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Article(id=1153429494040879618, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1153429493357203682, articleNumber=null, orderNo=null, doi=10.19812/j.cnki.jfsq11-5956/ts.20240627002, pmid=null, cstr=null, oa=null, hot=null, price=null, onlineType=0, articleFormat=0, articleType=null, articleTypeStr=null, receivedDate=1719417600000, receivedDateStr=2024-06-27, revisedDate=null, revisedDateStr=null, acceptedDate=null, acceptedDateStr=null, onlineDate=1752928621062, onlineDateStr=2025-07-19, pubDate=1741968000000, pubDateStr=2025-03-15, doiRegisterDate=null, doiRegisterDateStr=null, onlineIssueDate=1752928621062, onlineIssueDateStr=2025-07-19, onlineJustAcceptDate=null, onlineJustAcceptDateStr=null, onlineFirstDate=null, onlineFirstDateStr=null, sourceXml=null, magXml=null, createTime=1752928621062, creator=13701087609, updateTime=1752928621062, updator=13701087609, issue=Issue{id=1153429493357203682, tenantId=1146029695717560320, journalId=1149652044408987649, year='2025', volume='16', issue='5', pageStart='1', pageEnd='326', issueExtLink='null', onlineDate='null', pubDate='null', beforeIssueId=null, nextIssueId=null, price=null, status=1, issueComplete=1, articleOrder=1, issueType=-1, specialIssue=0, createTime=1752928620900, creator=13701087609, updateTime=1758690311058, updator=13701087609, preIssue=null, nextIssue=null, ext={EN=IssueExt(id=1177595773500932351, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1153429493357203682, language=EN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=), CN=IssueExt(id=1177595773500932352, tenantId=1146029695717560320, journalId=1149652044408987649, issueId=1153429493357203682, language=CN, specialIssueTitle=, coverIllustrator=null, specialIssueEditor=, specialIssueAbout=)}, issueFiles=null}, startPage=232, endPage=238, ext={EN=ArticleExt(id=1153429494414172676, articleId=1153429494040879618, tenantId=1146029695717560320, journalId=1149652044408987649, language=EN, title=Effects of storage methods on the quality safety of high-moisture harvested Zea mays L., columnId=1151895321388347923, journalTitle=Journal of Food Safety & Quality, columnName=Food Analysis and Detection, runingTitle=null, highlight=null, articleAbstract=

Objective To investigate the effects of different storage methods on the quality safety of high-moisture harvested Zea mays L.. Methods The Zhengdan 958 (ZD958) Zea mays L. variety was selected for the experiment, and the Zea mays L. harvested under the same moisture content was stored as ear Zea mays L. and grain Zea mays L.. Changes in fungi species, quantity, and mycotoxin content were measured. Results Predominant fungi during Zea mays L. storage were Fusarium, Aspergillus, and Penicillium. In the early stage of storage, the dominant fungal genus was Fusarium, and with the extension of storage time, the dominant fungal genera changed to Aspergillus and Penicillium; with the increase of storage days, the total number of fungi in Zea mays L. stored in ear and grain storage showed a trend of first increasing and then decreasing. The number of fungi in Zea mays L. stored in ears was significantly lower than that in Zea mays L. stored in grains (P<0.05). During the storage period of 10-100 days, the number of fungi in Zea mays L. stored in ears decreased by 90.3%-98.6% compared to Zea mays L. stored in grains; with the increase of storage days, except for gibberellin in Zea mays L. kernels stored in ear, the content of vomitoxin and gibberellin in Zea mays L. showed an increasing trend. The content of vomitoxin and gibberellin in Zea mays L. stored in ears was significantly lower than that in Zea mays L. stored in grains (P<0.05). And with the increase of storage days, the difference in the content of vomitoxin and gibberellin in Zea mays L. under the two storage methods increases. Conclusion After high-moisture harvesting, ear storage can reduce fungi quantity and mycotoxin content, ensuring the quality safety of Zea mays L..

, correspAuthors=Kui-Jie GONG, authorNote=null, correspAuthorsNote=null, copyrightStatement=null, copyrightOwner=null, extLink=null, articleAbsUrl=null, sourceXml=null, magXml=null, pdfUrl=null, pdf=null, pdfFileSize=null, pdfExtLink=null, richHtmlUrl=null, mobilePdfUrl=null, reviewReport=null, pdfFirstPage=null, abstractGraph=null, abstractGraphContent=null, abstractVideo=null, citation=null, cebUrl=null, magXmlContent=null, mapNumber=null, authorCompany=null, fund=null, authors=null, authorsList=Xing-Ya WANG, Shou-Mei ZHANG, Meng-Wen ZHANG, Li-Rong CHEN, Lin-Lin SUN, Yu-Qiu GUO, Kui-Jie GONG), CN=ArticleExt(id=1153429510096675481, articleId=1153429494040879618, tenantId=1146029695717560320, journalId=1149652044408987649, language=CN, title=储藏方法对高水分收获玉米质量安全的影响, columnId=1151895321958773274, journalTitle=食品安全质量检测学报, columnName=食品分析与检测, runingTitle=null, highlight=null, articleAbstract=

