Aspergillus flavus growth and aflatoxin production as influenced by total lipid content during growth and development of cottonseed

Aspergillus flavus infects several food and feed crops, such as corn (Zea mays L.), cotton (Gossypium hirsutum L.), peanuts (Arachis hypogaea L.), and tree nut crops and contaminates the seed with carcinogenic aflatoxins. These susceptible crops contain rich reserves of lipids and fatty acids. The nature of relationship between lipids and the ability of the fungus to infect and produce aflatoxins in mature cottonseed, a protein-rich animal feed, has been addressed previously. In this study, we tracked lipid accumulation in developing cottonseed (15–35 days post-anthesis [DPA]) and also the ability of an aflatoxigenic strain and an isogenic non-aflatoxigenic strain to grow and produce aflatoxins in planta. The aflatoxigenic strain Af-70 green fluorescent protein (GFP) and the isogenic, non-aflatoxigenic strain SRRC 1500 (fungal collections maintained at the Southern Regional Research Center) did not differ much in infection and colonization of cottonseed. The non-aflatoxigenic strain did not produce aflatoxin at all stages of cottonseed development, whereas the aflatoxigenic strain Af-70 GFP produced copious amounts of aflatoxin and it coincided with the increasing levels of lipids, especially in mature cottonseed (30–35 DPA). Fungal growth, as quantified by the GFP expression in the fungus, was highly correlated with toxin production.     

Identification of a Maize Kernel Pathogenesis-Related Protein and Evidence for Its Involvement in Resistance to Aspergillus flavus Infection and Aflatoxin Production

ABSTRACT Aflatoxins are carcinogens produced by Aspergillus flavus and A. parasiticus during infection of susceptible crops such as maize. Several aflatoxin-resistant maize genotypes have been identified and kernel proteins have been suggested to play an important role in resistance. In the present study, one protein (#717), which was expressed fivefold higher in three resistant lines compared with three susceptible ones, was identified using proteomics. This protein was sequenced and identified as a pathogenesis-related protein (PR-10) based on its sequence homology. To assess the involvement of this PR-10 protein (ZmPR-10) in host resistance of maize against fungal infection and aflatoxin production, the corresponding cDNA (pr-10) was cloned. It encodes a protein of 160 amino acids with a predicted molecular mass of 16.9 kDa and an iso-electric point of 5.38. The expression of pr-10 during kernel development increased fivefold between 7 and 22 days after pollination, and was induced upon A. flavus infection in the resistant but not in the susceptible genotype. The ZmPR-10 overexpressed in Escherichia coli exhibited a ribonucleolytic and antifungal activities. Leaf extracts of transgenic tobacco plants expressing maize pr-10 also demonstrated RNase activity and inhibited the growth of A. flavus. This evidence suggests that ZmPR-10 plays a role in kernel resistance by inhibiting fungal growth of A. flavus.     

Use of different chemometric approaches for an estimation of soil properties at field scale with near infrared spectroscopy

Visible and near infrared spectroscopy (vis‐NIRS) may be useful for an estimation of soil properties in arable fields, but the quality of results are often variable depending on the applied chemometric approach. Partial least squares regression (PLSR) may be replaced by approaches which employ supervised learning methods or variable selection procedures in order to increase the proportion of informative wavelengths used in the estimation procedure, to reduce the noise of the spectra and to find the best fitting solution. Objectives were (1) to compare the usefulness of PLSR with either PLSR combined with a genetic algorithm (GA‐PLSR) or support vector machine regression (SVMR) for an estimation of soil organic carbon (SOC), total nitrogen (N), pH, cation exchange capacity (CEC) and soil texture for surface soils (0–5 cm, n = 144) of an arable field in Bangalore (India) and (2) to test and optimize different calibration strategies for GA‐PLSR for an improved estimation of soil properties. PLSR was useful for an estimation of SOC, N, sand and clay. In the cross‐validation (n = 96), accuracies of estimated soil properties generally decreased in the order GA‐PLSR > SVMR > PLSR. However, the order of estimation accuracies for the random validation sample (n = 48) changed to SVMR > GA‐PLSR > PLSR for SOC, N, pH, and CEC, whereas for clay the order changed to SVMR > PLSR > GA‐PLSR. A sequential procedure, which used the most frequently selected wavelengths of the GA‐PLSR runs, proved to be useful for an improved estimation of SOC and N. Overall, SVMR especially improved estimations of SOC and clay, whereas GA‐PLSR was particularly useful for SOC and N and it was the only approach which successfully estimated CEC in cross‐validation and validation.     

