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棉花黄萎病菌毒素ELISA检测方法的建立及其应用 总被引：6，自引：1，他引：6
以纯化的棉花黄萎病菌毒素(PLPC)免疫新西兰白兔制备了PLPC特异性抗血清,建立了可特异性检测棉花黄萎病菌毒素的A蛋白双抗体夹心ELISA方法。应用建立的ELISA方法测定了病菌培养滤液、人工接种幼苗和大田病株不同组织中的毒素,结果表明:不同产毒能力菌株(VD 8和VD-5)在25℃下振荡培养3d后就能在培养滤液中检测到毒素,这比生物测定要早2～3d。将VD-8菌株的孢子以灌根方式接种泗棉3号幼苗5d后,就能从接种幼苗的茎杆和叶柄中检测到毒素,这比幼苗自然发病显症要早3～5d。在测定的30份大田病株茎杆、叶柄、叶脉样品中,阳性样品率为100%。这些结果表明建立的ELISA方法可用于菌株产毒能力的测定和大田棉花黄萎病的早期诊断。 相似文献
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棉花黄萎病菌毒素结合位点初探 总被引：6，自引：1，他引：5
以纯化的棉花黄萎病菌毒素(PLPC)免疫新西兰白兔制备了PLPC特异性抗血清。将PLPC(15μg·ml 1)用不同浓度的抗体(1～20μg·ml 1IgG)吸附后处理泗棉3号的切根苗,毒素所引起的症状都有不同程度的减轻。表明所制备的抗体在与毒素发生特异性免疫学反应的同时,可部分封闭毒素分子上与毒素受体结合的位点。利用竞争ELISA测定了泗棉3号幼苗子叶的质膜制剂与PLPC的结合活性。结果表明,质膜制剂与毒素结合后能部分阻断毒素与其抗体的免疫学反应,即质膜制剂中含有毒素的结合位点。分别用胰蛋白酶和煮沸处理质膜制剂后,质膜制剂对毒素与其抗体的反应的抑制作用消失,初步表明质膜制剂中与毒素结合的是蛋白质。 相似文献
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酶联免疫技术(ELISA)是在免疫学和细胞工程学基础上发展的一种微量检测技术,具有操作简便、灵敏性高等优点,已在食品农药残留、重金属污染、食品添加剂检测等诸多方面表现出了独特的优势,在食品安全检测领域有着广阔的应用前景。分析酶联免疫技术的原理和方法,从生物毒素、药物残留、致病微生物、重金属污染和食品的品质等方面评述其在食品安全检测中的应用。 相似文献
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黄曲霉毒素B1完全抗原合成及鼠源多抗血清的制备 总被引：3，自引：2，他引：1
为了合成黄曲霉毒素B1(AFB1)完全抗原,制备鼠源AFB1多克隆抗体血清。通过琥珀酰亚胺酯法在黄曲霉毒素B1(AFB1)分子上引入羧基,生成黄曲霉毒素B1肟(AFB1O)。用EDC法和DCC法合成AFB1-BSA和AFB1-OVA。采用薄层层析技术、紫外光谱技术(Uv)、SDS-PAGE鉴定完全抗原的合成。通过免疫BALB/c小鼠,获得鼠原多克隆血清,采用间接ELISA方法测定多抗血清的免疫效价,用竞争ELISA检测多克隆血清的敏感性和特异性。结果表明：免疫小鼠血清效价都在1:3200以上,其中6号鼠效价最高,到达1.28×10-4,敏感性好,半数抑制浓度IC50为22.268 ng/mL,交叉反应率低。本研究成功制备了AFB1完全抗原,获得了敏感的AFB1多克隆抗体血清,为以后制备AFB1单克隆抗体及免疫学快速检测方法奠定了基础。 相似文献
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史春悦 《农产品加工．学刊》2019,(14)
黄曲霉毒素主要是由黄曲霉、寄生曲霉产生的次级代谢产物,在自然界中尤其是湿热地区分布广泛,污染范围广,具有强致癌和致突变性。目前,用于检测黄曲霉毒素含量的技术主要包括大型仪器分析技术和免疫分析技术。对当前黄曲霉毒素检测技术的研究进展进行简要综述,旨在为食品中黄曲霉毒素的检测分析提供参考。 相似文献
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Fusarium graminearum (FG) and F. verticillioides (FV) produce the mycotoxins deoxynivalenol (DON), zearalenone (ZEA) and fumonisins (FUM), respectively, in maize. The EU
released limits for these toxins in food. Growing resistant varieties is one alternative to fulfill these limits. Quantification
of mycotoxin concentrations is expensive and time consuming. If indirect selection based on cost efficient and fast ear rot
rating is feasible, this could increase efficiency of selection. The objective of this study was to analyze correlations between
mycotoxin concentrations and ear rot rating by inoculating three maturity groups (early, mid-late, late) each comprising about
50 inbred lines tested in Central and Southern Europe. In the early maturity group flint lines were more susceptible in all
instances except ZEA than dent lines. Broad ranges and significant (P < 0.01) genotypic variances were detected, but also genotype × environment interaction variances were significant (P < 0.01). Heritabilities of ear rot rating were similar or higher than those of mycotoxin concentrations (0.61−0.93 and 0.56–0.89,
respectively). Although high genotypic correlations between FUM and DON or ZEA were found (0.77; 0.76, respectively), separate
testing of FV and FG and corresponding mycotoxins is necessary since genotypes resistant to FV were not necessarily resistant
to FG and vice versa. Medium to high heritabilities and high genotypic correlations between ear rot and corresponding mycotoxin
