Scientia Agricultura Sinica ›› 2013, Vol. 46 ›› Issue (4): 810-818.doi: 10.3864/j.issn.0578-1752.2013.04.015

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

Molecular Identification and rDNA-ITS Sequence Analysis of Fungal Pathogens in Kiwifruit During Storage

 DUAN  Ai-Li, LEI  Yu-Shan, SUN  Xiang-Yu, GAO  Gui-Tian, ZHAO  Jin-Mei, GU  Liu-Jie   

  1. 1.College of Food Engineering and Nutritional Science, Shaanxi Normal University, Xi’an 710062
    2.Rural Science and Technology Development Center of Shaanxi Province, Xi’an 710054
    3. Wuwei Industrial Enterprises Talent Service Center of Gansu Province, Wuwei 733000, Gansu
  • Received:2012-02-07 Online:2013-02-15 Published:2012-12-14

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Abstract: 【Objective】The fungal pathogens of ‘China optimal’, ‘Haiwode’ and ‘Qinmei’ kiwi fruit that were harvested in Zhouzhi County, Shaanxi Province, were isolated during storage and identified by rDNA-ITS sequences analysis.【Method】With rDNA-ITS analysis and morphological characterization, the species of Fungal pathogens were differentiated and identified. By means of the genetic relationship analysis and multiple comparison analysis of ITS sequences of fungal pathogens, the diversity of intra-species in the pathogens were investigated.【Result】Five strains were identified, e.g. Penicillium, Trichoderma, Alternaria, Mucor and Simplicillium. Penicillium was the dominant species, accounting for 63.3% of the tested strains. And Trichoderma was the second dominant species, accounting for 26.7%. There were little differences in ITS sequences among Trichoderm, approval ratings were 99%. But the differences of ITS sequences between the Penicillium were significant .Such as Penicillium sp., Penicillium purpurogenum, Penicillium chrysogenum, Penicillium paneum and Penicillium commune.【Conclusion】Penicillium are mainly fungal pathogens of ‘China optimal’, ‘Haiwode’ and ‘Qinmei’ kiwifruit that were harvested in Zhouzhi County, Shaanxi Province. The differences of rDNA-ITS sequences between Penicillium are obvious. So, rDNA-ITS sequence between ourtra-species and intra-species have a certain reference value.

Key words: kiwifruit , fungal , identification , rDNA-ITS sequence analysis

[1]唐艳. 1-MCP对猕猴桃采后生理生化变化及贮藏品质的影响[D]. 陕西杨凌: 西北农林科技大学, 2004.

Tang Y. Study on storage quality and related physiological and biochemical change of postharvest kiwifruit as influenced by 1-MCP[D]. Yangling, Shaanxi: Northwest Agriculture and Forestry University, 2004. (in Chinese)

[2]Hibbett D S. Ribosomal RNA and fungal systematics. Transactions of the Mycological Society of Japan, 1992, 33: 533-556.

[3]丁爱冬, 于梁, 石蕴莲. 猕猴桃采后病害鉴定和侵染规律研究.植物病理学报, 1993, 25(2): 149-153.

Ding A D, Yu L, Shi Y L. Study on disease and infection identified in kiwifruits. Acta Phytopathologica Sinica, 1993, 25(2): 149-153. (in Chinese)

[4]马松涛, 宋晓斌, 张学武, 刘艳玲. 猕猴桃花果病害研究现状及趋势.西北林学院学报, 2000,15 (3): 86-90.

Ma S T, Song X B, Zhang X W, Liu Y L. The disease research situation and trend in the flowers of kiwifruits. Journal of Northwest Forestry University, 2000, 15(3): 86-90. (in Chinese)

[5]Pantou M P, Mavidou 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: 159-174.

[6]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[D]. 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)

[7]周德庆. 微生物学教程. 北京:高等教育出版社, 1993.

Zhou D Q. Microbiology Tutorial. Beijing: Higher Education Press, 1993. (in Chinese)

[8]沈萍, 范秀容, 李广武. 微生物学实验. 北京:高等教育出版社, 1999.

Shen P, Fan X R, Li G W. Experimental Microbiology. Beijing: Higher  Education Press, 1999. (in Chinese)

[9]江汉湖. 食品微生物学. 北京:中国农业出版社, 2000.

Jiang H H. Food of Microbiology. Beijing: China Agriculture Press, 2000.(in Chinese)

[10]周颖, 张瑞, 郭颂, 怀晓, 范在丰, 周涛. 北京地区地黄花叶病病原的分子鉴定.植物保护学报, 2010, 37(5):447-452.

Zhou Y, Zhang R, Guo S, Huai X, Fan Z F, Zhou T. Molecular identification of viral pathogens inducing Rehmannia glutinosa mosaic disease in Beijing. Plant Protection, 2010, 37(5): 447-452. (in Chinese)

[11]孙祥瑞, 朱杰华, 刘蕊. 几种树皮腐烂病菌的分子鉴定. 河北农业大学学报, 2010, 33(6): 37-42.

Sun X R, Zhu J H, Liu R. Molecular identification of pathogens of several trees canker. Journal of Agricultural University of Hebei, 2010, 33(6):37-42. (in Chinese)

[12]Renske L, Paula L, Thomw K. Molecular identification of ectomycorrhizal mycelium in soil horizons. Applied and Environmental Microbiology, 2003, 69(1): 327-333.

[13]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)

[14]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.

