甘蓝型油菜抗菌核病研究进展

doi:10.11924/j.issn.1000-6850.casb15010180

摘要/Abstract

摘要： 油菜是主要油料作物之一，由核盘菌(Sclerotiniasclerotiorum(Lib.)deBary)引起的菌核病(sclerotiniastemrot)是油菜的主要病害。针对当前与菌核病相关研究的新进展，本研究从4个方面对其进行了概括：（1）核盘菌的侵染方式以及在侵染过程中核盘菌分泌的草酸与寄主中钙离子的动态关系；（2）油菜通过合成植保素、酚类化合物、木质素、几丁质酶和β-1,3-葡聚糖酶等来抵抗核盘菌入侵的抗病机理；（3）现有综合防治技术中抗性种质筛选、无花瓣育种和生物防治；（4）利用与草酸代谢相关基因、抗病相关基因、防御相关转录因子基因和抗菌肽基因开展的油菜基因工程研究成果。并且进一步提出了未来油菜抗菌核病研究的可能方向，这些总结与建议为今后油菜抗菌核病研究提供了有益参考。

关键词: 樱桃番茄, 樱桃番茄, 子叶, 再生体系

Abstract: Brassica napus was one of the major oil crops in the world. Sclerotinia stem rot was the main disease of Brassica napus caused by Sclerotinia sclerotiorum (Lib.) de Bary. In view of the current research progresses on sclerotinia stem rot, four related aspects were summarized in this paper: (1) the infection mode of Sclerotinia sclerotiorum and the dynamic relationship between oxalic acid secreted by Sclerotinia sclerotiorum and calcium ions in host; (2) the resistance mechanism of oilseed rape to sclerotinia stem rot by using synthesizing phytoalexin, phenolic compounds, lignin, chitinase and β- 1,3- glucanase; (3) resistant germplasm selection, apetalous breeding and microbial control in the comprehensive control techniques; (4) the research achievements in gene engineering by transforming oilseed rape with the genes relative to oxalic acid metabolism, pathogen resistance, defense- related transcription regulation factor and antibiotic peptide. Possible directions for future study on resistance to the sclerotinia stem rot of oilseed rape were further put forward and references were provided.

刘正立,刘春林. 甘蓝型油菜抗菌核病研究进展[J]. 中国农学通报, 2015, 31(15): 114-123.

参考文献

[1] Bolton M D, Thomma B P H J, Nelson B D. Sclerotinia sclerotiorum (Lib.) de Bary: biology and molecular traits of a cosmopolitan pathogen[J]. Molecular Plant Pathology, 2006, 7(1): 1- 16.
[2] Boland G J, Hall R. Index of plant hosts of Sclerotinia sclerotiorum[J]. Canadian Journal of Plant Pathology, 1994, 16(2): 93-108.
[3] Barbetti M J, Banga S K, Fu T D, et al. Comparative genotype reactions to Sclerotinia sclerotiorum within breeding populations of Brassica napus and B. juncea from India and China[J]. Euphytica, 2014,197(1):47-59.
[4] 费维新,李强生,吴新杰,等.利用栽培措施控制油菜菌核病的研究[J].中国油料作物学报,2002(03):47-49.
[5] McCartney H, Doughty K, Norton G, et al. A study of the effect of disease on seed quality parameters of oilseed rape[A]. Proc 10th Intern Rap Cong, Canberra, Australia, 1999.
[6] 张源,阮颖,彭琦,等.油菜菌核病致病机理研究进展[J].作物研究, 2006(05):549-551.
[7] 双桑,阮颖,彭冬平,等.油菜菌核病及抗病育种研究进展[J].作物研究,2006(05):552-556.
[8] Harel A, Bercovich S, Yarden O. Calcineurin Is Required for Sclerotial Development and Pathogenicity of Sclerotinia sclerotiorum in an Oxalic Acid- Independent Manner[J]. Molecular Plant-Microbe Interactions, 2006,19(6):682-693.
[9] Purdy L H. Sclerotinia sclerotiorum: History, Diseases and Symptomatology, Host Range, Geographic Distribution, and Impact[J]. Phytopathology, 1979,69(8):875.
[10] 王婧,刘泓利,宋超,等.甘蓝型油菜叶表皮蜡质组分及结构与菌核病抗性关系[J].植物生理学报,2012(10):958-964.
[11] 吴纯仁,刘后利.油菜菌核病致病机理的研究Ⅳ.病菌侵入途径和附着胞结构的观察[J].华中农业大学学报,1990(01):56-58.
