem fragments of Arctium lappa L., Helianthus annuus L., Glycine max (L.) Merr., and Xanthium italicum Moretti, growing in or nearby the same fields in Vojvodina. Results underscored the heterogeneity of the Phomopsis anamorphs in the D. arctii (Lasch) Trav, complex, Intra-specific vegetative incompatibility groups were distinguished among Phomopsis and/or Diaporthe populations from A. lappa. More than one phytopathogenic Phomopsis and/or Diaporthe species were identified on X. italicum. P. longicolla Hobbs was isolated for the first time in Yugoslavia from soybean stem debris. The holomorph Diaporthe (Phomopsis) helianthi Muntanola- Cvetkovic, Mihaljcevic et Petrov showed distinguishing characteristics with respect to other congeneric fungi.     

Diaporthe amygdali,a species complex or a complex species?

Delimitation of species boundaries within the fungal genus Diaporthe has been challenging, but the analyses of combined multilocus DNA sequences has become an important tool to infer phylogenetic relationships and to circumscribe species. However, analyses of congruence between individual gene genealogies and the application of the genealogical concordance principle have been somehow overlooked. We noted that a group of species including D. amygdali, D. garethjonesii, D. sterilis, D. kadsurae, D. ternstroemia, D. ovoicicola, D. fusicola, D. chongqingensis and D. mediterranea, commonly known as D. amygdali complex, occupy a monophyletic clade in Diaporthe phylogenies but the limits of all species within the complex are not entirely clear. To assess the boundaries of species within this complex we employed the Genealogical Concordance Phylogenetic Species Recognition principle (GCPSR) and coalescence-based models: General Mixed Yule-Coalescent (GMYC) and Poisson Tree Processes (PTP). The incongruence detected between individual gene phylogenies, as well as the results of coalescent methods do not support the recognition of lineages within the complex as distinct species. Moreover, results support the absence of reproductive isolation and barriers to gene flow in this complex, thus providing further evidence that the D. amygdali species complex constitutes a single species. This study highlights the relevance of the application of the GCPSR principle, showing that concatenation analysis of multilocus DNA sequences, although being a powerful tool, might lead to an erroneous definition of species limits. Additionally, it further shows that coalescent methods are useful tools to assist in a more robust delimitation of species boundaries in the genus Diaporthe.     

Possible phytotoxic metabolites in culture filtrates of Diaporthe phaseolorum var. sojae

Filter-sterile culture filtrates of an isolate of Diaporthe phaseolorum var. sojae (Phomopsis sojae), causal agent of pod and stem blight and seed decay of soybeans (Glycine max), grown on Czapeks-dox broth significantly (P = 0.05) inhibited germination of cabbage (Brassica oleracea), cantaloupe (Cucumis melo), onion (Allium cepa), soybean, and wheat (Triticum vulgare) seeds and wilted soybean seedling cuttings within 24 hr. Inhibition of seed germination and severity of soybean seedling wilt increased with increased concentrations of culture filtrates and increased incubation time of the cultures.     

3-Hydroxypropionic Acid as an Antibacterial Agent from Endophytic Fungi Diaporthe phaseolorum

Endophytic fungi are considered a rich source of active compounds resulting from their secondary metabolism. Fungi from marine environment grow in a habitat with unique conditions that can contribute to the activation of metabolic pathways of synthesis of different unknown molecules. The production of these compounds may support the adaptation and survival of the fungi in the marine ecosystem. Mangroves are ecosystems situated between land and sea. They are frequently found in tropical and subtropical areas and enclose approximately 18.1 million hectares of the planet. The great biodiversity found in these ecosystems shows the importance of researching them, including studies regarding new compounds derived from the endophytic fungi that inhabit these ecosystems. 3-hydroxypropionic acid (3-HPA) has been isolated from the mangrove endophytic fungus Diaporthe phaseolorum, which was obtained from branches of Laguncularia racemosa. The structure of this compound was elucidated by spectroscopic methods, mainly 1D and 2D NMR. In bioassays, 3-HPA showed antimicrobial activities against both Staphylococcus aureus and Salmonella typhi. The structure of this antibiotic was modified by the chemical reaction of Fischer-Speier esterification to evaluate the biologic activity of its chemical analog. The esterified product, 3-hydroxypropanoic ethyl ester, did not exhibit antibiotic activity, suggesting that the free carboxylic acid group is important to the pharmacological activity. The antibiotic-producing strain was identified with internal transcribed spacer sequence data. To the best of our knowledge, this is the first report of antibacterial activity by 3-HPA against the growth of medically important pathogens.     

Antioxidant compounds extracted from Diaporthe schini using supercritical CO2 plus cosolvent

Bioprocess and Biosystems Engineering - Endophytic fungi have been highlight in the production of secondary metabolites with different bioactive properties, such as in the production of the...     

海洋真菌Diaporthe sp.产真菌环氧二烯发酵条件的优化

从海洋木栖真菌HLY-2分离得到的真菌环氧二烯(Mycoepoxydiene)是新发现的高抗肿瘤活性化合物。通过正交设计,改良普通半海水PD培养基的配方,比较固体和液体两种发酵方法,得到了使该化合物产量大幅提高的最佳培养条件:在固体发酵条件下土豆250g/L,海水300mL/L,葡萄糖30g/L,乳糖50g/L,磷酸二氢钾0·65mmol/L,硫酸铵1g/L,产量高达543mg/L,比普通的PD培养基提高了43倍,也比液体发酵的最高产量提高了近15倍。同时还对固体发酵和液体发酵造成的产量差异进行了分析。并检验了发酵产物的抗肿瘤活性。     

Endophytic Diaporthe from Southeast China are genetically diverse based on multi-locus phylogeny analyses

Species of Diaporthe (anamorph Phomopsis) comprise a diverse and widely distributed group of phytopathogens, saprophytes and endophytes. However, the degree of genetic diversity of endophytic Diaporthe has not yet been fully investigated. In this paper, a survey of endophytes from 28 plants in southeast China yielded 116 Diaporthe isolates, out of which 64 haplotypes were determined using DnaSP ver. 5.1 based on alignment result of internal transcribed spacer of ribosomal nucleotide sequences (ITS rDNA). Many haplotypes turned out to be quite different from known species and displayed high diversity. Among them, 14 strains from 5 discriminating terminal clades were selected to go through further analysis according to partial sequence of translation elongation factor 1-α (tef1-α) and again they got separated from others. The following multi-gene phylogenetic analysis based on ITS rDNA, tef1-α, β tubulin and calmodulin grouped eight most discrepant strains into three distinctive clusters, cluster 1 (Rc001, Eu004 and Eu009), cluster 2 (ZJWCF252, Sjm001 and Ac001) and cluster 3 (Pcs013 and Sfp005) respectively with high support values. These clusters above represent three potentially novel species. This research provides strong evidence of high biodiversity and novelty of Diaporthe endophytes from southeast China, which is thus important not only for better resolving the taxonomy in this genus, but also for further utilization due to their multiple application.