Time course study of substrate utilization by Aspergillus flavus in medium simulating corn (Zea mays) kernels.

Utilization of the three major corn reserve materials, starch, triglycerides (refined corn oil), and zein (storage protein), by Aspergillus flavus was monitored in vitro over a 7-day fermentation. Medium composition in which proportions of reserve materials initially approximated proportions in mature corn kernels changed little over the first 18 h. Subsequently, hydrolysis of both starch and triglycerides occurred simultaneously, with peak concentrations of glucose and free fatty acids on day 2 of the fermentation period. Fatty acid concentrations dropped relatively rapidly after day 2 but increased again after day 6. Aflatoxin B(1) production increased after 36 h, with a peak at day 4. Aflatoxin B(1) production paralleled fungal biomass production during the exponential growth phase. A. flavus did not appear to preferentially utilize any of the released fatty acids. A number of fungus-specific metabolites were detected, including arabitol, erythritol, mannitol, trehalose, and kojic acid. Mannitol exceeded the other metabolites in concentration, and the timing of mannitol production closely paralleled that of aflatoxin B(1). Kojic acid concentrations peaked at day 6. In contrast to previously described selective use of simple carbohydrates by A. flavus, less discrimination was displayed when faced with utilization of complex substrates such as starch or triglycerides.     

Isoform patterns of chitinase and beta-1,3-glucanase in maturing corn kernels (Zea mays L.) associated with Aspergillus flavus milk stage infection

Isoform patterns of chitinase and beta-1,3-glucanase of maturing kernels of yellow dent corn (Pioneer 3394) infected with Aspergillus flavus at the milk stage were investigated through polyacrylamide gel electrophoresis (PAGE). Proteins on the sodium dodecyl sulfate (SDS) gel with an apparent molecular mass range of 23-46 kDa were differentially present in the kernels infected with both aflatoxin-producing and non-aflatoxin-producing strains of A. flavus. From in-gel (native PAGE) enzyme activity assays, three bands corresponding to chitinase isoforms and two bands corresponding to beta-1,3-glucanase isoforms were detected in the infected kernels. One chitinase isoform of 29 kDa was present only in the infected kernels, and another one of 28 kDa was present in both infected and noninfected kernels. They were judged to be acidic on the basis of their migration on an acrylamide isoelectric focusing (IEF) gel. For the beta-1,3-glucanase, one isoform of 35 kDa was present in both infected and noninfected kernels, but another one, a 33 kDa isoform, was present only in the infected kernels. Both acidic and basic beta-1,3-glucanase isoforms were detected in the IEF gel. The results of this study are the first to demonstrate patterns of enhanced or inducible proteins in maturing corn kernels in response to A. flavus infection at the milk stage. The results also indicate that only particular isoforms of the two hydrolytic enzymes are involved in the maturing corn kernels infected at the milk stage with A. flavus.     

Dynamics of mycotoxin and Aspergillus flavus levels in aging Bt and non-Bt corn residues under Mississippi no-till conditions

Mycotoxin and Aspergillus flavus levels in soil-surface corn debris left by no-till agriculture methods (stover, cobs, and cobs with grain) were determined during the December-March fallow period for near-isogenic Bt and non-Bt hybrid corn. By December, average mycotoxin levels in non-Bt corn were many times higher in cobs with grain than in grain harvested in September (total aflatoxins, 774 vs 211 ng/g; total fumonisins, 216 vs 3.5 microg/g; cyclopiazonic acid, 4102 vs 72.2 microg/g; zearalenone, 0.2 vs < 0.1 microg/g). No trichothecenes were detected. Levels of mycotoxins and A. flavus propagules were approximately 10- to 50-fold lower in cobs without grain and stover, respectively, for all mycotoxins except zearalenone. Mycotoxin levels in corn debris fractions decreased during winter but began to rise in March. Levels of all mycotoxins and A. flavus propagules were lower in harvested grain and debris from Bt than non-Bt corn, but differences were significant (p < 0.05) only for aflatoxins.     