[12] 杨谦.核盘菌侵入油菜超微结构及侵染机制的研究[J].植物病理学报,1994(03):245-249.
[13] Kim K S, Min J Y, Dickman M B. Oxalic Acid Is an Elicitor of Plant Programmed Cell Death during Sclerotinia sclerotiorum Disease Development[J]. Molecular Plant- Microbe Interactions, 2008, 21(5): 605-612.
[14] Bateman D, Beer S. Simultaneous production and synergistic action of oxalic acid and polygalacturonase during pathogenesis by Sclerotium rolfsii[J]. Phytopathology, 1965, 55: 204.
[15] Guimar?es R L, Stotz H U. Oxalate production by Sclerotinia sclerotiorum deregulates guard cells during infection[J]. Plant Physiology, 2004, 136(3): 3703-3711.
[16] Cessna S G, Sears V E, Dickman M B, et al. Oxalic Acid, a Pathogenicity Factor for Sclerotinia sclerotiorum, Suppresses the Oxidative Burst of the Host Plant[J]. The Plant Cell Online, 2000, 12(11): 2191-2199.
[17] 吕琳慧,徐幼平,任至玄,等.Ca2+信号通路对本氏烟叶位介导的核盘菌抗性的影响[J].浙江大学学报:农业与生命科学版,2014(06): 605-610.
[18] Heller A, Witt-Geiges T. Oxalic acid has an additional, detoxifying function in Sclerotinia sclerotiorum pathogenesis[J]. PLoS One, 2013, 8(8): e72292.
[19] Dixon R A, Lamb C J. Molecular Communication in Interactions Between Plants and Microbial Pathogens[J]. Annual Review of Plant Physiology and Plant Molecular Biology, 1990,41(1):339-367.
[20] Pedras M S, Ahiahonu P W. Phytotoxin production and phytoalexin elicitation by the phytopathogenic fungus Sclerotinia sclerotiorum[J]. Journal of Chemical Ecology, 2004,30(11):2163-2179.
[21] 赵建伟,肖玲,何凤仙,等.甘蓝型油菜远缘杂交新品系某些酶的活性与抗菌核病的关系[J].中国油料作物学报,1998(01):41-44.
[22] 姜伟丽.油菜抗菌核病品种的筛选和抗性机理分析[D].南京:南京农业大学,2008.
[23] Eynck C, Koopmann B, Karlovsky P, et al. Internal resistance in winter oilseed rape inhibits systemic spread of the vascular pathogen Verticillium longisporum[J]. Phytopathology,2009,99(7): 802-811.
[24] Broekaert W F, Terras F R, Cammue B P, et al. Plant defensins: novel antimicrobial peptides as components of the host defense system[J]. Plant Physiology, 1995, 108(4): 1353-1358.
[25] 杨向东.木质素合成调控及其与甘蓝型油菜抗菌核病和抗倒伏性关系研究[D].北京:中国农业科学院,2006.
[26] 刘丹.木质素相关基因表达调控对油菜木质素合成及抗菌核病和抗倒伏性的影响[D].北京:中国农业科学院,2008.
[27] Taira T, Ohnuma T, Yamagami T, et al. Antifungal activity of rye (Secale cereale) seed chitinases: the different binding manner of class I and class II chitinases to the fungal cell walls[J]. Biosci Biotechnol Biochem, 2002, 66(5): 970-977.
[28] Takenaka Y, Nakano S, Tamoi M, et al. Chitinase gene expression in response to environmental stresses in Arabidopsis thaliana: chitinase inhibitor allosamidin enhances stress tolerance[J]. Biosci Biotechnol Biochem, 2009, 73(5): 1066-1071.
[29] Lawton K, Ward E, Payne G, et al. Acidic and basic class III chitinase mRNA accumulation in response to TMV infection of tobacco[J]. Plant Molecular Biology, 1992, 19(5): 735-743.
[30] Ishisaki K, Honda Y, Taniguchi H, et al. Heterogonous expression and characterization of a plant class IV chitinase from the pitcher of the carnivorous plant Nepenthes alata[J]. Glycobiology, 2012, 22 (3): 345-351.
[31] Belchi-Navarro S, Almagro L, Sabater-Jara A B, et al. Induction of trans- resveratrol and extracellular pathogenesis- related proteins in elicited suspension cultured cells of Vitis vinifera cv Monastrell[J]. Journal of Plant Physiology, 2013, 170(3): 258-264.
[32] Ahmed N U, Park J I, Seo M S, et al. Identification and expression analysis of chitinase genes related to biotic stress resistance in Brassica[J]. Molecular Biology Reports, 2012, 39(4): 3649-3657.