目的 探索不同储藏方法对高水分收获玉米质量安全的影响。方法 选用郑单958 (ZD958)玉米为试验材料, 将同一水分下收获的玉米进行果穗储藏和籽粒储藏, 测定其真菌种类及数量、毒素含量的变化。结果 玉米储藏期间的优势真菌属为镰刀菌属(Fusarium)、曲霉属(Aspergillus)和青霉属(Penicillium), 其中储藏前期优势真菌属为镰刀菌属, 随着储藏时间的延长, 优势真菌属改变为曲霉属和青霉属; 随着储藏天数的增多, 果穗储藏和籽粒储藏玉米的真菌总数量均呈先增加后降低的趋势。果穗储藏玉米的真菌数量显著低于籽粒储藏玉米(P<0.05)。储藏10~100 d期间, 果穗储藏玉米的真菌数量比籽粒储藏玉米减少了90.3%~98.6%; 随着储藏天数的增多, 除果穗储藏玉米籽粒的赤霉烯酮外, 玉米的呕吐毒素和赤霉烯酮含量均呈现增加的趋势。果穗储藏玉米的呕吐毒素和赤霉烯酮含量显著低于籽粒储藏玉米(P<0.05)。且随着储藏天数的增多, 两种储藏方法下玉米的呕吐毒素和赤霉烯酮含量差异加大。结论 高水分收获玉米后, 果穗储藏能够降低玉米的真菌数量和毒素含量, 保证其质量安全。

, correspAuthors=龚魁杰, authorNote=null, correspAuthorsNote=
* 龚魁杰(1970—), 男, 博士, 研究员, 主要研究方向为谷物营养与质量安全。E-mail:
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王兴亚(1990—), 女, 博士, 助理研究员, 主要研究方向为谷物营养与质量安全。E-mail:

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王兴亚(1990—), 女, 博士, 助理研究员, 主要研究方向为谷物营养与质量安全。E-mail:

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王兴亚(1990—), 女, 博士, 助理研究员, 主要研究方向为谷物营养与质量安全。E-mail:

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注: 不同小写字母表示组间具有显著性差异, P<0.05。

, figureFileSmall=LahtwYI/jCy2pU1QYuBrUw==, figureFileBig=dL00L4YXMn7741jsYOGLgg==, tableContent=null), ArticleFig(id=1177619442902123203, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153429494040879618, language=EN, label=Table 1, caption=

Changes in dominant fungal species and quantities under different storage methods

, figureFileSmall=null, figureFileBig=null, tableContent=
储藏方法 储藏天数/d 真菌数量/(104 CFU/g)
镰刀菌属 曲霉属 青霉属 根霉属 链格孢属 篮状菌属 总计
果穗储藏 0 20b 5b 10a 0a 0b 0a 35b
10 15b 5b 0c 0a 5a 0a 25b
20 30a 10a 10a 0a 0b 0a 50a
30 10c 10a 10a 0a 5a 0a 35b
70 0d 5b 5b 0a 0b 0a 10c
100 0d 5b 5b 0a 0b 0a 10c
籽粒储藏 0 20f 5f 10e 0e 0e 0c 35e
10 165c 105e 85d 5d 15c 0c 375d
20 240b 160c 90d 0e 25b 0c 515c
30 380a 600a 410a 50a 35a 25a 1500a
70 110d 295b 225b 40b 20b 10b 700b
100 75e 130d 100c 25c 5d 15b 350d
), ArticleFig(id=1177619443090866884, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153429494040879618, language=CN, label=表1, caption=

不同储藏方法下优势真菌及真菌数量变化

, figureFileSmall=null, figureFileBig=null, tableContent=
储藏方法 储藏天数/d 真菌数量/(104 CFU/g)
镰刀菌属 曲霉属 青霉属 根霉属 链格孢属 篮状菌属 总计
果穗储藏 0 20b 5b 10a 0a 0b 0a 35b
10 15b 5b 0c 0a 5a 0a 25b
20 30a 10a 10a 0a 0b 0a 50a
30 10c 10a 10a 0a 5a 0a 35b
70 0d 5b 5b 0a 0b 0a 10c
100 0d 5b 5b 0a 0b 0a 10c
籽粒储藏 0 20f 5f 10e 0e 0e 0c 35e
10 165c 105e 85d 5d 15c 0c 375d
20 240b 160c 90d 0e 25b 0c 515c
30 380a 600a 410a 50a 35a 25a 1500a
70 110d 295b 225b 40b 20b 10b 700b
100 75e 130d 100c 25c 5d 15b 350d
), ArticleFig(id=1177619443321553605, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153429494040879618, language=EN, label=Table 2, caption=

