仓晓燕,李文凤,王晓燕,张荣跃,单红丽,黄应昆
问题探讨
检疫性病害甘蔗白条病的发生危害与防控对策
仓晓燕,李文凤,王晓燕,张荣跃,单红丽,黄应昆*
(云南省农业科学院甘蔗研究所/云南省甘蔗遗传改良重点实验室,开远661699)
甘蔗白条病的广泛传播性和毁灭性,给甘蔗产量和糖分带来损失,对其症状、致病病原、侵染途径和病原检测等进行了分析,并提出了今后的防控对策。
甘蔗;白条病;发生危害;防控对策
甘蔗白条黄单孢菌[Xanthoomonas albilineans(Ashby)Dowson)是引起甘蔗白条病的病原菌[1-2]。1911年在澳大利亚首次报道[3],在过去的几十年,病害在多米尼加岛(拉丁美洲岛屿)、厄瓜多尔(位于拉丁美洲)、危地马拉(拉丁美洲国家)、洪都拉斯(拉丁美洲国家)、美国路易斯安那州和德克萨斯州、墨西哥、扎伊尔(非洲中西部共和国)有报道,现有超过66个国家受影响[4-6]。另外,一些爆发也发生在美国佛罗里达州、瓜德罗普岛和毛里求斯,这些地方的病害发生是感病品种大面积种植的结果[7]。鉴于甘蔗白条病的广泛传播性和毁灭性,给甘蔗产量和糖分带来的损失,本文对其症状、致病病原、侵染途径和病原检测等进行了分析,并提出了今后的防控对策。
甘蔗白条病为系统性病害,有慢性型和急性型两种[[8-9]。慢性型主要危害叶片,在叶片上产生狭窄的白色条纹,沿维管束延伸破坏蔗株叶绿体发育,感病较重的病株蔗茎的节部长出许多侧芽和细小的分蘖,分蘖的叶片上也出现上述的白色条纹,纵剖蔗茎可见维管束变红,发病严重时蔗株茎内会出现空腔以及感病甘蔗品种的快速死亡。这些症状是病原物数量高时,木质部的新陈代谢受到阻碍的结果。急性型,蔗株叶片不表现任何外表症状,便突然整株枯萎死亡。在甘蔗生长旺季突遇干旱条件时,易发生急性型症状。其与生理性缺水枯萎不同之处,在于病茎再行分蘖时,其叶片又可表现慢性型病斑[10-11]。
病原物为甘蔗白条黄单孢菌[Xanthoomonas albilineans(Ashby)Dowson][12]。在蔗糖和蛋白胨培养基上,菌落颜色可从蜜黄色到浅黄色。革兰氏染色反应为阴性,极生单根鞭毛。黄单孢菌生长缓慢,生长需要氨基酸,有抗生素抗性[9]。自然条件下,病菌除侵染甘蔗外,还传染玉米、白茅和两耳草等,并存在致病性不同的菌株[13-17]。
田间主要通过种苗/砍蔗工具传播蔓延[10,18],在不同气候因子影响下,甘蔗白条黄单孢菌在甘蔗叶表面定殖,通过气孔或伤口进入叶片,然后在木质部扩展,蔗叶出现症状,甘蔗白条黄单孢菌从蔗叶迁移到蔗茎,影响甘蔗分蘖,表现为甘蔗白条病症状或侧枝增多[19-22]。Klett P等[23]在感病甘蔗根部发现病原物甘蔗白条黄单孢,推测有通过根部接触传播的可能。
病害发生的轻重同蔗田地势和土壤、栽培管理、品种等因素有关,蔗地低湿易积水易诱发此病,大面积种植感病品种为本病发生流行的主要条件。品种间抗性有差异,在中国台湾蔗区经过鉴定抗病的甘蔗品种有NCO310、F156、F160、F170和F173等。表现较抗病的品种有Q42、Q50、Q98、Q813、POJ36、POJ2725、CP807、CP29-116、Co290、Co301、Co331、Co421、B4908等[10]。
甘蔗白条黄单孢病原菌的潜伏期长,蔗株感染以后很长一段时间可不表现病状,病原物检测需灵敏的方法。血清学方法,包括酶联方法,斑点和组织免疫印痕法可以特异性地检测黄单孢菌,最低可检测到维管束提取物中浓度为105细胞/mL的病原物[24-27]。用于诊断黄单孢病害的核糖体DNA序列的PCR引物也已经得到运用[28-30],首次检测甘蔗白条黄单孢菌用的PCR引物是甘蔗黄单孢菌毒素基因,可检测到浓度为2×104细胞/mL的甘蔗白条黄单孢菌[31-32],特异性地扩增甘蔗白条黄单孢菌的ITS区DNA也有同样作用[33-34]。运用PCR方法,卢文洁等[35]对法国、泰国、澳大利亚、柬埔寨、菲律宾引进的32个甘蔗品种进行PCR检测,均未检测到甘蔗白条病病菌。
