番茄交链孢病原菌鉴定及产毒分析

目的 通过对番茄病果中链格孢病原菌的分离鉴定及产毒分析,为番茄采收后病害防控提供理论依据。方法 采集福建省5个县市番茄病果,通过切片和组织分离法从番茄病害部位分离纯化病原菌,结合形态学特征和分子生物学对病原菌进行鉴定。根据柯赫氏法则确定病原菌的致病性,并利用超高效液相串联质谱法对分离得到的交链格孢菌产毒能力进行检测分析。结果 在发病番茄病害部位共分离得到的 3株互隔链格孢菌株均具产毒能力,但毒素种类和产毒能力各有不同,其中NH06菌株产毒量最高,可产生细交链格孢酮酸(tenuazonic acid,TeA )、交链孢酚 (alternariol, AOH)、交链孢酚单甲醚(alternariol monomethyl ether, AME)、交链孢烯(altenuene,ALT )、腾毒素 (tentoxin,TEN ) 5种毒素,而NP002和NH07仅检出产生4种毒素。结论 福建省番茄种植产区内存在交链格孢菌的污染,且产毒种类多,因此,在番茄种植产区内病果的处理应引起关注和重视。     

水果制品中真菌毒素污染及控制研究进展

真菌侵染是造成水果腐烂和水果及其制品中真菌毒素残留的根本原因。全球由于真菌侵染而腐烂的水果损失约占新鲜水果的20%~30%,而真菌毒素也成为危害人体健康的潜在风险。本文首先综述了水果制品中常见的6种真菌毒素的来源、稳定性及毒性,包括展青霉素、赭曲霉毒素A、黄曲霉毒素B1、交链孢酚、交链孢酚单甲醚、细交链孢菌酮酸、赭曲霉毒素A和黄曲霉毒素B1展青霉素,并分析了各种真菌毒素在不同种类水果制品中的污染状况。其次,重点从控制霉菌生长、产毒和毒素消减3个层面,综述了目前在水果采前、采后和加工3个环节真菌毒素的防控措施,并提出真菌毒素防控领域尚需进一步开展的研究,明确安全、无污染和精准的绿色防控手段为防控真菌毒素污染的发展方向,采取传统控制手段结合生物防控等新技术有望实现对真菌毒素的有效控制。     

臭氧处理对交链孢菌生长及其毒素积累的抑制作用

交链孢菌（Alternaria spp.）易侵染农作物,引起农产品病害,而且能够代谢产生交链孢毒素,包括细交链孢酮酸（tenuazonic acid,TeA）、交链孢酚（alternariol,AOH）、交链孢酚单甲醚（alternariol monomethyl ether,AME）等,严重影响人体健康。因此,亟需高效、安全的方法用以防控交链孢菌及其毒素积累。本实验研究了臭氧处理对体外互隔交链孢（A. alternata）生长及其产毒能力的影响。结果表明,臭氧处理组的菌落直径显著低于对照组（P＜0.05）,且臭氧处理可显著抑制互隔交链孢产生TeA、AOH、AME这3 种交链孢毒素;利用扫描电子显微镜观察臭氧处理后互隔交链孢的微观形态,发现孢子和菌丝发生了凹陷、褶皱、断裂等异常现象;臭氧处理后的交链孢菌对番茄果实的致病力明显减弱,同时交链孢菌的产毒能力明显降低,20 mg/L臭氧处理条件下TeA、AOH、AME含量比对照组分别减少了36.1%、89.9%、93.2%。此外,臭氧对TeA、AOH、AME具有降解作用,降解率随着臭氧质量浓度的增加和处理时间的延长而显著提高,TeA经过20 mg/L臭氧处理30 min即可被完全降解,AOH及AME经20 mg/L臭氧处理120 min后降解率可达90%以上。综上,臭氧处理可以作为农产品及其制品中互隔交链孢及其毒素污染的防治手段。     

农产品及其制品中交链孢酚和交链孢酚单甲醚研究进展

交链孢菌(Alternaria spp.)是一类重要的植物病原体,能产生多种毒素,其中交链孢酚(alternariol,AOH)和交链孢酚单甲醚(alternariol monomethyl ether,AME)是目前从食物中检出的比较普遍和主要的2种交链孢毒素,可广泛污染蔬菜、水果及谷物等农产品及其制品。本文对AOH和AME的结构及理化性质、毒性、产生影响因素、生物及化学合成和污染现状等进行了综述,并对交链孢毒素限量标准的制定、快速检测方法等进行了展望。     