Correlation analysis between major toxigenic fungi and mycotoxins

, figureFileSmall=null, figureFileBig=null, tableContent=
真菌数量 毒素含量
呕吐毒素 黄曲霉毒素B1 赤霉烯酮
镰刀菌属 0.995** - 0.996**
黄曲霉菌 - -0.351 -
), ArticleFig(id=1177619443443188422, tenantId=1146029695717560320, journalId=1149652044408987649, articleId=1153429494040879618, language=CN, label=表2, caption=

主要产毒真菌与真菌毒素相关性分析

, figureFileSmall=null, figureFileBig=null, tableContent=
真菌数量 毒素含量
呕吐毒素 黄曲霉毒素B1 赤霉烯酮
镰刀菌属 0.995** - 0.996**
黄曲霉菌 - -0.351 -
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储藏方法对高水分收获玉米质量安全的影响
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王兴亚 1 , 张守梅 1 , 张孟雯 2 , 陈利容 1 , 孙琳琳 1 , 郭玉秋 1 , 龚魁杰 1, *
食品安全质量检测学报 | 食品分析与检测 2025,16(5): 232-238
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食品安全质量检测学报 | 食品分析与检测 2025, 16(5): 232-238
储藏方法对高水分收获玉米质量安全的影响
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王兴亚1 , 张守梅1, 张孟雯2, 陈利容1, 孙琳琳1, 郭玉秋1, 龚魁杰1, *
作者信息
  • 1.山东省农业科学院作物研究所, 济南 250100
  • 2.中国农业大学农学院, 北京 100193

    • 王兴亚(1990—), 女, 博士, 助理研究员, 主要研究方向为谷物营养与质量安全。E-mail:

通讯作者:

* 龚魁杰(1970—), 男, 博士, 研究员, 主要研究方向为谷物营养与质量安全。E-mail:
Effects of storage methods on the quality safety of high-moisture harvested Zea mays L.
Xing-Ya WANG1 , Shou-Mei ZHANG1, Meng-Wen ZHANG2, Li-Rong CHEN1, Lin-Lin SUN1, Yu-Qiu GUO1, Kui-Jie GONG1, *
Affiliations
  • 1. Crop Research Institute, Shandong Academy of Agricultural Sciences, Ji'nan 250100, China
  • 2. College of Agronomy and Biotechnology, China Agricultural University, Beijing 100193, China
出版时间: 2025-03-15 doi: 10.19812/j.cnki.jfsq11-5956/ts.20240627002
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目的 探索不同储藏方法对高水分收获玉米质量安全的影响。方法 选用郑单958 (ZD958)玉米为试验材料, 将同一水分下收获的玉米进行果穗储藏和籽粒储藏, 测定其真菌种类及数量、毒素含量的变化。结果 玉米储藏期间的优势真菌属为镰刀菌属(Fusarium)、曲霉属(Aspergillus)和青霉属(Penicillium), 其中储藏前期优势真菌属为镰刀菌属, 随着储藏时间的延长, 优势真菌属改变为曲霉属和青霉属; 随着储藏天数的增多, 果穗储藏和籽粒储藏玉米的真菌总数量均呈先增加后降低的趋势。果穗储藏玉米的真菌数量显著低于籽粒储藏玉米(P<0.05)。储藏10~100 d期间, 果穗储藏玉米的真菌数量比籽粒储藏玉米减少了90.3%~98.6%; 随着储藏天数的增多, 除果穗储藏玉米籽粒的赤霉烯酮外, 玉米的呕吐毒素和赤霉烯酮含量均呈现增加的趋势。果穗储藏玉米的呕吐毒素和赤霉烯酮含量显著低于籽粒储藏玉米(P<0.05)。且随着储藏天数的增多, 两种储藏方法下玉米的呕吐毒素和赤霉烯酮含量差异加大。结论 高水分收获玉米后, 果穗储藏能够降低玉米的真菌数量和毒素含量, 保证其质量安全。

玉米  /  储藏方法  /  真菌毒素  /  质量安全

Objective To investigate the effects of different storage methods on the quality safety of high-moisture harvested Zea mays L.. Methods The Zhengdan 958 (ZD958) Zea mays L. variety was selected for the experiment, and the Zea mays L. harvested under the same moisture content was stored as ear Zea mays L. and grain Zea mays L.. Changes in fungi species, quantity, and mycotoxin content were measured. Results Predominant fungi during Zea mays L. storage were Fusarium, Aspergillus, and Penicillium. In the early stage of storage, the dominant fungal genus was Fusarium, and with the extension of storage time, the dominant fungal genera changed to Aspergillus and Penicillium; with the increase of storage days, the total number of fungi in Zea mays L. stored in ear and grain storage showed a trend of first increasing and then decreasing. The number of fungi in Zea mays L. stored in ears was significantly lower than that in Zea mays L. stored in grains (P<0.05). During the storage period of 10-100 days, the number of fungi in Zea mays L. stored in ears decreased by 90.3%-98.6% compared to Zea mays L. stored in grains; with the increase of storage days, except for gibberellin in Zea mays L. kernels stored in ear, the content of vomitoxin and gibberellin in Zea mays L. showed an increasing trend. The content of vomitoxin and gibberellin in Zea mays L. stored in ears was significantly lower than that in Zea mays L. stored in grains (P<0.05). And with the increase of storage days, the difference in the content of vomitoxin and gibberellin in Zea mays L. under the two storage methods increases. Conclusion After high-moisture harvesting, ear storage can reduce fungi quantity and mycotoxin content, ensuring the quality safety of Zea mays L..

