Three previously undescribed metabolites from Cordyceps cicadae JXCH-1,an entomopathogenic fungus

Three previously undescribed compounds,cordycicadione(1),cordycicadin F(2),and 7-hydroxybassiatin(3),were isolated from the cultures of Cordyceps cicadae JXCH1,an entomopathogenic fungus.Their structures and relative configurations were elucidated primarily by NMR spectroscopic analysis.The absolute configurations of 1 and 2 were determined by ECD calculations.Single-crystal X-ray diffraction method was adopted to determine the absolute configuration of 3.Compound 2 is a polycyclic polyketide with an unusual enol ether moiety and a spiro ring.The compounds obtained in this study were subjected to screening their inhibition against the proliferation of the human lung cancer cell line A549 and the production of nitric oxide in murine macrophages RAW264.7.

Functional genomics in the rice blast fungus to unravel the fungal pathogenicity

A rapidly growing number of successful genome sequencing projects in plant pathogenic fungi greatly increase the demands for tools and methodologies to study fungal pathogenicity at genomic scale. Magnaporthe oryzae is an economically important plant pathogenic fungus whose genome is fully sequenced. Recently we have reported the development and application of functional genomics platform technologies in M. oryzae. This model approach would have many practical ramifications in design and implementation of upcoming functional genomics studies of filamentous fungi aimed at understanding fungal pathogenicity.

Studies on the Isolation, Identification and In Vitro Growth Rates of the Three Pathogenic Fungi from Panax notoginseng Cultivated in Wenshan Eparchy

Objective] The aim of this study was to simultaneously isolate and identify the main pathogenic fungi of the root rot, black spot and round spot from the Panax notoginseng plants cultivated in Wenshan Eparchy of Yunnan Province of China. [Method] The pathogenic fungi were isolated and purified by using potato dextrose agar （PDA） medium. The morphological identification was accomplished first according to the colony forms of the fungi when cultivated in vitro, then accord-ing to the symptom characteristics and colony forms of the re-isolated fungi in the reverse inoculation experiments. The molecular identification was performed accord-ing to the amplification and alignment of the internal transcribed space （ITS） se-quences of the fungi. The increases of the diameters and thickness of the colonies of the fungi cultivated in vitro were employed to indicate the growth rates of the fungi. [Results] The consistency of the colony forms and symptom characteristics and the 96%-99% similarities revealed in the ITS sequence alignments al proved that the main pathogenic fungi of the root rot, black spot and round spot of the P. notoginseng plants raised in Wenshan were Cylindrocarpon didymium, Alternaria panax and Mycocentrospora acerina, respectively. When cultivated in vitro in the same temperature, humidity and il umination, the increases of the colony diameters and thickness of C. didymium were the highest, fol owed by those of A. panax, then those of M. acerina. During different cultivation periods, the differences of the colony diameters and thickness of the three fungi al reached extremely significant level. However, at the same cultivation time, the differences of the diameters and thickness among the three fungi only reached significant level. [Conclusion] The main pathogenic fungi which result in the root rot, black spot and round spot of the P. notoginseng in Wenshan are C. didymium, A. panax and M. acerina, respec-tively. When these three diseases break out at the same time, the root rot wil spread fastest, fol owed orderly by the black spot and the round spot.

Isolation and Identification of Pandora neoaphidis and Its Pathogenicity against Turnip Aphid(Lipaphis erysimi)