[33] Floerl S, Druebert C, Majcherczyk A, et al. Defence reactions in the apoplastic proteome of oilseed rape (Brassica napus var. napus) attenuate Verticillium longisporum growth but not disease symptoms[J]. BMC Plant Biology, 2008, 8: 129.
[34] Li C X, Liu S Y, Sivasithamparam K, et al. New sources of resistance to Sclerotinia stem rot caused by Sclerotinia sclerotiorum in Chinese and Australian Brassica napus and B. juncea germplasm screened under Western Australian conditions[J]. Australasian Plant Pathology, 2009, 38(2): 149-152.
[35] 陈玉卿,张洁夫,伍贻美,等.芸薹属油菜种质资源抗(耐)菌核病、病毒病的鉴定[J].中国油料,1993(02):6-9.
[36] M.H D R, Petzoldt. Breeding for resistance to sclerotinia sclerotiorum in brassica oleracea[J]. Acta Hort. (ISHS), 1996,407: 103-108.
[37] 黄永菊,陈军,李云昌.甘蓝型油菜菌核病抗(耐)性的遗传研究Ⅰ. 抗性遗传属性与配合力分析[J].中国油料作物学报,2000(04):2-6.
[38] Mei J, Qian L, Disi J O, et al. Identification of resistant sources against Sclerotinia sclerotiorum in Brassica species with emphasis on B. oleracea[J]. Euphytica, 2011, 177(3): 393-399.
[39] 刘澄清,黄有菊,邹崇顺,等.中双二号双低高产多抗(耐)甘蓝型油菜新品种选育[J].中国油料,1990(04):63-67.
[40] Wang H, Liu G, Zheng Y, et al. Breeding of Brassica napus cultivar Zhongshuang No. 9 with resistance to Sclerotinia sclerotiorum[J]. Chinese journal of oil crop sciences,2002,24(4):71-73.
[41] 陈社员,官春云,王国槐,等.双低油菜新品种湘油 15号的选育[J].湖南农业大学学报:自然科学版,2003(2):103-105.
[42] 李强生,Hamc Cartney, AHeran,等.油菜菌核病病原菌侵染条件的研究[J].安徽农业科学,2000(3):314-315.
[43] Mclean D M. Role of dead flower parts in infection of certain crucifers by Sclerotinia sclerotiorum (Lib) de Bary[J]. Plant Dis. Reptr,1958,42(5):663-666.
[44] Jamaux I, Gelie B, Lamarque C. Early stages of infection of rapeseed petals and leaves by Sclerotinia sclerotiorum revealed by scanning electron microscopy[J]. Plant Pathology,1995,44(1):22-30.
[45] 傅寿仲.油菜高产抗病育种的新思路—无花瓣育种[J].上海农业学报,1990(03):76-77.
[46] 蔡明,刘贵华,秦选佑,等.甘蓝型油菜与埃塞俄比亚芥种间杂种的无花瓣性状表现[J].中国油料,1991(01):88-89.
[47] 张洁夫.甘蓝型油菜无花瓣和脂肪酸的遗传与分子标记[D].南京: 南京农业大学,2007.
[48] Fray J M, Evans, et al. Physiological assessment of apetalous flowers and erectophile pods in oilseed rape (Brassica napus)[M]. Cambridge, Royaume-Uni: Cambridge University Press,1996.
[49] 张瑞茂,李敏,陈大伦,等.甘蓝型无花瓣油菜的缺陷及克服途径研究[J].贵州农业科学,2010(04):12-16.
[50] 陈大伦,张瑞茂,李敏,等.甘蓝型无花瓣油菜对菌核病的抗(耐)性研究[J].贵州农业科学,2010(03):70-75.
[51] Jiang L. A Mutant with Apetalous Flowers in Oilseed Rape (Brassica napus): Mode of Inheritance and Influence on Crop Physiology and Sclerotinia Infection[D]. G?ttingen, Georg-AugustUniversity of G?ttingen,2001.
[52] 杨龙.生防菌盾壳霉防治油菜菌核病的生态学基础及其应用研究[D].武汉:华中农业大学,2009.
[53] 高俊明,马丽娜,李欣,等.盾壳霉对核盘菌的拮抗作用研究[J].植物保护,2006(06):66-70.
[54] 张姝,韩巨才.植病生防菌小盾壳霉的两种商品制剂[J].农药,2007 (12):846-856.
[55] 侯毅平,章四平,王建新,等.枯草芽胞杆菌 NJ-18对油菜菌核病的防治效果及其定殖动态[J].植物病理学报,2013(04):411-417.
