Isolation and identification of main pathogenic fungus from post-harvest Akesu Fuji apple fruits
-
摘要: 为了明确阿克苏红富士苹果采后腐烂的主要病原菌种类。通过对阿克苏红富士苹果采后常温和低温条件下自然发病的果实进行病原菌的分离筛选,采用经典微生物分离筛选方法,结合形态学观察、基因间隔序列(Internal transcribed spacer,ITS)、28S核糖体大亚基序列(large subunit r DNA,r DNA-LSU)及微管蛋白B(Beta tubulin,β-tubulin)序列的测定方法,并进行系统进化树的构建;同时,进行菌株的回接及病斑形态的验证。最终从腐烂的阿克苏红富士苹果病斑中分离出2类真菌,由形态学观察初步确定为链格孢霉(Alternaria sp.)和青霉(Penicillium sp.),通过进一步的多基因序列分析及菌株的回接及病斑的观察鉴定2种真菌分别为交链格孢霉(Alternaria alternata)和扩展青霉(Penicillium expansum)。本研究结果可为新疆阿克苏红富士采后病原真菌的快速鉴定及病害的防治提供理论依据。
-
关键词:
- 苹果 /
- 采后病害 /
- 微管蛋白B /
- 基因间隔序列 /
- 28S核糖体大亚基序列
Abstract: The objective of this project was to clarify the main pathogenic fungus on harvested Akesu Fuji apple after rotten.The harvested Akesu Fuji apple was used for the isolation and screening of pathogenic bacteria under the condition of room temperature and low temperature.The method was the classical microorganism of isolation and screening with the morphological observation,the internal transcribed spacer,( ITS) 28 S ribosomal( large subunit r DNA,r DNA- LSU) and testing the sequence of microtubulin B( Beta tubulin,β- tubulin) as well as the structure of phylogenetic tree.Meanwhile the verification of tie- back of bacterial strain and the lesion were conducted.The lesion was separated into two fungus from the storage of red Fuji apple,and also the two fungus were identified as Alternaria sp.and Penicillium sp.through the morphological observation.Further sequence analysis using the tie- back of bacterial train and lesion confirmed that,the two types of Fuji were Alternaria alternata and Penicillium expansum.This study provided the theoretical basis for the research of rapid identification of pathogenic bacteria in harvested Akesu red Fuji apple in Xinjiang.-
Keywords:
- apple /
- pathogens after harvest /
- β-tubulin /
- ITS /
- LSU
-
[1] 新疆阿克苏地区苹果产业发展探讨.区域行业经济[M].新疆:新疆区域发展出版社,2015:25-26. [2] 新疆维吾尔自治区统计局.新疆统计年鉴[M].北京:中国统计出版社,2014:335-336. [3] 姚子鹏,王友升,郭晓敏,等.桃果实贮藏期间病原真菌的ITS r DNA序列分析与鉴定[J].中国食品学报,2011,11(4):172-178. [4] 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].杨凌:西北农林科技大学,2009. [5] WHITETJ,BRUNST,LEES.Analysis of phytogenetic relationship sbyam-plification and direct sequencing of ribosomal RNA genes[C].INNIS MA.PCRProtocols:AGuideto Methods and Applications.New York Academic,1990:315-322.
[6] Baldauf SL,Palmer JD.Animals and fungi are each other’s closest relatives:congruent evidence from multiple proteins[J].Proc Natl Acad Sci USA,1993,90:11538-11562.
[7] O’Donnell KE,Cigelnik,Nirenberg HI.Molecular systematics and phylogeography of the Gibberellafujikumi species complex[J].Mycologia,1998,90:465-493.
[8] Yli-Mattilat T,Mach RL,Alekhinaia IA.Phylogenetic relationship of Fusarium langsethiaeto Fusarium poae and Fusarium sporotrichioidesas and inferred by IGS,ITS,β-tubulin sequences and UP-PCR hybridization analysis[J].International Journal of Food Microbiology,2004,95:267-285.
[9] Bichard.Penicilliumviticola,a new species isolated from a grape in Japan[J].Mycoscience,2011,52(5):338-343.
[10] Erpenbeck D,Nichols SA,Voigt O,et al.Phylogenetic analysesunder secondary structure-specific substitution models outperform traditional approaches:case studies with diploblast LSU[J].J Mol Evol,2007,64(5):543-557.
[11] Friedrich M,Tautz D.Ribosomal DNA phylogeny of the majorextant arthropod classes and the evolution of myriapods[J].Nature,1995,376(6536):165-167.
[12] Pryor B M,Gilbertson R L.Molecular phylogenetic relationships amongst Alternaria species and related fungi based upon analysis of nuclear LSU and mt SSU r DNA sequences[J].Mycological Research,2000,104(11):1312-1321.
[13] 魏景超.真菌鉴定手册[M].上海:上海科学技术出版,1979:130-133. [14] 张天宇.中国真菌志[M].北京:北京科学出版社,1883:136-139. [15] Tamura K,Peterson D,Peterson N,et al.MEGA5:molecular evolutionary genetics analysis using maximum likelihood,evolutionary distance,and maximum parsimony methods[J].Molecular Biology&Evolution,2011,28(10):2731-2739.
[16] 柳涛,童军茂,马伟荣,等.哈密瓜采后冷藏中主要病原菌的分离鉴定[J].现代食品科技,2013,29(8):2030-2034. [17] 曹娜,湛东锐,李连强,等.分子生物学与形态学相结合鉴定盐生海芦笋内生真菌Salicorn15[J].食品科学,2012,33(23):180-183. [18] 杜小琴,李杰,秦文,等.甜樱桃果实采后病原菌的分离鉴定及其生物学特性研究[J].食品工业科技,2015,4(18):197-202. [19] 王友升,陈玉娟,张燕.李果实贮藏期间4株病原真菌的分离、鉴定及碳源代谢指纹图谱分析[J].食品科学,2012,33(13):235-239. [20] Igual M,Garcia-Martinez E,Martin-Esparza M E,et al.Effect of processing on the drying kinetics and function value of dried apricot[J].Food Research International,2012,47(2):284-290.
[21] 吴思雅,阿衣木古丽·艾赛提,郑灿龙,等.新疆赛买提杏采后主要病原真菌的鉴定[J].食品工业科技,2016,37(1):149-152.
计量
- 文章访问数: 131
- HTML全文浏览量: 17
- PDF下载量: 316