光照对交链孢毒素合成的调控

目的 明确光调控交链孢（Alternaria）产真菌毒素的作用。方法 在光/暗培养条件下,比较分析光照对2种交链孢菌株（ATCC 66981和Pear-3）的菌丝生长、孢子形成以及毒素合成的影响。结果 持续白光照射对交链孢菌丝生长影响不显著,而显著抑制其孢子形成。光照刺激交链孢菌ATCC 66981中交链孢酚（alternariol, AOH）、交链孢酚单甲醚（alternariol monomethyl ether, AME）和腾毒素（tentoxin, TEN）3种毒素的产生。菌株持续光照培养30天,AOH、AME和TEN毒素浓度分别为120、182和173 g/L;较之黑暗培养,分别增加了1.1、5.9和9.6倍。光照抑制交链孢菌Pear-3中细交链孢菌酮酸（tenuazonic acid, TeA）毒素的合成。菌株光照培养6天,TeA毒素浓度即已达142 g/L;而黑暗培养甚至高达325 g/L,毒素产率反而增加了1.3倍。结论 光照不影响交链孢菌丝生长,而对孢子形成影响显著;持续白光照射能够促进交链孢中AOH、AME和TEN毒素的产生,而抑制TeA毒素的产生。     

白光光照对交链孢毒素合成的调控研究

目的研究白光光照对交链孢(Alternaria)毒素合成的影响。方法在光/暗培养条件下,比较分析白光光照对2种交链孢菌株(ATCC 66981和Pear-3)的菌丝生长、孢子形成以及毒素合成的影响。结果持续白光照射对交链孢菌丝生长影响不显著,而显著抑制其孢子形成。光照刺激交链孢菌ATCC66981中交链孢酚(alternariol, AOH)、交链孢酚单甲醚(alternariol monomethyl ether, AME)和腾毒素(tentoxin, TEN)3种毒素的产生。菌株持续光照培养30d,AOH、AME和TEN毒素浓度分别为120、182和173?g/L;较之黑暗培养,分别增加了1.1、5.9和9.6倍。光照抑制交链孢菌Pear-3中细交链孢菌酮酸(tenuazonic acid, TeA)毒素的合成。菌株光照培养6 d, TeA毒素浓度达到142?g/L;而黑暗培养后其浓度高达325?g/L,毒素产率增加了1.3倍。结论光照不影响交链孢菌丝生长,而对孢子形成影响显著;持续白光照射能够促进交链孢中AOH、AME和TEN毒素的产生,而抑制TeA毒素的产生。     

重要产毒镰刀菌合成白僵菌素和恩镰孢菌素研究进展

白僵菌素(beauvericin,BEA)和恩镰孢菌素(enniatins,ENNs)是由镰刀菌属的多种真菌产生的六酯肽类真菌毒素,该类毒素对上皮细胞、免疫细胞、卵巢细胞等具有很强的毒性作用。本文主要针对能产生BEA和ENNs的重要产毒镰刀菌的形态学特征、分子遗传学特征以及影响BEA和ENNs产生的环境条件如温度、底物等因素进行概述。重点阐述了产毒镰刀菌在两类毒素合成酶的基因水平和氨基酸水平的差异及影响产毒等主要因素,为BEA和ENNs及其产毒镰刀菌的预防和控制提供理论依据。     

继代培养对链格孢菌生长及产毒的影响

摘 要：目的 探究连续继代培养对链格孢菌(Alternaria alternata)菌株的生长以及产毒的影响。方法 将10株链格孢菌在马铃薯葡萄糖琼脂培养基(potato dextrose agar，PDA)上连续继代培养，分析菌株的生长、产孢情况；利用超高效液相色谱-串联质谱法(ultra performance liquid chromatography-tandem mass spectrometry，UPLC-MS/MS)检测并分析主要链格孢霉毒素(包括交链孢酚(alternariol，AOH)、交链孢酚单甲醚(alternariol monomethyl ether，AME)、交链孢烯(altenuene，ALT)、细交链格孢菌酮酸(tenuazonicacid，TeA))的含量变化。结果 在PDA培养基中，连续继代培养链格孢菌极易出现角变、菌丝变白、菌丝生长速率加快等菌株退化现象。随着继代培养次数的增加，菌株生长速度加快，菌落直径变大，角变率增大；10株链格孢菌中有7株(MY-4、MY-12、MY-15、MY-73、MY-80、MY-103、MY-117)产孢量随着继代次数的增加先增加后降低，3株(MY-65、MY-67和MY-108)产孢量随着继代次数的增加显著降低；10株链格孢菌中有4株可以产生TeA毒素，且TeA毒素的含量随着继代次数的增加先上升后下降，第五代菌株(F5)产毒能力最强。继代培养对ALT毒素含量影响较大，除MY-103以外，随着继代次数的增加，ALT毒素的含量显著降低(达91.13%~99.85%)。AOH和AME毒素含量随着继代次数的增加呈下降或者呈先上升后下降的趋势。结论 连续继代培养5代对链格孢菌的生长、产孢、产毒等均会产生较大影响。在菌株的实际应用过程中，应该避免菌株过多次的继代培养，尽量使用5代以内的链格孢菌。     