6.1加强检疫
鉴于甘蔗白条病的广泛传播性和毁灭性,甘蔗检疫检测技术是甘蔗品种/材料交换的基础,利用国际标准的检疫程序和检测技术,应用符合国际标准的规范化的检疫程序及完善的甘蔗病害分子检测体系对引进的甘蔗品种/材料进行准确的疫情监测和病害检测,是甘蔗安全生产和品种交换的前提和保证[10,35]。
6.2选用无病种苗做种
6.2.1 选用抗病品种、健康种苗因地制宜选育并种植抗病甘蔗品种是防治该病害最有效的措施[36-37]。从无病地区调运蔗种,或在轻病蔗田选择外表健康的甘蔗做种[38]。在发生区引种必须认真检查,确保引进的种苗无病,是防止甘蔗白条病传播、发生的最好措施[10]。
6.2.2 种苗温水处理种苗播种前,采用流动水预浸泡48h,然后再用50℃的温水处理2h,可达到95%的防治效果[10,40],宜采用成熟但不太老的中间节段做种苗,以2~3芽苗为好。
6.2.3 建立无病苗圃将经过温水处理的种苗集中种植,建立脱毒种苗基地一级、二级、三级种苗圃、并实施耕作刀具的隔离和消毒,为大面积生产提供无病种苗[10]。
6.3加强栽培管理
田间发病株及时拔除烧毁,杜绝种苗带病;科学施肥,合理排灌,防止干旱,雨后及时开沟排水,增强蔗株的抗病力,减轻病害发生。
7.1发病机理
甘蔗品种和生长环境的不同,潜伏期病原物浓度不同,侵染为什么可以潜伏几周或几月,然后急性发展,值得分子植物病理学进一步深入研究。明确致病机理,可采取新的抗病策略,包括开发新的抗甘蔗白条病的抗病基因。
甘蔗白条黄单孢菌基因组测序的目的在于,从基因组角度明确甘蔗白条黄单孢菌致病相关的基因组特征。甘蔗白条黄单孢菌的高致病性的菌株GPEPC73基因组(3.8M)(NCBI参考序列号NC_013722.1)比已测序其它黄单孢菌菌株基因组(约5M)小。甘蔗白条黄单孢菌基因组特征有一些不同,如没有致病的细菌Ⅲ型分泌系统(Hrp-T3SS),在基因组中发现两个CRISPR(中间短回文的重复序列)插入位点,这些基因组的特征和木质部定殖特征相关[39]。
甘蔗白条黄单孢菌已知的致病因子是阻断叶绿体分化的白条黄单孢毒素(albicidin)。白条黄单孢毒素干扰寄主的抗性机制,导致系统性的侵染。另外,甘蔗白条黄单孢菌可利用细胞壁降解酶(CWDEs)降解细胞壁产物作为碳源[40-43],细胞壁降解酶(CWDEs)被认为是致病因子[44-47]。Fleites L A等[48]报道OmpA1基因影响甘蔗白条黄单孢菌致病性以及在叶部的定殖。Pieretti I等[49]结合比较基因组学和SSH研究表明,可能的致病基因还包括ABC转运蛋白基因、甲基化受体蛋白基因及氧化还原酶基因等。
7.2转基因方法导入蔗株提高抗性
澳大利亚研究者从生物防治甘蔗白条病的细菌中克隆到白条病解毒基因(albD)[50],虽然育成品种对甘蔗白条病均有一定的抗性,可在一定程度上减轻由甘蔗白条病引起的蔗株叶片灼烧症状,但并未得到完全抗甘蔗白条病的品种。生物技术是提高甘蔗抗病性的重要途径,今后应有更多运用。
[1]Saumtally A S,Dookun-Saumtally A,Rao G P,et al.Leaf scald of sugarcane:a disease of worldwide importance[M].France:CIRAD-ISSCT,2004.