[Objective] The study was to isolate and identify the pathogen of Pandora neoaphidis and to determine its pathogenicity against turnip aphid （Lipaphis erysimi）.[Method] Based on the morphological characteristics,the species of pathogenic fungus was identified.Spore shower method was used to carry out bioassay on the pathogen against turnip aphid.[Result] Primary conidia were ovoid,bitunicate and uninucleate,（24.7±1.4）μm×（10.7±0.9）μm,L/D=2.3±0.2.Secondary conidia had the similar shape with the primary ones,（18.6±2.1）μm×（13.3±1.3）μm,L/D=1.4±0.2.Hyphal body was like mycelium with the diameter of （10.6±0.8）μm.Conidiophores had palmate branch with the diameter of （10.0±0.9）μm.Pseudocystidia was not branched,which had rough base with the diameter of （19.2±1.7）μm,and gradually became more angular towards the apex with the diameter of （8.0±0.9）μm at tips.Rhizoid was like monohyphal shape with the diameter of （21.0±3.0）μm at base,the terminal apex had regular discoid holdfast.No resting spores were observed.The lethal dose of the pathogen against turnip aphid was 18.2/mm2.[Conclusion] The entomopathogenic fungus against turnip aphid was identified to be Pandora neoaphidis,and the pathogen was confirmed to have strong pathogenicity against turnip aphid.

Understanding the lifestyles and pathogenicity mechanisms of obligate biotrophic fungi in wheat:The emerging genomics era

Obligate biotrophic fungi cause serious and widespread diseases of crop plants, but are challenging to investigate because they cannot be cultured in vitro. The two economically important groups of biotrophic fungi parasitizing wheat are the rust and powdery mildew pathogens, but their obligate biotrophic lifestyles and pathogenicity mechanisms are not well understood at the molecular level. With the advent of next generation sequencing technology, increasing numbers of pathogen genomes are becoming available. Research in plant pathology has entered a new genomics era. This review summarizes recent progress in understanding the biology and pathogenesis of biotrophic fungal pathogens attacking wheat based on pathogen genomics. We particularly focus on the three wheat rust and the powdery mildew fungi in regard to genome sequencing, avirulence gene cloning, effector discovery, and pathogenomics. We predict that coordinated study of both wheat and its pathogens should reveal new insights in biotrophic adaptation, pathogenicity mechanisms,and population dynamics of these fungi that will assist in development of new strategies for breeding wheat varieties with durable resistance.

Analysis of the Abnormal Segregation of Pathogenicity in Magnaporthe grisea by Using a Genetic Cross of Oryza and Eleusine Isolates

A genetic cross between Oryza isolate Y93-164a-1 and Eleusine isolate SA98-4 was established, and the pathogenicity of 151 F1 progeny isolates was investigated on both host plants rice and finger millet. Results showed that the segregation of pathogenicity in this genetic cross was abnormal, i.e., most of the progeny isolates were nonpathogenic on both host plants. However, no abnormal segregation was observed when middle repetitive sequence MGR586 and 31 single-copy RFLP markers from all of the chromosomes were genetically analyzed. At the same time, comparison of the chromosomal organization among two pairs of parental isolates did not find any genomic abnormity. These results suggested that the ＂abnormal＂ inheritance of pathogenicity in this cross was most likely due to the reassortment of numerous host species specificity genes but not the biased segregation of the host species specificity genes. The host species specificities in M. grisea were likely to be multigenically controlled, at least in the genetic cross involving rice pathogen and the grasses pathogen other than rice.

A LAMP-assay-based specific microbiota analysis reveals community dynamics and potential interactions of 13 major soybean root pathogens

Soybean root diseases are associated with numerous fungal and oomycete pathogens;however,the community dynamics and interactions of these pathogens are largely unknown.We performed 13 loop-mediated isothermal amplification(LAMP)assays that targeted specific soybean root pathogens,and traditional isolation assays.A total of 159 samples were collected from three locations in the Huang-Huai-Hai region of China at three soybean growth stages(30,60,and 90 days after planting)in 2016.In LAMP results,we found that pathogen communities differed slightly among locations,but changed dramatically between soybean growth stages.Phytophthora sojae,Rhizoctonia solani,and Fusarium oxysporum were most frequently detected at the early stage,whereas Phomopsis longicolla,Fusarium equiseti,and Fusarium virguliforme were most common in the later stages.Most samples(86%)contained two to six pathogen species.Interestingly,the less detectable species tended to exist in the samples containing more detected species,and some pathogens preferentially co-occurred in diseased tissue,including P.sojae–R.solani–F.oxysporum and F.virguliforme–Calonectria ilicicola,implying potential interactions during infection.The LAMP detection results were confirmed by traditional isolation methods.The isolated strains exhibited different virulence to soybean,further implying a beneficial interaction among some pathogens.

