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).     

Airborne Penicillium in the grain shops of Nagpur (India)

The airborne Penicillium spp. and total airborne fungal spore concentration was investigated in the grain shops of Nagpur city, India, using a volumetric Hi‐Air sampler system Mark II (Hi Media Laboratories Ltd., India). The mycotoxins were analysed from the Penicillium isolates obtained from the seeds by thin layer chromatography.

The mean concentration of the total fungi isolated from different grain shops ranged from 7.8×10² to 1.1×10³ CFU/m³. The mean concentration of Penicillium isolated from the air of grain shops ranged from 8.6×10¹ CFU/m³ (10.8%) to 1.7×10² CFU/m³ (19.9%). Among the 13 species of Penicillium which were isolated, P. citrinum Thom was the most prevalent species (24.2%), followed by P. oxalicum Currie & Thom (16.5), P. digitatum Saccardo (8.9%), P. janthinellum Biourge (8.7%), P. funiculosum Thom (8.3%), P. chrysogenum Thom (6.4%), P. purpurogenum Stoll (6.2%), P. brevicompactum Dierckx (4.8%), P. frequentans Westling (4.2%), P. italicum Wehmer (3.8%), P. rubrum Stoll (3.4%), P. expansum Link (2.9%) and P. cyclopium Westling (1.6%).

Penicillium species were also isolated from seeds such as wheat, maize, soybean, and groundnut. The mycotoxins roquefortin C, citrinin, rubratoxin B, cyclopiazonic acid, verrucosidin, mitorubrinic acid and two unknown metabolites were isolated from Penicillium isolates.     

Phosphorus sorption and extractability in Andic soil incubated with plant residues of variable P content

Sodium acetate extractable P and P sorption were measured in a high P-sorbing Andept soil (Mission series) following incubation with Winter wheat (Triticum aestivum (L.)Nugaines) and red clover (Trifolium pratense (L.)Florex) plant residue. The effects of plant residue P content, moisture regime, and quantity added were evaluated over a twelve week incubation period. Increased extractable P and decreased P sorption occured when the P content of residue exceeded 0.1% P for both winter wheat and red clover. The extractable P decreased and the P sorbed increased for all treatments and the check as the incubation time progressed. Extractable P increased and P sorption decreased more under air dry moisture conditions than at –0.03 MPa or saturation. Extractable P levels increased and P sorption decreased with increasing amounts of added residue. The effects of the residue additions decreased with increasing incubation time.     

Three-dimensional temporal and spatial distribution of adult Rhyzopertha dominica in stored wheat and corn under different temperatures, moisture contents, and adult densities

Three-dimensional temporal and spatial distributions of adult Rhyzopertha dominica (F.) at adult densities of 1.0, 5.0, and 10.0 adults per kg grain and at 20 +/- 1, 25 +/- 1, and 30 +/- 1 degrees C were determined in 1.5 t bins filled with wheat (Triticum aestivum L.) with 11.0 +/- 0.8, 13.0 +/- 0.6, and 15.0 +/- 0.5% moisture content (wet basis) or corn (Zea mays L.) with 13.0 +/- 0.2% moisture content (wet basis). At each of five sampled locations, grain was separated into three 15-kg vertical layers, and adult numbers in each layer were counted. Inside both corn and wheat, adults did not prefer any location in the same layer except at high introduced insect density in wheat. The adults were recovered from any layer of the corn and >12, 65, and 45% of adults were recovered in the bottom layer of the corn at 20, 25, and 30 degrees C; respectively. However, <1% of adults were recovered in the bottom layer of wheat. Numbers of adults correlated with those in adjacent locations in both vertical and horizontal directions, and the temporal continuous property existed in both wheat and corn. Adults had highly clumped distribution at any grain temperature and moisture content. This aggregation behavior decreased with the increase of adult density and redistribution speed. Grain type influenced their redistribution speed, and this resulted in the different redistribution patterns inside wheat and corn bulks. These characterized distribution patterns could be used to develop sampling plans and integrated pest management programs in stored grain bins.     

Mycological contamination of Ogiri (fermented sesame seeds) during production in Sierra Leone

Ogiri (fermented sesame seeds) was prepared in the laboratory and at home by selected participants. No fungal participation was observed during the normal fermentation process. However, fungal contamination was observed when improper handling techniques were employed. Out of the 49 samples examined, 41 fungal isolates were made. Aspergillus spp were the dominant group irrespective of the locality. Toxigenic Aspergillus included A. flavus Link ex Fries, A. tamarii Kita and A. ochraceus Wilhelm. P. citrinum Thom was the only toxigenic Penicillium isolated. The role of the different types of wraps used for the fermentation, marketing and storage of the product was also determined.     