Inhibition of aflatoxin biosynthesis in Aspergillus flavus by diferuloylputrescine and p-coumaroylferuloylputrescine

A mixture of diferuloylputrescine/p-coumaroylferuloylputrescine (85:15, w/w) demonstrated inhibitory activity against aflatoxin B1 biosynthesis in Aspergillus flavus isolate AF13. Inhibition was concentration dependent, with a 50% effective dose of 30 microg of diferuloylputrescine/p-coumaroylferuloylputrescine per milliliter of medium. Aflatoxin inhibition levels of up to 93% were achieved using this conjugated polyamine material. This diconjugated polyamine mixture did not display inhibitory effects on A. flavus growth (mycelial weight) at any of the concentrations tested. A survey of hand-dissected corn (Zea mays) kernel tissues, including endosperm, germ, pericarp, and wax, revealed that the highest concentrations of these conjugated polyamines were localized in the pericarp of the seed. Analysis of a number of corn accessions did not reveal a correlation between diferuloylputrescine/ p-coumaroylferuloylputrescine concentration and resistance/susceptibility to A. flavus infection. The localization of these diconjugated polyamine components in the pericarp, which functions as a physical barrier and surrounds the internal food storage reserves, suggests a defensive role for these materials.     

Aflatoxin variation among corn samples with varying ratios of Aspergillus flavus-inoculated/noninoculated kernels.

Aflatoxin levels and physical properties of corn kernels inoculated with Aspergillus flavus during development and noninoculated kernels were compared in samples with various proportions of the 2 kernel types. The relationship between mean toxin levels and associated standard deviations of 5 samples demonstrated a linear association from the lowest toxin in noninoculated corn through a mixture of 60% inoculated/40% noninoculated. However, at the highest toxin level in the 100% inoculated material, a reduction in sample variation was observed. Examination of individual kernal weights showed that inoculated kernels were distinctly lighter than noninoculated seed. A uniform grinding procedure of the samples yielded heterogeneous particle sizes based on the starting corn. The large particle fraction (greater than 500 micrometers) decreased from 100% noninoculated kernels through the mixtures to the 100% inoculated seed; particles below 150 micrometers were most abundant in the ground samples from inoculated kernels. In addition, the density of particles within a size category varied; lower densities were observed in samples obtained from A. flavus-inoculated kernels.     

Some features of stelar tissues isolated from corn roots

In relation to radial ion transport to xylem vessels in higher plant roots, some features of stele, cortex, and root tip freshly isolated from 2-day-old corn roots grown under moistened or submerged condition were investigated. The contents of P, K, and Mg in primary roots decreased gradually from the apical to the basa1 region axially. The stele contained P, Na, Cu, Fe, and Mn in higher level than the cortex. The O₂ uptake rate of root tip was much higher and that of freshly isolated stele was as high as that of fresh cortex. In stele, more CN sensitive respiration was observed than in cortex. Although isolated stele absorbed tess ³²p than cortex and root tip, the incorporation of ³²p into organic compounds in stele was as much as in root tip. These results show that the freshly isolated stele has high metabolic activity and must play an inherent role in radial and selective ion transport in situ.     

Physical characteristics of corn kernels as influenced by nitrogen fertilization

Abstract

Physical characteristics of corn (Zea mays L.) kernels influence end‐use. Agronomic practices including nitrogen (N) fertilization may influence kernel hardness. The objective of this study was to determine if com endosperm characteristics influence the effect of N fertilization on kernel hardness and breakage susceptibility. In 1988, six corn hybrids differing for endosperm were grown at two locations in Ohio and with two N rates (34 and 200 kg/ha). Kernels of the waxy hybrid were denser less susceptible to breakage than its near‐isogeneic normal counterpart. Soft kernels weighted less and were less dense than hard kernels, but these two hybrid classifications did not differ for breakage susceptibility. Hard kernels possessed a greater resistance to grinding than soft kernels. Increasing N fertilizer rate increased resistance to grinding and reduced susceptibility to breakage of both soft and hard endosperm types. Fertilization with N not only affects corn grain yield, but also affects physical characteristics that are important to end‐users.     

主要生态因子对贮藏玉米中黄曲霉生长影响的模拟

为了模拟贮藏玉米中主要生态因子对黄曲霉生长的影响,利用Baranyi和Roberts函数拟合了玉米中黄曲霉的生长数据,得到了不同温度和水分活度下玉米中黄曲霉的生长动力学模型。应用一个二次多项式函数分别建立了描述水分活度（aw）和温度对菌落生长的组合影响的模型。对模型的有效性分别进行了验证,其偏离因子分别为0.896和0.963。精确因子都小于1.15。结果证明构建的二次多项式预测模型可以很好的预报aw和温度对黄曲霉生长的比生长速率和迟滞期的组合影响。得出的方程可以用于预测贮藏玉米中黄曲霉生长情况。     

