-
真菌病害是植物病害中数量最大的一类,占植物病害总数的70%~80%。许多危害重、分布广的作物病害,如锈病、黑粉病、霜霉病、白粉病等都是由真菌引起的。与形态观察、次生代谢产物分析等常规检测技术不同,病原真菌的早期检测技术检测周期短、灵敏度高,能帮助快速检测到侵染初期或潜伏期的病原菌,并获悉其生长状态和发病阶段,以备人们有充分的时间采取科学合理的防治措施,最大限度地减少经济损失。因此,植物病原菌早期检测技术是防治病害大面积暴发的有效手段。从传统的组成物质观察和气相色谱检测到免疫学方法的建立,再到分子检测手段的不断成熟,病原菌早期检测技术越来越简便和精准,为建立植物病害的早期诊断方法和预测预报模型奠定了良好基础。
橡胶树(Hevea brasiliensis)是天然橡胶的主要来源,天然橡胶是一种重要的战略资源,约占全球橡胶消费总量的40%,世界每年对天然橡胶的需求不断增长[1-2]。炭疽菌(Colletotrichum)侵染橡胶树引起的炭疽病(Colletotrichum leaf disease,CLD)是亚洲天然橡胶产量下降的主要原因[3-4],该病害可导致橡胶树叶片变形和坏死,进而发生次生性落叶,严重影响胶乳产量[5]。由于炭疽菌具有潜伏侵染能力,橡胶树炭疽病的预测预报一直存在较大的技术难点。橡胶树炭疽菌典型的作用方式为半活体营养型侵染,在侵染初期通过与宿主细胞共生逃避抗性机制,同时满足自身营养和能源需求,随后进行坏死型营养生长,迅速扩散并杀死宿主细胞[6-7]。炭疽菌具有潜伏期的存在和作用方式转换的特点,因此不易被发现;在气候条件适宜的情况下,极短的时间内即可暴发成灾;预测难度大,给炭疽病防治造成困难。因此,建立潜伏侵染状态下的炭疽菌检测技术成为了橡胶树炭疽病早期诊断和流行预测的关键所在。笔者综述了常用的植物病原真菌早期检测技术,包括基于菌体结构组成的检测技术和基于核酸序列PCR扩增的检测技术,比较了各种早期检测技术的优缺点,论述了实时荧光定量PCR早期检测技术建立的关键步骤及在橡胶树炭疽病预测预报模型中的应用潜力,为橡胶树炭疽菌早期检测技术和预测预报模型的建立奠定基础。
Research progress on early detection technologies for plant fungi and their application in forecasting of rubber tree anthracnose
-
摘要:真菌是一类重要的植物病原菌,真菌引起的病害占全部植物病害的三分之二,建立快速、准确的植物真菌病害早期检测技术是解决植物病原真菌预测预报难题、防止病害大面积暴发和流行的关键。笔者综述了常用植物病原真菌早期检测技术的检测原理、应用现状和存在问题,炭疽菌荧光定量PCR早期检测技术的建立及其在橡胶树炭疽病预测预报模型构建中的应用前景,为橡胶树炭疽病早期检测及预测预报提供技术支持。Abstract:Fungi are one class of important plant pathogens, which accounts for two thirds of all plant diseases. The rapid and accurate early detection technologies for plant fungal diseases are the key to disease prediction and prevention of disease prevalence. The detection principle, application status and existing problems of common early detection technologies for fungal diseases were reviewed. The detection system of fluorescence quantitative PCR and its application prospect in the prediction model for rubber tree anthracnose were summarized, which provides reference for the early detection and prediction of rubber tree anthracnose.