Zea mays L.  /  storage methods  /  mycotoxin  /  quality safety
王兴亚, 张守梅, 张孟雯, 陈利容, 孙琳琳, 郭玉秋, 龚魁杰. 储藏方法对高水分收获玉米质量安全的影响. 食品安全质量检测学报, 2025 , 16 (5) : 232 -238 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20240627002
Xing-Ya WANG, Shou-Mei ZHANG, Meng-Wen ZHANG, Li-Rong CHEN, Lin-Lin SUN, Yu-Qiu GUO, Kui-Jie GONG. Effects of storage methods on the quality safety of high-moisture harvested Zea mays L.[J]. Journal of Food Safety & Quality, 2025 , 16 (5) : 232 -238 . DOI: 10.19812/j.cnki.jfsq11-5956/ts.20240627002
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0 引言
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玉米作为全球最重要的粮食作物之一, 其质量安全对全球粮食供应和人类饮食健康至关重要[1-2]。在中国, 玉米收获时的籽粒含水量一般在30%~40%之间, 高水分玉米收获后的储藏问题一直是制约玉米质量安全的关键因素[3]。以往我国小农户玉米收获后主要采用露天果穗储藏的方法进行储藏, 但近年来随着机械化粒收的推进, 高水分玉米收粒后直接储藏的比例将会增加[4-5]。因此, 研究不同储藏方法对质量安全的影响具有重要意义。
以往对于玉米储藏的研究多集中在储藏技术的优化、储藏环境的控制以及储藏期间品质的变化等方面[6-8]。研究表明, 使用新型的储粮技术, 比如低温储藏、气调储藏等, 配合适宜的环境条件, 能延长玉米的保质期, 减少储藏过程中的损耗[9-10]。但目前的研究尚缺乏对玉米果穗储藏和籽粒储藏两种不同储藏方法的深入探讨。现有的研究表明, 玉米采用果穗储藏, 穗轴内的营养物质可以继续运送到籽粒, 增加玉米粒的饱满程度, 促进籽粒后熟。并且, 果穗储藏孔隙度较大, 空气流通性好, 水分降低较快, 能够减少霉变和虫害的发生[11-12]。而籽粒储藏节省储藏空间, 但是容易呼吸作用放热, 引起粮堆内部温度升高, 加剧玉米的霉变和腐烂[13-14]。但是, 以往的研究并未对玉米果穗储藏和籽粒储藏两种储藏方法下的质量安全进行深入、量化的探究, 并且缺乏高水分收获条件下的研究, 需要进一步的探讨。
因此, 鉴于中国玉米生产普遍面临着高水分收获与储藏的挑战, 本研究模拟中国小农户对高水分玉米收获后的果穗储藏和籽粒储藏的方法, 重点研究两种不同储藏方法下玉米质量安全的相关指标, 分析不同储藏方法下玉米质量安全的变化。此研究不仅可以丰富玉米储藏科学的理论体系, 更为中国广大小农户提供科学储粮的参考依据, 对保障粮食安全和促进农业可持续发展具有重要意义。
1 材料与方法
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1.1 材料
储藏玉米品种选用郑单958 (ZD958), 于2022年玉米季在山东省农业科学院枣园试验基地(117.5°E, 36.7°N)种植。播种日期为6月20日, 收获日期为10月18日。收获后的高水分玉米(籽粒含水量为31.1%)放置于塑料网袋中, 采用果穗和籽粒两种方法进行储藏。试验共有2个处理, 每个处理设置3次重复。分别在储藏0、10、20、30、70和100 d取样。每个重复采集2份样品, 采集的样品全部放入无菌自封袋。其中一份样品用于进行真菌的分离和鉴定, 另外一份样品在75 ℃烘箱中烘至恒重, 利用小型磨粉机磨样后测定玉米呕吐毒素、黄曲霉毒素B1和赤霉烯酮的含量。
1.2 仪器设备与试剂
