Mycoflora and mycotoxins of peanut (Arachis hypogaea L.) seeds in Egypt. III. Cellulose-decomposing and mycotoxin-producing fungi

From 40 peanut seed samples collected in Egypt, forty-three species and one variety of fungi, belonging to 16 genera, were collected. The most dominant genera were Aspergillus (11 species + one variety), Penicillium (11 species) and Fusarium (4 species). From the preceding genera A. fumigatus, A. flavus, A. niger, P. chrysogenum and F. oxysporum were the most frequent species.Forty-nine isolates belonging to 12 species and one variety were tested for production of mycotoxins, after growth on liquid medium containing two carbon sources (sucrose or cellulose). Thin layer chromatographic analysis revealed that the quality and quantity of mycotoxins was higher on sucrose than cellulose. Mycotoxins identified were aflatoxins B₁, B₂, G₁ & G₂, citrinin; fumagillin; diacetoxyscirpenol T-2 toxin; satratoxin H; and zearalenone.     

Survey of the mycoflora and mycotoxins of cotton seeds and cotton seed products in Egypt

Thirty-nine species and 16 fungal genera were isolated from Egyptian cotton seeds, cotton seed meal and cotton seed cake on 1% glucose-Czapek's agar medium incubated at 28 °C. Aspergillus was the most frequent genus and it emerged in 87–100% of the samples contributing 70–98% of total fungi in the three substrates tested. The most common species were A. niger, A. flavus, A. fumigatus, A. terreus and Rhizopus stolonifer; A. niger, A. fumigatus and Penicillium corylophilum; and A. niger, A. flavus, A. terreus, A. nidulans and Rhizopus stolonifer, respectively. Cotton seeds and cotton seed products were naturally contaminated by aflatoxin B₁ and B₂. About 16% of the different substrates tested were positive for aflatoxin contamination. No citrinin, ochratoxin A, patulin, sterigmatocystin, diacetoxyscirpenol, T-2 toxin or zearalenone were detected in the samples assayed.     

Boron mobility in peanut (Arachis hypogaea L.)

In most plant families, boron (B) is phloem immobile. For plants such as peanut which bury their fruit, the mechanism for B delivery and the B source for fruit and seed growth remains enigmatic. Therefore, this study aimed to establish evidence of B retranslocation in peanut and to identify its importance in plant development. In a sand culture experiment, the increase in B contents in new organs after B withdrawal and the corresponding decline in B contents in older organs was evidence of B redistribution. In a foliar ¹⁰B experiment, the ¹⁰B abundance of treated-leaves decreased and ¹⁰B was detected in leaves and flowers formed after the application of foliar B. Application of ¹⁰B to the roots for a period also provided evidence for the retranslocation of ¹⁰B accumulated during the first growth period. The ¹⁰B abundance in older plant parts declined and ¹⁰B appeared in new organs (flowers, pegs, leaves) that had developed after the ¹⁰B supply had been replaced by ¹¹B. In the fourth experiment, foliar application of B reduced hollow heart, a symptom of B deficiency in seeds, in cv. TAG 24 from 39 to 8% and in Tainan 9 from 63 to 18%. These experiments all provide evidence for B retranslocation in peanut, but further work on the relative importance of the xylem and phloem pathways for B loading into the fruit is needed.     

Mycoflora and mycotoxins of Brazilian cashew kernels

Kernel samples of common and dwarf Brazilian cashew nuts were highly contaminated with field and storage fungi in comparison to healthy ones. In general, dwarf cashews were more contaminated than common. A total of 37 fungal species were identified. Aspergillus niger was the dominant species with more colonies being isolated from dwarf kernels. A. flavus was the next most frequently isolated species. Penicillium brevicompactum, and P. glabrum were the most frequently isolated penicillia, with higher contamination recorded from dwarf kernels. Chaetomium globosum was recorded at a high level. Nine species were recorded from cashew kernels for the first time. Multimycotoxin analysis by tlc and hplc were positive for mycotoxins and other secondary metabolites particularly from the infected samples. Hplc was only carried out on dwarf cashews. Aflatoxins were not detected by quantitative high performance thin layer chromatography. This revised version was published online in June 2006 with corrections to the Cover Date.     

