Scientia Agricultura Sinica ›› 2016, Vol. 49 ›› Issue (7): 1371-1381.doi: 10.3864/j.issn.0578-1752.2016.07.013

• STORAGE·FRESH-KEEPING·PROCESSING • Previous Articles     Next Articles

Fungal Diversity of Huaidao No. 5 Rice and the Dominant Culturable Fungal Strains During Storage

DU Li-hui, HE Xiao-ying, LIU Ling-ping, YUAN Jian, JU Xing-rong   

  1. College of Food Science and Engineering, Nanjing University of Finance and Economics/Collaborative Innovation Center for Modern Grain Circulation and Safety/Key Laboratory of Grains and Oils Quality Control and Processing, Nanjing 210023
  • Received:2015-09-10 Online:2016-04-01 Published:2016-04-01

RichHTML

2

PDF

804

Cited

1

Abstract: 【Objective】The objective of this study is to determine the fungal genus in 2013 Huaidao No. 5 rice by traditional morphological observation and molecular biology method, as well as to investigate the dominant fungal strains on Huaidao No.5 rice under different storage conditions. Results of this study could provide a scientific basis for the fungal genus identification and control of fungi growth in Huaidao No. 5 rice. 【Method】The culturable fungal strains were obtained by using both Rose Bengal and potato dextrose agar (PDA) media, following the instruction of GB 4789.15-2010. The Huaidao No. 5 rice collected from 4 different regions were mixed and then the moisture content were adjusted to 14.5% and 18.5% respectively. The samples were stored in the artificial climate equipment under 25℃. The culturable dominant fungi were determined by traditional culturing of the rice samples monthly removed from the simulated storage conditions. The fungal genus were identified by morphological observation of colonies, hyphae and spores by referencing to the . At the same time, the internal transcribed spacer (ITS) and 28S rDNA were amplified by PCR, and then ligated into T vector and sequenced. The fungal species were identified by constructing phylogenetic trees.【Result】In terms of fungal diversity in Huaidao No. 5 rice: 33 strains of fungi were isolated from four regions of the 2013 Huaidao No. 5 rice, mainly for Penicillium, Aspergillus and Fusarium, including 16 strains of Penicillium and 15 strains of Aspergillus, accounting for the separated strains of 48.5% and 45.5%, respectively, indicating that Penicillium and Aspergillus are the dominant fungi genus in 2013 Huaidao No. 5 rice. There are only 2 strains of Fusarium, which accounted for only 6% of the isolates. Simulation storage experiment results showed that Penicillium aurantiogriseum appeared to be the dominant strain from 6 month in moisture content of 14.5% for Huaidao No. 5 rice under 25℃ storage, and this strain persisted to be dominant in 7 and 8 months storage. While for the Huaidao No. 5 rice with a moisture content of 18.5%, a yeast strain appeared to be the dominant from 7 month and this strain persisted to be dominant at 8 month under 25℃ storage.【Conclusion】 33 strains of fungi were separated and identified from 2013 Huaidao No. 5 rice, including 16 strains of Penicillium and 15 strains of Aspergillus, respectively, indicating that Penicillium and Aspergillus are the dominant fungi genus in 2013 Huaidao No. 5 rice. And under different simulated storage conditions, different cultural fungal strains dominated the Huaidao No. 5 rice with different moisture content.