Studies on the fungus flora of three grains in Egypt

In wheat, corn and sorghum grains twenty-six genera and seventy-seven species including sixteen ofAspergillus and twenty-one ofPenicillium were identified.In grain samples adjusted to various moisture levels (up to 18.8 % on a dry-weight basis) and stored for 4 months at 8, 20 and 30 C seed-borne fungi were monthly identified and counted, and the germinability of the grains was tested. In the control samples (moisture content 7.1–8.2 %) temperature had no effect on the fungal counts and on the precentage germination. With the rise of temperature and moisture content the fungal counts markedly increased and the germinability declined.The list of fungi dominant in one or more of the experiments, included eight species ofAspergillus, six ofPenicillium, two ofFusarium and one each ofAlternaria andCurvularia. The order of dominance of these fungi varied according to the type of grain, the length of storage and the treatment.A. niger was the dominant organism in the control samples of the three grains. It could grow successfully at high moisture contents but above 15 %, it was usually overgrown by other fungi. In wheatP. citrinum andA. sydowii, in cornP. citrinum andA. terreus, and in sorghumA. terreus andA. niger were the dominant fungi at moisture contents above 15 % after four months storage at 30 C. When stored at 8 CPenicillium species tended to predominate over other fungi in grains with high moisture contents.In surface-sterilized grains adjusted to 15 % moisture content, inoculated with six dominant fungi separately and incubated at 30 C,A. niger, P. citrinum andP. variabile caused severe deterioration in the three grains;A. ochraceus in wheat only andF. moniliforme in wheat and corn;C. spicifer was slightly injurious to the three grains.     

Germination-associated events and the desiccation sensitivity of recalcitrant seeds — a study on three unrelated species

The storage behaviour of recalcitrant seeds was assessed using three diverse species: a gymnosperm, Araucaria angustifolia (Bert.) O. Kuntze; a herbaceous monocotyledon, Scadoxus membranaceus (Bak.) Friis Nordal; and a woody dicotyledon, Landolphia kirkii Dyer. Seeds were stored under conditions of high relative humidities that maintained seed moisture content and under low relative humidities that caused drying. At regular intervals moisture content was determined, germinability assessed and the ultrastructure of radicle meristem cells examined. Under storage at high relative humidity, seed moisture content was maintained at the original level and subcellular germination events were initiated in the short-term. Such seeds showed enhanced rates of germination when removed from storage and planted. Long-term storage under these conditions resulted in the initiation of subcellular damage which intensified with time and ultimately resulted in the loss of viability. The rate at which germination events proceeded varied among the three species, and could be directly correlated with the period of viability retention under humid storage conditions. Storage under desiccating conditions resulted in subcellular damage and rapid loss of viability. The rate at which the seeds dried varied among the three species. The proportion of water loss tolerated by the different species before loss of viability, correlated with the rate of drying. The storage behaviour of the seeds of these three species is discussed in terms of a previously described model.     

Storage behavior of Zygophyllum xanthoxylon (Bge.) Maxim seeds at low moisture contents

This research aimed to determine whether low moisture content storage improves the longevity and storability of Zygophyllum xanthoxylon (Bge.) Maxim seeds. The seeds of Z. xanthoxylon were dried from 11.43 to 4.81%, 3.81 and 2.41% moisture content in a desiccating container with silica gel. After storage for 24 months their level of vigor was measured. After the ageing process, some physiology indices, including the relative electrical conductivity, SOD, PER, CAT, volatile aldehydes and malondialdehyde were tested. The results indicated that SOD, PER and CAT activities of the low moisture content seeds were higher than the control seeds while volatile aldehydes and malondialdehyde were lower than the control group. The electrical conductivity showed that there was obvious statistical difference between low moisture content seeds with the control group. All the results showed that low moisture content storage is beneficial for maintaining the vigor of Z. xanthoxylon seeds. Thus Z. xanthoxylon seeds can be stored with low moisture content at ambient temperature condition for a long time.     