HNY-100B恒温振荡培养器(天津欧诺仪器股份有限公司); SW-CJ-2FD双人单面净化工作台(苏州净化设备有限公司); IX51倒置显微镜(日本OLYMPUS公司); GENIOS Plus酶标仪(瑞士TECAN公司); LDZX-50KBS立式高压灭菌器(上海申安医疗器械厂); DTC-100恒温金属浴(杭州米欧仪器有限公司); Sorval ST8R台式冷冻离心机(美国Thermo Fisher Scientific公司); 960型聚合酶链式反应(polymerase chain reaction, PCR)仪(杭州晶格科学仪器有限公司); PowerPac Basic电泳仪(伯乐生命医学产品有限公司); XW-80A旋涡混合仪(宁波新芝生物科技股份有限公司); DX-55不锈钢磨粉机(广州市大祥电子机械设备有限公司); GL124-1SCN万分之一电子天平(德国Sartorius公司); GZX-9240MBE电热鼓风干燥箱(上海博迅仪器有限公司)。
马铃薯葡萄糖琼脂(potato dextrose agar, PDA)培养基(青岛海博生物技术有限公司); 青链霉素(青霉素1000 mg/mL; 链霉素10 mg/mL)(以色列Biological Industries公司); 2×Taq DNA Master Mix、ITS通用引物、DNA Marker、6×DNA Loading Buffer、DNA Dye 10000(北京擎科生物技术有限公司); 琼脂糖(美国Thermo Fisher Scientific公司); 甲醇(南京晶格化学科技有限公司); 磷酸二氢钠、乙二胺四乙酸、甘油(上海阿拉丁生化科技股份有限公司); 毒素酶联免疫检测试剂盒(北京华安麦科生物技术有限公司)。
1.3 方法
1.3.1 真菌的分离和纯化
称取10 g样品放入装有90 mL无菌水的三角瓶中, 密封后置于恒温振荡培养器中以150 r/min的速度振荡20 min, 随后静置5 min。对上清液进行系列浓度梯度稀释。依次制备10-2、10-3和10-4浓度的稀释液。稀释完成后, 用移液枪准确吸取100 μL浓度为10-3和10-4的稀释液涂布于含有青链霉素的PDA培养基上, 每个浓度梯度重复3次。培养3~5 d后进行计数, 最终确定真菌数目。
观察菌落形态, 将不同菌落形态的真菌纯化于PDA培养基上, 在室温下培养3~5 d, 观察菌落大小、生长速度、颜色、纹理形态、菌丝状态和可溶性色素的产生情况等。挑取少量菌丝, 在光学显微镜下观察菌丝和孢子形态。
1.3.2 真菌的分子生物学鉴定
根据真菌形态学观察结果, 对不同形态的菌株编号, 并进行真菌分子生物学鉴定。步骤如下:
(1)真菌DNA组提取
采用热裂解法进行。首先, 用无菌接种针挑选微量菌丝放入含有100 μL超纯水的1.5 mL离心管中, 涡旋振荡1 min使其充分混合。随后, 以10000 r/min的转速离心30 s, 去除上清液。向离心管中加入100 μL裂解液(50 mmol/L的磷酸二氢钠, 1 mmol/L的乙二胺四乙酸和5%甘油, 调节pH到7.4, 使用前121 ℃高压蒸汽灭菌20 min, 4 ℃储藏备用), 于85 ℃的金属浴中加热20~30 min, 之后12000 r/min离心10 s, 所得产物即为真菌DNA组[15]。
(2)核糖体DNA中ITS序列扩增
采用基于18S核糖体DNA的ITS通用引物进行DNA片段扩增[16]。设置通用引物ITS1 (5’-TCCGTAGGTGAACCTGCGG-3’)和ITS4 (5’-TCCTCCGCTTATTGATATGC-3’)。反应体系为25 μL, 其中2×Taq DNA Master Mix 12.5 μL, 模板DNA 1 μL, ITS1 1 μL, ITS4 1 μL, 用超纯水补足体系。反应条件设置为: 首先在95 ℃下预变性3 min; 随后进行35个循环, 每一循环均遵循以下步骤: 第一, 在94 ℃下进行DNA变性, 持续1 min; 第二, 温度降至55 ℃以实现引物与模板的退火, 持续1 min; 第三, 在72 ℃下进行DNA链的延伸反应, 持续1 min; 最后, 完成所有预设循环后, 进行一次终末延伸, 即在72 ℃下延伸10 min。
(3) PCR产物验证
使用1%的琼脂糖凝胶电泳检测是否出现预期目标条带。随后, 将提取成功的真菌DNA组送至青岛擎科测序公司进行测序分析。
(4)系统发育树的构建
测序结果返回后, 使用Chromas 2.6.5软件进行必要序列剪接, 将处理好的序列提交至美国国家生物技术信息中心(National Center for Biotechnology Information, NCBI)数据库, 进行序列比对, 利用Mega 7.0软件选取邻接法构建系统发育树。
1.3.3 真菌毒素的测定