Key words: rice, storage, fungi, ITS, 28S rDNA, flora

[1]    Wen P C, Hinrichs S H, Rupp M E. Utilization of the internal transcribed spacer regions as molecular targets to detect and identify human fungal pathogens. Medical Mycology, 2002, 40(1): 87-109.
[2]    赵国柱, 张天字, 张猛. 核糖体基因簇在真菌系统学研究中的意义. 生命的化学, 2002, 22(1): 13-15.
Zhao G Z, Zhang T Y, Zhang M. Significance in the study of ribosomal gene cluster in fungal systematic. Chemistry of Life, 2002, 22(1): 13-15. (in Chinese)
[3]    Belloch C, Fernández-Espinar T, Querol A, Garcia M D, Barrio E. An analysis of inter- and intraspecific genetic variabilities in the Kluyveromyces marxianus group of yeast species for the reconsideration of the K. lactis taxon. Yeast, 2002, 19(3): 257-268.
[4]    章初龙, 徐同. Trichoderma harzianum及其近缘种的分子系统学研究. 生物多样性, 2003, 11(1): 10-19.
Zhang C L, Xu T. Molecular phylogenetic analysis of Trichoderma harzianum and its related species. Biodiversity Science, 2003, 11(1): 10-19. (in Chinese)
[5]    陈敏, 廖万清, 吴绍熙, 姚志荣, 陈江汉, 徐红, 陈裕充. 新生隐球菌变种间ITS序列差异的研究. 中华皮肤科杂态, 2007, 40(4): 230-233.
Chen M, Liao W Q, Wu S X, Yao Z R, Chen J H, Xu H, Chen Y C. Sequence diversity of ribosomal internal transcribed spacer (ITS) region among Cryptococcus neoformans varieties. Chinese Journal of Dermatology, 2007, 40(4): 230-233. (in Chinese)
[6]    Sugita T, Nishikawa A. Fungal identification method based on DNA sequence analysis: reassessment of the methods of the pharmaceutical society of Japan and the Japanese Pharmacopoeia. Journal of Health Science Tokyo, 2003, 49(6): 531-533.
[7]    Magan N, Lacey J. Effect of water activity, temperature and substrate on interactions between field and storage fungi. Transactions of the British Mycological Society, 1984, 82(1): 83-93.
[8]    Ayerst G. The effects of moisture and temperature on growth and spore germination in some fungi. Journal of Stored Products Research, 1969, 5(2): 127-141.
[9]    Barron G L, Lichtwardt R W. Quantitative estimations of the fungi associated with deterioration of stored corn in Iowa. Iowa State College Journal of Science, 1959, 34(2): 147-155.
[10]   Johanson A, Turner H C, McKay G J, Brown A E. A PCR-based method to distinguish fungi of the rice sheath-blight complex, Rhizoctonia solani, R. oryzae and R. oryzae-sativae. FEMS Microbiology Letters, 1998, 162(2): 289-294.
[11]   岳晓禹, 张恒业, 郭小华, 刘相东, 李自刚, 马丽卿. 贮藏稻谷中一株优势腐败霉菌的分离与鉴定. 现代食品科技. 2010, 26(4): 410-414.
Yue X Y, Zhang H Y, Guo X H, Liu X D, Li Z G, Ma L Q. Isolation and identification of a predominant spoiling mold strain from stored rice. Modern Food Science and Technology, 2010, 26(4): 410-414. (in Chinese)
[12]   周建新, 鞠兴荣, 孙肖东, 金浩, 姚明兰, 申海进. 不同储藏条件下稻谷霉菌区系演替的研究. 中国粮油学报, 2008, 23(5): 133-136.
Zhou J X, Ju X R, Sun X D, Hin H, Yao M L, Shen H J. Succession of mould flora for paddy in different storage conditions. Chinese Journal of Grain and Oil, 2008, 23(5): 133-136. (in Chinese)
[13]   Pantou M P, Mavridou A, Typas M A. IGS sequence variation, group-I introns and the complete nuclear ribosomal DNA of the entomopathogenic fungus Metarhizium: excellent tools for isolate detection and phylogenetic analysis. Fungal Genetics and Biology, 2003, 38(2): 159-174.
[14]   刘盼红. 贮藏苹果中展青霉素产生菌的分离鉴定和生长代谢特征研究. 陕西杨凌: 西北农林科技大学, 2008.
Liu P H. Research on isolation and identification of Patulin-Producing strains and their growth metabolic characteristics[D]. Yangling, Shaanxi: Northwest Agriculture and Forestry University, 2008. (in Chinese)
[15]   匡治州, 许杨. 核糖体rDNA ITS序列在真菌学研究中的应用. 生命的化学, 2004, 24(2): 120-122.
Kuang Z Z, Xu Y. The application of ribosomal gene rDNA in fungal research. Chemistry of Life, 2004, 24(2): 120-122. (in Chinese)