-
[1] CAI Z, LI G, LIN C, et al. Identifying pathogenicity genes in the rubber tree anthracnose fungusColletotrichum gloeosporioidesthrough random insertional mutagenesis [J]. Microbiological Research, 2013, 168(6): 340 − 350.doi:10.1016/j.micres.2013.01.005 [2] LIEBEREI R. South American leaf blight of the rubber tree (Heveaspp.): New steps in plant domestication using physiological features and molecular markers [J]. Annals of Botany, 2007, 100(6): 1125 − 1142.doi:10.1093/aob/mcm133 [3] LIU X, LI B, CAI J, et al.Colletotrichumspecies causing anthracnose of rubber trees in China [J]. Scientific Reports, 2018, 8(1): 10435 − 10449.doi:10.1038/s41598-018-28166-7 [4] THAMBUGALA T A D P and DESHAPPRIYA N. The role ofColletotrichumspecies on theColletotrichumleaf disease ofHevea brasiliensis- a preliminary study [J]. Journal of the National Science Foundation of Sri Lanka, 2009, 37(2): 135 − 138.doi:10.4038/jnsfsr.v37i2.1070 [5] GUYOT J, OMANDA E N, NDOUTOUME A, et al. Effect of controlling Colletotrichum leaf fall of rubber tree on epidemic development and rubber production [J]. Crop Protection, 2001, 20(7): 581 − 590.doi:10.1016/S0261-2194(01)00027-8 [6] DAUCH A L, AHN B and WATSON A K. Molecular monitoring of wild-type and genetically engineeredColletotrichum coccodesbiocontrol strains in planta [J]. Plant Disease, 2006, 90(12): 1504 − 1510.doi:10.1094/PD-90-1504 [7] WHARTON P S, DIÉGUEZ-URIBEONDO J. The biology ofColletotrichum acutatum[J]. Anales del Jardín Botánico de Madrid, 2004, 61(1): 3 − 22. [8] PEI M H, RUIZ C, HARRIS J, et al. Quantitative inoculations of poplars withMelampsora larici-populina[J]. European Journal of Plant Pathology, 2003, 109(3): 269 − 276.doi:10.1023/A:1022822503139 [9] SCHMITZ O, DANNEBERG G, HUNDESHAGEN B, et al. Quantification of vesicular-arbuscular mycorrhiza by biochemical parameters [J]. Journal of Plant Physiology, 1991, 139(1): 106 − 114.doi:10.1016/S0176-1617(11)80174-4 [10] SHANKAL M, GREGORY A, KALKHOVEN M J, et al. A competitive ELISA for detecting resistance to latent stem infection byDiaporthe toxicain narrow-leafed lupins [J]. Australasian Plant Pathology, 1998, 27(4): 251 − 258.doi:10.1071/AP98028 [11] GABLER J, KAČERGIUS A AND JOVAIŠIENĖ Z. Detection ofPhomopsis vacciniion blueberry and cranberry in Europe by direct tissue blot immunoassay and plate-trapped antigen ELISA [J]. Journal of Phytopathology, 2004, 152(11): 630 − 632. [12] BRILL L M. Analysis of two ELISA formats and antigen preparations using polyclonal antibodies againstPhomopsis longicolla[J]. Phytopathology, 1994, 84(2): 1047 − 1056. [13] LOMMEL S A, MCCAIN A H AND MORRIS T J. Evaluation of indirect enzyme-linked immunosorbent assay for the detection of plant viruses [J]. Phytopathology, 1982, 72(8): 1018 − 1022.doi:10.1094/Phyto-72-1018 [14] LISTER R M AND ROCHOW W F. Detection of barley yellow dwarf virus by enzyme-linked immunosorbent assay [J]. Phytopathology, 1979, 69(6): 649 − 654.doi:10.1094/Phyto-69-649 [15] HANSE B, RAAIJMAKERS E E M, SCHOONE A H L, et al.Stemphyliumspp., the cause of yellow leaf spot disease in sugar beet (Beta vulgarisL.) in