采用毒素试剂盒测定不同储藏方法和储藏天数下玉米中呕吐毒素、黄曲霉毒素B1和赤霉烯酮毒素的含量。具体步骤如下: 称取5 g烘干样品, 置于含有25 mL蒸馏水或者60%甲醇的三角瓶中, 密封后置于恒温培养振荡器中, 以200 r/min的速度振荡10 min。然后将振荡后的混合液体转移至10 mL离心管中, 以4000 r/min的速度离心5 min, 取上清液并用去离子水稀释。根据试剂盒说明书, 逐步加入标准品、样品和试剂盒中的各种试剂, 进行反应。最后, 利用酶标仪在450 nm和630 nm两个波长下测定各孔的吸光度值, 百分吸光率计算公式如式(1):
$\text { 百分吸光率 } / \%=\frac{B}{B_{0}} \times 100 \%$
其中, B为标准品或样品的吸光度值; B0为0 μg/kg标准品的吸光度值。
根据计算的百分吸光率和相应的标准品浓度绘制标准曲线。然后根据此标准曲线和测定样品的吸光度值, 结合样品稀释的倍数, 计算毒素含量。
1.4 数据处理
采用Excel 2019和SPSS 25.0软件对试验数据进行统计和分析, 显著水平设置为0.05, 运用Excel 2019作图。
2 结果与分析
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2.1 不同储藏方法下污染真菌的分离与鉴定
2.1.1 形态鉴定
参考《真菌鉴定手册》, 对玉米果穗储藏和籽粒储藏处理下主要污染真菌的菌落颜色、质地和分生孢子形态进行初步鉴定。在PDA培养基上, 镰刀菌产生大量棉絮状气生菌丝, 并产生紫红色色素, 孢子形态多样, 大型分生孢子具有多细胞结构, 其形态两端尖而弯曲, 呈典型的镰刀形状(图1A~C); 黄曲霉菌菌落生长较快, 结构疏松, 初期菌落略带黄色, 随后逐渐转变为黄绿色, 最终成熟时呈现褐色。其分生孢子头呈辐射状排列, 形态为半球形或近球形。分生孢子为圆形, 呈串珠状排列(图1D); 黑曲霉菌生长较为迅速, 其菌落最初呈白色, 通常带有鲜黄色区域, 菌丝生长密集, 后来逐渐转变为黑色, 菌落背面一般为无色或中央部分略带褐色。其分生孢子头开始为球形, 后逐渐变为放射状, 呈黑褐色, 小分生孢子为球形(图1E); 青霉菌的菌丝体为无色或淡色, 菌落呈现绿色、蓝色或黄色。其分生孢子梗长而直立, 顶端以特殊的对称或不对称的扫帚状分支, 分生孢子梗最后形成许多瓶梗, 瓶梗上着生分生孢子链。其分生孢子为球形或卵形(图1F); 根霉菌菌落生长迅速, 其形态呈现“棉花糖”状, 初始为白色, 随后逐渐变成灰色或黄棕色。菌丝宽大, 其上附有假根, 自假根处向上, 丛生着直立且不分枝的孢囊梗。孢囊梗的顶端显著膨大, 形成圆形的孢子囊, 囊内产生孢囊孢子(图1G); 链格孢菌的菌落生长速度中等, 菌落呈绒毛状, 颜色从灰黑至黑色。菌丝有隔膜, 分生孢子梗由菌丝顶端生成, 孢子脱落后有明显的孢痕。分生孢子一般为褐色、倒棒状, 表面具有横隔和纵隔, 呈壁砖状结构, 末端有短喙, 排列成较长的直链或斜链(图1H); 篮状菌菌落开始呈黄色, 一周后慢慢变成灰绿色, 表面粗糙, 边界呈规则圆状, 中间凸起, 显微镜下形态呈典型的青霉菌样, 具有帚状分生孢子梗(图1I)。
2.1.2 分子鉴定
根据形态学初步鉴定结果, 选择具有代表性的17株菌株, 分别编号M01~M17, 进行真菌的分子生物学鉴定, 根据鉴定结果构建系统进化树(图2)。从图2中可以看出, 高水分玉米收获后储藏期间表面优势真菌隶属6个属, 9个种。M01和M02可鉴定为镰刀菌属尖孢镰刀菌(Fusarium oxysporum), M03和M04可鉴定为镰刀菌属轮枝镰刀菌(Fusarium verticillioides), M05和M06可鉴定为镰刀菌属藤仓镰刀菌(Fusarium fujikuroi), M07和M08可鉴定为青霉属草酸青霉(Penicillium oxalicum), M09和M10可鉴定为曲霉属黄曲霉(Aspergillus flavus), M11和M12可鉴定为曲霉属黑曲霉(Aspergillus niger), M13和M14可鉴定为链格孢属链格孢菌(Alternaria alternata), M15和M16可鉴定为根霉属少根根霉(Rhizopus arrhizus), M17可鉴定为篮状菌属篮状菌(Talaromyces wortmannii)。
2.2 不同储藏方法对玉米籽粒真菌种类及数量的影响