Mycotoxin production on different cultivars and lines of broad bean (Vicia faba L.) seeds in Egypt

One hundred different cultivars and lines of broad bean (Vicia faba L.) seed samples were inoculated with Aspergillus flavus Link (CMI 102135) to determine varietal differences which may support or resist aflatoxin production. Thin-layer chromatographic analysis of the chloroform extracts of the different seed samples revealed that 11 cultivars/lines were highly resistant to seed invasion and aflatoxin production while 9 cultivars/lines showed partial resistance. The remaining 80 samples were susceptible to the establishment of A. flavus and aflatoxin accumulation. All the resistant cultivars/lines seed samples were inoculated also with three local isolates of fungi namely; Stachybotrys chartarum (Ehrenb. ex Link) Hughes, Aspergillus ochraceus Wilhelm, and Fusarium oxysporum Schlecht. The resistant seed samples were also resistant for colonization with these fungi and mycotoxin formation.     

花生镉污染研究进展

花生既是世界主要的油料作物,又是重要的植物蛋白来源和食品加工原料．随着花生直接食用和食品加工的不断增加,国际上对花生籽粒Cd含量问题越来越关注．我国是世界上重要的花生生产国和出口国．近年来,花生Cd含量偏高已经成为制约我国出口贸易的重要因素．本文从花生籽粒Cd富集能力、花生Cd含量的种内差异、籽粒中Cd的分布规律、影响花生籽粒Cd积累的机制和降低花生籽粒Cd含量技术等方面,对花生Cd污染研究的现状与问题进行了论述．指出在花生cd污染控制方面有2种策略可以考虑,一是降低花生对土壤Cd的吸收;二是控制Cd向籽粒的迁移富集．为此需要从3个方面加强对花生籽粒Cd积累机制的研究,即花生根系活性特征参数及其与籽粒Cd积累的关系;花生果荚Cd吸收机制及其对籽粒Cd含量的贡献;花生植株体内Cd迁移机制及其与籽粒Cd含量的关系．     

Fungal populations in the rhizosphere of peanut (Arachis hypogaea L.)

Summary A comparison of fungal populations in the rhizospheres of eight varieties of peanut grown in a red lateritic soil amended with farmyard manure was made by the dilution-plate technique. There was a marked increase in fungi in the rhizospheres of TMV 2, TMV 4, Pollachi Red and EC 1698, the increase was smaller in Spanish Improved and RS 1 while very little rhizosphere effect was shown by TMV 3 and Pondicherry 8. Age of the plant had a significant influence on numbers of fungi in the rhizosphere. High R/S ratios were obtained when the plants were 30 days old, at which time attained maximum vegetative growth and started to flower. The ratios gradually decreased after that age until the plants were three months old when there was again a small increase. This later rise in fungal populations is interpreted to be due to an increase in microbial activity around dead or senescent roots. No correlation could be established between numbers of root nodules produced by a variety and its rhizosphere effect. Preferential stimulation of certain fungi in the rhizosphere of some of the varieties was noticed.     

Utility of EST-derived SSR in cultivated peanut (Arachis hypogaea L.) and Arachis wild species

Background  

Lack of sufficient molecular markers hinders current genetic research in peanuts (Arachis hypogaea L.). It is necessary to develop more molecular markers for potential use in peanut genetic research. With the development of peanut EST projects, a vast amount of available EST sequence data has been generated. These data offered an opportunity to identify SSR in ESTs by data mining.     

Effects of vesicular-arbuscular mycorrhizae on growth and phosphorus and zinc nutrition of peanut (Arachis hypogaea L.) in an Oxisol from subtropical Australia

Peanut plants (cv. Shulamit) were grown in an Oxisol soil in pots in the glasshouse to assess effects of soil sterilization and inoculation with spores of vesicular-arbuscular mycorrhizal fungi (VAMF) on the response to five rates of phosphorus (0 to 240 kg P ha^–1) and two rates of zinc (0 and 10 kg Zn ha^–1) fertilizers.Both P and Zn nutrition were affected by VAMF activity but the dominant role of VAMF in this soil type was in uptake of P. In the absence of VAMF there was a clear threshold level of P application (60 kg P ha^–1) below which plants grew poorly, which resulted in a sigmoidal response of dry matter to applied P. The maximum response was not fully defined because dry matter production continued to increase up to 240 kg P ha^–1. Tissue P concentration of non-mycorrhizal plants increased linearly with P rate and was always significantly less than that in mycorrhizal plants.Mycorrhizal plants responded without threshold to increasing P rate, attaining maximum dry matter at 120 kg P ha^–1 in inoculated sterilized soil and at 30 kg P ha^–1 in nonsterile soil. These differences in maximal P rates and in the greater dry matter produced in sterile soil at high P rates were attributed to the negative effects of the root-knot nematodeMeloidogyne hapla in nonsterile soil.Plant weight did not respond to zinc fertilizer but tissue Zn concentration increased with applied Zn. Tissue Zn concentration and uptake were increased by VAMF.     

