湖北省典型花生种植区土壤中黄曲霉菌分布及产毒力研究
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摘要
为掌握湖北省花生种植区土壤中黄曲霉菌的分布和产毒特征,从罗田、红安、钟祥、襄阳四个典型花生种植区采集土壤样品40份,并进行黄曲霉菌分离、鉴定和产毒力研究。研究结果表明:湖北省不同花生种植区共分离鉴定到黄曲霉菌51株,土壤中黄曲霉菌落数为127.5cfu/g。不同种植区土壤中黄曲霉菌分布存在显著差异,钟祥土壤中黄曲霉菌落数最高,罗田最低;鉴定获得黄曲霉菌株中产毒菌株占96%,产毒量范围0～227.81μg/L,不产毒菌株占4%;产毒菌株分为只产AFB_1、产AFB_1+AFB_2、产AFB_1+AFB_2+AFG_1和产AFB_1+AFB_2+AFG_1+AFG_2毒素4种类型,其中以产AFB_1+AFB_2的菌株占比最高,为65%;不同种植区黄曲霉菌株产毒力研究发现,钟祥每克土壤中黄曲霉菌产AFB_1的量最高,达11 679.70μg/L。本研究可为湖北花生黄曲霉毒素污染预警和防控提供理论依据。
To study the occurence and distribution of aflatoxin by Aspergillus flavus in soils, samples from the typical peanut areas in Hubei Province were collected. In total 40 soil samples from Luotian, Hongan, Zhongxiang and Xiangyang were collected for A.flavus isolation, identification and aflatoxin detection. Results showed that 51 strains were isolated from the 4 areas. The population density of A.flavus was 127.5 cfu/g in one gram soil. But the population of A.flavus in soils of different peanut areas were significantly different. The highest populations of A.flavus colonies were found in Zhongxiang, and those from Luotian were the lowest. Among the strains, aflatoxigenic strains accounted for 96%, with AFT content of 0-227.81 μg/L, and atoxigenic strains was 4%. There were 4 types of aflatoxigenic strains, which were strains only producing AFB_1, strains producing AFB_1+AFB_2, strains producing AFB_1+AFB_2+AFG_1 and strains producing AFB_1+AFB_2+AFG_1+AFG_2. Among them, strains producing AFB_1+ AFB_2 accounted for the highest proportion,reaching 65%. Studies on aflatoxin production in different planting areas found that the highest amount of AFB_1 contamination per gram of soil was in Zhongxiang,reaching 11 679. 70μg/L. This study provided a theoretical base for predicting and controlling of aflatoxin contamination in typical peanut planting areas in Hubei Province.
To study the occurence and distribution of aflatoxin by Aspergillus flavus in soils, samples from the typical peanut areas in Hubei Province were collected. In total 40 soil samples from Luotian, Hongan, Zhongxiang and Xiangyang were collected for A.flavus isolation, identification and aflatoxin detection. Results showed that 51 strains were isolated from the 4 areas. The population density of A.flavus was 127.5 cfu/g in one gram soil. But the population of A.flavus in soils of different peanut areas were significantly different. The highest populations of A.flavus colonies were found in Zhongxiang, and those from Luotian were the lowest. Among the strains, aflatoxigenic strains accounted for 96%, with AFT content of 0-227.81 μg/L, and atoxigenic strains was 4%. There were 4 types of aflatoxigenic strains, which were strains only producing AFB_1, strains producing AFB_1+AFB_2, strains producing AFB_1+AFB_2+AFG_1 and strains producing AFB_1+AFB_2+AFG_1+AFG_2. Among them, strains producing AFB_1+ AFB_2 accounted for the highest proportion,reaching 65%. Studies on aflatoxin production in different planting areas found that the highest amount of AFB_1 contamination per gram of soil was in Zhongxiang,reaching 11 679. 70μg/L. This study provided a theoretical base for predicting and controlling of aflatoxin contamination in typical peanut planting areas in Hubei Province.
引文
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[24] Yin Y,Lou T,Yan L,et al.Molecular characterization of toxigenic and atoxigenic Aspergillus flavus isolates,collected from peanut fields in China[J].J Appl Microbiol,2009,107(6):1 857-1 865.
[25] Donner M,Atehnkeng J,Sikora R,et al.Molecular characterization of atoxigenic strains for biological control of aflatoxins in Nigeria[J].Food Add Contamin,2010,27(5):576-590.