Influence of long-term fertilisation and crop rotation on changes in fungal and bacterial residues in a tropical rice-field soil

Amino sugars, as a microbial residue biomarker, are highly involved in microbial-mediated soil organic matter formation. However, accumulation of microbial biomass and responses of bacterial and fungal residues to the management practices are different and poorly characterized in rice soils. The objectives of this study were to evaluate the effects of mineral fertiliser (MIN), farmyard manure (FYM) and groundnut oil cake (GOC) on crop yield and co-accumulation of microbial residues and microbial biomass under rice-monoculture (RRR) and rice–legume–rice (RLR) systems. In the organic fertiliser treatments and RLR, rice grain yield and stocks of soil and microbial nutrients were significantly higher than those of the MIN treatment and RRR, respectively. The increased presence of saprotrophic fungi in the organic fertiliser treatments and RRR was indicated by significantly increased ergosterol/C_mic ratio and extractable sulphur. In both crop rotation systems, the long-term application of FYM and GOC led to increased bacterial residues as indicated by greater accumulation of muramic acid. In contrast, the higher fungal C/bacterial C ratio and lower ergosterol/C_mic ratio in the MIN treatment, is likely caused by a shift within the fungal community structure towards ergosterol-free arbuscular mycorrhizal fungi (AMF). The organic fertiliser treatments contributed 22 % more microbial residual C to soil organic C compared to the MIN treatment. Our results suggest that the negative relationship between the ratios ergosterol/C_mic and fungal C/bacterial C encourages studying responses of both saprotrophic fungi and AMF when assessing management effects on the soil microbial community.     

Vulnerability of low temperature induced needle retention in balsam fir (Abies balsamea L.) to vapor pressure deficits

Low temperature (LT) exposure has been shown to delay postharvest needle abscission and senescence (NAS) in balsam fir and changes in vapor pressure deficit (VPD) are expected to alter these processes. Two and half year-old seedlings were exposed to a LT of 5°C for 15 days while the control (CT) group was maintained at 22°C. Seedlings were then exposed to four different VPD (0.22, 0.87, 1.3 and 1.86?kPa) and observed for relative water content (RWC), xylem pressure potential (XPP), membrane injury index (MII), stem capacitance (SC) and NAS. An interactive effect of LT and VPD was observed in RWC, XPP, MII and NAS. Low VPD (0.22 and 0.87?kPa) resulted in less negative XPP, lower electrolyte leakage, higher SC and ultimately higher NAS than those at high VPD. Maximum NAS was recorded at 0.22?kPa. At 1.86?kPa, LT had 5× lower RWC (13%), 3× more negative XPP (?1.1?MPa), 1.8× higher membrane damage and 35% lower NAS (47 days) than CT. The SC declined with an increase in VPD with no effect of LT. The XPP and RWC of LT seedlings showed a positive relationship with NAS with R² values of 0.54 and 0.59, respectively. LT offered no benefits to NAS at high VPD environments.     

Antifungal and peroxidative activities of nonheme chloroperoxidase in relation to transgenic plant protection

Nonheme chloroperoxidase (CPO-P) of Pseudomonas pyrrocinia catalyzes the oxidation of alkyl acids to peracids by hydrogen peroxide. Alkyl peracids possess potent antifungal activity as found with peracetate: 50% killing (LD(50)) of Aspergillus flavus occurred at 25 microM compared to 3.0 mM for the hydrogen peroxide substrate. To evaluate whether CPO-P could protect plants from fungal infection, tobacco was transformed with a gene for CPO-P from P. pyrrocinia and assayed for antifungal activity. Leaf extracts from transformed plants inhibited growth of A. flavus by up to 100%, and levels of inhibition were quantitatively correlated to the amounts of CPO-P activity expressed in leaves. To clarify if the peroxidative activity of CPO-P could be the basis for the increased resistance, the antifungal activity of the purified enzyme was investigated. The LD(50) of hydrogen peroxide combined with CPO-P occurred at 2.0 mM against A. flavus. Because this value was too small to account for the enhanced antifungal activity of transgenic plants, the kinetics of the enzyme reaction was examined and it was found that the concentration of hydrogen peroxide needed for enzyme saturation (K(m) = 5.9 mM) was already lethal. Thus, the peroxidative activity of CPO-P is not the basis for antifungal activity or enhanced resistance in transgenic plants expressing the gene.     

Isoflavone composition, phenol content, and antioxidant activity of soybean seeds from India and Bulgaria

Isoflavone levels and isoflavone chemical composition in 11 cultivars of soybean, including 4 Indian and 7 genotypes of soybean grown in Bulgaria, were analyzed as determined by C 18 reversed phase high-performance liquid chromatography coupled with a photodiode array detector. Antioxidant activity of soybean extracts was assessed using the 2,2-diphenyl-1-picrylhydrazyl (DPPH) radical scavenging activity, and total phenolic compounds (TPC) were determined by using Folin-Ciocalteu reagent. The range of total isoflavones (TI) was 558.2-1048.6 microg g (-1) of soy in Indian cultivars, and it was 627.9-1716.9 microg g (-1) of soy in the case of Bulgarian cultivars. The highest and lowest total isoflavone contents were observed for Maus-2 (1048.6 microg g (-1) of soy) and Hardee (558.2 microg g (-1) of soy), respectively, for the Indian cultivars, and they were observed for Boryara (1716.9 microg g (-1) of soy) and Line 5 (627.9 microg g (-1) of soy) for the Bulgarian genotypes. DPPH radical scavenging activity did not differ significantly among the cultivars and did not correlate with TI, whereas TPC correlated well with TI and weakly with DPPH. Malonylglucoside of all the aglycones, total genistein (TGin), and total daidzein (TDin) showed strong correlation with total isoflavones, whereas acetylglucoside and aglycone levels did not significantly correlate with total isoflavone. Profiling of soybean isoflavone is helpful in understanding the regulation of isoflavone biosynthesis for greater improved resistance of crops to disease and greater health benefits for humans. This comparative study of soybean cultivars grown in India and Bulgaria throws light on their composition and nutraceutical value.