Agrobacterium tumefaciens mediated gene transfer in peanut (Arachis hypogaea L.)

Transgenic peanut plants were produced using Agrobacterium mediated gene transfer. Primary leaf explants of peanut were co-cultivated with Agrobacterium tumefaciens LBA 4404 harbouring the binary plasmid pBI 121 (conferring -glucuronidase activity and resistance to kanamycin) and cultured on regeneration medium supplemented with kanamycin to select putatively transformed shoots. They were rooted and plants were transferred to soil. Stable integration and expression of the transgenes were confirmed by NPT II assay, Southern blot hybridization and GUS assay.Abbreviations BA 6-benzyladenine - GUS -glucuronidase - IAA indole-3-acetic acid - NAA -naphthaleneacetic acid - NOS nopaline synthase - NPT II neomycin phosphotransferase II - SDS Lauryl sulfate     

Plant regeneration through somatic embryogenesis in peanut (Arachis hypogaea L.)

Somatic embryos were induced from immature cotyledons and immature embryonal axis ofArachis hypogaea L. on L-6 basal medium supplemented with NAA, picloram or 2,4-D at 5–50 mg 1^-1. Immature embryonal axis produced a higher number of somatic embryos in comparison with immature cotyledons. The highest number of responding cultures was produced on medium supplemented with NAA (50 mg 1^-1), while the highest average number of somatic embryos per culture was produced on medium with 2,4-D (10 or 20 mg 1^-1) and picloram (30 mg 1^-1) from cotyledons. The somatic embryos developed into plants on basal medium supplemented with activated charcoal and about 100 plants were successfully transferred to the field. Acknowledgement: The authors wish to thank Nuclear Agriculture Division, BARC for supplyingA. hypogaea seeds and Mr. R.M. Mudliar for photography.     

Diniconazole's effect on peanut (Arachis hypogaea L.) growth and development

Greenhouse nutrient solution studies demonstrated that diniconazole will decrease peanut (Arachis hypogaea L.) shoot growth when either root or shoot applied. Root growth and development were decreased by root and, to a lesser extent, by shoot uptake of diniconazole. Diniconazole is apparently xylem translocated, but not phloem translocated. Concentrations of 200 ppb ES isomer of diniconazole in nutrient solution (root uptake) increased specific leaf weight and starch deposits in the leaf. Field applications of 193 g ES isomer ha^–1 of diniconazole reduced main stem height by 33%, leaf area index by 16%, and total vegetative dry weight by 19%, but had no effect on average leaf size. Decreased germination of seeds from plants treated with 1435 g ha^–1 diaminozide was associated with increased seed dormancy. Seed dormancy was counteracted by either ethylene gas or storage for 150 days after harvest. Soil applications of diniconazole were more effective than foliar appliations in reducing vine growth. Diniconazole's ER isomer is a broad spectrum fungicide that reduced damage (when compared to the control) bySclerotium rolfsii andRhizoctonia solani. The reduced damage by these diseases was thought to be the primary reason for the significant pod yield increase (when compared to the control) observed with the diniconazole treatments. In drought-stressed plots, populations of the two-spotted spider mite (Tetranychus urticae) were increased by diniconazole.Mention of a trademark, proprietary product, or vendor does not constitute a guarantee by the University of Georgia or the U.S. Department of Agriculture and does not imply UGA or USDA approval to the exclusion of other products or vendors that also may be suitable.     

RFLP variability in peanut (Arachis hypogaea L.) cultivars and wild species

Summary RFLP variability was studied in eight U.S. peanut cultivars, representing the four market types, and in 14 wild Arachis species accessions, using random genomic clones from a PstI library. Very low levels of RFLP variability were found among the allotetraploids, which included the U.S. cultivars and Arachis monticola, a wild species. The diploid wild species were very diverse, however. RFLP patterns of the allotetraploids were more complex than the diploids, and the two constituent genomes could usually be distinguished. On the basis of RFLP band sharing, A. ipaensis, A. duranensis, and A. spegazzinii appeared most closely related to the diploid progenitor species of the allotetraploids. A dendrogram of relationships among the diploid wild species was constructed based on band sharing.     