[26] 陈茹,刘钟滨.黄曲霉菌aflR基因启动子序列变异与黄曲霉毒素产生相关联[J].细胞生物学杂志,2006,28(6):912-916.
[2] 都晓慧,丁小霞,周海燕,等.产后花生黄曲霉毒素污染监测抽样方法研究[J].中国油料作物学报,2015,37(6):876-880.
[3] 王后苗.花生抗黄曲霉菌产毒机制研究[D] .北京:中国农业科学院,2016.
[4] Blount W P,J Br.Turkey “X” disease[J].Turk Fed,1961,9:52-55.
[5] 王后苗,廖伯寿.农作物收获前黄曲霉毒素污染与控制措施[J].作物学报,2012,38(1):1-9.
[6] 武琳霞,都晓慧,丁小霞,等.花生黄曲霉毒素污染预警技术研究进展[J] .中国油料作物学报,2016,38(1):120-126.
[7] Klich M A.Aspergillus flavus:the major producer of aflatoxin[J].Mol Plant Pathol,2007,8(6):713-722.
[8] 姬宁.花生油中黄曲霉毒素B1碱炼脱毒及安全性评价[D] .泰安:山东农业大学,2015.
[9] 丁小霞.中国产后花生黄曲霉毒素污染与风险评估方法研究[D].北京:中国农业科学院,2011.
[10] Amaike S,Keller N P.Aspergillus flavus[J].Annu Rev Phytopathol,2011,49:107-133.
[11] 秦文彦.潜藏性产黄曲霉毒素真菌的多重PCR检测体系构建及真菌DNA提取技术的改进[D].杭州:浙江大学,2007.
[12] 戴显红.典型区果际土壤微生态环境对花生黄曲霉毒素污染影响的初步研究[D].北京:中国农业科学院,2017.
[13] Henry S H,Bosch F X.Aflatoxin,hepatitis and worldwide liver cancer risks[J].Adv Exp Med Biol,2002:504(48),229-233.
[14] 李秉鸿,李筠.去除饲料黄曲霉毒素的方法[J].中国饲料,1997,21:33-34.
[15] Ding X X,Wu L X,Li P W,et al.Risk assessment on dietary exposure to aflatoxin B1 in post-harvest peanuts in the Yangtze River Ecological Region[J].Toxins,2015,7:4 157-4 174.
[16] 2017中国统计年鉴[OL].http://www.stats.gov.cn/tjsj/ndsj/2017/indexch.htm.
[17] 杨生瑞.黄曲霉菌株分离、鉴定及产毒能力分析[J] .中国粮油学报,2015,27(6):110-114.
[18] 张初署.中国四个生态区花生土壤中黄曲霉菌分布、产毒特征及遗传多样性研究[D].北京:中国农业科学院,2013.
[19] Klich M A.Aspergillus flavus:The major producer of aflatoxin[J].Mol Plant Pathol,2007,8(6):713-722.
[20] Hedayati M T,Pasqualotto A C,Warn P A,et al.Aspergillus flavus:human pathogen,allergen and mycotoxin producer[J].Microbiology,2007,153:1 677-1 692.
[21] Zhang C S,Selvaraj J N,Yang Q L,et al.A survey of aflatoxin-producing Aspergillus sp.from peanut field soils in four agroecological zones of China[J].Toxins,2017,9(1):40.
[22] Bhatnagar D,Ehrlich K C,Cleveland T E,et al.Molecular genetic analysis and regulation of aflatoxin biosynthesis[J].Appl Microbiol Biotechnol,2003,61(2):83-93.
[23] Wei D D,Zhou L,Selvaraj J N,et al.Molecular characterization of atoxigenic Aspergillus flavus isolates collected in China[J].J Microbiol,2014,52(7):559-565.
[24] Yin Y,Lou T,Yan L,et al.Molecular characterization of toxigenic and atoxigenic Aspergillus flavus isolates,collected from peanut fields in China[J].J Appl Microbiol,2009,107(6):1 857-1 865.
[25] Donner M,Atehnkeng J,Sikora R,et al.Molecular characterization of atoxigenic strains for biological control of aflatoxins in Nigeria[J].Food Add Contamin,2010,27(5):576-590.
[26] 陈茹,刘钟滨.黄曲霉菌aflR基因启动子序列变异与黄曲霉毒素产生相关联[J].细胞生物学杂志,2006,28(6):912-916.