the Netherlands [J]. European Journal of Plant Pathology, 2015, 142(2): 319 − 330.doi:10.1007/s10658-015-0617-8 [16] SAKAMOTO S, PUTALUN W, VIMOLMANGKANG S, et al. Enzyme-linked immunosorbent assay for the quantitative/qualitative analysis of plant secondary metabolites [J]. Journal of Natural Medicines, 2017, 72(2): 32 − 42. [17] KITISRIPANYA T, KRITTANAI S, UDOMSIN O, et al. Development of an enzyme-linked immunosorbent assay for determination of miroestrol using an anti-miroestrol monoclonal antibody [J]. Planta Medica, 2017, 83(10): 855 − 861.doi:10.1055/s-0043-102689 [18] GREEN H AND JENSEN D F A. Tool for monitoringTrichoderma hanianum: Ⅱ. The use of a GUS transformant for ecological studies in the rhizosphere [J]. Phytopalhology, 1995, 85: 1436 − 1440.doi:10.1094/Phyto-85-1436 [19] FREEMAN S, MAIMON M AND PINKAS Y. Use of GUS transformants ofFusarium subglutinansfor determining etiology of mango malformation disease [J]. Phytopathology, 1999, 89(6): 456 − 461.doi:10.1094/PHYTO.1999.89.6.456 [20] CHALFIE M T Y, EUSKIRCHEN G, WARD W W, et al. Green fluorescent protein as marker for gene expression [J]. Science, 1994, 263(11): 802 − 805. [21] OLIVAIN C, HUMBERT C, NAHALKOVA J, et al. Colonization of tomato root by pathogenic and nonpathogenicFusarium oxysporumstrains inoculated together and separately into the soil [J]. Applied and Environmental Microbiology, 2006, 72(2): 1523 − 1531.doi:10.1128/AEM.72.2.1523-1531.2006 [22] CHEN N, HSIANG T AND GOODWIN P H. Use of green fluorescent protein to quantify the growth ofColletotrichumduring infection of tobacco [J]. Journal of Microbiological Methods, 2003, 53(1): 113 − 122.doi:10.1016/S0167-7012(02)00234-8 [23] SIGAL H, STANLEY F AND AMIR S. Use of green fluorescent protein-transgenic strains to study pathogenic and nonpathogenic lifestyles inColletotrichum acutatum[J]. Phytopathology, 2002, 92(7): 743 − 749.doi:10.1094/PHYTO.2002.92.7.743 [24] PARIKKA P AND LEMMETTY A. Tracing latent infection ofColletotrichum acutatumon strawberry by PCR [J]. European Journal of Plant Pathology, 2004, 110(4): 393 − 398.doi:10.1023/B:EJPP.0000021073.67137.d2 [25] PARAN I AND MICHELMORE R W. Development of reliable PCR-based markers linked to downy mildew resistance genes in lettuce [J]. Theoretical and Applied Genetics, 1993, 85(8): 985 − 993.doi:10.1007/BF00215038 [26] MERLIER D D, CHANDELIER A, DEBRUXELLES N, et al. Characterization of AlderPhytophthoraisolates from Wallonia and development of SCAR primers for their specific detection [J]. Journal of Phytopathology, 2005, 153(2): 99 − 107.doi:10.1111/j.1439-0434.2005.00936.x [27] SPADARO D, PELLEGRINO C, GARIBALDI A, et al. Development of SCAR primers for the detection ofCadophora luteo-olivaceaon kiwifruit and pome fruit and ofCadophora malorumon pome fruit [J]. Phytopathologia Mediterranea, 2011, 50(3): 430 − 441. [28] NITHYA K, BUKHARI K A I M, VALLUVAPARIDASAN V, et al. Molecular detection ofColletotrichum falcatumcausing red rot disease of sugarcane (Saccharum officinarum) using a SCAR marker [J]. Annals of Applied Biology, 2012, 160(2): 168 − 173.doi:10.1111/j.1744-7348.2011.00529.x [29] PÉREZ-HERNÁNDEZ O, NAM M H, GLEASON M L, et al. Development of a nested polymerase chain reaction assay for detection ofColletotrichum acutatumon symptomless strawberry leaves [J]. Plant Disease, 2008, 92(12): 1655 − 1661.doi:10.1094/PDIS-92-12-1655 [30] 张磊, 常有宏, 刘邮洲, 等. 