Desiccation and storage studies on Dipterocarpm seeds

The relationship of seed moisture content (fresh weight basis) to germination, and the effect on viability of various storage conditions were examined for five species of the tropical forest tree genus Dipterocarpus. It was shown that seeds fall into two groups with regard to desiccation tolerance. Firstly, D. obtusifolius and D. turbinatus cannot be dried below about 45% moisture content without damage; a sigmoid curve was found to fit the relationship between germination and moisture content for the latter species. Secondly, D. intricatus, D. tuberculatus and D. alatus can be safely dried to 10%, 12% and 17% moisture contents respectively, but desiccation to near 7% moisture content reduced viability by at least a half. Storage studies showed that seed of D. intricatus and D. tuberculatus possessed increased longevity as moisture contents were reduced within the range 6–20%. It was concluded that seeds in the first group are ‘recalcitrant’ and that those in the second group are ‘orthodox’ in their storage physiology, according to the categories described by Roberts (1973). Wide differences between species in seed desiccation rates were observed. In 15% relative humidity D. intricatus dried to 7% moisture content within a week, whilst D. obtusifolius retained 30% moisture content even after 5 wk; other species had intermediate desiccation rates. Seed size and structure may partly account for the differences observed. Correlations were observed between seed storage physiology and other factors which were investigated. ‘Orthodox’ seeds had quicker desiccation rates, were derived from drier habitats, and had smaller embryos than those of ‘recalcitrant’ seeds. ‘Orthodox’ seeds, with the possible exception of D. alatus, should be kept at 0–3°C with about 12% moisture content in the short term and, provided less than 10% germination is lost on freezing, at-18°C with about 8% moisture content in the long term. ‘Recalcitrant’ seeds should be stored in ventilated containers at 21°C and with moisture contents above 45–50%.     

Effect of temperature and moisture content on Egyptian peanut seed-borne fungi

In peanut seeds adjusted to various levels of moisture content (8.5 %, 13,5 %, 17.5 % and 21 % on a dry-weight basis) and stored for six months at 5°, 15°, 28° and 45 °C, seed-borne fungi were monthly identified and counted using the dilution plate method, and the germinability of seeds was tested. The total count of seed-borne fungi (recovered at 28 °C) increased regularly and the germinability declined with the rise moisture content and with the lengthening of the storage period. At 5°, 15° and 28 °C,A. fumigatus was almost the most dominant fungal species followed by several fungi such asAspergillus flavus, A. niger, A. terreus, Penicillium funiculosum, P. notatum, Pyrenochaeta decipians andScopulariopsis brevicaulis. The degree of dominance of each fungal species depended upon the conditions of storage and the length of storage period.A. flavus gained its highest count in seeds adjusted to 13.5 % stored at 15 °C for 1 month.Penicillium predominated at 17.5 % and 21 %, and at 13.5 % and 17.5 % when the seeds were stored for long periods at 5° and 15 °C respectively.The total count of thermophilic fungi (recovered at 45 °C) significantly increased with the rise of moisture content and with the lengthening of the storage period between 1 and 2 months and regularly declined after 4 months.A. fumigatus was so extremely dominant that it was the main component of the total fungal flora. Several fungi truly thermophilic were isolated also. They were,Mucor pusillus, Humicola grisea var.thermoidae, H. insolens, H. lanuginosa (Thermomyces lanuginosus) andSporotrichum thermophile.     

The Effect of Glumes on Fungal Infection of Germinating Seed of Spelt (Triticum spelta L.) in Comparison to Wheat (Triticum aestivum L.)

The influence of fungi on seedling emergence of naked and hulled spelt (Triticum spelta L.) and winter wheat (Triticum aestivum L.) was investigated. Seeds were sown in flat trays and placed in a growth chamber under stress conditions (low temperature and water logging) for four weeks, followed by favourable growth conditions. At weekly intervals, 150 seeds were removed, surface sterilized, and investigated for fungi. Pythium aristosporum Vanterpool, a causal agent of damping-off, was found to be best adapted to the unfavourable conditions and to be a major cause of pre-emergence damping-off. The frequency of seed-borne fungi decreased during the stress period, whereas soil-borne fungi survived and could be isolated from seeds and glumes in the following period with more favourable growth conditions. Seedlings from hulled seeds of spelt emerged more frequently than from naked kernels of spelt and both emerged at a higher rate than from seeds of winter wheat. This is good evidence that glumes protect seeds against fungal colonization and therefore increase the fitness of spelt under unfavourable germinating conditions.     

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.     