Brocaeloid D,a novel compound isolated from a wheat pathogenic fungus, Microdochium majus 99049

Microbes serve as the most important resource for drug discovery.During our screening for bioactive compounds from our natural products library,a pathogenic fungus,Microdochium majus strain 99049,from wheat was selected for further investigation.A new alkaloid named brocaeloid D(1),together with six previously characterized compounds(2–7)were identified.Compound 1 belongs to 4-oxoquinoline with C-2 reversed prenylation and a succinimide substructure.All the structures of these newly isolated compounds were determined by different means in spectroscopic experiments.The absolute configurations of 1 was further deduced from comparison of its CD spectrum with that of known compound 2.The bioactivities of these identified compounds were evaluated against several pathogenic microorganisms and cancer cell lines.Compounds 1–5 showed activity against HUH-7 human hepatoma cells with IC50 values of 80μg/mL.Compound 6 showed mild activity against HeLa cells(IC50=51.9μg/mL),weak anti-MTB activity(MIC=80μg/mL),and moderate anti-MRSA activity(MIC=25μg/mL),and compound 7 showed weak anti-MRSA activity(MIC=100μg/mL).

Two novel aliphatic unsaturated alcohols isolated from a pathogenic fungus Fusarium proliferatum

Phytopathogenic fungi have attracted great attention as a promising source for new drug discovery.In the progress of our ongoing study for bioactive natural products from an in-house phytopathogenic fungi library,a pathogenic fungus,Fusarium proliferatum strain 13294(FP13294),was selected for chemical investigation.Two novel aliphatic unsaturated alcohols named fusariumnols A and B(1 and 2),together with one previously characterized sesquiterpenoid lignoren(3)were identified.Structures of 1-3 were assigned by mass spectrometry and NMR spectroscopy.Their bioactivities were assessed against Staphylococcus epidermidis,S.aureus,and Methicillin-resistant S.aureus(MRSA).Compounds 1 and 2 exhibited weak antibacterial activity against S.epidermidis(MIC=100μM).

A Resistance Bioassay for Rhizoctonia Root and Crown Rot and Damping-off Caused by the Anastomosis Groups AG 2-2111B and AG 4 in Sugar Beet

In order to determine the level of resistance of sugar beet varieties against Rhizoctonia solani AG 2-21IIB and AG 4, a methodology was implemented under greenhouse conditions that contemplated the most important criteria regarding to plant-pathogen interaction. The effect of plant growth stage on the development of the disease was evaluated. Seven sugar beet varieties were tested for resistance to R. solani AG 2-2IIIB and AG 4. To detect differences in leaf temperature between/L solani inoculated plants and non-infected plants, an infrared （IR） camera was tested. High incidence of R. solani AG 2-2IIIB and AG 4 in sugar beet plants was evident when the fungal inoculum was applied to two and four weeks old plants. At four weeks after sowing, it was the optimum time to inoculate sugar beet plants in order to generate R. solani infection, since at this time all plants were infected. Significant differences were detected regarding disease incidence between sugar beet varieties inoculated with different anastomosis groups. Leaf temperature was significant different between inoculated and non-inoculated plants, demonstrated that this technique could be a new tool for breeders to screen for resistance of new varieties.

Stem Histopathology of Sesame Seedlings Infected with Alternaria alternata

In the present study, histopathology of three varieties of sesame TS 3, TS 5 and SG 27 infected with Alternaria alternata was carried out to understand the mechanism of fungal infection and penetration in sesame plant as well as to determine the histological manifestation in sesame cells by light microscopy. Fungus was identified in infected tissues as a dark bluish black with toluidine blue O staining. Light microscopic examination of sesame stem showed that the fungus was present in epidermis, hypodermis and cortical parenchyma tissue as the symptoms became visible by naked eye ten days after inoculation (DAI). As the disease progress, the fungus moved from cortical parenchyma to vascular bundle, xylem and phloem. Later on, it completely overlapped the vascular bundle and entered in pith. When necrotic lesion appeared, fungus was present abundantly in epidermis, hypodermis, cortical parenchyma, vascular bundles and in pith. Due to its excessive growth and complete overlapping of cells, disorganization or destruction of cells of sesame took place. It was concluded that the Alternaria alternata was not a tissue limited pathogen instead of this it spread in to all tissues of stem from epidermis to pith.