番茄中交链孢菌及其产毒的防治技术研究进展

交链孢属真菌是导致番茄采后病害的主要病原菌,腐烂的番茄不仅造成巨大的经济损失,而且会在其腐烂部位及其周围健康组织中积累大量的真菌毒素。目前加工工艺的限制导致番茄制品也可能受到真菌毒素的污染,进而对人类和动物健康造成潜在的威胁(包括致癌、致畸、致突变等作用)。因此,本文综述了国内外番茄及其制品中交链孢毒素的污染情况,以及交链孢菌及其毒素污染防治技术的研究进展,以期为降低番茄及其制品的风险隐患提供参考。     

小麦中呕吐毒素研究进展

小麦是全球三大谷物之一。由镰刀菌引起的小麦赤霉病是我国最主要的小麦病害之一,能造成小麦严重减产、品质下降,而且镰刀菌还能产生多种真菌毒素,进一步危害小麦及其制品的质量安全,其中,呕吐毒素(deoxynivalenol, DON)是小麦中检出率最高、危害最严重的真菌毒素之一,已成为关系小麦及其制品食用安全的重要问题。DON在小麦等谷物及其制品中的污染和防控是全球面临的严峻挑战,对DON进行定期检测和污染分析,研究高效、安全的降解技术,确保消费者安全,已成为目前世界各国政府和公众广泛关注和高度重视的热点问题。本文综述了DON的理化性质及毒性、影响产生的因素、污染现状及防控与脱毒方面的研究进展情况,以期为小麦中DON的风险评估及防控策略的制定提供有利参考。     

Toxigenic Alternaria species from Argentinean blueberries

Blueberries are traditionally consumed in North America, some European countries and Japan. In Argentina, the blueberry crop is profitable because production starts in November, when the northern hemisphere lacks fresh fruit. Fungal contaminants can grow and produce mycotoxins in fresh fruit. The aims of this work were to identify the main genera of the mycobiota of blueberries grown in Argentina and to determine the toxicogenic potential, pathogenicity and host specificity of the species isolated. The genus Alternaria was the main component of the blueberry mycobiota (95%); minor proportions of Phoma spp. (4%) and Penicillium spp. (1%) were also isolated. According to their sporulation patterns, 127 Alternaria isolates belonged to the Alternaria tenuissima species-group, 5 to the Alternaria alternata species-group and 2 to the Alternaria arborescens species-group. The last mentioned species-group was not isolated at 5°C. Of the 134 isolates, 61% were toxicogenic in autoclaved rice; 97% of these produced alternariol (AOH) in a range from 0.14 to 119.18 mg/kg, 95% produced alternariol methylether (AME) in a range from 1.23 to 901.74 mg/kg and 65% produced tenuazonic acid (TA) in a range from 0.13 to 2778 mg/kg. Fifty two isolates co-produced the three mycotoxins. According to the size of the lesion that they caused on blueberries, the isolates were classified as slightly pathogenic, moderately pathogenic and very pathogenic. No significant differences in pathogenicity were found on different blueberry varieties. In this work, high incidence and toxicogenic potential of the Alternaria isolates from blueberries were demonstrated. Thus, more studies should be done to evaluate the health risk posed by the presence of the Alternaria toxins in blueberries and in the manufactured by-products.     

Mycotoxin production by Alternaria strains isolated from Argentinean wheat

The toxigenic potential of Alternaria strains isolated from Argentinean wheat was investigated. A total of 123 strains were assayed for the production of tenuazonic acid (TA), alternariol (AOH) and alternariol monomethyl ether (AME). All but one of the isolates were able to produce at least one of the three mycotoxins. TA was produced by 72% of the strains (1-14782 mg/kg), AOH by 87% (4-622 mg/kg) and AME by 91% (7-2625 mg/kg). The average level of TA detected for all strains (1757 mg/kg) was higher than the average level of both alternariols (162 mg/kg for AOH and 620 mg/kg for AME). TA was the toxin produced at the highest concentration but in lower frequency. Most of the strains were able to synthesize more than one toxin: 74 isolates (60%) were positive for all three toxins, 30 (24%) for both AOH and AME, 5 (4%) for both TA and AME, and 2 (2%) for TA and AOH. The widespread occurrence of Alternaria in wheat and its ability to produce mycotoxins suggests the possible occurrence of its toxins in wheat naturally infected with this fungus.     

