读书报告12-磷酸对天竺葵青枯病的控制课件.ppt
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1、Control of Bacterial Wilt of Geranium with Phosphorous Acid D.J.Norman 磷酸对天竺葵青枯病的控制主要内容:一:前言 二:材料与方法 三:结果与讨论一、前言Once it becomes established in a susceptible crop,southern bacterial wilt,caused by Ralstonia solanacearum is very hard to eradicate.There are two primary avenues by which R.solanacearum m
2、oves and gains access to crops:one is via water,and the other is through infected propagation materials.由青枯菌引起的南方青枯病一旦侵染敏感作物,就很难根除。青枯菌侵染作物的两种途径是:通过水流侵染通过被侵染物质传播。To combat bacterial wilt,resistant cultivars have been recommended in areas of the world where the pathogen is endemic.In susceptible green
3、housegrown crops,only strict sanitation has been successful in prohibiting plant infestation.为了抵抗地方性青枯病,世界上很多地方都推荐种植抗性作物。种植在温室的敏感作物只有控制环境卫生才能抑制其侵染。Geraniums are susceptible to races 1 and 3 of R.solanacearum.Most geraniums produced in the world are vegetatively propagated in Guatemala,Costa Rica,Col
4、umbia,China,and Kenya.In all these locations,endemic populations of R.solanacearum exist.There are no known treatments that are effective in protecting geranium plants.天竺葵对于青枯菌属1和3是敏感的,天竺葵广泛种植在世界各地如危地马拉,哥斯达黎加,哥伦比亚,中国和肯尼亚。但这些地方都存在地方性青枯病,目前还没有有效的方法保护天竺葵作物。Thus,the objective of this research was to det
5、ermine if geranium plants could be protected from infection with applications of selected bactericides and chemicals.因此本实验的目的就是对天竺葵作物施以筛选过的细菌和化学药品来验证是否能保护作物免受侵染。二、材料与方法1 Screening of products.A race 1(biovar 1)strain(R1B1)isolated in Florida(P673)from pothos(Epipremnum aureum(Linden&Andre)Blunt.)was
6、 used in the initial screening of prospective products.The isolates from pothos are of Central American origin and possess a broader host range than strains endemic in Florida.在弗罗里达的黄金葛里分离出来的R1B1(P673)被用于开始预期产品的筛选。它是在在美国中部分离出来的菌属比弗罗里达的地方性菌属更原始,且具有更广泛的寄主。Promising products were later tested in enviro
7、nmental chambers with a race 3(biovar 2)strain(R3B2)isolated from geranium(UW551),provided by C.Allen,University of Wis-consin.更多的产品将会用R3B2(UW551)在环境检测箱中进行检测,R3B2有威斯康辛大学的艾伦提供。材料包括:三异丙基乙黄酰,苯并噻二唑,硫酸铜,氢氧化铜,铜锡,铜盐,双氧水,糖醛,苯烷基二甲基氯化铵,奥索利酸(Starner),枯草芽孢杆菌,磷酸钾盐。The products were mixed in water at the highest
8、labeled rates recommended by the manufacturers for plant production.Products are not labeled for control of Ralstonia.These products were applied as a drench through potting medium at 50 ml per 9-cm pot.In the initial screening,products were applied every 7 days for a total of four applications.Plan
9、ts were inoculated with R.solanacearum 3 days after the first application.Products were not retested unless plants were protected from infection.这些物质以浸液浇到盆栽介质中,每盆50 ml,浇到9cm深处。在开始的筛选中每7天施一次,总共4次。第一次施用之后3天开始接种青枯菌。这些物质不再测定除非作物被侵染。Products were screened with zonal geranium plants.Rooted geranium cuttin
10、gs were transplanted into 9-cm-diameter plastic pots containing 85 g of Vergo Container Mix A(60%Canadian peat,20%vermiculite,and 20%perlite by volume,pH 6.5)and fertilized with 1.5 g of Osmocote 15-9-12 with micronutrients per pot.产品的筛选用带状天竺葵,其根要移栽到直径为9cm的塑料盆中,盆中有85g介质(60%碳泥,20%蛭石,and 20%珍珠岩,pH 6.5
11、)和1.5g奥绿肥(15-9-12)。Plants were grown for a minimum of 4 weeks before treatments.Experiments were conducted in greenhouses with temperatures maintained between 18 and 32C and light levels between 285 and 380 mol.m-2.s-1.Plants were arranged in a completely randomized design with 10 plants per treatme
12、nt.在处理之前,作物最少生长4周,该实验在温室中进行,温度:18 到 32C,光照:285 到 380 mol.m-2.s-1.实验完全随机区组,每个处理10株。Two additional treatments were included in each test:a noninoculated control(saline,8.5 g NaCl/liter applied without R.solanacearum)and a disease control(no product applied,inoculated with pathogen).每个实验增加两个处理:不接种控制(含有
