Mycoflora and aflatoxin production in pigeon pea stored in jute sacks and iron bins

The mycoflora, moisture content and aflatoxin contamination of pigeon pea (Cajanus cajan (L.) Millisp) stored in jute sacks and iron bins were determined at monthly intervals for a year. The predominant fungi on freshly harvested seeds wereAlternaria spp.,Botryodiplodia theobromae, Fusarium spp. andPhoma spp. These fungi gradually disappeared from stored seeds with time and by 5–6 months, most were not isolated. The fungi that succeeded the initially dominant ones were mainly members of the generaAspergillus, Penicillium andRhizopus. Population of these fungi increased up to the end of one year storage. Higher incidence of mycoflora andAspergillus flavus were recorded in jute-sack samples throughout the storage period. The moisture content of stored seeds was found to fluctuate with the prevailing weather condition, being low during the dry season and slightly high during the wet season. The stored seeds were free of aflatoxins for 3 and 5 months in jute sacks and iron bins respectively. The level of aflatoxins detected in jute-sack storage system was considerably higher than that occurring in the iron bin system. Of 196 isolates ofA. flavus screened, 48% were toxigenic in liquid culture (54% from jute sacks and 41% from iron bins).     

Fungal bioremediation of polyethylene: Challenges and perspectives

Plastics have become ubiquitous in both their adoption as materials and as environmental contaminants. Widespread pollution of these versatile, man-made and largely petroleum-derived polymers has resulted from their long-term mass production, inappropriate disposal and inadequate end of life management. Polyethylene (PE) is at the forefront of this problem, accounting for one-third of plastic demand in Europe in part due to its extensive use in packaging. Current recycling and incineration processes do not represent sustainable solutions to tackle plastic waste, especially once it becomes littered, and the development of new waste-management and remediation technologies are needed. Mycoremediation (fungal-based biodegradation) of PE has been the topic of several studies over the last two decades. The utility of these studies is limited by an inconclusive definition of biodegradation and a lack of knowledge regarding the biological systems responsible. This review highlights relevant features of fungi as potential bioremediation agents, before discussing the evidence for fungal biodegradation of both high- and low-density PE. An up-to-date perspective on mycoremediation as a future solution to PE waste is provided.     

Deteriorative changes of maize,groundnut and soybean seeds by fungi in storage

This paper deals with investigations on fungal infection, moisture content, germinability and deterioration of three seeds, viz., maize (starchy), groundnut (oily) and soybean (proteinaceous) in storage at the locality of Santiniketan, West Bengal, India, under natural condition for 1 year. The airspora of storage environment was trapped using culture plate method. Different species of Aspergillus (A. candidus, A. flavus, A. niger, A. terreus, and A. ruber) were dominant followed by Rhizopus, Penicillium, Curvularia, Fusarium, Alternaria, etc. Seed moisture was maximum in the rainy season followed by a gradual decrease during longer storage. A gradual decrease in field fungi with simultaneous increase in storage fungi accompanied by a reduction in germinability occurred in all seeds as storage proceeded. A gradual loss of carbohydrate (both soluble and insoluble) content in all the seeds were recorded. A loss of protein content was recorded followed by a small increase. Oil content decreased in prolonged storage with simultaneous increase in fatty acid. This revised version was published online in June 2006 with corrections to the Cover Date.     

Prevalence and control of wheat (Triticum aestivum L.) seed borne fungi in farmer-saved seeds

High incidence of diseases associated with the use of seeds saved from previous harvests as desire of maintaining local varieties with special attributes is of increased concern in wheat industry worldwide. Prevalent of seed-borne fungi in farmer-saved seeds and seed dressing fungicides to prevent infection from seeds to seedlings was studied in Northern Tanzania. One hundred and thirty five untreated farmer-saved seed lots were collected randomly from farmers. Alternaria alternata, Bipolaris sorokiniana, Drechslera tritici, Fusarium graminearum, Fusarium moniliforme, Aspergillus flavus, Cladosporium sphaerospermum, Epicoccum purpurascens, Pyricularia oryzae and Penicillium corylophilum were fungi isolated in farmer-saved seeds. Mean seed infection was 29% causing average grain yield loss of 1.2 mt/ha^?1. Seed dressing with Metalaxy plus (Methyl carboxaitide), Mancozeb (Manganase-zinc salt) and Baytan (Chlorophenoxy ethanol) increased seed germination by 14, 13 and 17%, respectively, and grain yield by 28, 20 and 18%, respectively. Farmer-saved seeds were heavily infected by fungi with low grain yield performance.     