Identification of chemical components of corn kernel pericarp wax associated with resistance to Aspergillus flavus infection and aflatoxin production

Kernel pericarp wax of the corn breeding population GT-MAS:gk has been associated with resistance to Aspergillus flavus infection and aflatoxin production. GT-MAS:gk wax, previously compared to waxes of three susceptible genotypes, was presently compared to wax of a different, and more numerous, group of susceptible lines. Wax separation by TLC confirmed previous findings, demonstrating a unique GT-MAS:gk band and a unique "susceptible" band. Only GT-MAS:gk wax inhibited the growth of A. flavus; however, no association was established, as before, between kernel wax abundance and resistance. Gas chromatography-mass spectroscopy (GC-MS) analysis of kernel whole wax showed a higher percentage of phenol-like compounds in wax from GT-MAS:gk than in waxes from the susceptible lines. The GT-MAS:gk unique band contained phenol-like compounds and ethyl-hexadecanoate; butyl-hexadecanoate was preeminent in most of the "susceptible bands". Alkylresorcinol (phenolic compounds) content was dramatically higher in GT-MAS:gk wax than in the wax of susceptible lines. An alkylresorcinol, 5-methylresorcinol, also inhibited in vitro growth of A. flavus. These and other phenolic compounds may contribute to kernel wax inhibition of A. flavus infection/aflatoxin production. Further investigation is needed to confirm a role for them in GT-MAS:gk resistance.     

Changes in free amino acid and sugar levels of dried figs during aflatoxin B1 production by Aspergillus flavus and Aspergillus parasiticus

An aqueous slurry of gamma-irradiated sterilized dried figs was inoculated with toxigenic strains of Aspergillus flavus and Aspergillus parasiticus. During incubation at 28 degrees C, pH, fructose, glucose, and free amino acids were determined by high-performance liquid chromatography and liquid chromatography (LC)/mass spectrometry, respectively, over 13 time points (1-20 days). At the same 13 time points using a LC/time-of-flight mass spectrometry screening method, aflatoxin B 1 and other secondary metabolites were simultaneously monitored. During the course of incubation, the pH significantly decreased and aflatoxin B 1 formation correlated with a reduction in proline content for both fungi. Of the 22 free amino acids that were monitored, only proline and cystine were found to be critical in supporting aflatoxin production. Levels of fructose and glucose steadily declined during incubation, until glucose was almost exhausted after 21 days. These time-course experiments confirmed the need for carbon and nitrogen sources for aflatoxin production in dried figs, and the favorable composition of figs as a fungal growth medium.     

Aflatoxin production in six peanut (Arachis hypogaea L.) genotypes infected with Aspergillus flavus and Aspergillus parasiticus, isolated from peanut production areas of Cordoba, Argentina

Aflatoxin contamination is one of the main factors affecting peanut seed quality. One of the strategies to decrease the risk of peanut aflatoxin contamination is the use of genotypes with resistance to Aspergillus infection. This laboratory study reports the resistance to Aspergillus infection and aflatoxin contamination of six peanut genotypes inoculated with 21 Aspergillus isolates obtained from the peanut production region of Cordoba, Argentina. The resistance was investigated in the seed coat and cotyledons of three resistant genotypes (J11, PI 337394, and PI 337409) and three breeding lines (Manfredi 68, Colorado Irradiado, and Florman INTA) developed at the Instituto Nacional de Tecnologia Agropecuaria (INTA), Manfredi Experimental Station, Cordoba, Argentina. Resistance to fungal colonization and aflatoxin contamination was found to be associated with seed coat integrity in the PI 337394, PI 337409, and J11 genotypes, whereas the INTA breeding lines such as Colorado Irradiado showed a moderate resistance and the Manfredi 68 and Florman INTA genotypes the least resistance. Furthermore, another type of resistance associated with cotyledons was found only in the PI 337394 genotype.     