Partial characterization of an allergenic glycoprotein from peanut (Arachis hypogaea L.)

A concanavalin A-reactive glycoprotein allergen has been isolated from peanut (Arachis hypogaea). The allergen was separated by affinity chromatography and purified by gel permeation and ion-exchange chromatography. The monomeric molecular weight is 65,000 and the pI is 4.6. The presence of one cysteine residue per molecule results in some dimer formation. Concanavalin A-reactive glycoprotein is a potent allergen for peanut-sensitive patients in both in vivo and in vitro tests. It is allergenically stable, on in vitro examination, at temperatures of up to 100 degrees C and over the pH range 2.8-10. Removal of the carbohydrate moiety failed to eliminate the allergenicity. Concanavalin A-reactive glycoprotein is identified in the crossed immunoelectrophoretic pattern as a major antigen of peanut protein extract but its structural characteristics indicate that it is probably not a component of the major storage-protein complex, arachin.     

Somatic embryogenesis from the axillary meristems of peanut (Arachis hypogaea L.)

Developmental anomalies in the plumule meristem of peanut (Arachis hypogaea L.) somatic embryos resulted in poor shoot differentiation and reduced plant recovery. Existing meristems with caulogenic potential have never been tested for embryogenesis in peanut. The present experiment was designed to test the mature zygotic embryo axis derived plumule with three meristems for somatic embryogenesis. Embryogenic masses and embryos developed from the caulogenic meristems in the axils. Exposure of 2 weeks in primary medium with 90.5 μM 2,4-D suppressed the shoot tip differentiation temporarily which then regained the ability to form the shoot on withdrawal of 2,4-D. Exposure of 4 weeks in primary medium with 90.5 μM 2,4-D suppressed the shoot tip differentiation irreversibly. No shoot formation was noted from the tips in any of the cultures which were in secondary medium with 13.6 μM 2,4-D. Development of somatic embryos directly from axillary meristems was confirmed histologically. Conversion frequency of these embryos was 11%. Thus, in this report, we describe a method to obtain somatic embryos from the determined organogenic buds of the axillary meristem, by culturing the nodal explant vertically on embryo induction medium. It also displays the possibility of obtaining both embryogenic and organogenic potential in two parts of the same explant simultaneously. The possibility of extending this approach for genetic transformation in in vivo system through direct DNA delivery or Agrobacterium injection in meristems can also be explored. Using Agrobacterium rhizogenes, we have demonstrated the possibility of gene transfer in the axillary meristems of seed-derived plumule explant.     

Effects of desiccation on peanut (Arachis hypogaea L.) seed protein composition

Response of peanut to desiccation was studied by monitoring changes in the seed protein content and composition during a 14 day desiccation period using a combination of electrophoretic and immunochemical techniques. Following desiccation, the protein content of ‘white’ (most immature) and ‘orange’ seed increased, while that of the ‘brown’ (mature) seed was not affected. Sodium dodecyl sulphate polyacrylamide gel electrophoresis (SDS-PAGE) showed no major qualitative differences in the protein composition during desiccation of the samples. However, immunoblotting with anti-dehydrin antisera revealed the presence of several new proteins in the desiccated samples compared with the controls. One of the dehydrin-like proteins, band ‘c’ was found to be related to water-stress, while the other proteins appeared to be the storage proteins accumulated as the seed matured in vitro. Capillary electrophoresis (CE) showed major changes in the protein quantity and quality of ‘white’ seed during the 0–14 days of desiccation. In contrast, in the ‘orange’ and ‘brown’ seeds changes in protein composition were less significant. Results indicated that there are several dehydrin-like proteins expressed in peanuts, however, not all of them are related to water stress.     