[56] 高小宁,陈亚菲,韩青梅,等.内生细菌 EDR2的鉴定及其对油菜菌核病的防治[J].西北农林科技大学学报:自然科学版,2013(02): 175-188.
[57] 陈士云,杨宝玉,高梅影,等.一株抑制油菜核盘菌菌核形成的解淀粉芽孢杆菌[J].应用与环境生物学报,2005(03):373-376.
[58] Fernando W G D, Nakkeeran S, Zhang Y, et al. Biological control of Sclerotinia sclerotiorum (Lib.) de Bary by Pseudomonas and Bacillus species on canola petals[J]. Crop Protection,2007,26(2): 100-107.
[59] Li H, Li H B, Bai Y, et al. The use of Pseudomonas fluorescens P13 to control sclerotinia stem rot (Sclerotinia sclerotiorum) of oilseed rape[J]. Journal of Microbiology,2011,49(6):884-889.
[60] Yu X, Li B, Fu Y P, et al. A geminivirus- related DNA mycovirus that confers hypovirulence to a plant pathogenic fungus[J]. Proceedings of the National Academy of Sciences of the United States of America,2010,107(18):8387-8392.
[61] Yu X, Li B, Fu Y P, et al. Extracellular transmission of a DNA mycovirus and its use as a natural fungicide[J]. Proceedings of the National Academy of Sciences of the United States of America, 2013,110(4):1452-1457.
[62] Dong X, Ji R, Guo X, et al. Expressing a gene encoding wheat oxalate oxidase enhances resistance to Sclerotinia sclerotiorum in oilseed rape (Brassica napus)[J]. Planta, 2008, 228(2): 331-340.
[63] Thompson C, Dunwell J, Johnstone C, et al. Degradation of oxalic acid by transgenic oilseed rape plants expressing oxalate oxidase[J]. Euphytica,1995,85(1-3):169-172.
[64] Lan H Y, Wang C H, Zhang L H, et al. Studies on transgenic oilseed rape (Brassica napus) plants transformed with beta- 1,3- glucanase and chitinase genes and its resistance to Sclerotinia sclerotiorium[J]. Sheng Wu Gong Cheng Xue Bao,2000,16(2):142-146.
[65] Grison R, Grezes- Besset B, Schneider M, et al. Field tolerance to fungal pathogens of Brassica napus constitutively expressing a chimeric chitinase gene[J]. Nature Biotechnology,1996,14(5):643- 646.
[66] Mondal K K, Chatterjee S C, Viswakarma N, et al. ChitinaseMediated Inhibitory Activity of Brassica Transgenic on Growth of Alternaria brassicae[J]. Current Microbiology,2003,47(3):171-173.
[67] Mondal K K, Bhattacharya R C, Koundal K R, et al. Transgenic Indian mustard (Brassica juncea) expressing tomato glucanase leads to arrested growth of Alternaria brassicae[J]. Plant Cell Reports, 2007, 26(2): 247-252.
[68] 陈雁,饶勇强,孟金陵.转双价广谱抗病基因创造甘蓝型油菜抗菌核病新品种的研究[J].分子植物育种,2003(04):457-463.
[69] Liu H, Guo X, Naeem M S, et al. Transgenic Brassica napus L. lines carrying a two gene construct demonstrate enhanced resistance against Plutella xylostella and Sclerotinia sclerotiorum[J]. Plant Cell, Tissue and Organ Culture (PCTOC), 2011, 106(1): 143-151.
[70] 王新发.转几丁质酶基因和β-1,3-葡聚糖酶基因的甘蓝型油菜及其对菌核病抗性的研究[D].北京:中国农业科学院,2003.
[71] Melander M, Kamnert I, Happstadius I, et al. Stability of transgene integration and expression in subsequent generations of doubled haploid oilseed rape transformed with chitinase and beta- 1,3- glucanase genes in a double-gene construct[J]. Plant Cell Reports, 2006,25(9):942-952.
[72] 张学江,黄军艳,付浩,等.过表达甘蓝型油菜BnNRR基因增强了油菜对菌核病的感病性[C].中国植物病理学会 2009年学术年会,中国云南昆明,2009:1.
[73] 陈丽玲.在油菜中超量表达拟南芥 NPR1基因对油菜抗病性的影响[D].武汉:湖北大学,2011.
[74] Wang Z, Fang H, Chen Y, et al. Overexpression of BnWRKY33 in oilseed rape enhances resistance to Sclerotinia sclerotiorum[J]. Molecular Plant Pathology,2014,15(7):677-689.