[15]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)

[16]Thompson J D, Gibson T J, Plewniak F, Jeanmougin F, Higgins D G. The CLUSTAL-X windows interface: flexible strategies for multiple sequence alignment aided by quality analysis tools. Nucleic Acids Research, 1997, 25(24): 4876-4882.

[17]Koichiro T, Joel D, Masatoshi N,  Sudhir K. MEGA4: molecular evolutionarygenetics analysis (MEGA) software version 4.0. Molecular Biology and Evolution, 2007, 24(8): 1596-1599.

[18]杨合同, 唐文华, 徐砚珂, 王加宁, 姚碗生.木霉菌属的定义及其属下分类.山东科学, 2002, 15(2): 15-22.

Yang H T, Tang W H, Xu Y K, Wang J N, Yao W S. Del ineation of the genus Trichoderma and its sub-genus division. Shandong Science, 2002, 15(2): 15-22. ( in Chinese)

[19]Ninet B, Jan I, Bontems O, Lechenne B, Jousson O, Lew D, Schrenzel J, Panizzon RG, Monod M. Molecular identification of Fusarium species in Onychomycoses. Dermatology, 2005, 210: 21-25.

[20]Lievens B, Claes L, Vakalounakis D J, Vanachter A C, Thomma B P. A robust identification and detection assay to discriminate the cucumber pathogens Fusarium oxysporum f. sp. cucumerinum and f. sp. radicis-cucumerinum. Environmental  Microbiology, 2007, 9(9): 2145-2161.

[21]Vakalounakis D J, Fragkiadakis G A. Genetic diversity of Fusarium oxysporum isolates from cucumber: differentiation by pathogenicity, vegetative compatibility and RAPD fingerprinting. Phytopathology, 1999, 89(2): 161-168.

[22]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)

[23]李海波, 吴学谦, 王立武, 付立忠, 魏海龙, 吴庆其. 青藏高原黄绿蜜环菌纯培养菌种的分离培养及分子鉴定.菌物学报, 2008, 27(6): 873-883.

Li H B, Wu X Q, Wang L W, Fu L Z, Wei H L, Wu Q Q. Pure culture isolation cultivation and molecular identification of Armillaria luteovirens from Tibet Plateau. Mycosystema, 2008, 27(6): 873-883. (in Chinese)

[24]王建波, 张文驹, 陈家宽. 核rDNA的ITS序列在被子植物系统与进化研究中的应用.植物分类学报, 1999, 37(4):407-416.

Wang J B, Zhang W J, Chen J K. Application of ITS sequences of nuclear rDNA in phylogenetic and evolutionary studies of angiosperms. Acta Phytotaxonomica Sinica, 1999, 37(4): 407-416. (in Chinese)

[25]唐建辉,王伟.西瓜炭疽菌核糖体基因ITS区段的克隆及序列分析.安徽农业科学, 2007, 35(9): 2566-2568.

Tang J H, Wang W. Cloning and sequencing of ribosomal RNA gene ITS in Colletotrichum orbiculare. Journal Anhui Agricultural Sciences, 2007, 35(9): 2566-2568. (in Chinese)

[26]郑雪芳, 蓝江林, 曹宜, 肖荣凤, 葛慈斌, 刘波.瓜类作物枯萎病病原菌的分类鉴定及ITS序列差异性分析.中国农业科学, 2008, 41(12): 4077-4084.

Zheng X F, Lan J L, Cao Y, Xiao R F, Ge C B, Liu B. Identification and ITS sequence analysis of Fusarium wilt pathogens from the cucurbits crops. Scientia Agricultura Sinica, 2008, 41(12): 4077-4084. (in Chinese)

[27]Pritsch K, Boyle H, Munch J C. Characterization and identification  of black alder ectomycorrhizas by PCR／RFLP analyses of the  rDNA internal transcribed spacer(ITS). New Phytologist, 1997, 137: 357-369.

[28]Karen O, Hogborg N, Dalllberg A. Inter and intruspecific variation in the ITS region of rDNA of ectomycorrhizal fungi in Fennoscandia as detected by endonuclease analysis. New Phytologist, 1997, (136): 313-325.

[29]Cooke D E L, Drenth A, Duncan J M, Wagels G, Brasier C M. A molecular phylogeny of phytophthora and related Oomycetos. Fungal Genetics and Biology, 2000, 30: 17-32.

[30]Huang, W Y, Cai Y Z, Surveswara S, Hyde K D, Cork  H, Sun M. Molecular phylogenetic identification of endophytic fungi isolated from three artemisia species. Fungal Diversity, 2009, 30: 69-88.

[31]Laurene L, Lisa A C, Lori M. Internal transcribed spacer- sequence-based phylogeny and polymerase chain reaction-restriction fragment length polymorphism differentiation of Tilletia walkeri and T. Indica Phytopathology, 2001, 91(10): 935-940.

[32]鲁绯, 高丽华, 张金兰. 一株丹贝中分离霉菌的鉴定.中国酿造, 2010, 219(6): 35-39.

Lu F, Gao L H, Zhang J L. Identification of a mould strain in tempe. Brewing in China, 2010, 219(6): 35-39. (in Chinese)

[33]Constantino R, Gonzalo P, Geraid F B. Isolation and characterization of melanized fungi from limestone formations in Mallorca. Mycological Progress, 2005, 4(1): 23-38.

[34]张广志,文成敬.木霉对玉米纹枯病的生物防治.植物保护学报,  2005, 32(4): 1-4.

Zhang G Z, Wen C J. Biocontrol of maize sheath blight with Trichoderma spp. Acta Phytophylacica Sinica, 2005, 32(4): 1-4. (in Chinese)
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