[2]Zhang L,Birch R.The gene for albicidin detoxification from Pantoea dispersa encodes an esterase and attenuates pathogenicity of Xanthomonas albilineans[J].Proceedings of the National Academy of Sciences,USA,1997,94:9984-9989.
[3]North D S.Colonial Sugar Refining Company.Leaf-Scald,A Bacterial Disease of Sugarcane[M].Australia Sydney Colonial Sugar Refining Company,1926.
[4]Rott P,Davis M.Leaf scald.In A guide to sugarcane diseases[M].Montpellier:CIRAD-ISSCT,2000:339.
[5]Martin J.Robinson P.Leaf scald In Sugar-Cane Diseases of the World[M].Elsevier Publishing Company.Amsterdam,TheNetherlands,1961:79-101.
[6]Saumtally A S,Dookun-Saumtally A,Rao G P,et al.Leaf scald of sugarcane:a disease of worldwide importance[M]. Phytopathology,2006b,96:1081-1091.
[7]Davis M J,Rott P,Warmuth CJ et al.Intraspecific genomic variation within Xanthomonas albilineans,the sugarcane leaf scald pathogen[J].Phytopathology,1997,87(3):316-324.
[8]Pieretti I,Royer M,Barbe V,et al.The complete genome of Xanthomonas albilineans provides new insights into the reductive genome evolution of the xylem-limited Xanthomonadaceae[J].BMC Genomics,2009,10(1):7428-7436.
[9]Orian,G.Artificial hosts of the sugarcane leaf scald organism[J].Rev.Agric.Sucr.Ile Maurice,1942,21:285-304.
[10]黄应昆,李文凤等.甘蔗主要病虫草害原色图谱[M].昆明:云南科技出版社,2002.
[11]Ryan C C.CHAPTER III-Leaf scald.Diseases of Sugarcane[M].Amsterdam:Elsevier Besloten Vennootschap,1989:39-58.
[12]Birch R G.Xanthomonas albilineans and the antipathogenesis approach to disease control[J].Molecular Plant Pathology,2001, 2(2):1-11.
[13]Alvarez A M,Schenck S,Benedict A A.Differentiation of Xanthomonas albilineans strains with monoclonal antibody reaction patterns and DNA fingerprints[J].Plant Pathology,1996,45(2):358-366.
[14]Dabbas K M,Ferro M I T,Barros N M D,et al.Diferential gene expression in sugarcane infected with Xanthomonas albilineans, causal agent of leaf scald[J].Summa Phytopathologica,2006,32(4):328-338.
[15]Autrey,Saumtally,Dookun A,et al.Studieson variation in the leaf scald pathogen,Xanthomonas albilineans[J].Combined Proceedings of International Plant Propagators'Society Sugarcane Technol,1995,21:485-497.
[16]Mohamed IS,Rott P,Davis,et al.Differentiation of Xanthomonas albilineans strains based on multiplication of the pathogen in sugarcane varieties[J].Process Internal of Socical Sugrcane,1996,22:486-492.
[17]Persley G J.Pathogenic variation in Xanthomonas albilineans(Ashby)Dowson,the causal agent of leaf scald disease of sugarcane[J]. Australian Journal of Biological Sciences,1973,26(4):781-786.