Phosphorus benefits of different legume crops to subsequent wheat grown in different soils of Western Australia

Certain legume crops, including white lupin (Lupinus albus L.), mobilise soil-bound phosphorus (P) through root exudates. The changes in the rhizosphere enhance P availability to these crops, and possibly to subsequent crops growing in the same soil. We conducted a pot experiment to compare phosphorus acquisition of three legume species with that of wheat, and to determine whether the legume crops influence growth and P uptake of a subsequent wheat crop. Field pea (Pisum sativum L.), faba bean (Vicia faba L.), white lupin (Lupinus albus L.) and wheat (Triticum aestivum L.) were grown in three different soils to which we added no or 20 mg P kg^–1 soil (P0, P20). Growth, P content and rhizosphere carboxylates varied significantly amongst crops, soils and P levels. Total P content of the plants was increased with applied phosphorus. Phosphorus content of faba bean was 3.9 and 8.8 mg/pot, at P0 and P20, respectively, which was about double that of all other species at the respective P levels. Field pea and white lupin had large amounts of rhizosphere carboxylates, whereas wheat and faba bean had negligible amounts in all three soils at both P levels. Wheat grew better after legumes than after wheat in all three soils. The effect of the previous plant species was greater when these previous species had received P fertiliser. All the legumes increased plant biomass of subsequent wheat significantly over the unplanted pots in all the soils. Faba bean was unparalleled in promoting subsequent wheat growth on all fertilised soils. This experiment clearly demonstrated a residual benefit of the legume crops on the growth of the subsequent wheat crop due to enhanced P uptake. Faba bean appeared to be a suitable P-mobilising legume crop plant for use in rotations with wheat.     

Biological,physical and chemical changes in stored wheat

Six lots of wheat from six farmers' bins in Manitoba were adjusted to a range of 5 moisture contents, held at 6 temperatures and sampled at 6 times during storage. After sampling, seeds were surface sterilized with mercuric chloride and subsamples plated on Czapek's agar, on filter paper moistened with water, or on filter paper moistened with a 7.5% sodium chloride solution. The microflora on the seed was determined, germination counts were made, and the seed was examined by government inspectors for its condition and grade. A total of 1192 samples were examined. In addition, 180 samples were subjected to a fat acidity test. Deterioration for each combination of temperature and moisture, the fungi involved, and the consequent effect on condition, grade and fatty acid content were observed. The effect of high moisture content (>20%) and low temperatures (3–10 ° C) on infection of seed by Penicillium and the consequent effect on germination, condition, grade and fatty acid value are stressed. The interrelationship among fungi, and among fungi and temperature, moisture, storage time, fat acidity values and germination are indicated by correlation coefficient matrices.Deceased July 25, 1982.     

The content of N-acylphosphatidylethanolamines in the seeds of cultivated plants and grain products

The content of the minor class of phospholipids, N-acylphosphatidylethanolamines (NAPEs) in mature seeds of cultivated plants: kidney bean (Phaseolus vulgaris L.), soybean (Glycine max (L.) Merr.), soft spring wheat (Triticum aestivum L. emend. Fiori et Paol), barley (Hordeum vulgare L.), and oat (Avena sativa L.), and the products of technological processing of grain cultures (floor, bran) was studied. Reliable NAPE identification was performed by a comparison of their chromatographic mobility with the marker NAPE sample from wheat flour and synthetic phosphatidylmethanol, and also using specific reagents. Kidney bean cv. Shchedraya seeds contained the highest amount of NAPEs. In legumes, the content of NAPEs varied not only in different species but also in different cultivars of a single species. In cereals, the highest NAPE content was detected in the barley seeds and best quality wheat flour. The content of NAPEs in cereal seeds was less variable than in legumes. NAPE quantification showed that accumulation of this phospholipid class occurred in parallel with accumulation of total phospholipids in seeds. The relation between NAPE content and the processes of seed development and their lipid composition is discussed. The seeds and products best suitable for NAPE isolation are recommended.     

The impact of LED illumination on antioxidant properties of sprouted seeds

The objective of this study was to evaluate the effect of the light emitting diode (LED) spectra on the antioxidant properties of sprouted wheat (Triticum aestivum L.), radish (Raphanus sativus L.), and lentil (Lens esculenta Moenh.) seeds. Lighting experiments were performed under controlled conditions (PPFD - 100 μmol m⁻² s⁻¹; 12 h photoperiod; 27°C). The LED conditions used were: L1 - 638 nm; L2 - 455 nm, 638 nm, 669 nm, 731 nm (basal components); L3 - basal + 385 nm; L4 - basal + 510 nm and L5 - basal + 595 nm. Wheat and lentil sprouts were shown to accumulate less phenolic compounds and were more sensitive to light spectral differences when compared to radish sprouts. The antioxidant properties and contents of antioxidant compounds in seeds germinated in the dark were significantly lower than LED treated seeds. The higher content of total phenols and significant increase in alpha-tocopherol and vitamin C concentration resulted in altered DPPH free-radical scavenging capacity. Therefore we conclude that the LED spectra, based on basal components supplemented with green (510 nm) light can improve the antioxidant properties of sprouted seeds of lentil and wheat. The highest antioxidant properties of radish seeds were caused by radiation with supplemental amber (595 nm) light.     

