山东省玉米穗腐病病原菌的分离鉴定及优势种的系统发育分析

doi:10.7668/hbnxb.201751508

摘要/Abstract

摘要： 旨在明确山东省玉米穗腐病病原菌组成及优势病原菌的流行趋势。对2015-2017年在山东省采集的138份玉米穗腐病样品进行了病原菌的分离，采用形态学和分子生物学方法对分离物进行鉴定，并对优势病原菌进行系统发育分析，明确其发生发展规律。分离鉴定结果表明，分离到的真菌共有18种，其中镰孢属的拟轮枝镰孢菌分离频率为67.39%，为山东省优势病原菌；其次为层出镰孢菌，分离频率为10.87%；禾谷镰孢复合种、木贼镰孢菌和尖孢镰孢菌的分离频率分别为7.97%，5.80%，0.72%；木霉属的哈茨木霉分离频率为7.91%；青霉属的草酸青霉分离频率为6.52%；曲霉属黄曲霉和黑曲霉的分离频率分别为2.17%和5.07%。基于EF-1α基因序列构建了山东省2015-2017年拟轮枝镰孢菌的系统发育树，并分析了山东省菌株与河北省菌株的遗传进化关系。系统发育分析结果发现，89个拟轮枝镰孢菌菌株可以分为2个大群，其遗传距离较小，为0.002~0.016，表明山东省2015-2017年的菌株之间遗传关系较近；河北省和山东省拟轮枝镰孢菌菌株的遗传距离较小，相似性较高，亲缘关系较近，存在较为频繁的基因交流。研究结果为深入开展拟轮枝镰孢菌穗腐病的抗性育种、发生发展规律以及综合防治提供科学依据。

关键词: 玉米穗腐病, 拟轮枝镰孢菌, 分离频率, 遗传关系

Abstract: The objective of this study was to determine the composition of pathogen causing Maize ear rot in Shandong Province and to analyze the epidemic trend of dominant pathogen. A total of 138 Maize ear rot samples were collected from Shandong Province during 2015-2017 and were used for pathogen isolation.The isolates were identified by morphological and molecular biology methods, and the occurrence regularity of dominant pathogen was clarified by phylogenetic analysis. The results of isolation and identification showed that a total of 18 fungi were obtained. The isolation frequency of Fusarium verticillioides was 67.39%, which was the dominant pathogen in Shandong Province, followed by F. proliferatum, F.graminearum, F.equiseti and F.oxysporium with separation frequencies of 10.87%, 7.97%, 5.80% and 0.72%, respectively; And the separation frequencies of Trichoderma harzianum, Penicillium oxalicum, Aspergillus flavus and A. niger were 7.91%, 6.52%, 2.17% and 5.07%, respectively. Based on the EF-1α gene sequence, the phylogenetic tree of F. verticillioides isolates in Shandong Province from 2015 to 2017 was constructed, and the genetic evolution relationship of isolates between Shandong Province and Hebei Province was analyzed. Phylogenetic analysis showed that 89 isolates of F. verticillioides could be divided into two large groups with a small genetic distance of 0.002-0.016, indicating that the genetic relationship between these isolates was relatively close. The genetic distance of F. verticillioides isolates in Shandong Province and Hebei Province was small and showed a close genetic relationship, indicating that there were frequent gene exchanges. The results of this study would provide a scientific basis for resistance breeding and integrated control of F. verticillioides.

Key words: Maize ear rot, Fusarium verticillioides, Separation frequency, Genetic relationship

中图分类号:

S435.131

丁梦军, 杨扬, 孙华, 马红霞, 刘树森, 石洁. 山东省玉米穗腐病病原菌的分离鉴定及优势种的系统发育分析[J]. 华北农学报, 2019, 34(5): 216-223. doi: 10.7668/hbnxb.201751508.