[18]Birch R G.Xanthomonas albilineans and the antipathogenesis approach to disease control[J].Molecular Plant Pathology,2001, 2(2):1-11.
[19]Autrey L,Saumtally S,Dookun A,et al.Aerial transmission of the leaf scald pathogen,Xanthomonas albilineans[J].Process Internal of Socical Sugrcane Technolg.,1995,21:508-526.
[20]Daugrois J H,Dumont V,Champoiseau P,et al.Aerial contamination of sugarcane in Guadeloupe by two strains of Xanthomonas albilineans[J].European journal of plant pathology,2003,109(5):445-458.
[21]Champoiseau P,Rott P,Daugrois J H.Epiphytic populations of Xanthomonas albilineans and subsequent sugarcane stalk infection are linked to rainfall in Guadeloupe[J].Plant Dis.2009,93(93):339-346.
[22]Rott P C,Costet L,Davis M J,et al.At least two separate gene clusters are involved in albicidin production by Xanthomonas albilineans[J].Journal of Bacteriology,1996,178(15):4590-4596.
[23]Klett P,Rott P.Inoculum sources for the spread of leaf scald disease of sugarcane caused by Xanthomonas albilineans in Guadeloupe[J].Journal of Phytopathology,1994,142(3):283-291.
[24]Rott P,Davis M,Baudin P.Serological variability in Xanthomonas albilineans,causal agent of leaf scald disease of sugarcane[J]. Plant Pathol.,1994,43(2):344-349.
[25]Alvarez A M,Schenck S,Benedict A A.Differentiation of Xanthomonas albilineans strains with monoclonal antibody reaction patterns and DNA fingerprints[J].Plant Pathology,1996,45(2):358-366.
[26]Davis M J,Rott P,Baudin P,et al.Evaluation of selective media and immunoassays for detection of Xanthomonas albilineans, causal agent of sugarcane leaf scald disease[J].Plant Disease,1994,78(1):78-82.
[27]Comstock J C,Irey M S.Detection of the sugarcane leaf scald pathogen,Xanthomonas albilineans,using tissue blot immunoassay, ELISA,and isolation techniques[J].Plant Disease,1992,76(10):1033-1035.
[28]Pan Y B,Grisham M P,Burner D M.A polymerase chain reaction protocol for the detection of Xanthomonas albilineans,the causal agent of sugarcane leaf scald disease[J].Plant Disease,1997,81(2):189-194.
[29]Pan Y B,Grisham M P,Burner D M et al.Detecting sugarcane disease pathogens by DNA-based technology[M].Process of Inter-American Sugar Cane Seminars,1998:224-237.
[30]Pan Y B,Grisham M P,Burner D M,et al.Distribution of the leaf scald pathogen in infected sugarcane stalks[J].Journal of American Society Sugarcane Technology,1997,92(6):23-29.
[31]Jensen M A,Webster J A,Straus N.Rapid identification of bacteria on the basis of polymerase chain reaction-amplified ribosomal DNA spacer polymorphisms[J].Applied&Environmental Microbiology,1993,59(4):945-952.
[32]Davis M J,Rott P,Astua M G.Multiplex polymerase chain reaction(PCR)for diagnosis of leaf scald and ratoon stunting diseases[J]. Sugar Y Azucar,1997,92(6):26.
[33]Honeycutt R J,Sobral B W,Mcclelland M.tRNA intergenic spacers reveal polymorphisms diagnostic for Xanthomonas albilineans[J]. Microbiology,1995,141(12):3229-3239.
[34]Pan Y B,Grisham M P,Burner D M,et al.Development of polymerase chain reaction primers highly specific for Xanthomonas albilineans,the causal bacterium of sugarcane leaf scald disease[J].Plant Disease,1999,83(3):218-222.
[35]卢文洁,李文凤,黄应昆,等.国外甘蔗品种主要病虫害检疫及分子检测[J].云南农业大学学报:自然科学版,2009,24(6):804-808.