[75] Wu J, Wu L T, Liu Z B, et al. A plant defensin gene from Orychophragmus violaceus can improve Brassica napus' resistance to Sclerotinia sclerotiorum[J]. African Journal of Biotechnology, 2009,8(22):6101-6109.
[76] Hennin C, Hofte M, Diederichsen E. Functional expression of Cf9 and Avr9 genes in Brassica napus induces enhanced resistance to Leptosphaeria maculans[J]. Mol Plant Microbe Interact,2001,14(9): 1075-1085.
[77] Wang Y, Fristensky B. Transgenic canola lines expressing pea defense gene DRR206 have resistance to aggressive blackleg isolates and to Rhizoctonia solani[J]. Molecular Breeding,2001,8(3): 263-271.
[78] Yajima W, Verma S S, Shah S, et al. Expression of anti-sclerotinia scFv in transgenic Brassica napus enhances tolerance against stem rot[J]. New biotechnology,2010,27(6):816-821.
[79] Verma S S, Yajima W R, Rahman M H, et al. A cysteine- rich antimicrobial peptide from Pinus monticola (PmAMP1) confers resistance to multiple fungal pathogens in canola (Brassica napus)[J]. Plant Molecular Biology,2012,79(1-2):61-74.
[80] Fan Y, Du K, Gao Y, et al. Transformation of LTP gene into Brassica napus to enhance its resistance to Sclerotinia sclerotiorum[J]. Russian Journal of Genetics, 2013, 49(4): 380-387.
[81] 汪小福.抗菌肽 Thanatin基因载体构建和表达及提高植物对菌核病抗性的研究[D].合肥:安徽农业大学,2007.
[82] Ma L L, Huo R, Gao X W, et al. Transgenic Rape with hrf2 Gene Encoding HarpinXooc Resistant to Sclerotinia sclerotinorium[J]. Agricultural Sciences in China,2008,7(4):455-461.
[83] 王淑文,陆俊杏,万华方,等.BnMAPK1超量表达提高甘蓝型油菜菌核病抗性[J].作物学报,2014(04):745-750.
[84] Wang Z, Mao H, Dong C, et al. Overexpression of Brassica napus MPK4 enhances resistance to Sclerotinia sclerotiorum in oilseed rape[J]. Molecular Plant- Microbe Interactions, 2009, 22(3): 235- 244.
[85] Jiang Y Z, Fu X L, Wen M L, et al. Overexpression of an nsLTPslike antimicrobial protein gene (LJAMP2) from motherwort (Leonurus japonicus) enhances resistance to Sclerotinia sclerotiorum in oilseed rape (Brassica napus)[J]. Physiological and Molecular Plant Pathology,2013,82:81-87.
[86] Guo X L D, Hu CH, Liu X J, et al. Transformation of oilseed rape with a novel gene from a bacterial strain to improve resistance to sclerotinia sclerotiorum[J]. Acta horticulturae,2006, 706: 265-268.
[87] 舒佳宾.草酸脱羧酶对油菜抗菌核病的研究[D].长沙:湖南农业大学,2012.
[88] Livingstone D M, Hampton J L, Phipps P M, et al. Enhancing resistance to Sclerotinia minor in peanut by expressing a barley oxalate oxidase gene[J]. Plant Physiology, 2005, 137(4): 1354-1362.
[89] Cunha W G, Tinoco M L P, Pancoti H L, et al. High resistance to Sclerotinia sclerotiorum in transgenic soybean plants transformed to express an oxalate decarboxylase gene[J]. Plant Pathology,2010, 59(4):654-660.
[90] Kesarwani M, Azam M, Natarajan K, et al. Oxalate decarboxylase from Collybia velutipes. Molecular cloning and its overexpression to confer resistance to fungal infection in transgenic tobacco and tomato[J]. Journal of Biological Chemistry, 2000, 275(10): 7230- 7238.
[91] Liu S, Liu Y, Yang X, et al. The Brassica oleracea genome reveals the asymmetrical evolution of polyploid genomes[J]. Nat Commun, 2014(5):3930.
[92] Wang X, Wang H, Wang J, et al. The genome of the mesopolyploid crop species Brassica rapa[J]. Nature Genetics, 2011, 43(10): 1035- 1039.
[93] Amselem J, Cuomo C A, van Kan J A, et al. Genomic analysis of the necrotrophic fungal pathogens Sclerotinia sclerotiorum and Botrytis cinerea[J]. PLoS Genetics, 2011, 7(8): e1002230.