Effects of 1-methylcyclopropene alone and in combination with polyethylene bags on the postharvest life of mango fruit

Experiments were conducted to determine how 1‐methylcyclopropene (1‐MCP) treatments influence ethylene‐stimulated ripening of harvested mango cv. Zihua fruit at 20°C. The ripening response of fungicide (prochloraz) treated fruit was characterised following various 1‐MCP treatments in sealed jars followed by storage in polyethylene bags and/or subsequent ethephon (ethylene) exposure. Exposure of fruit to increasing concentrations of 1‐MCP for 12 h resulted in the reduced softening of produce when subsequently held in air for 7 days after ethephon treatment. Application levels of between 1 and 100 μl litre^?1 1‐MCP had increasing impact, while 200 μl litre^?1 1‐MCP apparently began to approach response saturation. Exposure of fruit to 50 or 100 μl litre^?1 concentrations of 1‐MCP for periods from 1 to 24 h subsequently resulted in reduced softening of produce when held in air for 7 days after ethephon treatment. Increasing periods of exposure from 1 to 12 h had increasing impact, while exposure times greater that 12 h appeared to reach saturation. In the absence of ethephon‐stimulation, the natural ripening of mangoes held in polyethylene bags was delayed by prior exposure to 100 μl litre^?1 1‐MCP for 12 h. Extended holding of 1‐MCP treated and non‐1‐MCP treated control fruit in polyethyene bags encouraged physiological and pathological deterioration. Following exposure to 100 μl litre^?1 1‐MCP for 12 h, mango fruit held for 10 days in polyethylene bags showed a delay in the onset of ripening relative to bagged but non‐1‐MCP treated control fruit. Treatment with 1‐MCP allowed storage of mango fruit in plastic bags at 20°C for 30 days. Observations suggest that 1‐MCP treatments do not adversely influence the quality of the post‐storage ethephon‐ripened fruit. Thus, application of 1‐MCP in combination with the use of polyethylene bags can extend the postharvest life of mango fruit at ambient temperature. Treatments that extend postharvest life are important in developing countries, such as China, where the cold chain infrastructure is often lacking.     

瓜实蝇的有效积温及其在中国的年发生代数预测

瓜实蝇Bactrocera cucurbitae Coquillett是世界上重要的检疫性害虫,通过在14、18、22、26、30、34℃温度下孵化和饲养实验,测定该虫卵、幼虫、蛹、产卵前期、世代的发育起点温度和有效积温分别是8．0982℃、4．5245℃、4．5245℃、7．4751℃、10．0019℃和28．8506、162．7462、145．7249、166．5126、503．8343日度。根据该虫的致死温度和有效积温模型,运用数学分析和地理信息系统技术,预测了瓜实蝇在中国的适生区和年发生代数,结果表明：该虫可在中国48．96％地区发生（气象站点所代表的地区）,1年发生2-12代,以4～6代为主。     

Spatial and temporal nature of reactive oxygen species production and programmed cell death in elm (Ulmus pumila L.) seeds during controlled deterioration