Increased Resistance to Penicillium Seed Rot in Transgenic Wheat Over‐expressing Puroindolines

Puroindolines (PINs) are the main components of the wheat grain hardness locus (Ha) and have in vitro antimicrobial activity against bacteria and fungi. Here, we examined the effect of variation in PINA and/or PINB content upon Penicillium sp. seed fungal growth inhibition. The Penicillium sp. assays were germination assays performed after incubating seeds in Penicillium sp. contaminated soil. The first set of wheat genotypes consisted of two sets of transgenic isolines created in the varieties ‘Bobwhite’ and ‘Hi‐Line’ having over‐expression of PINA and/or PINB. The second set of genotypes consisted of near‐isogenic lines (NILs) varying for mutations in PINA or PINB created in the varieties ‘Explorer’ and ‘Hank’. After incubation in Penicillium sp.‐infected soil, transgenic wheat seeds over‐expressing PINA in both ‘Hi‐Line’ and ‘Bobwhite’ and both PINs in ‘Hi‐Line’ exhibited significantly reduced fungal infection and increased germination. No significant differences in Penicillium sp. infection or germination rates were observed in seeds of the NILs. The results indicate that puroindolines native role in seeds is to increase seed viability and that when over‐expressed as transgenes, the puroindolines are effective antifungal proteins.     

Rapid change of AM fungal community in a rain-fed wheat field with short-term plastic film mulching practice

Plastic film mulching (PFM) is a widely used agricultural practice in the temperate semi-arid Loess Plateau of China. However, how beneficial soil microbes, arbuscular mycorrhizal (AM) fungi in particular, respond to the PFM practice is not known. Here, a field experiment was performed to study the effects of a 3-month short-term PFM practice on AM fungi in plots planted with spring wheat (Triticum aestivum L. cv. Dingxi-2) in the Loess Plateau. AM colonization, spore density, wheat spike weight, and grain phosphorus (P) content were significantly increased in the PFM treatments, and these changes were mainly attributable to changes in soil properties such as available P and soil moisture. Alkaline phosphatase activity was significantly higher in PFM soils, but levels of AM fungal-related glomalin were similar between treatments. A total of nine AM fungal phylotypes were detected in root samples based on AM fungal SSU rDNA analyses, with six and five phylotypes in PFM and no-PFM plots, respectively. Although AM fungal phylotype richness was not statistically different between treatments, the community compositions were different, with four and three specific phylotypes in the PFM and no-PFM plots, respectively. A significant and rapid change in AM fungal, wheat, and soil variables following PFM suggested that the functioning of the AM symbiosis had been changed in the wheat field under PFM. Future studies are needed to investigate whether PFM applied over a longer term has a similar effect on the AM fungal community and their functioning in an agricultural ecosystem.     

Characterisation of Soybean and Wheat Seeds by Nuclear Magnetic Resonance Spectroscopy

The effects of equilibration under different air relative humidities (RH, 1 – 90 %) and temperatures (35 and 45 °C) on soybean (Glycine max) and wheat (Triticum aestivum) seeds were studied using different techniques. Seed moisture content, electrical conductivity (EC) of seed leachate and per cent seed germination were measured following standard procedures, and compared with nuclear magnetic resonance spin-spin relaxation time (T₂) measurements. Moisture contents of soybean and wheat seeds, following the reverse sigmoidal trend, were greater at 35 than at 45 °C at any particular RH. Changes in T₂ were related to the changes in germination percentage and leachate EC of both soybean and wheat seeds. Equilibrating soybean seeds at RH 11 % decreased germination percentage with corresponding decrease in T₂. On the contrary, EC of seed leachate increased. In wheat seeds equilibrated at 45 °C, T² was maximal at RH 5.5 %. T₂ declined in seeds equilibrated at high RH (> 80 %) together with low germination percentage.