concentrations (0.87–0.99) suggest frequent identification of lines with reduced mycotoxin concentrations by ear rot rating.
Assuming fixed budgets we conclude that indirect selection by applying cost efficient ear rot rating could increase selection
intensity and therefore is more effective than direct selection for reduced mycotoxin concentrations. 相似文献
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Katashi Kubo Naoyuki Kawada Takashi Nakajima Kazuyuki Hirayae Masaya Fujita 《Euphytica》2014,200(1):81-93
Fusarium head blight (FHB) remains a serious problem that causes yield and grain quality losses, and mycotoxin accumulation in wheat production in western Japan. A 3-year field trial with artificial FHB inoculation was conducted to evaluate varietal characteristics of FHB resistance among 31 wheat cultivars/lines cultivated in western Japan, including one standard line. Severity of FHB, frequency of Fusarium-damaged kernels (FDK), deoxynivalenol concentration (DON), nivalenol concentration (NIV), and grain yield showed significant differences among years and among cultivars/lines. Interaction between years and cultivars/lines was also significant in these traits, but F values were larger for cultivars/lines than for the interaction. Correlation analysis showed that cultivars/lines with lower FHB severities tended to have lower FDK, DON and NIV, and a higher yield. Resistance to kernel infection (RKI), residuals calculated by regressing FDK against FHB severity, and resistance to mycotoxin accumulation (RTA), residuals calculated by regressing DON + NIV against FDK, also differed significantly among cultivars/lines. These results indicated that varietal differences in response to FHB symptom development, RKI and RTA exist among wheat cultivars/lines in western Japan. Such information is important to aid producers in controlling the disease and for breeders to improve FHB resistance and reduce mycotoxin accumulation in commercial wheat cultivars. 相似文献
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T. Miedaner C. Reinbrecht U. Lauber M. Schollenberger H. H. Geiger 《Plant Breeding》2001,120(2):97-105
Fusarium culmorum is one of the most important Fusarium species causing head blight infections in wheat, rye, and triticale. It is known as a potent mycotoxin producer with deoxynivalenol (DON), 3‐acetyl deoxynivalenol (3‐ADON), and nivalenol (NIV) being the most prevalent toxins. In this study, the effect of winter cereal species, host genotype, and environment on DON accumulation and Fusarium head blight (FHB) was analysed by inoculating 12 rye, eight wheat, and six triticale genotypes of different resistance levels with a DON‐producing isolate at three locations in 2 years (six environments). Seven resistance traits were assessed, including head blight rating and relative plot yield. In addition, ergosterol, DON and 3‐ADON contents in the grain were determined. A growth‐chamber experiment with an artificially synchronized flowering date was also conducted with a subset of two rye, wheat and triticale genotypes. Although rye genotypes were, on average, affected by Fusarium infections much the same as wheat genotypes, wheat accumulated twice as