Seed deterioration is poorly understood and remains an active area for research. Seeds of elm (Ulmus pumila L.) were aged at 37 °C above water [controlled deterioration treatment (CDT)] for various lengths of time to assess programmed cell death (PCD) and reactive oxygen species (ROS) product in embryonic tissues during a 5 d period. The hallmarks of PCD were identified in the elm seeds during CDT including TUNEL experiments, DNA laddering, cytochrome c (cyt c) leakage and enzymatic activities. These analyses indicated that PCD occurred systematically and progressively in deteriorated elm seeds. Cyt c release and increase in caspase‐3‐like/DEVDase activity occurred during CDT, which could be suppressed by ascorbic acid (AsA) and caspase‐3 inhibitor Ac‐DEVD‐CHO, respectively. In situ localization of ROS production indicated that the distinct spatial‐temporal signature of ROS during CDT coincided with the changes in PCD hallmark features. Multiple antioxidant elements were activated during the first few days of CDT, but were subsequently depleted as PCD progressed. Taken together, our findings identify PCD as a key mechanism that occurs asymmetrically during elm seeds CDT and suggest an important role for PCD in seeds deterioration.     

Fungal infections of the phytoplankton: seasonality, minimal host density, and specificity in a mesotrophic lake

Phytoplankton infections by fungal parasites in the upper, mixed layer of a mesotrophic northern temperate lake were analysed according to the following parameters: host and parasite species, host population density and prevalence of infection, resting spore formation by the parasite, and the lowest host density at which parasites appeared. The phytoplankton taxa recorded included the Cyanobacteria, Dinomastigota, Chrysophyceae, Bacillariophyceae, Chlorophyceae, Cryptophyceae and Haptophyceae, but infection was never found in the last two classes. The parasites belonged almost exclusively to the monocentric Chytridiomycetes. Fungal epidemics occurred at all times of the year. Parasites appeared at population densities as low as about 1 cell ml⁻¹ in some host species, with infection prevalence sometimes exceeding 80%. The proportion of the total phytoplankton biovolume infected by fungi was usually much <1%, but occasionally reached 10%. Parasitism proved to be highly species-specific, with one parasite species usually infecting only one host species. In the case of Zygorhizidium planktonicum , which infected both Asterionella formosa and Synedra acus , there is evidence that two species-specific formae speciales , each infecting only one of these two host species, are present in the lake.     

Community dynamics of evergreen broadleaf forests in southwestern Japan. II. Species composition and density of seeds buried in the soil of a climax evergreen oak forest

The species composition and density of seeds in the soil of a climax evergeen oak forest in the Kasugayama Forest Reserve, Nara, Japan were studied by directly extracting seeds from soil samples and using soil samples in planting boxes as the basis of germination tests. A total of 33 species were identified from all seeds collected in 6 stands: 11 evergreen broadleaf, 15 deciduous broadleaf, 2 coniferous, 2 liana, 1 herb, and 2 grass species. Species producing sap fruits and dry fruits accounted for 60% and 40% of the total number of species, respectively. The species composition of all the seeds was independent of the species composition of the forest vegetation. The mean density of the seeds was 22,134 seeds/m²·5 cm.Eurya japonica, of which seeds were found in all soil samples, was the most abundant species, followed byCryptomeria japonica, Ilex micrococca, andBoehmeria longispica. Pioneer species such asMallotus japonicus, Zanthoxylum ailanthoides, andAralia elata were found in all soil samples in spite of the paucity of adult trees in the forest. Seeds of evergreen oaks were relatively aboundant but no viable seeds were found. ViableE. japonica, I. micrococca, Symplocos prunifolia, andB. longispica seeds were abundant.     

Fungal metabolism of polycyclic aromatic hydrocarbons: past, present and future applications in bioremediation

This article examines the importance of non-ligninolytic and ligninolytic fungi in the bioremediation of polycyclic aromatic hydrocarbon contaminated wastes. The research from the initial studies in Dave Gibson’s laboratory to the present are discussed. Received 10 August 1997/ Accepted in revised form 15 August 1997     

Proteinase activities in resting and germinating seeds of Scots pine, Pinus sylvestris