Phytochemical inhibition of aflatoxigenicity in Aspergillus flavus by constituents of walnut (Juglans regia)

Tulare walnut, a cultivar highly resistant to aflatoxin formation, was investigated for endogenous phytochemical constituents capable of inhibiting aflatoxigenesis in Aspergillus flavus. The activity, located entirely in the pellicle (seed coat), was extractable to various degrees with polar solvents, although some activity remained unextractable, indicating that the bioactivity resided in a complex of hydrolyzable tannins. These tannins can be hydrolyzed by a fungal tannase present in A. flavus, yielding gallic acid and ellagic acid, testing of which showed that only gallic acid had potent inhibitory activity toward aflatoxin biosynthesis. Comparison of the gallic and ellagic acid content in the pellicle of Tulare and Chico cultivars, over the 2002 and 2003 growing seasons, showed that the gallic acid content increased rapidly with maturation of the nut and was 1.5-2 times higher in Tulare than in Chico. Gallic acid content in the pellicle at maturity of a series of commercial English walnut cultivars, and two black walnut species, was determined as an indicator of potential for inhibition of aflatoxigenesis. Regulation of gallic acid levels in the hydrolyzable tannins of walnuts by conventional breeding or genetic manipulation has the potential to provide new cultivars with high resistance to aflatoxigenesis.     

基于仿生脱粒的鸡喙啄取玉米果穗引起籽粒离散过程分析

在先离散后脱粒工艺基础上,为探究玉米果穗的低损伤离散,该文利用玉米离散试验系统对鸡喙啄取玉米籽粒的过程进行试验研究。结果表明,鸡喙接触玉米籽粒后,籽粒离鸡喙越近,推力的水平分力越大,籽粒运动越明显,越容易从果穗上分离下来,籽粒离鸡喙越远,推力的水平分力越小,籽粒越难与果穗分离;离散过程中,籽粒遵循"组砌规律"进行力的传递,传递的范围近似为"塔形",脱离果穗的籽粒的运动类似于斜抛运动;玉米果穗在果穗切线方向上的受力最大,其次是玉米果穗轴线方向上的受力,垂直于试验台方向上的受力最小;验证试验结果:玉米果穗的平均离散率为67.53%,平均离散损伤率为0.16%,表明模仿鸡喙的离散辊对玉米果穗有较好的离散效果,且损伤率低。该研究对低损伤玉米脱粒系统的设计提供了仿生学思路。     

一株黄曲霉拮抗菌的筛选、鉴定及特性研究

为获得拮抗黄曲霉和降解黄曲霉毒素的生物防治菌株,以烟台黄曲霉污染区域土壤为材料,采用稀释分离法和琼脂扩散法,筛选到一株对黄曲霉有抑制作用且可降解黄曲霉毒素的菌株Y-17-3。根据菌株的形态学特征、生理生化试验结合16S rDNA基因序列分析和系统发育树构建,确定菌株 Y-17-3 为空气芽孢杆菌(Bacillus aerius)。该菌株发酵液经硫酸铵沉淀的抗菌活性及稳定性试验,发现70%饱和硫酸铵沉淀的蛋白粗提物抑制黄曲霉活性最强,可降低黄曲霉孢子的萌发率,抑制菌丝的生长和菌丝球的形成,且该蛋白粗提物对温度、pH值和蛋白酶敏感。菌株Y-17-3能够降解黄曲霉毒素B₁,培养6 d时的降解率达90%。综上,所分离的空气芽孢杆菌Y-17-3菌株能够通过产生拮抗蛋白来抑制黄曲霉生长,且具有较强的黄曲霉毒素降解能力,在农业生物防治领域具有一定的应用前景。     

Cyclopiazonic acid production by cultures of Aspergillus and Penicillium species isolated from dried beans, corn meal, macaroni, and pecans

Ninety-five isolates of Aspergillus and Penicillium species from selected dried foods were examined for their ability to produce cyclopiazonic acid (CPA). The isolates were grown in sterile synthetic liquid medium at 28 degrees C for 8 days in the dark. The medium and mold mycelia were then extracted with chloroform. CPA was semiquantitatively determined by thin layer chromatography through visual comparison with standards. The cultures of A. flavus were also examined for their ability to produce aflatoxin. One A. tamarii and all 13 P. urticae isolates produced CPA, whereas only 19 of the 31 (61%) A. flavus isolates produced CPA, and 6 (19%) A. flavus produced aflatoxin. All 13 P. urticae isolates also produced patulin and griseofulvin. CPA-producing A. flavus was found in all food types but not in all samples. CPA-producing P. urticae was found only in dried beans and macaroni.     