高水分玉米收获后至储藏100 d, 两种储藏方法下玉米籽粒表面共分离出728株优势真菌(10-3浓度下), 联合传统形态学鉴定和基于18S核糖体DNA的ITS序列分析, 确定玉米储藏期间表面优势真菌分别为镰刀菌属Fusarium oxysporum、Fusarium verticillioides、Fusarium fujikuroi, 曲霉属Aspergillus flavus、Aspergillus niger、青霉属Penicillium oxalicum, 根霉属Rhizopus arrhizus, 链格孢属Alternaria alternata, 篮状菌属Talaromyces wortmannii(表1)。整个储藏期, 以镰刀菌属、曲霉属和青霉属为主, 其次为根霉属、链格孢属和篮状菌属。但是, 随着储藏时间的推进, 优势菌属发生改变。储藏初期(0~20 d)霉菌优势属为镰刀菌属,储藏30 d时果穗储藏的优势菌属为镰刀菌属、曲霉属和青霉属, 籽粒储藏的优势菌属为曲霉属、青霉属和镰刀菌属, 储藏30 d后优势菌属转变为曲霉属和青霉属。这与他人的研究结果是一致的, 如张守梅等[17]的研究表明, 在储藏初期, 玉米的优势污染菌为镰刀菌属, 但是储藏2个月后, 优势菌属发生改变, 青霉属和曲霉属占据主导地位。这可能是因为青霉属和曲霉属均能产生大量的孢子, 在外界环境适宜的情况下, 孢子容易散播, 造成真菌的大量繁殖。
储藏方法和储藏时间对真菌总数量均有显著影响(P<0.05)。同一储藏方法下, 随着储藏天数的增多, 真菌总数量整体上呈先增加后降低的趋势。果穗储藏和籽粒储藏玉米真菌总数量分别在储藏后20 d和30 d最多。不同的储藏方法下, 果穗储藏的真菌数量显著低于籽粒储藏(P<0.05), 储藏10~100 d期间, 果穗储藏的真菌数量比籽粒储藏玉米减少了90.3%~98.6%。
2.3 不同储藏方法对玉米籽粒真菌毒素含量的影响
如图3所示, 除果穗储藏玉米籽粒的赤霉烯酮外, 储藏方法和储藏天数对玉米籽粒中呕吐毒素和赤霉烯酮含量均产生显著影响(P<0.05)。果穗储藏方法下, 储藏0~30 d时, 玉米籽粒中呕吐毒素无显著变化, 储藏70~100 d时呕吐毒素含量显著增加, 分别较储藏0 d时增加了30.5%和99.5%。籽粒储藏方法下, 随着储藏天数的增多, 玉米籽粒中呕吐毒素和赤霉烯酮含量均呈现显著增加的趋势, 储藏100 d时呕吐毒素和赤霉烯酮含量分别为575.6 μg/kg和717.0 μg/kg, 分别是储藏0 d的29.5倍和177.9倍。不同的储藏方法下, 果穗储藏玉米籽粒的呕吐毒素和赤霉烯酮含量显著低于籽粒储藏(P<0.05)。此外, 随着储藏天数的增多, 两种储藏方法下玉米籽粒的呕吐毒素和赤霉烯酮含量差异加大, 储藏10~100 d, 果穗储藏较籽粒储藏玉米籽粒的呕吐毒素含量降低幅度由58.4%增加至93.2%, 赤霉烯酮含量的降低幅度由87.8%增加至99.4%。在整个储藏期间, 果穗储藏和籽粒储藏的黄曲霉毒素B1含量均较低, 在0.19~0.40 μg/kg之间, 且储藏方法和储藏天数对玉米籽粒中黄曲霉毒素B1含量均无显著影响。在本研究中, 真菌数量呈现先增加后降低的趋势, 而随着储藏天数的增多, 果穗储藏方法下玉米籽粒中呕吐毒素含量和籽粒储藏方法下呕吐毒素和赤霉烯酮含量均呈现显著增加的趋势。这主要是因为霉菌的数量受储藏条件的影响, 但是霉菌寄附在粮食的表面和内部, 产生诸如呕吐毒素和玉米赤霉烯酮等有毒代谢物质, 这些毒素不易分解, 会随着时间的推移逐渐累积[18-19]。
2.4 储藏期间玉米中主要真菌与真菌毒素相关性分析
呕吐毒素和赤霉烯酮的产生主要由镰刀菌属引起, 而黄曲霉毒素B1的产生则主要由黄曲霉菌引起。相关性分析显示(表2), 呕吐毒素和赤霉烯酮的含量与镰刀菌属的数量之间存在极显著的正相关, 相关系数分别为0.995和0.996。黄曲霉毒素B1含量与黄曲霉菌数量无显著相关性。这表明, 该研究中分离的镰刀菌属多为产毒菌, 而分离出的黄曲霉菌大部分均为不产毒菌。
3 讨论与结论
收起
为了确保未来玉米的质量安全, 选择合适的储藏方法至关重要。近年来的研究主要集中在如何优化大粮仓中玉米的储藏技术上, 对农户储粮方法的研究相对不足[20-22]。鉴于我国农业以小农户生产模式为主, 深入探究玉米收获后的储藏方法以有效维护其质量安全, 具有极高的现实意义与紧迫性。本研究模拟了中国小农户对玉米高水分收获后的储藏方法, 分别进行了露天果穗储藏和籽粒储藏, 通过对玉米中真菌数量和毒素含量进行分析, 探讨了不同储藏方法对玉米质量安全的影响。