[16]   Schmidt O, Moreth U. Data bank of rDNA-ITS sequences from building-rot fungi for their identification. Wood Science and Technology, 2002, 36: 429-433.
[17]   袁晖. 腐烂苹果中真菌分离鉴定和PCR快速检测及条件优化[D]. 陕西杨凌: 西北农林科技大学, 2009.
Yuan H. Separation and identification of fungi in rot apple and rapid detection and optimization of PCR [D]. Yangling, Shaanxi:Northwest Agriculture and Forestry University, 2009. (in Chinese)
[18]   Vakalounakis D J, Fragkiadakis G A. Genetic diversity of Fusariumoxysporum isolates from cucumber: differentiation by pathogenicity, vegetative compatibility and RAPD fingerprinting. Phytopathology, 1999, 89(2): 161-168.
[19]   王琢, 闫培生. 真菌毒素产生菌的分子鉴定研究进展. 中国农业科技导报, 2010, 12(5): 42-50.
Wang Z, Yan P S. Research progress on molecular identification of mycotoxin producing fungi. Journal of Agricultural Science and Technology, 2010, 12(5): 42-50. (in Chinese)
[20]   殷蔚申 张耀东 庄桂. 我国稻谷真菌区系调查及其演替规律的研究. 郑州粮食学院学报. 1986(3): 3-17.
Yin S W, Zhang Y D, Zhuang G. The study on grain mycoflora and succession regulation of Chinese rice. Journal of Zhengzhou Institute of Technology, 1986(3): 3-17. (in Chinese)
[21]   段爱莉, 雷玉山, 孙翔宇, 高贵田, 赵金梅, 谷留杰. 猕猴桃果实贮藏期主要真菌病害的 rDNA-ITS 鉴定及序列分析. 中国农业科学, 2012, 46(4): 810-818.
Duan A L, Lei Y S, Sun X Y, Gao G T, Zhao J M, Gu L J. Molecular identification and rDNA-ITS sequence analysis of fungal pathogens in kiwifruit during storage. Scientia Agricultura Sinica, 2012, 46(4): 810-818. (in Chinese)
[22]   Lichtwardt R W, Barron G L, Tiffany L H. Mold flora associated with shelled corn in Iowa. Iowa State College Journal of Science, 1958, 33(1): 1-11.
[23]   黄坊英, 廖权辉, 毛朝安, 黄伯爱, 陈曼玲, 何淑英, 陈绪嫦. 早造稻谷田间真菌区系的初步调查研究. 粮食储藏, 1982, 3: 35-41.
Huang F Y, Liao Q H, Mao C A, Huang B A, Chen M L, He S Y, Chen X C. Preliminary investigation of mycoflora on Zaodao rice. Grain Storage, 1982, 3: 35-41. (in Chinese)
[24]   李从虎, 郑佳, 谢菲, 陈欣, 周荣清. 郫县豆瓣曲曲霉的筛选、分子鉴定及其酶学性质的研究. 中国酿造, 2010, 29(5): 30-34.
Li C H, Zheng J, Xie F, Chen X, Zhou R Q. Screening, molecular classification of Aspergillus in Pixian’ horse-bean and its enzymatic characteristics.China Brewing, 2010, 29(5): 30-34. (in Chinese)
[25]   燕勇, 李卫平, 高雯洁, 沈志英, 王恒辉, 陈黎霞. rDNA-ITS序列分析在真菌鉴定中的应用. 中国卫生检验杂志, 2008, 18(10): 1958-1961.
Yan Y, Li W P, Gao W J, Shen Z Y, Wang H H, Chen L X. Application of rDNA-ITS sequences in fungi identification. Chinses Journal of Health Laboratory Technology, 2008, 18(10): 1958-1961. (in Chinese)
[26]   郑雪松, 杨虹, 李道棠, 韩文卿. 基因间隔序列 (ITS) 在细菌分类鉴定和种群分析中的应用. 应用与环境生物学报, 2003, 9(6): 678-684.
Zheng X S, Yang H, Li D T, Han W Q. Use of 16S-23S intergenic transcribed spacer in identification and community analysis of bacteria. Chinese Journal of Applied and Environmental Biology, 2003, 9(6): 678-684. (in Chinese)
[1] PENG TingShen, LU JiuYan, WU MeiLin, YAN YuXin, LIU HongZhou, NAN WenBin, QIN XiaoJian, LI Ming, GONG JunYi, LIANG YongShu. QTL Analysis of Yield-Related Traits in Both Huangnuo2# and Changbai7# of Perennial Chinese Rice [J]. Scientia Agricultura Sinica, 2026, 59(7): 1361-1379.
[2] CUI JieHao, ZHANG Meng, WANG Qin, YU JiaYan, LIN Kun, LI ShangZe, LAN Heng, GENG YanQiu, ZHANG Qiang, GUO LiYing, SHAO XiWen. Evaluation of Lodging Resistance and Its Physiological Mechanisms in Japonica Rice Resources [J]. Scientia Agricultura Sinica, 2026, 59(7): 1420-1438.
[3] YUAN HaoLiang, NIE Jun, LI Peng, LU YanHong, LIAO YuLin, XU ChangXu, LI ZhongYi, CAO WeiDong, ZHANG JiangLin. Effects of Co-Utilization of Chinese Milk Vetch and Rice Straw on Soil Phosphorus Composition and Phosphorus Activation of Paddy Field in Southern China [J]. Scientia Agricultura Sinica, 2026, 59(7): 1480-1491.