much DON as rye. Triticale was least affected and the grain contained slightly more DON than rye. In the growth‐chamber experiment, wheat and rye again showed similar head blight ratings, but rye had a somewhat lower relative head weight and a DON content nine times lower than wheat (3.9 vs. 35.3 mg/kg). Triticale was least susceptible with a five times lower DON content than wheat. Significant (P = 0.01) genotypic variation for DON accumulation existed in wheat and rye. The differences between and within cereal species in the field experiments were highly influenced by environment for resistance traits and mycotoxin contents. Nevertheless, mean mycotoxin content of the grain could not be associated with general weather conditions in the individual environments. Strong genotype‐environment interactions were found for all cereal species. This was mainly due to three wheat varieties and one rye genotype being environmentally extremely unstable. The more resistant entries, however, showed a higher environmental stability of FHB resistance and tolerance to DON accumulation. Correlations between resistance traits and DON content were high in wheat (P = 0.01), with the most resistant varieties also accumulating less DON, but with variability in rye. In conclusion, the medium to large genotypic variation in wheat and rye offers good possibilities for reducing DON content in the grains by resistance selection. Large confounding effects caused by the environment will require multiple locations and/or years to evaluate FHB resistance and mycotoxin accumulation. 相似文献
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D. A. Presello A. O. Pereyra J. Iglesias C. M. Fauguel D. A. Sampietro G. H. Eyhérabide 《Euphytica》2011,178(1):23-29
Random S5 inbreds derived from three F2 maize (Zea mays L.) populations (L1934 × LP918, LP915 × LP2541 and L7310 × L7266) were selected for ear rot resistance after inoculation
with a low-fumonisin producing isolate belonging to F. proliferatum. The four less susceptible and the four most susceptible inbreds from each population were crossed and F1 seeds were pooled. Resistant and susceptible pools from each population were evaluated for disease severity (percentage of
the ear visibly diseased) after inoculation with the isolate used for selection, and high toxigenic isolates belonging to
F.
verticillioides and F. graminearum. Grain mycotoxin concentration was assessed by ELISA. Differences in disease resistance to each fungus were observed between
resistant and susceptible pools in most populations and environments indicating that selection after inoculation with a single
species might be effective to develop broad-based resistance to Fusarium. Resistant pools exhibited, after inoculation with F. verticillioides, low grain fumonisin concentrations in most populations and years. Positive genotypic correlations between disease severity
and fumonisin concentration (0.89 < rg < 0.98, depending on fungal species and year) indicate that selection for disease severity accounted for most of the variability
for field fumonisin accumulation. Selection seemed to be also effective to reduce grain deoxynivalenol and zearalenone concentrations