花生对镉胁迫的生理响应及品种间差异

深入研究花生镉毒害机理对于筛选和利用抗镉污染花生种质资源具有重要的理论和实际意义.本文以14个花生品种为材料,以花期花生功能叶叶绿素含量、根系和叶片丙二醛含量、细胞膜透性和根系氧化活力等生理参数为指标,利用人工气候箱砂培试验,研究了6种Cd浓度胁迫下花生植株的生理毒害反应及其品种间遗传差异.结果表明: 在0～60 mg Cd ·L^-1范围内,随着营养液添加Cd浓度的增加,花生叶绿素含量和根氧化活力极显著降低,根、叶细胞膜透性和根、叶丙二醛含量极显著增加,且品种与Cd浓度间具有显著交互作用; 花生根、叶细胞膜透性是对镉胁迫响应最为敏感的生理参数,而叶绿素含量对镉胁迫响应相对不敏感; 各生理参数与营养液Cd浓度关系的线性回归方程的斜率(b)与截距(a)之比值的绝对值︱b/a︱能够较好地描述不同花生品种对镉胁迫的敏感性.对6个生理参数的︱b/a︱值进行综合赋值及敏感性五级聚类分析得知,在供试花生品种中,“中花4号”、“湘农55号”和“湘农3010-w”等3个品种对镉反应高度敏感(Ⅰ级);“莱农29”、“湘农小果w2-7”、“丰花2号”、“莱农13”、“豫花15号”和“丰花3号”等6个品种反应敏感(Ⅱ级);“湘农312”、“祁阳小籽”和“平度01”等3个品种反应一般(Ⅲ级);“花育23 ”和“花育20”等两个品种反应钝感(Ⅳ级).     

Genetic and phenetic analyses of Bradyrhizobium strains nodulating peanut (Arachis hypogaea L.) roots.

Seventeen Bradyrhizobium sp. strains and one Azorhizobium strain were compared on the basis of five genetic and phenetic features: (i) partial sequence analyses of the 16S rRNA gene (rDNA), (ii) randomly amplified DNA polymorphisms (RAPD) using three oligonucleotide primers, (iii) total cellular protein profiles, (iv) utilization of 21 aliphatic and 22 aromatic substrates, and (v) intrinsic resistances to seven antibiotics. Partial 16S rDNA analysis revealed the presence of only two rDNA homology (i.e., identity) groups among the 17 Bradyrhizobium strains. The partial 16S rDNA sequences of Bradyrhizobium sp. strains form a tight similarity (> 95%) cluster with Rhodopseudomonas palustris, Nitrobacter species, Afipia species, and Blastobacter denitrificans but were less similar to other members of the alpha-Proteobacteria, including other members of the Rhizobiaceae family. Clustering the Bradyrhizobium sp. strains for their RAPD profiles, protein profiles, and substrate utilization data revealed more diversity than rDNA analysis. Intrinsic antibiotic resistance yielded strain-specific patterns that could not be clustered. High rDNA similarity appeared to be a prerequisite, but it did not necessarily lead to high similarity values between RAPD profiles, protein profiles, and substrate utilization. The various relationship structures, coming forth from each of the studied features, had low compatibilities, casting doubt on the usefulness of a polyphasic approach in rhizobial taxonomy.     

Fungal populations and mycotoxins in silage in Assiut and Sohag governorates in Egypt,with a special reference to characteristic Aspergilli toxins

Forty silage samples were collected from Assiut and Sohag governorates in Egypt to measure the presence of fungal population in silage. Forty-three species and 2 species varieties belonging to 17 genera were isolated using glucose Czapeks and Sabourauds dextrose agar media at 28 ^°C. The most prevalent genera were Aspergillus (57.5 and 100 of the samples), Penicillium (100 and 55%) on the two mentioned media, respectively. Also, Fusarium oxysporum and Gibberella fujikurori were recovered in moderate incidences. Mycotoxin profiles were also determined in these samples: Aflatoxins showed the highest incidence rates of occurrence, it occurred in 22.5% of all samples analyzed. Other mycotoxins were detected from all samples (T₂ toxins and sterigmatocystin at incidence of 7.5 and 5%, respectively). The screening of the characteristics mycotoxins of different isolates of Aspergillus isolated from silage samples was tested. The results clarified that some mycotoxins (aflatoxins – aspergillic acid – beta nitro propionic acid – cyclopiazonic acid– kojic acid and sterigmatocystin) were produced by some isolates of A. flavus. Some isolates of A.fumigatus could produce gliotoxin and verrucologen. All of A. niger isolates tested were able to produce kojic acid. One isolate of A. ochraceous formed ochratoxin A and other isolate produced penicillic acid. Concerning A. terreus isolates, the results showed that 5 isolates were able to produce citrinin and 4 isolates had ability to produce patulin. A. versicolor isolates showed the ability to produce ochratoxin A.