[4] XU YangHaoJun, CHEN LiMing, YANG ShiQi, TANG YiFan, TAN XueMing, ZENG YongJun, PAN XiaoHua, ZENG YanHua. Effects of Long-Term Different Straw Returning Methods on Soil Organic Carbon, Nutrients and Aggregate Formation in Different Soil Layers of Double Cropping Rice Field [J]. Scientia Agricultura Sinica, 2026, 59(7): 1492-1506.
[5] SU YiFan, YANG ZhanXu, WANG Di, MAO JunCheng, WEI MengMeng, CHEN Ze, BAI XinRan, CHU TianGe, MA ChangNing, QIAO MingFei, SUN Quan, HU DaGang. Effects of 2, 4-Epibrassinolide on Postharvest Storage Quality and Physiological Performance of Apple [J]. Scientia Agricultura Sinica, 2026, 59(7): 1536-1551.
[6] YE MeJin, WU Lei, MD NAHIBUZZAMAN Lohani, YIN Li, HU XinRong, LIU YaXi, JIANG YunFeng, CHEN GuoYue, PU ZhiEn, LI Yang, LI Ting, ZOU YaYa, WU JiaYi, MA Jian. Genome-Wide Association Study-Based Identification of Loci Controlling Mature Embryo Size in Chinese Wheat Landraces and Their Genetic Effects Analysis [J]. Scientia Agricultura Sinica, 2026, 59(6): 1157-1171.
[7] LI XingYu, HUANG Rong, XIAN YiMing, TIAN JiaoJiao, MA XiaoJin, YANG QiaoXi, LI Bing, WANG ChangQuan. Characteristics of Organic Carbon Fractions and Carbon Dioxide Emissions of Different Size Aggregates in Rice Field Soils in Response to Long-Term Fertilization [J]. Scientia Agricultura Sinica, 2026, 59(6): 1255-1271.
[8] MA ZhaoHui, QUAN ChengZhe, CHENG HaiTao, YANG KanJie, LI XinRui, LÜ WenYan. The Breeding Goals and Strategies of Northeast Japonica Rice Under the Background of Zhongke Fa No.5 [J]. Scientia Agricultura Sinica, 2026, 59(5): 927-936.
[9] ZHANG WeiJian, YAN ShengJi, SHANG ZiYin, TANG ZhiWei, WU LiuGe, LI JiaRui, CHEN HaoTian, DENG AiXing, ZHANG Jun, ZHANG Xin, ZHENG ChengYan, SONG ZhenWei. Methane Emissions from Paddy Fields: Not Entirely Attributable to Rice Cultivation [J]. Scientia Agricultura Sinica, 2026, 59(4): 824-833.
[10] CHEN Min, JIAO ZiLan, QIAO ChengBin, XU Hao, ZHANG Bi, MA DongHua, KONG WeiRu, WANG JingWen, SONG JiaWei, LUO ChengKe, LI PeiFu, TIAN Lei. Morpho-Physiological Responses and Adaptive Strategies of Rice Germplasm Accessions from Different Subspecies Under Salt Stress [J]. Scientia Agricultura Sinica, 2026, 59(4): 705-722.
[11] GUO FuCheng, TANG HaiJiang, HAO XinYi, MA GuoLin, YANG JiuJu, HUANG LinFeng, TIAN Lei, WANG Bin, LUO ChengKe. Effects of Different Irrigation Methods on Water-Salt Transport, Rice Yield, and Water Use Efficiency in Saline Soil in Ningxia [J]. Scientia Agricultura Sinica, 2026, 59(4): 750-764.
[12] LUO Wei, YU Hong, YUAN LiXin, WANG LingLing, ZHAO JinPeng, YIN Wei, WANG MingTian, WANG RuLin. Change of Geographic Distributions of Ratoon Rice in Sichuan- Chongqing Under Global Climate Change [J]. Scientia Agricultura Sinica, 2026, 59(4): 765-780.
[13] ZHU Shu, GUO ZhiPeng, SUN Ying. Functional Analysis of Rice Target of Rapamycin OsTOR in Regulating Root Elongation [J]. Scientia Agricultura Sinica, 2026, 59(3): 475-485.
[14] LÜ WenYan, CHENG HaiTao, MA ZhaoHui, TIAN ShuHua. Discussion on Hybridization Breeding Technology and Strategy of Rice in the New Era of Breeding [J]. Scientia Agricultura Sinica, 2026, 59(2): 233-238.
[15] LIAO TingLu, SHI YaFei, XIAO DongHao, SHE YangMengFei, GUO FuCheng, YANG JiuJu, TANG HaiJiang, LUO ChengKe. The Effect of Exogenous Nitroprusside on Sugar Metabolism in Rice Seedlings Under Alkaline Stress [J]. Scientia Agricultura Sinica, 2026, 59(2): 265-277.
Viewed
Full text


Abstract

Cited
  Shared   
  Discussed   
No Suggested Reading articles found!
京ICP备09089781号-30
Copyright 2018 © Editorial office of Scientia Agricultura Sinica
Tel: 86-010-82106279    E-mail: zgnykx@mail.caas.net.cn
Supported by Beijing Magtech Co.Ltd