Scientia Agricultura Sinica ›› 2014, Vol. 47 ›› Issue (12): 2335-2347.doi: 10.3864/j.issn.0578-1752.2014.12.006

• PLANT PROTECTION • Previous Articles     Next Articles

Genetic Diversity of Botrytis cinerea Isolates from Broad Bean

 HUANG  Yan-1, 2 , ZHU  Zhen-Dong-1, DUAN  Can-Xing-1, WU  Xiao-Fei-1, DONG  Fang-Yang-2   

  1. 1、Institute of Crop Sciences, Chinese Academy of Agricultural Sciences/National Key Facility for Crop Gene Resources and Genetic Improvement, Beijing 100081;
    2、College of Life Science and Technology, Hebei Normal University of Science & Technology,Changli 066600, Hebei
  • Received:2013-11-06 Online:2014-06-15 Published:2013-12-09

RichHTML

1

PDF

789

Cited

5

Abstract: 【Objective】Botrytis cinerea is one of the most important pathogens causing chocolate spot, blossom blight and pod rot in broad bean. B. cinerea is a species complex that comprises two cryptic species, Group I and Group II, and exhibits a great morphological and genetic diversity. However, the status of B. cinerea species complex from broad bean as well as their genetic characteristics have been poorly understood. Therefore, this study aims to determine the status and investigate the genetic diversity and genetic differentiation of isolates of B. cinerea complex from broad bean obtained from six different geographic regions. 【Method】The B. cinerea isolates were observed for cultural characteristics on potato dextrose agar. Transposons types of the isolates were detected using Flipper- and Boty-specific primers. Group determination of the isolates was analyzed by PCR-RFLP of Bc-hch gene, and their genetic characteristics was analyzed using SSR markers. 【Result】 All the 100 isolates were identified as B. cinerea group II. Three colony types, including mycelial type, sclerotial type and conidial type, were produced, and 76 isolates belonged to sclerotial type. Three transposon genotypes were identified in 92 isolates, which contained Boty-only, Fliper-only and Boty + Fliper genotypes. The most prevalent transposon genotype was Boty-only which was detected in isolates from Chongqing, Sichuan, Gansu and Hebei, while genotype Fliper-only was only found in isolates from Jiangsu. SSR analysis detected 92 multilocus genotypes (MLGs) among the 100 isolates with averaged gene and genotypic diversity of 0.5140 and 0.9982, respectively. Analysis of AMOVA revealed that genetic variation within populations accounted for 86.70% of the total genetic variation. The standardized index of association (rD) was estimated to range from 0.0312 to 0.1261 with the average value of 0.0491. In all comparsions, the values of fixation index (FST) ranged from 0.0085-0.2650 with the average value of 0.1330. All 100 isolates were divided into 10 genetic groups by UPGMA clustering analysis, and most of genetic groups included isolates from different geographic regions. In addition, two genetic groups were inferred in 92 MLGs by Bayesian analysis, and both populations from Chongqing and Sichuan belonged to one group, the population from Qinghai belonged to the other group, while popultions from Jiangsu, Hebei and Gansu were admixture of two genetic groups.【Conclusion】The populations of B. cinerea complex from broad bean made up of only B. cinerea group II. The sclerotial type was main colony morphology. Most of the isolates belonged to transposon genotypes whose distribution existed distinct geographic difference. Great genetic diversity was found in all six populations, and genetic variation mainly presented within populations. Sexual reproduction was the main mode of reproduction. Moderate to large genetic differentiation was detected among most populations.

Key words: Vicia faba , chocolate spot , Botrytis cinerea , transposon , genetic diversity

[1]Alghamdi S S, Migdadi H M, Ammar M H, Paull J G, Siddique K H M. Faba bean genomics: current status and future prospects. Euphytica, 2012, 186(3): 609-624.

[2]Harrison J G. The biology of Botrytis spp. on Vicia beans and chocolate spot disease-a review. Plant Pathology, 1988, 37(2): 168-201.

[3]俞大绂. 蚕豆病害. 北京: 科学出版社, 1979.

Yu D F. Diseases of Broad Bean. Beijing: Science Press, 1979. (in Chinese)

[4]王昆, 沈友康, 郑江, 沈亚萍, 王其礼, 刘锦生. 蚕豆赤斑病流行趋势预报的模糊识别模式研究. 植物病理学报, 1995, 25(3): 279-284.

Wang K, Shen Y K, Zheng J, Shen Y P, Wang Q L, Liu J S. On the fuzzy recognition models of epidemic trend forecasting of chocolate spot of broad bean (Botrytis fabae Sardina). Acta Phytopathologica Sinica, 1995, 25(3): 279-284. (in Chinese)

[5]黄燕, 段灿星, 陆鸣, 东方阳, 朱振东. 蚕豆赤斑病病原菌鉴定. 植物保护, 2012, 38(6): 115-118.