Resting seeds of Scots pine contained a moderate amount of acid proteinase activity, about 90% of which was inhibited by pepstatin A and about 10% by p-hydroxymer-curibenzoate. In gel chromatography on Sephacryl S-200 the proteinase activity showed a complex elution pattern with poorly separated peaks at positions corresponding to mol. wts. 100,000 and 30,000 and several shoulders. The results suggested that pine proteinases I and II, which are the main proteinases in the endosperms of germinating seeds (Salmia 1981: Physiol. Plant. 51: 253–258), were not present in the resting seeds.—Seedling extracts showed a low level of acid proteinase activity, which separated into several peaks in chromatography on Sephacryl S-200. As none of the peaks had the catalytic properties of proteinase I or II, it seems that these endospermal enzymes are also lacking in the seedling tissues.—In the endosperms of germinating seeds the activity of the pepstatin-sensitive acid proteinase(s) remained at a constant level throughout the period of reserve protein mobilization (lasting up to the stage when the length of dark-grown seedlings was 60 mm). Proteinases I and II were absent from resting seeds, showed a small increase up to the 20-mm stage, and then increased rapidly up to the 60-mm stage.—Resting embryos contained relatively higher acid proteinase activity than resting endosperms, and again about 90% of it was inhibited by pepstatin A and about 10% by p-hy-droxymercuribenzoate. During germination the former activity decreased, the latter activity remained at approximately the same level, and the activity of the other acid proteinases increased continuously with the growth of the seedling.—It is concluded that the pepstatin-sensitive proteinase(s), which is not affected by endogenous proteinase inhibitors, plays a central role in the initiation of reserve protein mobilization in both the embryo and the endosperm. Proteinases I and II, on the other hand, seem to account for the greater part of reserve protein breakdown in the main protein storage tissue, the endosperm.     

Spectrophotometric measurement of carotenes, xanthophylls, and chlorophylls in extracts from plant seeds

The method of spectrophotometric measurement of carotenoid and chlorophyll content in extracts from plant seeds was modified. The pigments were extracted with a mixture of petroleum ether (PE) and tetrahydrofuran (THF) (PE to THF ratio 4: 1). Equations for calculations of β-carotene, lutein, chlorophyll a and b content in PE: THF mixture were obtained using specific absorption coefficients of solutions of particular pigments. It was shown that subtraction of chlorophyll contribution from the absorption spectrum of seed extracts yields spectra, which could be used in some cases for calculation of carotene and xanthophyll contents.     

Fungal occurrence,disease incidence and severity,and yield of maize symptomatic for seedling disease in Mississippi

A study was conducted in Mississippi from 1995 to 1997 comparing soil rhizosphere fungi isolated from Pioneer 3167 hybrid maize (Zea mays L.) planted on Brooksville silty clay and Memphis silt loam soils. Maize seedlings were collected over four sampling dates from conventional and no-tillage plots. Eleven fungal genera consisting of nineteen species were isolated from these plants; Trichoderma spp. were most frequently isolated, followed by Fusarium spp. The highest disease incidence occurred in tilled plots of the latest planting date on Brooksville silty clay when samples were collected 17 days after planting. Root disease was most severe in 1996 from seedlings planted on the last planting date in tilled plots sampled 17 days after planting. Yields were significantly (P ≤ 0.05) higher on Brooksville silty clay soil than on Memphis silt loam in both 1995 and 1996. Yields were highest from no-tillage plots and from maize planted on the earliest date. There was a significant correlation between incidence of root infection and disease severity. There was no correlation between the incidence of root infection and yield or between disease severity and yield at either location. This revised version was published online in June 2006 with corrections to the Cover Date.     

Deterioration of western redcedar (Thuja plicata Donn ex D. Don) seeds: protein oxidation and in vivo NMR monitoring of storage oils