DING Mengjun, YANG Yang, SUN Hua, MA Hongxia, LIU Shusen, SHI Jie. Isolation and Identification of Maize ear rot Pathogens and Phylogenetic Analysis of Dominant Species in Shandong Province[J]. ACTA AGRICULTURAE BOREALI-SINICA, 2019, 34(5): 216-223. doi: 10.7668/hbnxb.201751508.

http://www.hbnxb.net/CN/Y2019/V34/I5/216

参考文献

[1] 任旭. 我国玉米穗腐病主要致病镰孢菌多样性研究[D]. 北京:中国农业科学院,2011.doi:10.7666/d.Y1932401.
Rn X. Diwersitu analyses of Fusarium spp., the main causal agents of maize ear rot in China[D]. Beijing:Chinese Academy of Agricultural Sciences, 2011.
[2] Mesterházy á, Lemmens M, Reid L M. Breeding for resistance to ear rots caused by Fusarium spp. in maize-a review[J]. Plant Breeding, 2012, 131(1):1-19. doi:10.1111/j.1439-0523.2011.01936.x.
[3] 段灿星,王晓鸣,宋凤景,孙素丽,周丹妮,朱振东.玉米抗穗腐病研究进展[J].中国农业科学,2015,48(11):2152-2164. doi:10.3864/j.issn.0578-1752.2015.11.007.
Dan C X, Wang X M, Song F J, Sun S L, Zhou D N, Zhu Z D. Advances in research on maize resistance to ear rot[J].Scientia Agricultura Sinica,2015,48(11):2152-2164.
[4] 胡韬纲.玉米穗腐病研究进展[J].粮食科技与经济,2015,40(3):50-52.doi:10.16465/j.gste.cn431252ts.20150315.
H T G. Research progress on maize ear rot[J]. Grain Science and Technology and Economy, 2015, 40(3):50-52.
[5] 席靖豪. 黄淮海夏玉米穗腐病病原多样性分析及玉米新品种抗病性鉴定[D].郑州:河南农业大学,2018.
X J H. Pathogen diversity of summer maize ear rot in Huang-Huai-Hai region and resistance identification of new maize cultivars[D]. Zhengzhou:Henan Agricultural University, 2018.
[6] Stumpf R,dos Santos J,Gomes L B,Silva C N,Tessmann D J,Ferreira F D,Junior M M,Del Ponte E M. Fusarium species and fumonisins associated with maize kernels produced in Rio Grande do Sul State for the 2008/09 and 2009/10 growing seasons[J].Brazilian Journal of Microbiology,2013,44(1):89-95. doi:10.1590/S1517-83822013000100012.
[7] Görtz A, Oerke E C, Steiner U, Waalwijk C,Vries I D,Dehne H W. Biodiversity of Fusarium species causing ear rot of maize in Germany[J]. Cereal Research Communications, 2008, 36(S6):617-622.doi:10.1556/CRC.36.2008.Suppl.B.51.
[8] 秦子惠,任旭,江凯,武小菲,杨知还,王晓鸣.我国玉米穗腐病致病镰孢种群及禾谷镰孢复合种的鉴定[J].植物保护学报,2014,41(5):589-596. doi:10.13802/j.cnki.zwbhxb.2014.05.032.
Qn Z H, Ren X, Jiang K, Wu X F, Yang Z H, Wang X M. Identification of Fusarium species and F.graminearum species complex causing maize ear rot in China[J]. Acta Phytophylacica Sinica,2014,41(5):589-596.
[9] 燕玉超,张福平,刘筱,雷声剑.河南省气候变化特征及其对旱涝的影响[J].中国沙漠,2016,36(3):777-786.doi:10.7522/j.issn.1000-694X.2015.00080.
Yn Y C, Zhang F P, Liu X, Lei S J. Climate change and its influence on drought and flood in Henan, China[J]. Journal of Desert Research, 2016, 36(3):777-786.
[10] 朱峰,孙兆明, 李树超.山东省玉米生产现状与发展对策分析[J].农业科技通讯,2015(3):6-9,12.doi:10.3969/j.issn.1000-6400.2015.03.002.
Zu F, Sun Z M, Li S C. Analysis of corn production status and development countermeasures in Shandong Province[J]. Bulletin of Agricultural Science and Technology, 2015(3):6-9,12.
[11] 魏景超. 真菌鉴定手册[M]. 上海:上海科学技术出版社,1979.
Wi J C. Fungal identification manual[M].Shanghai:Shanghai Science and Technology Press, 1979.