Huang Y, Duan C X, Lu M, Dongfang Y, Zhu Z D. Identification of pathogens causing chocolate spot on broad bean. Plant Protection, 2012, 38(6): 115-118. (in Chinese)

[6]Zhang J, Wu M D, Li G Q, Yang L, Yu L, Jiang D H, Huang H C, Zhuang W Y. Botrytis fabiopsis, a new species causing chocolate spot of broad bean in central China. Mycologia, 2010, 102(5): 1114-1126.

[7]Williamson B, Tudzynski B, Tudzynski P, van Kan J A. Botrytis cinerea: the cause of grey mould disease. Molecular Plant Pathology, 2007, 8(5): 561-580.

[8]Mirzaei S, Mohammadi Goltapeh E, Shams-Bakhsh M, Safaie N, Chaichi M. Genetic and phenotypic diversity among Botrytis cinerea isolates in Iran. Journal of Phytopathology, 2009, 157: 474-482.

[9]Esterio M, Muñoz G, Ramos C, Cofré G, Estévez R, Salinas A, Auger J. Characterization of Botrytis cinerea isolates present in Thompson seedless table grapes in the Central Valley of Chile. Plant Disease, 2011, 95(6): 683-690.

[10]Giraud T, Fortini D, Levis C, Leroux P, Brygoo Y. RFLP markers show genetic recombination in Botryotinia fuckeliana (Botrytis cinerea) and transposable elements reveal two sympatric species. Molecular Biology and Evolution, 1997, 14(11): 1177-1185.

[11]Giraud T, Fortini D, Levis C, Lamarque C, Leroux P, LoBuglio K, Brygoo Y. Two sibling species of the Botrytis cinerea complex, transposa and vacuma, are found in sympatry on numerous host plants. Phytopathology, 1999, 89(10): 967-973.

[12]Fournier E, Levis C, Fortini D, Giraud T, Leroux P, Brygoo Y. Characterization of Bc-hch, the Botrytis cinerea homolog of the Neurospora crassa het-c vegetative incompatibility locus, and its use as a population marker. Mycologia, 2003, 95(2): 951-961.

[13]Fournier E, Giraud T, Albertini C, Brygoo Y. Partition of the Botrytis cinerea complex in France using multiple gene genealogies. Mycologia, 2005, 97(6): 1251-1267.

[14]Ma Z H, Michailides T J. Genetic structure of Botrytis cinerea populations from different host plants in California. Plant Disease, 2005, 89(10): 1083-1089.

[15]Kretschmer M, Hahn M. Fungicide resistance and genetic diversity of Botrytis cinerea isolates from a vineyard in Germany. Journal of Plant Diseases and Protection, 2008, 115(5): 214-219.

[16]Leroch M, Plesken C, Weber R W, Kauff F, Scalliet G, Hahn M. Gray mold populations in German strawberry fields are resistant to multiple fungicides and dominated by a novel clade closely related to Botrytis cinerea. Applied and Environmental Microbiology, 2013, 79(1): 159-167.

[17]Fekete É, Fekete E, Irinyi L, Karaffa L, Árnyasi M, Asadollahi M, Sándro E. Genetic diversity of a Botrytis cinerea cryptic species complex in Hungary. Microbiological Research, 2012, 167(5): 283-291.

[18]Wessels B A, Lamprecht S C, Linde C C, Fourie P H, Mostert L. Characterization of the genetic variation and fungicide resistance in Botrytis cinerea populations on rooibos seedlings in the Western Cape of South Africa. European Journal of Plant Pathology, 2013, 136(2): 407-417.

[19]Walker A S, Gautier A, Confais J, Martinho D, Viaud M, Le Pêcheur P, Dupont J, Fournier E. Botrytis pseudocinerea, a new cryptic species causing gray mold in French vineyards in sympatry with Botrytis cinerea. Phytopathology, 2011, 101(12): 1433-1445.

[20]Fournier E, Giraud T. Sympatric genetic differentiation of a generalist pathogenic fungus, Botrytis cinerea, on two different host plants, grapevine and bramble. Journal of Evolutionary Biology, 2008, 21(1): 122-132.

[21]Isenegger D A, Ades P K, Ford R, Taylor P W J. Status of the Botrytis cinerea species complex and microsatellite analysis of transposon types in South Asia and Australia. Fungal Diversity, 2008, 29: 17-26.

[22]Möller E M, Bahnweg G, Sandermann H, Geiger H H. A simple and efficient protocol for isolation of high molecular weight DNA from filamentous fungi, fruit bodies and infected plant tissues. Nucleic Acids Research, 1992, 20(22): 6115-6116.

[23]Levis C, Fortini D, Brygoo Y. Flipper, a mobile Fot1-like transposable element in Botrytis cinerea. Molecular and General Genetics, 1997, 254(6): 674-680.