Use of Waste Vegetable Biomasses Treated by Steam Explosion for the Horticultural Crop Protection

The purpose of this study was to assess the suppressive effect of Waste Vegetable Biomasses （WVBs） treated by the Steam Explosion technique in a continuous plant, against soil-borne plant pathogens. In order to assess their disease suppression, five WVBs （Miscanthus biomass, durum wheat straw, rice straw, corn stalk and wood shaving） and commercial compost were tested in vivo at three different doses （10, 20 and 30% of potting mix） on seven horticultural pathosystems plant/fungus： tomato/Phytophthora nicotianae, cucumber/Pythium ultimum, lettuce/Fusariurn oxysporum f. sp. lactucae, melordFusariurn oxysporum f. sp. melonis, bearffRhizoctonia solani, eggplant/Verticillium dahlie and fennel/Sclerotinia sclerotiorum. The results showed that the corn stalk was more efficient respect to Miscanthus, compost, wheat straw, rice straw and wood shaving in all the patbosystems and at all the doses tested. The corn stalk suppression ranged from 97% in eggplant/F, dahliae to 35% in lettuce/F, oxysporum f. sp. lactucae, and it was significantly higher with respect to the other substrates. In general, the wheat straw, rice straw and wood shaving were statistically found less efficient as suppressive substrate with respect to corn stalk, Miscanthus and compost at the 30% dose in four pathosystems In particular, the wood shaving suppressiveness ranged from 48% in eggplant/V, dahliae to 12% in lettuce/F, oxysporum f. sp. lactucae. The different suppressiveness observed could be attributed to different concentration of the microbial inhibitory substances （furfurals, organic acids and lignosulfonates） produced during the processing of fresh biomass.

安徽药用牡丹炭疽病病原菌鉴定

牡丹(Paeonia suffruticosa Andr.)是芍药科植物,为多年生落叶灌木,具有很高的观赏和药用价值。因其风姿华丽、色彩艳丽、品种繁多、深受大家喜爱,是中国传统名花和世界著名花卉之一。其根皮也被称为丹皮,具有抗炎、抑制血小板、止痛、清热等作用,是六味地黄丸等著名中成药的主要配方,也是饮片中的常用品[1-2]。近年来,随着牡丹种植面积的不断扩大,造成牡丹病害的大面积发生,已给牡丹生产带来严重影响。

Biochar for Sustainable Soil Health:A Review of Prospects and Concerns

Biochar as a soil amendment is confronted with the challenge that it must benefit soil health as it can be by no means separated from soils once it is added. The available literature even though sparse and mostly based on short-term studies has been encouraging and the trend obtained so far has raised many hopes. Biochar has been reported to positively impact an array of soil processes ranging from benefiting soil biology, controlling soil-borne pathogens, enhancing nitrogen fixation, improving soil physical and chemical properties,decreasing nitrate(NO-3) leaching and nitrous oxide(N2O) emission to remediation of contaminated soils. However, very little biochar is still utilized as soil amendment mainly because these benefits are yet to be quantified, and also the mechanisms by which the soil health is improved are poorly understood. Due to the infancy of research regarding this subject, there are still more questions than answers. The future research efforts must focus on carrying out long-term experiments and uncover the mechanisms underlying these processes so that key concerns surrounding the use of biochar are addressed before its large scale application is recommended.