玉米全生长期叶面积指数收获测量法的改进

农作物全生长期冠层表现不同的结构,常规的叶面积指数测量仪器不能适用于全生长期的叶面积指数测量,提出改进的收获测量法可进行玉米全生长期叶面积指数的测量,并且测量结果也具有可比性,该法在减少常规直接测量法工作量的同时也减少了对玉米的破坏。通过对比不同生长期单株样本叶面积计算的两种方法,得出二元二次回归法比常规的形状因子法计算精度高的结论。同时,分析不同生长期玉米秆所占总面积比例的规律,得出进行叶面积指数的准确测量,玉米秆的表面积必须进行准确考虑。该研究可为同类作物叶面积指数测量提供参考,可以有效推动叶面积指数的准确快速测量及遥感反演的验证工作。     

Inhibition of aflatoxin B(1) biosynthesis in Aspergillus flavus by anthocyanidins and related flavonoids.

Anthocyanidins and precursors or related flavonoids were tested at concentrations from 0.3 to 9.7 mM ( approximately 0.1-3.0 mg/mL) for activity against growth and aflatoxin B(1) biosynthesis by Aspergillus flavus Link:Fr. NRRL 3357. Aflatoxin B(1) production was inhibited by all anthocyanidins tested, and 3-hydroxy compounds were more active than 3-deoxy forms. Monoglycosides of cyanidin were 40% less inhibitory than the aglycon, whereas a monoglucoside and a diglucoside of pelargonidin were 80 and 5%, respectively, as active as the aglycon. Of eight flavonoids tested, only kaempferol was moderately active, whereas luteolin and catechin were weakly inhibitory. Binary combinations of delphinidin and three other aflatoxin inhibitors acted independently of each other. Results with an aflatoxin pathway mutant indicated that anthocyanidin inhibition occurred before norsolorinic acid synthesis.     

分层破胚剥皮玉米不同部位营养成分富集特征

为明确玉米籽粒营养成分的分布差异及不同部位富集特征,应用快速缓苏、微量着水半湿法分层破胚剥皮技术,结合靶向代谢组学方法,对郑单958玉米不同部位的营养成分及基础代谢物质进行分析与比较。结果表明玉米籽粒不同部位的淀粉、脂肪、矿物元素和膳食纤维等营养物质含量存在显著差异（P<0.05）。该研究中的玉米内皮层可能主要由种皮、糊粉层及部分外胚乳构成,该部位营养成分的种类及含量均较为丰富,其中水溶性膳食纤维含量显著高于其他部位（P<0.05）,可作为玉米水溶性膳食纤维的提取分离来源。K、P和Mg元素是玉米中含量最高的矿物元素,主要存在于胚芽中,Fe、Zn、Mn和Cu元素在胚芽和玉米皮层中均有较多分布,精制加工会导致这些矿物元素的损失。玉米胚芽中水解氨基酸种类较其他部位丰富且含量较高（P<0.05）,甜味氨基酸占总游离氨基酸含量的24.49%,高于玉米皮层部位、显著高于胚乳部位。研究结果为玉米营养健康食品的创制、玉米精深加工及相关专用装备的研发提供参考。     

异硫氰酸苄酯对于黄曲霉生长速率和产毒情况的影响

为明确异硫氰酸苄酯(BITC)在不同条件下对黄曲霉的抑制效果,本研究以熏蒸法,在28℃培养条件下,分别以花生和玉米为培养基质,研究不同浓度(0、5、10、15、20 mg·L-1)异硫氰酸苄酯在不同水分活度(aw)(0.930、0.960、0.980、0.995)下对黄曲霉(Aspergillus flavus)生长和...     

Identification and effects of maize silk volatiles on cultures of Aspergillus flavus

Volatiles generated from corn silks of individual genotypes of maize were found to exhibit differences in biological activities when the volatiles were exposed to 5-day solid cultures of Aspergillus flavus. In inverted potato dextrose-agar Petri plate bioassays, it was found that volatiles emitted from silks of the different maize genotypes had a profound effect on the growth of the fungus and, consequently, aflatoxin production. To determine the underlying cause for this bioactivity, volatiles emitted from the maize silks were trapped on Tenax glass columns and were analyzed by GC-MS. Aflatoxin field-resistant maize genotypes exhibited a larger relative concentration of the antifungal aldehyde, furfural (2-furancarboxaldehyde), when compared to the relative concentrations of the field-susceptible varieties tested. In a closed-container 5-day study, it was observed that fresh 1- and 4-day-old corn silk samples of aflatoxin-resistant maize genotypes emitted higher concentrations of furfural compared to those from susceptible genotypes. This observation probably explains the reason for the bioactivity observed in the in vitro bioassays, and the presence of furfural appears to contribute to a defense mechanism for protecting the developing maize kernel from fungal attack.