[24]Diolez A, Marches F, Fortini D, Brygoo Y. Boty, a long terminal repeat retroelement in the phytopathoenic fungus Botrytis cinerea. Applied and Environmental Microbiology, 1995, 61(1): 103-108.

[25]Fournier E, Giraud T, Loiseau A, Vautrin D, Estoup A, Solignac M, Cornuet J M, Brygoo Y. Characterization of nine polymorphic microsatellite loci inthe fungus Botrytis cinerea (Ascomycota). Molecular Ecology Notes, 2002, 2(3): 253-255.

[26]Yeh F, Yang R C, Boyle T B J, Ye Z H, Mao J X. POPGENE, the user-friendly shareware for population genetic analysis. Molecular Biology and Biotechnology Center, University of Alberta, Canada, 1999.

[27]Nei M, Li W H. Mathematical model for studying genetic variation in terms of restriction endonucleases. Proceedings of the National Academy of Sciences of the United States of America, 1979, 76(10): 5269-5273.

[28]Rohlf F J. NTSYSpc Version 2.1. Numerical Taxonomy and Multivariate Analysis System. Exeter Software, Setauket, New York, 1997.

[29]Excoffier L, Laval S, Schneider G. Arlequin version 3.0: an integrated software package for population genetics data analysis. Evolutionary Bioinformatics Online, 2005, 1: 47-50.

[30]Agapow P M, Burt A. Indices of multilocus linkage disequilibrium. Molecular Ecology Notes, 2001, 1(1/2): 101-102.

[31]Pritchard J K, Stephens M, Donelly P. Inference of population structure using multilocus genotype data. Genetics, 2000, 155: 945-959.

[32]Evanno G, Regnaut S, Goudet J. Detecting the number of clusters of individuals using the software STRUCTURE: a simulation study. Molecular Ecology, 2005, 14(8): 2611-2620.

[33]Earl D A, vonHoldt B M. STRUCTURE HARVESTER: a website and program for visualizing structure output and implementing the Evanno method. Conservation Genetics Resources, 2012, 4(2): 359-361.

[34]Karchani-Balma S, Gautier A, Raies A, Fournier E. Geography, plants, and growing systems shape the genetic structure of Tunisian Botrytis cinerea populations. Phytopathology, 2008, 98(12): 1271-1279.

[35]Muñoz C, Gómez Talquenca S, Oriolani E, Combina M. Genetic characterization of grapevine-infecting Botrytis cinerea isolates from Argentina. Revista Iberoamericana de Micología, 2010, 27(2): 66-70.

[36]Grindle M. Phenotypic differences between natural and induced variants of Botrytis cinerea. Journal of General Microbiology, 1979, 111(1): 109-120.

[37]Martinez F, Blancard D, Lecomte P, Levis C, Dubos B, Fermaud M. Phenotypic differences between vacuma and transposa subpopulations of Botrytis cinerea. European Journal of Plant Pathology, 2003, 109(5): 479-488.

[38]张静. 湖北省灰霉病病菌区系和灰葡萄孢菌多样性研究[D]. 武汉: 华中农业大学, 2010.

Zhang J. Study on taxonomy of Botrytis species in Hubei province and diversity of B. cinerea[D]. Wuhan: Huazhong Agricultural University, 2010. (in Chinese)

[39]Váczy K Z, Sándor E, Karaffa L, Fekete E, Fekete É, Árnyasi M, Czégledi L, Kovics G J, Druzhinina I S, Kubicek C P. Sexual recombination in the Botrytis cinerea populations in Hungarian vineyards. Phytopathology, 2008, 98(12): 1312-1319.