梨轮纹病和炭疽病病原菌PCR检测[J]. 江苏农业学报, 2012, 28(2): 415 − 420.doi:10.3969/j.issn.1000-4440.2012.02.032 [31] CHEN Y Y, CONNER R L, GILLARD C L, et al. A specific and sensitive method for the detection ofColletotrichum lindemuthianumin dry bean tissue [J]. Plant Disease, 2007, 91(10): 1271 − 1276.doi:10.1094/PDIS-91-10-1271 [32] TOMITA N, MORI Y, KANDA H, et al. Loop-mediated isothermal amplification (LAMP) of gene sequences and simple visual detection of products [J]. Nature Protocols, 2008, 3(5): 877 − 882.doi:10.1038/nprot.2008.57 [33] KHAN M, LI B, JIANG Y, et al. Evaluation of different PCR-based assays and LAMP method for rapid detection ofPhytophthora infestansby targeting theYpt1gene [J]. Frontiers in Microbiology, 2017, 8: 1920 − 1931.doi:10.3389/fmicb.2017.01920 [34] SI AMMOUR M, BILODEAU G J, TREMBLAY D M, et al. Development of real-time isothermal amplification assays for on-site detection ofPhytophthora infestansin potato leaves [J]. Plant Disease, 2017, 101(7): 1269 − 1277.doi:10.1094/PDIS-12-16-1780-RE [35] TIAN Q, LU C, WANG S, et al. Rapid diagnosis of soybean anthracnose caused byColletotrichum truncatumusing a loop-mediated isothermal amplification (LAMP) assay [J]. European Journal of Plant Pathology, 2016, 148(4): 785 − 793. [36] KHAN M, WANG R, LI B, et al. Comparative evaluation of the LAMP assay and PCR-based assays for the rapid detection ofAlternaria solani[J]. Front Microbiol, 2018, 9: 2089 − 2100.doi:10.3389/fmicb.2018.02089 [37] DEBODE J, VAN HEMELRIJCK W, BAEYEN S, et al. Quantitative detection and monitoring ofColletotrichum acutatumin strawberry leaves using real-time PCR [J]. Plant Pathology, 2009, 58(3): 504 − 514.doi:10.1111/j.1365-3059.2008.01987.x [38] CHEN Y Y, CONNER R L, GILLARD C L, et al. A quantitative real-time PCR assay for detection ofColletotrichum lindemuthianuminnavy bean seeds [J]. Plant Pathology, 2013, 62(4): 900 − 907.doi:10.1111/j.1365-3059.2012.02692.x [39] SRINIVASAN M, KOTHANDARAMAN S V, VAIKUNTAVASAN P, et al. Development of conventional and real-time PCR protocols for specific and sensitive detection ofColletotrichum capsiciin chilli (Capsicum annuumL) [J]. Phytoparasitica, 2014, 42(4): 437 − 444.doi:10.1007/s12600-013-0380-3 [40] OO M M, LIM G, JANG H A, et al. Characterization and pathogenicity of new record of anthracnose on various chili varieties caused byColletotrichum scovilleiin Korea [J]. Mycobiology, 2017, 45(3): 184 − 191.doi:10.5941/MYCO.2017.45.3.184 [41] DIAO Y Z, ZHANG C, LIU F, et al.Colletotrichumspecies causing anthracnose disease of chili in China [J]. Persoonia Molecular Phylogeny & Evolution of Fungi, 2017, 38: 20 − 37. [42] WANG Y C, HAO X Y, WANG L, et al. DiverseColletotrichumspecies cause anthracnose of tea plants (Camellia sinensis(L) OKuntze) in China [J]. Scientific Reports, 2016, 6: 