[36]Walker D I T.Breeding for resistance Sugarcane Improvement through Breeding[J].Elsevier Science Publishers B.V.Amsterdam, 1987:445-502.
[37]Flynn J L,Anderlini T A.Disease incidence and yield performance of tissue culture generated seedcane over the crop cycle in Louisiana[J].Journal of American Society of Sugar Cane Technologists,1990,10:113.
[38]Lee T S G.Micropropagation of sugarcane(Saccharum,spp.)[J].Plant Cell,Tissue and Organ Culture(PCTOC),1987,10(1):47-55.
[39]Pieretti I,Royer M,Barbe V,et al.Genomic insights into strategies used by Xanthomonas albilineans with its reduced artillery to spread within sugarcane xylem vessels[J].BMC Genomics,2012,13(1):1-23.
[40]Birch R G,Patil S S.Preliminary characterization of an antibiotic produced by Xanthomonas albilineans which inhibits DNA synthesis in Escherichia coli[J].Gene.Microbiol,1985,131(5):1069-1075.
[41]Birch R G,Patil S S.Correlation between albicidin production and chlorosis induction by Xanthomonas albilineans,the sugarcaneleaf scald pathogen[J].Physiolg Molecular Plant Pathology,1987a,30(2):199-206.
[42]Birch R G,Patil S S.Evidence that an albicidin-like phytotoxin induces chlorosis in sugarcane leaf scald disease by blocking plastid DNA replication[J].Physiolg Molecular Plant Pathology,1987b,30(2):207-214.
[43]Hashimi S M,Wall M K,Smith A B,et al.The phytotoxin albicidin is a novel inhibitor of DNA gyrase[J].Antimicrobal Agents Chemotherapy,2007,51(1):181-187.
[44]Roper M C,Greve L C,Warren J G,et al.Xylella fastidiosa requires polygalacturonase for colonization and pathogenicity in Vitis vinifera grapevines[J].Mol.Plant Microbe Interact,2007,20(4):411-419.
[45]Chatterjee S,Almeida R P P,Lindow,S.Living in two worlds:the plant and insect lifestyles of Xylella fastidiosa[J].Annual Review of Phytopathology,2008,46(46):243-271.
[46]Pérez-Donoso A G,Sun Q,Roper M C,et al.Cell wall-degrading enzymes enlarge the pore size of intervessel pit membranes in healthy and Xylella fastidiosa-infected grapevines[J].Plant Physiol.,2010,152(3):1748-1759.
[47]Rott P,FleitesL,Marlow G,et al.Identificationof new candidate pathogenicity factors in the xylem-invading pathogen Xanthomonas albilineans by transposon mutagenesis[J].Molecule of Plant Microbe Interact,2011,24(5):594-605.
[48]Fleites L A,Mensi I,Gargani D,et al.Xanthomonas albilineans OmpA1 appears to be functionally modular and both the OMC and C-like domains are necessary for leaf scald disease of sugarcane[J].Molecular Plant-Microbe Interactions,2013,26(10):1200-1210.
[49]Pieretti I,Pesic A,Petras D,et al.What makes Xanthomonas albilineans unique amongst xanthomonads?[J].Frontiers in Plant Science,2015,6:289.
[50]雷凯健,刘子铎,洪玉枝.甘蔗叶灼病解毒蛋白-AlbDS40位点饱和突变分析[J].河南大学学报:自然版,2008,38(4):397-401.
S435.661
:B
:1007-2624(2017)03-0069-04
10.13570/j.cnki.scc.2017.03.024
2016-12-20
现代农业产业技术体系建设专项资金(CARS-20-2-2);云南省现代农业产业技术体系建设专项资金。
仓晓燕(1983-),女,研究实习员,主要从事甘蔗病害研究。E-mail:67454340@163.com
黄应昆(1964-),男,研究员,主要从事甘蔗病虫害防控研究。Tel:0873-7227017,E-mail:huangyk64@163.com