研究结果表明, 在果穗储藏和籽粒储藏两种储藏方法下, 随着储藏天数的增加, 真菌数量整体上呈现先增加后降低的趋势。这种现象的出现主要受到温度和水分两方面因素的影响。温度和水分是影响微生物生长的关键因素。当温度较高且水分含量较大时, 微生物活动旺盛, 菌落生长迅速。反之, 当温度和水分含量较低时, 微生物的生长会受到抑制[23-25]。以往研究均表明, 控制环境的水分条件, 保持干燥与低温储藏, 是粮食防霉的关键[26-28]。玉米储藏0~30 d期间正值黄淮海地区的10—11月份, 这段时间内平均温度相对较高, 同时新收获的玉米水分含量也较高, 这为真菌的繁殖提供了有利条件。然而, 随着12月份的到来, 温度逐渐降低, 且玉米籽粒的含水量也下降至10%以下。在这种环境下, 不利于真菌的生长, 因此真菌数量呈现出先增加后减少的趋势。
不同的储藏方法下, 果穗储藏的玉米真菌数量与呕吐毒素和赤霉烯酮毒素含量均显著低于籽粒储藏, 且随着储藏时间的延长, 这种差异变得更加明显。主要有以下3方面的原因: 首先, 黄淮海地区热量资源相对较少, 能用于脱水的积温有限, 玉米籽粒收获时含水量较高, 导致收获时破损率偏高, 破碎的玉米籽粒容易遭受真菌的侵染[29-31]。而果穗储藏则避免这一环节, 保持了玉米籽粒的完整性, 减少损伤和破碎率, 从而降低真菌侵染的概率。其次, 在储藏过程中, 果穗储藏玉米籽粒紧密地嵌在穗轴上, 这种结构不仅有利于降低籽粒的含水量, 还能减少虫蛀和霉菌感染的风险[32]。第三, 果穗储藏孔隙度较大, 空气流通性好, 水分降低较快, 能够减少霉变和虫害的发生[11-12]。而籽粒储藏节省储藏空间, 但是水分降低较慢, 并且容易呼吸作用放热, 引起粮堆内部温度升高, 加剧玉米的霉变和腐烂。因此, 果穗储藏在保持玉米质量安全方面表现出明显的优势。
综上所述, 高水分玉米收获后, 果穗储藏较籽粒储藏能显著降低真菌数量和毒素含量, 保证玉米的质量安全。
基金
收起
  • 山东省自然科学基金青年项目(ZR2021QC012)
  • 山东省现代农业产业技术体系玉米创新团队项目(SDAIT-02-11)
文献
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doi: 10.19812/j.cnki.jfsq11-5956/ts.20240627002
  • 接收时间：2024-06-27
  • 首发时间：2025-07-19
  • 出版时间：2025-03-15
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  • 收稿日期：2024-06-27
基金
山东省自然科学基金青年项目(ZR2021QC012)
山东省现代农业产业技术体系玉米创新团队项目(SDAIT-02-11)
作者信息
    1.山东省农业科学院作物研究所, 济南 250100
    2.中国农业大学农学院, 北京 100193

通讯作者:

* 龚魁杰(1970—), 男, 博士, 研究员, 主要研究方向为谷物营养与质量安全。E-mail:
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2种不同金属材料的力学参数
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Family		 属数
Number of
genus		 种数
Number of
species		 占总种数比例
Percentage of
total species (%)		 属
Genus		 种数
Number of
species		 占总种数比例
Percentage of total
species (%)
鹅膏菌科Amanitaceae 2 11 5.26 鹅膏菌属 Amanita 10 4.78
小菇科 Mycenaceae 2 12 5.74 丝盖伞属 Inocybe 5 2.39
多孔菌科 Polyporaceae 8 14 6.70 蜡蘑属 Laccaria 5 2.39
红菇科 Russulaceae 3 23 11.00 小皮伞属 Marasmius 6 2.87
小菇属 Mycena 11 5.26
光柄菇属 Pluteus 5 2.39
红菇属 Russula 17 8.13
栓菌属 Trametes 5 2.39
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