黑老虎枝枯病病原鉴定及其生物学特性

黑老虎[Kadsura coccinea(Lem.)A.C.Smith]又名冷饭团、布福娜,果实含丰富的Vc、Ve及多种微量元素,是山区野果珍品[1]。上世纪末,人们从野外引种试栽,目前在黔东南地区实现了规模化栽培。笔者于2017和2018年调查发现,黑老虎枝枯病在黔东南州各种植基地普遍发生,造成严重损失,但关于黑老虎病害的报道甚少[2,3]。

Mycorrhizal Symbioses of Cotton Grown on Sodic Soils:A Review from an Australian Perspective

The majority of terrestrial plants form some type of mycorrhizal symbiosis.This established symbiosis therefore exists in most commercially important crops, which includes cotton.Arbuscular mycorrhizal fungus(AMF) can colonise 50%–90% of cotton root length under field and controlled conditions.Mycorrhization improves growth and nutrient uptake(especially phosphorus) of cotton,particularly at the early growth stages.Mycorrhizal symbioses help plants to counter the stresses imposed by physical and chemical soil constraints; however, adverse environmental conditions may restrict the mycorrhizal associations and consequently may reduce nutrient uptake and impair plant growth.In Australia, cotton is mainly grown on sodic soils that contain more than 6% of the total cations as exchangeable sodium.High levels of sodium in the soil create adverse physical and chemical soil conditions that may negatively affect mycorrhizal symbioses of cotton.This review discusses the cotton mycorrhizal colonisation, plant growth, and disease protection effects, potential negative effects of physical and chemical properties of sodic soils, and influences of some agronomic management practices.In addition, the research gaps were identified and some practical applications of the research outcomes were suggested.

Biofumigation effects of brassicaceous cover crops on soil health in cucurbit agroecosystems in Hawaii, USA

Brassicaceous cover crops, such as brown mustard (Brassica juncea) and oil radish (Raphanus sativus), are commonly used for biofumigation, a process that utilizes isothiocyanates (ITCs) generated from the hydrolysis of glucosinolates in Brassica plants to suppress soil-borne pathogens, including plant-parasitic nematodes. Given the biocidal nature of ITCs, limited information is available on the non-target effects of biofumigation on free-living nematodes, which are reliable soil health indicators. The objectives of this study were to determine if biofumigation methods effective against plant-parasitic nematodes would have non-target effects on free-living nematodes, and to examine the relationships between biofumigation indicators and nematode communities. Three field trials were conducted to examine whether different biofumigation methods would affect free-living nematodes. Tissue maceration of biofumigant crops, soil tillage, and black plastic mulching were adopted singly or in combination to generate different regimes of biofumigation efficacy. Termination of biofumigant crops by tissue maceration and soil tillage followed by black plastic mulching for one week was most effective in suppressing plant-parasitic nematodes and enhancing bacterial decomposition. However, these effects did not last through the subsequent zucchini (Cucurbita pepo) crop cycle. When comparing changes in soil glucose and sulfate concentrations as indicators of biofumigation efficacy, we found that soil sulfate was a better indicator of biofumigation efficacy than soil glucose, owing to the more stable state of sulfate in soil. Canonical correspondence analysis between soil sulfate as a biofumigation indicator and nematode soil health indicators revealed strong positive correlations of sulfate level with the abundances of bacterivorous and carnivorous nematodes, enrichment index, brown mustard biomass, and soil temperature. However, biofumigation did not affect the nematode community structure. This study demonstrated that biofumigation can suppress plant-parasitic nematodes without compromising soil health.