[12] 方中达. 植病研究法[M]. 3版.北京:中国农业出版社,1998.
Fng Z D. Plant disease research method[M].The third edition. Beijing:China Agriculture Press, 1998.
[13] White T J, Bruns T, Lee S, Taylor J. Amplification and direct sequencing of fungal ribosomal RNA genes for phylogenetics[J]. PCR Protocols:a Guide to Methods and Applications, 1990, 18:315-322.doi:10.1016/B978-0-12-372180-8.50042-1.
[14] Nicholson P, Simpson D R, Weston G, Rezanoor H N,Lees A K,Parry D W,Joyce D. Detection and quantification of Fusarium culmorum and Fusarium graminearum in cereals using PCR assays[J]. Physiological and Molecular Plant Pathology, 1998, 53(1):17-37.doi:10.1006/pmpp.1998.0170.
[15] Mulè G, Susca A, Stea G, Moretti A. A species-specific PCR assay based on the calmodulin partial gene for identification of Fusarium verticillioides, F. proliferatum and F. subglutinans[J]. European Journal of Plant Pathology, 2004, 110(5/6):495-502.doi:10.1023/b:ejpp.0000032389.84048.71.
[16] O'Donnell K, Ward T J, Aberra D, Kistlercd H C,Aokie T,Orwiga N,Kimuraf M,Bjørnstadb Å,Klemsdalb S S.Multilocus genotyping and molecular phylogenetics resolve a novel head blight pathogen within the Fusarium graminearum species complex from Ethiopia[J]. Fungal Genetics and Biology, 2008, 45(11):1514-1522.doi:10.1016/j.fgb.2008.09.002.
[17] 石琳,段霞瑜,高洁,周益林,曹世勤,张勃.甘肃南部小麦白粉菌群体的毒性及多基因片段分析[J].植物病理学报,2014,44(4):414-421. doi:10.13926/j.cnki.apps.2014.04.010.
Si L, Duan X Y, Gao J, Zhou Y L, Cao S Q, Zhang B. Virulence and multigene analyses of the wheat powdery mildew population in Southern Gansu[J]. Acta Phytopathologica Sinica, 2014, 44(4):414-421.
[18] 孙华,张海剑,郭宁,石洁,陈丹,马红霞.黄淮海夏玉米主产区穗腐病病原菌的分离鉴定[J].植物保护学报,2017,44(5):796-802.doi:10.13802/j.cnki.zwbhxb.2017.2016078.
Sn H, Zhang H J, Guo N, Shi J, Chen D, Ma H X. Isolation and identification of pathogens causing maize ear rot in Huang-Huai-Hai summer corn region[J]. Journal of Plant Protection, 2017,44(5):796-802.
[19] 秦子惠. 玉米穗腐病致病镰孢特征及玉米-小麦致病镰孢关系研究[D].北京:中国农业科学院,2014.
Qn Z H. Characterists of Fusarium species causing maize ear rot and relationship of pathogenic Fusarium spp. between maize and wheat[D]. Beijing:Chinese Academy of Agricultural Sciences Dissertation, 2014.
[20] 郭成,魏宏玉,郭满库,何苏琴,金社林,陈红梅,王晓鸣,郭建国.甘肃玉米穗腐病样品中轮枝镰孢菌的分离鉴定及生物学特性[J].植物病理学报,2014,44(1):17-25. doi:10.13926/j.cnki.apps.2014.01.003.
Go C,Wei H Y,Guo M K,He S Q,Jin S L,Chen H M,Wang X M,Guo J G. Isolation,identification and biological characteristics of Fusarium verticillioides from maize ear rot sam ples in Gansu Province[J].Acta Phytopathologica Sinica, 2014, 44(1):17-25.
[21] 周丹妮,王晓鸣,李丹丹,杨洋,陈国康,段灿星.重庆及周边地区玉米穗腐病致病镰孢菌的分离与鉴定[J].植物保护学报,2016,43(5):782-788. doi:10.13802/j.cnki.zwbhxb.2016.05.011.
Zou D N,Wang X M, Li D D, Yang Y,Chen G K, Duan C X. Isolation and identification of Fusarium species causing maize ear rot in Chongqing and its vicinity[J].Journal of Plant Protection,2016,43(5):782-788.
[22] 肖淑芹,许佳宁,闫丽斌,隋韵涵,薛春生,陈捷.辽宁省玉米镰孢穗腐病病原菌的鉴定与分布[J].植物保护学报,2017,44(5):803-808. doi:10.13802/j.cnki.zwbhxb.2017.2016056.