Toxigenic profile of Alternaria alternata and Alternaria radicina occurring on umbelliferous plants.

Alternaria alternata and Alternaria radicina are fungal species that occur in several food crops and may produce mycotoxins and phytotoxins. The toxigenic profile of A. alternata and A. radicina isolated from carrot and other umbelliferous plants was determined by growing the fungus on rice and carrot discs. Most of the tested isolates of A. alternata produced the mycotoxins tenuazonic acid, alternariol, alternariol methyl ether and altertoxin-I on rice. Only alternariol and alternariol methyl ether were produced on carrot discs. When cultured on rice, none of the isolates of A. alternata from umbelliferous plants produced AAL toxins and fumonisins. AAL toxins, but not fumonisins, were instead produced by A. alternata f. sp. lycopersici isolate NRRL 18822 isolated from tomato. A. radicina produced the phytotoxic compounds radicinin, epi-radicinol and radicinol on carrot discs, whereas it produced radicinin and radicinol on rice. Although A. alternata has been frequently found in organic carrots, none of the above mycotoxins was detected in carrot roots or in carrot commercial products. The reduction of alternariol and alternariol methyl ether during carrot juice processing at laboratory scale was estimated to be >98%. Based on these findings and previous reports, it can be concluded that Alternaria mycotoxins in carrots do not represent a hazard for consumers.     

Alternaria toxins in wheat during the 2004 to 2005 Argentinean harvest

The natural occurrence of Alternaria mycotoxins in Argentinean wheat from the zone 5 South during the 2004 to 2005 harvest was investigated in 64 wheat samples. All samples were highly contaminated with a wide range of fungal species. Alternaria was found as the main component of the mycota, with an infection percentage of 100%. Three mycotoxins produced by species of Alternaria were determined in wheat: alternariol, alternariol monomethyl ether, and tenuazonic acid. Alternariol was detected in 4 (6%) of 64 samples, with a range of 645 to 1,388 microg/kg (mean of 1,054 microg/kg); alternariol monomethyl ether, with a range of 566 to 7,451 microg/kg (mean of 2,118 microg/kg) in 15 (23%) of 64 samples; and tenuazonic acid in 12 (19%) of 64 samples, with a range of 1,001 to 8,814 microg/kg (mean, 2,313 microg/kg). Alternariol monomethyl ether was the predominant toxin, but tenuazonic acid was detected in higher concentrations. Alternariol was present in fewer samples and in lower levels than were the other toxins. Tenuazonic acid and alternariol monomethyl ether occurred together in four samples, while tenuazonic acid and alternariol co-occurred in one sample. This the first report of the natural occurrence of Alternaria mycotoxins in Argentinean wheat. Toxin levels were high, probably due to the heavy infection with Alternaria species found in the samples.     

链格孢霉毒素检测方法研究进展

链格孢霉菌属于丝状真菌,是一种普遍存在于环境中的病原体和腐生菌,是低温环境下导致水果、蔬菜等农产品腐烂变质的主要微生物。链格孢霉毒素中有70多种具有明显毒性,对人或牲畜具有诱变性、致癌性和致畸性等慢性或急性毒性作用,因此引起了科学家们的广泛关注。但到目前为止,国内外现行的食品中真菌毒素的限量标准中尚不包括链格孢霉毒素,并且通常采用液相、液相-质谱、气相、气相-质谱等相关的检测方法进行检测。本文首先对链格孢霉菌的3种主要的次级代谢产物——链格孢酚、链格孢酚单甲醚和细交链孢菌酮酸的分类及毒性性质进行介绍;其次,对其主要的传统检测方法和快速检测方法进行综述;最后,对链格孢霉毒素检测的新技术新方法进行了展望。     

Epidemiology of mycotoxigenic fungi associated with Fusarium ear blight and apple blue mould: A review

Research on the epidemiology of plant diseases is briefly reviewed, focusing on two diseases caused by mycotoxin-producing pathogens: Fusarium ear (or head) blight (FEB or FHB) of small grain cereals and apple blue mould (Penicillium expansum). Pathogen development during its key life cycles is discussed in relation to important environmental factors and host resistance. Current control methods are also reviewed, focusing on cultural and biological methods. The future challenge is to understand the relationships between disease severity, fungal biomass and the production of associated mycotoxins in order to minimize risks of both disease damage to crop yields and threat to human health posed by mycotoxins.