13、盐分,NaCl,无青枯菌);病害控制(不施其他物质,但接种病菌)。For inoculum production,R.solanacearum strains were grown on triphenyltetrazolium chloride(TZC)medium(11)for 48 h,and cells were harvested and spectrophotometrically adjusted(A600)in saline to 5 108CFU/ml.接种青枯菌株的要在氯化三苯基四氮唑(TZC)介质中生长48小时,达到5 108CFU/ml.的时候收获菌株。Wilt sym
14、ptoms were recorded as they occurred.Bacteria were reisolated 2 weeks after last treatment(6 weeks after inoculation)from plants without wilt symptoms.A cross-section of stem 1 cm in length was removed approximately 0.5 cm above the soil line of each geranium plant.萎蔫症状一出现就要记录下来,最后一次处理之后两周,细菌就要从没有萎蔫
15、症状的植物中分离。取1cm茎,从离地面0.5cm开始取。通过处理把悬浮液加到TZC介质中培养然后进行检测。2 Further testing of promising products.Geranium plants were found not to be systemically infected by R.solanacearum when treated with either K-Phite(a.i.53%mono-and di-potassium salts of phosphorous acid)or Starner(a.i.20%oxolinic acid)at the ini
16、tial test rates.在初始的检测中,当施以53%的磷酸一钾或二钾或 Starner,天竺葵不能被青枯菌系统性侵染。To determine effective application rates and intervals for these products,four rates were tested(vol/vol,0.25,0.5,0.75,1.0%)at three application regimes(one application;two applications at 14-day interval;and four applications at 7-day i
17、nterval).Bactericides were applied 3 days before pathogen inoculation as in the initial product screen.为了测定这些物质的有效性和周期,用4种比率(0.25,0.5,0.75,1.0%)测定,用三种施用方式(一次性施用,两次施用间隔为14天,四次施用间隔为7天),杀菌剂在接种病原菌之前三天施用。Each treatment again contained 10 replicates.Using the R1B1 strain,four tests were conducted with two
18、 tests at each inoculum concentration:5 108 CFU/ml(3 108 CFU/gram soil),and 1 107CFU/ml(6 106 CFU/gram soil).每个处理10个重复,用R1B1菌株,进行了两个接种测试:5 108 CFU/ml(3 108 CFU/gram soil),1 107CFU/ml(6 106 CFU/gram soil).Two additional controls(five plants each)were added at the lower inoculum rate to monitor concen
19、trations of R.solanacearum within the potting medium.These controls consisted of:(i)pots containing potting medium without a geranium plant,yet inoculated with R.solanacearum,and(ii)pots containing potting medium without plant or R.solanacearum inoculation.Additional controls were watered like all o
20、ther treatments.在接种较低的盆栽介质中增加了两个处理,每个处理5株,只有盆栽介质(盆栽介质+天竺葵,盆栽介质+接种)。与其它处理一样浇水。Six weeks after first chemical application,a cork borer was used to retrieve a 1-g sample of potting medium from the first five replications of all treatments.A dilution series of the soil sample was done in SDW and replica
21、plated onto modified semiselective medium.Typical R.solanacearum colonies were counted after 48 h growth at 28C.Bacteria were reisolated from all geranium plants 6 weeks after inoculation,as previously described.Colonies from plates with typical R.solanacearum growth pattern were suspended in saline
22、(8.5 g/liter NaCl)and injected into parenchymatous tissue of tobacco for hypersensitive response(HR)following protocols of Lozano and Sequeira(12).在28C下培养48小时后计算青枯菌群,接种6周后从天竺葵中分离细菌,如前面所述得到的悬浮液注入马铃薯的薄壁组织中来测其敏感性。分离茎和土壤,把马铃薯种到每个盆子中来验证青枯菌是否足够病害发展。马铃薯对于R1B1 和R3B2菌属是十分敏感的,最少4周后就会出现症状,与前面相同,出现症状就进行分离。The R
23、3B2 strain was tested in environmental chambers set on a 12-h day/night cycle(19C night,24C day,310 mol.m-2.s-1).Two tests were conducted at an inoculum concentration of 5 108CFU/ml(3 108 CFU/gram soil).All other experimental parameters and controls were the same as previously described.R3B2在环境检测箱内放
24、12h后测定,两个实验的接种浓度为5 108CFU/ml 其它的实验参数和控制条件都与前面相同。Antibacterial efficacy of compounds containing phosphorus Many products containing P are currently sold in the ornamental plant industry as fertilizers or fungicides.Common ingredients include P2O5,H3PO3,and H3PO4 with mono-and/or di-potassium.We teste
25、d the efficacy for controlling bacterial wilt by P2O5,H3PO3,H3PO4,and KCl.很多含磷产品在工厂都以肥料或菌剂来买,一般有P2O5,H3PO3,kH2PO4,k2HPO4。我们用P2O5,H3PO3,H3PO4,和 KCl来测它们对青枯菌的控制效率。Reagent grade KCl and the three P chemicals buffered with CaCO3 were used in the first test,and the P chemicals only buffered with KOH were
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