Xao S Q, Xu J N, Yan L B, Sui Y H, Xue C S, Chen J. Identification and distribution of Fusarium species causing maize ear rot in Liaoning Province[J]. Journal of Plant Protection, 2017, 44(5):803-808.
[23] 孙华,丁梦军,张家齐,石洁,郭宁,李坡.河北省穗腐病病原菌鉴定及潜在产伏马毒素镰孢菌系统发育分析[J].植物病理学报, 2019, 49(2):151-159.doi:10.13926/j.cnki.apps.000284.
Sn H, Ding M J, Zhang J Q, Shi J, Guo N, Li P. Identification of pathogens causing maize ear rot and the phylogenetic analysis of fumonisins-producing Fusarium species in Hebei Province[J]. Acta Phytopathologica Sinica, 2019, 49(2):151-159.
[24] 许大凤,张海珊,李廷春,齐耀程,姜超强,周本国.安徽凤阳玉米穗腐病病原菌的分离鉴定[J].安徽农业科学,2017,45(25):145-148,150. doi:10.13989/J.cnki.0517-6611.2017.25.046.
X D F,Zhang H S,Li T C,Qi Y C,Jiang C Q,Zhou B G. Isolation and identification of the pathogen of maize ear rot in Fengyang,Anhui Province[J]. Journal of Anhui Agricultural Sciences, 2017, 45(25):145-148,150.
[25] 张婷,孙晓东,吕国忠.我国东北地区玉米穗腐镰孢菌的种类及其分离频率[J].菌物研究,2011,9(1):9-14,36. doi:10.13341/j.jfr.2011.01.006.
Zang T, Sun X D, Lü G Z. Fusarium species and its isolation frequency from rot ears of maize in Northeast China[J]. Journal of Fungal Research, 2011, 9(1):9-14,36.
[26] 高艳,李艳荣,王秀丽. 开封市1980-2009年9月份降雨情况对玉米品质的影响[J]. 河南农业,2011(1):54. doi:10.3969/j.issn.1006-950X.2011.01.047.
Go Y, Li Y R, Wang Y L. Effects on maize quality of rainfall from 1980 to September 2009 in Kaifeng City[J]. Agriculture of Henan, 2011(1):54.
[27] Bottalico A. Fusarium diseases of cereals:species complex and related mycotoxin profiles, in Europe[J]. Journal of Plant Pathology, 1998, 80(2):85-103. doi:10.2307/41997909.
[28] Logrieco A,Mulè G,Moretti A, Bottalico A. Toxigenic Fusarium species and mycotoxins associated with maize ear rot in Europe[J].European Journal of Plant Pathology, 2002,108(7):597-609. doi:10.1023/A:1020679029993.
[29] 宋嘉佳,王娜,伯忠凯,康桂红.2015年秋季(9-11月)山东天气评述[J].山东气象,2015,35(4):50-52.doi:10.19513/j.cnki.issn1005-0582.2015.04.011.
Sng J J, Wang N, Bo Z K, Kang G H. A review of the weather in Shandong during the autumn of 2015(September-November)[J]. Journal of Shandong Meteorology, 2015, 35(4):50-52.
[30] 王文青,万明波,孟祥新,康桂红,李静.2015年夏季(6-8月)山东天气评述[J].山东气象,2015,35(3):52-55. doi:10.19513/j.cnki.issn1005-0582.2015.03.013.
Wng W Q, Wan M B, Meng X X, Kang G H, Li J. A review of the weather in Shandong during the summer of 2015(June-August)[J]. Weather of Shandong,2015,35(3):50-52.
[31] Chulze S N, Ramirez M L, Torres A, Leslie J F. Genetic variation in Fusarium section Liseola from no-till maize in Argentina[J]. Applied and Environmental Microbiology, 2000, 66(12):5312-5315. doi:10.1128/AEM.66.12.5312-5315.2000.
[32] 任旭,朱振东,李洪杰,段灿星,王晓鸣.轮枝镰孢SSR标记开发及在玉米分离群体遗传多样性分析中的应用[J].中国农业科学,2012,45(1):52-66. doi:10.3864/j.issn.0578-1752.2012.01.007.
Rn X, Zhu Z D, Li H J, Duan C X, Wang X M. SSR Marker development and analysis of genetic diversity of Fusarium verticillioides isolated from maize in China[J]. Scientia Agricultura Sinica, 2012,45(1):52-66.

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