New insights in the battle between wheat and Puccinia striiformis

Wheat stripe rust caused by Puccinia striiformis f.sp.tritici(Pst)poses a great threat to wheat production worldwide.The rapid change in virulence of Pst leads to a loss of resistance in currently resistant wheat cultivars,which results in frequent disease epidemics.Therefore,a major focus is currently placed on investigating the molecular mechanisms underlying this rapid variation of pathogenicity and coevolving wheat resistance.Limited by the lack of a system for stable transformation of Pst and the difficulties in wheat transformation,it is not easy to generate deeper insights into the wheat-Pst interaction using established genetic methods.Nevertheless,considerable effort has been made to unravel the wheat-Pst interaction and significant progress is being made.Histology and cytology have revealed basic details of infection strategies and defense responses during wheat-Pst interactions,identified cellular components involved in wheat-Pst interactions,and have helped to elucidate their role in the infection process or in plant defense responses.Transcriptome and genome sequencing has revealed the molecular features and dynamics of the wheat-Pst pathosystem.Extensive molecular analyses have led to the identification of major components in the wheat resistance response and in Pst virulence.Studies of wheat-Pst interactions have now entered a new phase in which cellular and molecular approaches are being used.This review focuses on the cellular biology of wheat-Pst interactions and integrates the emerging data from molecular analyses with the histocytological observations.

Secretome profiling reveals temperature-dependent growth of Aspergillus fumigatus

Aspergillus fumigatus is a ubiquitous opportunistic fungus. In this study, systematic analyses were carried out to study the temperature adaptability of A. fumigatus. A total of 241 glycoside hydrolases and 69 proteases in the secretome revealed the strong capability of A. fumigatus to degrade plant biomass and protein substrates. In total, 129 pathogenesis-related proteins detected in the secretome were strongly correlated with glycoside hydrolases and proteases. The variety and abundance of proteins remained at temperatures of 34°C–45°C. The percentage of endo-1,4-xylanase increased when the temperature was lowered to 20°C, while the percentage of cellobiohydrolase increased as temperature was increased, suggesting that the strain obtains carbon mainly by degrading xylan and cellulose, and the main types of proteases in the secretome were aminopeptidases and carboxypeptidases. Only half of the proteins were retained and their abundance declined to 9.7% at 55°C. The activities of the remaining β-glycosidases and proteases were merely 35% and 24%, respectively, when the secretome was treated at 60°C for 2 h. Therefore, temperatures >60°C restrict the growth of A. fumigatus.

The genus Pythium in Taiwan,China(1)-a synoptic review

The genus Pythium,with slightly over 280 described species,has been classified traditionally with other filamentous,coenocytic,sporangia-producing fungi as“Phycomyetes”.However,with recent advances in chemical,ultrastructural and molecular studies,Pythium spp.are now considered as“fungus-like organisms”or“pseudo-fungi”and are placed in the Kingdom Chromista or Kingdom Straminopila,distinct from the true fungi of the Kingdom Fungi or Kingdom Mycota.They are widely distributed throughout the world as soil saprophytes or plant pathogens.Because of the warm moist maritime climate,Taiwan,China,is especially rich in Pythium species.To date,48 species of Pythium have been reported from Taiwan,China,with the dominant species being Py.vexans,Py.spinosum,Py.splendens,Py.aphanidermatum,Py.dissotocum and Py.acanthicum.There is no definite geographical distribution of Pythium spp.in Taiwan,China.Twenty nine species of Pythium have proven to be plant pathogens attacking a wide variety of woody and herbaceous plants primarily causing pre-and post-emergence seedling damping-off,root rot,stem rot and rotting of fruits,tubers and ginger rhizomes,resulting in serious economic losses.The most important plant pathogenic species include Py.aphanidermatum and Py.Myriotylum,which are most active during the hot and wet summer months;whereas Py.splendens,Py.spinosum,Py.ultimum and Py.irregulare cause the greatest damage in the cool winter.Most Pythium spp.are non-specific pathogens,infecting mainly juvenile or succulent tissues.This review attempts to assess the taxonomic position of the genus Pythium and provide details of the historical development of the study of Pythium as pathogens in Taiwan,China,causing diseases of sugarcane,trees,vegetables,fruits,specialty crops and flowering plants,as well as measures to control these diseases.Of special note is the introduction of the S-H mixture which,when used as soil amendment,effectively controls many soil-borne Pythium diseases during the early stages of plant growth.The diversity of Pythium species in Taiwan,China,is discussed in comparison with the situation in the mainland of China and suggestions are made to fully utilize Pythium spp.as agents for biological control,in industry and medicine.