35287 − 35300.doi:10.1038/srep35287 [43] HYDE K D. Colletotrichum: a catalogue of confusion [J]. Fungal Diversity, 2009, 39: 1 − 17. [44] NIU X, GAO H, QI J, et al.Colletotrichumspecies associated with jute (Corchorus capsularisL) anthracnose in southeastern China [J]. Scientific Reports, 2016, 6: 25179 − 25188.doi:10.1038/srep25179 [45] MARTINEZ-CULEBRAS P V, BARRIO E, GARCIA M D, et al. Identification ofColletotrichumspecies responsible for anthracnose of strawberry based on the internal transcribed spacers of the ribosomal region [J]. FEMS Microbiol Lett, 2000, 189(1): 97 − 101.doi:10.1111/j.1574-6968.2000.tb09213.x [46] SCHOCH C L, SEIFERT K A, HUHNDORF S, et al. Nuclear ribosomal internal transcribed spacer (ITS) region as a universal DNA barcode marker for Fungi [J]. Proceedings of the National Academy of Sciences of the United States of America, 2012, 109(16): 6241 − 6246.doi:10.1073/pnas.1117018109 [47] SREENIVASAPRASAD S, TALHINHAS P. Genotypic and phenotypic diversity inColletotrichum acutatum, a cosmopolitan pathogen causing anthracnose on a wide range of hosts [J]. Molecular Plant Pathology, 2005, 6(4): 361 − 378.doi:10.1111/j.1364-3703.2005.00291.x [48] ZHU Z X, ZHENG L, HSIANG T, et al. Detection and quantification ofFusarium communeinhost tissue and infected soil using real-time PCR [J]. Plant Pathology, 2016, 65: 218 − 226.doi:10.1111/ppa.12412 [49] ZHANG X, HARRINGTON T C, BATZER J C, et al. Detection ofColletotrichum acutatumsensu lato on strawberry by loop-mediated isothermal amplification [J]. Plant Disease, 2016, 100(9): 1804 − 1812.doi:10.1094/PDIS-09-15-1013-RE [50] SCARLETT K, TESORIERO L, DANIEL R, et al. Detection and quantification ofFusarium oxysporumf spcucumerinumin environmental samples using a specific quantitative PCR assay [J]. European Journal of Plant Pathology, 2013, 137(2): 315 − 324.doi:10.1007/s10658-013-0244-1 [51] NATH V S, HEGDE V M, JEEVA M L, et al. Rapid and sensitive detection ofPhytophthora colocasiaeresponsible for the taro leaf blight using conventional and real-time PCR assay [J]. FEMS Microbiol Lett, 2014, 352(2): 174 − 183.doi:10.1111/1574-6968.12395 [52] JIMÉNEZ-FERNÁNDEZ D, MONTES-BORREGO M, NAVAS-CORTÉS J A, et al. Identification and quantification ofFusarium oxysporumin planta and soil by means of an improved specific and quantitative PCR assay [J]. Applied Soil Ecology, 2010, 46(3): 372 − 382.doi:10.1016/j.apsoil.2010.10.001 [53] YANG H C, HAUDENSHIELD J S AND HARTMAN G L. Multiplex real-time PCR detection and differentiation ofColletotrichumspecies infecting soybean [J]. Plant Disease, 2015, 99(11): 1559 − 1568.doi:10.1094/PDIS-11-14-1189-RE [54] LIU L, YAN Y, HUANG J, et al. A novel MFS transporter geneChMfs1is important for hyphal morphology, conidiation, and pathogenicity inColletotrichum higginsianum[J]. Frontiers in Microbiology, 2017, 8: 1953 − 1964.doi:10.3389/fmicb.2017.01953 [55] LIU F, TANG G, ZHENG X, et al. Molecular and phenotypic characterization ofColletotrichumspecies associated with anthracnose disease in peppers from Sichuan province, China [J]. Scientific Reports, 2016, 6: 32761 − 32778.doi:10.1038/srep32761 [56] PHOTITA W, LUM S, MCKENZIE E H C, et al. Are some endophytes ofMusa acuminatalatent pathogens? [J]. Fungal Diversity, 2004, 16: 131 − 140. [57] BARCELOS Q L, PINTO J M, VAILLANCOURT L J, et al. Characterization ofGlomerellastrains recovered from anthracnose lesions on common bean plants in Brazil [J]. PLoS One, 2014, 9(3): e90910.doi:10.1371/journal.pone.0090910 [58] WANG F, QIN G, SUI Z, et al. Improved method for assaying maize plant resistance to maize rough dwarf disease by artificial inoculation and real-time RT-PCR [J]. European Journal of Plant Pathology, 2006, 116(4): 289 − 300.doi:10.1007/s10658-006-9060-1 [59] HOSSAIN M M, SULTANA F, MIYAZAWA M, et al. Plant growth-promoting fungusPenicilliumspp. GP15-1 enhances growth and confers protection against damping-off and anthracnose in the cucumber [J]. Journal of Oleo Science, 2014, 63(4): 391 − 400.doi:10.5650/jos.ess13143 [60] JIMÉNEZ-FERNÁNDEZ D, MONTES-BORREGO M, JIMÉNEZ-DÍAZ R M. In planta and soil quantification ofFusarium oxysporumf spcicerisand evaluation of Fusarium wilt resistance in chickpea with a newly developed quantitative polymerase chain reaction assay [J]. Phytopathology, 2011, 101: 250 − 262.doi:10.1094/PHYTO-07-10-0190 [61] KOOMEN I, JEFFRIES P. Effects of antagonistic microorganisms on the postharvest development ofColletotrichum gloeosporioideson mango [J]. Plant Pathology, 1993, 42(2): 230 − 237.doi:10.1111/j.1365-3059.1993.tb01495.x [62] LEANDRO L F, GLEASON M L, NUTTER F W, et al. Influence of temperature and wetness duration on conidia and appressoria ofColletotrichum acutatumon symptomless strawberry leaves [J]. Phytopathology, 2003, 93(4): 513 − 520.doi:10.1094/PHYTO.2003.93.4.513 [63] LEANDRO L F S, GLEASON M L, WEGULO S N, et al. Germination and sporulation ofColletotrichum acutatumon symptomless strawberry leaves [J]. Phytopathology, 2001, 91(7): 659 − 664.doi:10.1094/PHYTO.2001.91.7.659 [64] HYRE R A. Progress in forecasting late blight of potato and tomato [J]. Plant Disease, 1954, 38: 245 − 253. [65] WALLIN J R. Forecasting tomato and potato late blight in north-central region (Abs) [J]. Phytopathology, 1951, 47: 37 − 38. [66] 杨信东, 李葵花, 高洁, 等. 烟草野火病“天气促病指数”表解模型的建立[J]. 吉林农业大学学报, 2002, 2(2): 154 − 157. [67] FURUTA K, NAGASHIMA S, INUKAI T, et al. Construction of a system for the strawberry nursery production towards elimination of latent infection of anthracnose fungi by a combination of PCR and microtube hybridization [J]. The Plant Pathology Journal, 2017, 33(1): 80 − 86.doi:10.5423/PPJ.NT.05.2016.0132 [68] ELIAS L M, FORTKAMP D, SARTORI S B, et al. The potential of compounds isolated fromXylariaspp. as antifungal agents against anthracnose [J]. Biotechnology and Industrial Microbiology, 2018, 49(4): 840 − 847. [69] QIN B, ZHENG F AND ZHANG Y. Molecular cloning and characterization of aMlogene in rubber tree (Hevea brasiliensis) [J]. Journal of Plant Physiology, 2015, 175: 78 − 85.doi:10.1016/j.jplph.2014.10.019 [70] 范会雄, 李德威, 黄宏积, 等. 橡胶树炭疽病发生流行规律及防治研究[J]. 植物保护, 1996, 22(5): 31 − 32.
计量
- 文章访问数:392
- HTML全文浏览量:74
- PDF下载量:31
- 被引次数:0