[40]Isenegger D A, Macleod W J, Ford R, Taylor P W J. Genotypic diversity and migration of clonal lineages of Botrytis cinerea from chickpea fields of Bangladesh inferred by microsatellite markers. Plant Pathology, 2008, 57: 967-973.
[1] ZHANG DongMei, ZHOU XinXin, XIAO GuiLin, ZENG XiangGuo, WANG ChunYan, WANG ZeXian, HAN YongChao. Phenotypic Characteristics of Strawberry Floral Organs in Response to Botrytis cinerea Infection and Methods for Gray Mold Resistance Evaluation [J]. Scientia Agricultura Sinica, 2026, 59(7): 1456-1466.
[2] ZHAO ZiJie, SONG Hao, DONG XiaoOu, WAN JianMin. Progress in Transposable Element-Assisted Targeted Insertion of Large DNA Fragments [J]. Scientia Agricultura Sinica, 2026, 59(6): 1141-1156.
[3] HE ZhiLin, SUN CuiXia, YUE HongLi, TAN YueXia, ZHANG YaoHai, WANG FuSheng, LIU SiTao, JIANG Dong. Genetic Diversity Analysis and GWAS of Alloocimene Based on Resequencing of Citron, Lemon Germplasm Resources [J]. Scientia Agricultura Sinica, 2026, 59(2): 386-401.
[4] ZHANG TianYu, LI Bai, ZANG JinPing, CAO HongZhe, DONG JinGao, XING JiHong, ZHANG Kang. Genome-Wide Identification and Expression Analysis of HMG Family Genes in Botrytis cinerea [J]. Scientia Agricultura Sinica, 2025, 58(4): 704-718.
[5] XU YuJuan, ZHANG Jie, WANG TianYi, CHEN HaoYang, ZHAO JiaJia, WU BangBang, HAO YuQiong, LI XiaoHua, ZHENG XingWei, ZUO JingJing, ZHENG Jun. Identification of Glu-A3 and Glu-B3 of Low-Molecular-Weight Glutenin in Shanxi Wheat and Its Effect on Quality [J]. Scientia Agricultura Sinica, 2025, 58(24): 5110-5127.
[6] CHEN CaiJin, MA Lin, JIANG QingXue, LIU JinHui, MIAO Tong, ZHANG ZhiPeng, MENG Xiang, MA XiaoRan, ZHOU XinYue, ZHANG Jian, LIU WenHui, WANG XueMin. Genetic Diversity Analysis of Phenotypic Traits of 244 Forage Oat Germplasm Resources [J]. Scientia Agricultura Sinica, 2025, 58(23): 4825-4836.
[7] WANG Fan, LIU ChenWei, LU HongChen, XU RenChao, BIAN XiaoChun. Transcriptome Analysis of Vicia faba Response to Alternaria alternata Infection and Validation of the Disease Resistance Function of VfPR4 [J]. Scientia Agricultura Sinica, 2025, 58(22): 4656-4672.
[8] WEI YiMin, ZHOU MeiLiang, TANG Yu. Origin, Evolution and Spread of Crop Buckwheat [J]. Scientia Agricultura Sinica, 2025, 58(21): 4305-4316.
[9] LIU XiaoXu, ZHONG ZeXin, QIU JiaRen, YANG ChunXiao, ZHANG YongJun, XIE Wen, ZHANG YouJun, PAN HuiPeng. GENETIC DIVERSITY OF MTCO1 IN DIFFERENT GEOGRAPHICAL POPULATIONS OF MEGALUROTHRIPS USITATUS [J]. Scientia Agricultura Sinica, 2025, 58(21): 4361-4371.
[10] GUO MengZe, ZHANG Lei, SUN PingPing, JIANG Biao, YAN JinQiang, LI ZhengNan. Molecular Characterization and Evolutionary Dynamics of Tomato Leaf Curl New Delhi Virus Isolate from Wax Gourd (Benincasa hispida) in Guangdong [J]. Scientia Agricultura Sinica, 2025, 58(19): 3890-3904.
[11] LUO JiaRui, WU SanLing, GUO Fu, LIU Zhen, SONG JingHan, TAN YuanYuan, SHU QingYao. Identification and Characterization of Retrotransposon Tos17 in the Genomes of Indica Rice [J]. Scientia Agricultura Sinica, 2025, 58(15): 2933-2947.
[12] LIU PengPeng, LI JiangBo, XU HongJun, NIE YingBin, HAN XinNian, KONG DeZhen, SANG Wei. Genetic Diversity Analysis of Protein Fractions and Quality in Xinjiang Winter Wheat Cultivar Resources [J]. Scientia Agricultura Sinica, 2025, 58(15): 2948-2959.
[13] WU CuiCui, WANG RuoYu, MA Chi, HE MeiYue, YIN XiaoKang, FENG JiaYi, ZHOU BinHan, JIANG YuFan, JIN HanBing, ZHAO LiLi, SUN Ji, FANG ZhengWu, CHENG Ling, ZHU ZhanWang, LIU YiKe, ZHANG YingXin, WANG ShuPing. Screening of Active Centromeric Retrotransposons of Rye and Their Response to Stress [J]. Scientia Agricultura Sinica, 2025, 58(14): 2720-2738.
[14] ZHAO YuLei, XIN JiaLu, LI ChengNan, LI Shan, XIE XuFei, YIN Xiao. Screening of Target Genes Downstream of VviERF045, a Transcription Factor Associated with Gray Mold Resistance in Vitis vinifera [J]. Scientia Agricultura Sinica, 2025, 58(13): 2578-2590.
[15] WANG Hui, DING BaoPeng, LI YuXian, REN QuanRu, ZHOU Hai, ZHAO JunLiang, HU HaiFei. Research Progress and Prospects on Crop Pan-Genomics [J]. Scientia Agricultura Sinica, 2025, 58(11): 2045-2061.
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