Deterioration of conifer seeds during prolonged storage hasa negative impact on reforestation and gene conservation efforts.Western redcedar (Thuja plicata Donn ex D. Don) is a speciesof tremendous value to the forest industry. The seeds of thisspecies are particularly prone to viability losses during long-termstorage. Reliable tools to assess losses in seed viability duringstorage and their underlying causes, as well as the developmentof methods to prevent storage-related deterioration of seedsare needed by the forest industry. In this work, various imagingmethods and biochemical analyses were applied to study deteriorationof western redcedar seeds. Seedlots that exhibited poor germinationperformance, i.e. those that had experienced the greatest lossesof viability during prolonged storage, exhibited greater abundanceof oxidized proteins, detected by protein oxidation assays,and more pronounced changes in their in vivo ¹³C NMR spectra,most likely due to storage oil oxidation. The proportion ofoxidized proteins also increased when seeds were subjected toaccelerated ageing treatments. Detection of oxidized oils andproteins may constitute a reliable and useful tool for the forestindustry. Key words: Conifer seeds, in vivo NMR spectroscopy, MRI, oil peroxidation, protein carbonylation, seed deterioration, seed storage, storage lipids, western redcedar Received 23 May 2007; Revised 28 November 2007 Accepted 17 December 2007     

Fungal communities in mycorrhizal roots of conifer seedlings in forest nurseries under different cultivation systems, assessed by morphotyping, direct sequencing and mycelial isolation

Fungi colonising root tips of Pinus sylvestris and Picea abies grown under four different seedling cultivation systems were assessed by morphotyping, direct sequencing and isolation methods. Roots were morphotyped using two approaches: (1) 10% of the whole root system from 30 seedlings of each species and (2) 20 randomly selected tips per plant from 300 seedlings of each species. The first approach yielded 15 morphotypes, the second yielded 27, including 18 new morphotypes. The overall community consisted of 33 morphotypes. The level of mycorrhizal colonisation of roots determined by each approach was about 50%. The cultivation system had a marked effect on the level of mycorrhizal colonisation. In pine, the highest level of colonisation (48%) was observed in bare-root systems, while in spruce, colonisation was highest in polyethylene rolls (71%). Direct internal transcribed spacer ribosomal DNA sequencing and isolation detected a total of 93 fungal taxa, including 27 mycorrhizal. A total of 71 (76.3%) fungi were identified at least to a genus level. The overlap between the two methods was low. Only 13 (13.9%) of taxa were both sequenced and isolated, 47 (50.5%) were detected exclusively by sequencing and 33 (35.5%) exclusively by isolation. All isolated mycorrhizal fungi were also detected by direct sequencing. Characteristic mycorrhizas were Phialophora finlandia, Amphinema byssoides, Rhizopogon rubescens, Suillus luteus and Thelephora terrestris. There was a moderate similarity in mycorrhizal communities between pine and spruce and among different cultivation systems.Electronic supplementary material Supplementary material is available for this article at and accessible for authorised users     

Fungal Community Analyses of Endophytic Fungi from Two Oak Species,Quercus mongolica and Quercus serrata,in Korea

Fungal endophytes have been recorded in various plant species with a richness of diversity, and their presence plays an essential role in host plant protection against biotic and abiotic stresses. This study applied the Illumina MiSeq sequencing platform based on the amplification of fungal ribosomal ITS2 region to analyze fungal endophytic communities of two oak species (Quercus mongolica and Q. serrata) with different oak wilt disease susceptibilities in Korea. The results showed a total of 230,768 sequencing reads were obtained and clustered at a 97% similarity threshold into 709 operational taxonomic units (OTUs). The OTUs of Q. serrata were higher than that of Q. mongolica with the number of 617 OTUs and 512 OTUs, respectively. Shannon index also showed that Q. serrata had a significantly higher level of fungal diversity than Q. mongolica. Total of OTUs were assigned into 5 fungal phyla, 17 classes, 60 orders, 133 families, 195 genera, and 280 species. Ascomycota was the dominant phylum with 75.11% relative abundance, followed by Basidiomycota with 5.28%. Leptosillia, Aureobasidium and Acanthostigma were the most abundant genera detected in Q. serrata with the average relative abundance of 2.85, 2.76, and 2.19%, respectively. On the other hand, Peltaster, Cladosporium and Monochaetia were the most common genera detected in Q. mongolica with the average relative abundance of 4.83, 3.03, and 2.87%, respectively. Our results indicated that fungal endophytic communities were significantly different between two oak species and these differences could influence responses of host trees to oak wilt disease caused by Raffaelea quercus-mongolicae.