after inoculation with F. graminearum. Ratios between grain deoxynivalenol concentration and disease severity were lower in L7310 × L7266 than those observed in
the other populations suggesting that mechanisms affecting mycotoxin accumulation might exist in this population and additional
responses should be feasible if including deoxynivalenol concentration as another selection parameter. 相似文献
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真菌毒素(mycotoxin)是小麦及面制食品重要的安全风险之一。为明确真菌毒素在小麦磨粉及食品加工链条中的变化规律,解决目前我国尚缺少面粉和面制食品真菌毒素限量标准的问题,开展小麦真菌毒素污染风险评估,以受玉米赤霉烯酮(ZEN)污染的小麦为材料,用超高效液相色谱串联质谱法(UPLC-MS/MS)检测了不同磨粉组分及馒头、面包、面条加工过程中不同环节加工品中ZEN的含量。结果表明,小麦磨粉后粗麸皮和细麸皮中ZEN的含量显著升高,达到小麦籽粒的2倍以上,小麦粉中ZEN的含量平均降低70%以上;小麦粉加工成馒头和面包后,ZEN的含量分别增高1.8倍和1.0倍,加工成面条后因ZEN在煮制过程中部分溶于水中而降低30%以上。研究结果表明,加工过程对小麦及面制食品中的毒素水平有显著影响,对小麦、面粉及不同的小麦制品分别制定ZEN限量更为科学合理。 相似文献
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赭曲霉毒素A检测方法的研究进展 总被引：1，自引：0，他引：1
赭曲霉毒素A是由曲霉属和青霉属菌株产生的一种有毒真菌次级代谢产物。毒理学研究表明,赭曲霉毒素A具有很强的肝脏毒性和肾脏毒性,并有致畸、致突变和致癌作用。赭曲霉毒素A广泛存在于谷物及谷物产品、香料和咖啡豆等产品中,严重危害人类健康。许多国家建立了赭曲霉毒素A的限量标准,因此有必要研究准确、灵敏的赭曲霉毒素A的检测方法。综述了赭曲霉毒素A的理化性质及其检测分析方法的研究进展。 相似文献
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Marine Ollier Vincent Talle Anne‐Laure Brisset Zo Le Bihan Simon Duerr Marc Lemmens Ellen Goudemand Olivier Robert Jean‐Louis Hilbert Hermann Buerstmayr 《Plant Breeding》2019,138(1):69-81
Fusarium head blight (FHB) is a cereal disease of major importance responsible for yield losses and mycotoxin contaminations in grains. Here, we introduce a new measurement approach to quantify FHB severity on grains based on the evaluation of the whitened kernel surface (WKS) using digital image analysis. The applicability of WKS was assessed on two bread wheat and one triticale grain sample sets (265 samples). Pearson correlation coefficients between Fusarium‐damaged kernels (FDK) and WKS range from r = 0.77 to r = 0.81 and from r = 0.61 to r = 0.86 for the correlation between deoxynivalenol (DON) content and WKS. This new scoring method facilitates fast and reliable assessment of the resistance to kernel infection and shows significant correlation with mycotoxin content. WKS can be automated and does not suffer from the “human factor” inherent to visual scorings. As a low‐cost and fast approach, this method appears particularly attractive for breeding and genetic analysis of FHB resistance where typically large numbers of experimental lines need to be evaluated, and for which WKS is suggested as an alternative to visual FDK scorings. 相似文献
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辐照技术在绿色储粮中的应用研究 总被引：1，自引：0，他引：1
全面综述了辐照技术在储粮害虫防治、病茵抑制与真菌毒素降解中的广泛应用及其对粮食品质的影响。随着绿色储粮工程的深入实施,辐照技术将有广阔的应用前景。 相似文献
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由赤霉菌引起的小麦赤霉病是小麦生产的主要病害之一,小麦赤霉病抗源十分贫乏,而且多数表现为多基因遗传,在小麦育种中较难应用。从小麦近缘属种中寻求赤霉病抗性基因并把它引入栽培品种中,有重要的学术意义和经济价值。本研究以来自抗病小麦材料PI481521和栽培小麦Langdon杂交得到的14个4倍体小麦单条染色体代换系为试验材料,进行室内赤霉病的抗性鉴定,同时用脱氧雪腐镰刀菌烯醇(DON)试剂盒来测定接种后籽粒中的毒素含量。表型鉴定结果表明,14个代换系的抗病性存在较大的差异,其中代换系LDN(PI481521-3A)和LDN(PI481521-7B)分别在感染小穗数和小穗感染百分率上与抗性亲本无显著性差异,不存在抗病性高于抗性亲本的类型;毒素含量分析表明,2个亲本的毒素含量没有显著差异,各代换系的毒素含量均高于抗性亲本PI481521、LDN(PI481521-1A)、LDN(PI481521-2A)及LDN(PI481521-6A)的毒素含量较高,与2个亲本存在显著差异。这一鉴定结果为下一步抗性基因和毒素基因的定位研究提供了基础材料,对赤霉病抗性指标进行相关性分析,发现小穗感染百分率与麦粒毒素含量呈显著正相关。因此,在田间进行抗赤霉病种质筛选及抗病育种后代选择时,可以小穗感染百分率作为衡量指标以简化操作程序。 相似文献