讀書報告12磷酸對天竺葵青枯病的控制課件VIP

1、Control of Bacterial Wilt of Geranium with Phosphorous Acid D. J. Norman 磷酸對天竺葵青枯病的控制 讀書報告12磷酸對天竺葵青枯病的控制2 主要內(nèi)容： 一：前言 二：材料與方法 三：結(jié)果與討論 讀書報告12磷酸對天竺葵青枯病的控制3 一、前言 Once it becomes established in a susceptible crop,southern bacterial wilt, caused by Ralstonia solanacearum is very hard to eradicate. There a

2、re two primary avenues by which R. solanacearum moves 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，就很難根除。青枯菌 侵染作物的兩種途徑是：通過水流侵染通過被侵染物質(zhì)傳播。 To combat bacterial wilt, resistant cultivars have been recommended in areas of the w

3、orld where the pathogen is endemic .In susceptible greenhousegrown crops, only strict sanitation has been successful in prohibiting plant infestation. 為了抵抗地方性青枯病，世界上很多地方都推薦種植抗性作物。種植 在溫室的敏感作物只有控制環(huán)境衛(wèi)生才能抑制其侵染。 讀書報告12磷酸對天竺葵青枯病的控制4 Geraniums are susceptible to races 1 and 3 of R. solanacearum . Most gera

4、niums produced in the world are vegetatively propagated in Guatemala, Costa Rica,Columbia, 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. 天竺葵對于青枯菌屬1和3是敏感的，天竺葵廣泛種植在世界各地如危地 馬拉，哥斯達(dá)黎

5、加，哥倫比亞，中國和肯尼亞。但這些地方都存在地方 性青枯病，目前還沒有有效的方法保護(hù)天竺葵作物。 Thus, the objective of this research was to determine if geranium plants could be protected from infection with applications of selected bactericides and chemicals. 因此本實驗的目的就是對天竺葵作物施以篩選過的細(xì)菌和化學(xué)藥品來驗 證是否能保護(hù)作物免受侵染。 讀書報告12磷酸對天竺葵青枯病的控制5 二、材料與方法 1 Screening

6、of products. A race 1 (biovar 1) strain (R1B1) isolated in Florida (P673) from pothos (Epipremnum aureum (Linden 病害控制（不施其他物質(zhì)，但接種病菌）。 For inoculum production, R. solanacearum strains were grown on triphenyltetrazolium chloride (TZC) medium (11) for 48 h, and cells were harvested and spectrophotometri

7、cally adjusted (A600) in saline to 5 108CFU/ml. 接種青枯菌株的要在氯化三苯基四氮唑（TZC）介質(zhì)中生長48小時， 達(dá)到5 108CFU/ml.的時候收獲菌株。 讀書報告12磷酸對天竺葵青枯病的控制9 Wilt symptoms were recorded as they occurred. Bacteria were reisolated 2 weeks after last treatment (6 weeks after inoculation) from plants without wilt symptoms. A cross-secti

8、on of stem 1 cm in length was removed approximately 0.5 cm above the soil line of each geranium plant. 萎蔫癥狀一出現(xiàn)就要記錄下來，最后一次處理之后兩周，細(xì)菌就要從 沒有萎蔫癥狀的植物中分離。取1cm莖，從離地面0.5cm開始取。通過 處理把懸浮液加到TZC介質(zhì)中培養(yǎng)然后進(jìn)行檢測。 2 Further testing of promising products. Geranium plants were found not to be systemically infected by R. s

9、olanacearum 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 initial test rates. 在初始的檢測中， 當(dāng)施以53%的磷酸一鉀或二鉀或 Starner，天竺葵不 能被青枯菌系統(tǒng)性侵染。 讀書報告12磷酸對天竺葵青枯病的控制10 To determine effective application rates and intervals for t

10、hese 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 interval). Bactericides were applied 3 days before pathogen inoculation as in the initial product screen. 為了測定這些物質(zhì)的

11、有效性和周期，用4種比率（0.25, 0.5, 0.75, 1.0%） 測定，用三種施用方式（一次性施用，兩次施用間隔為14天，四次施 用間隔為7天)，殺菌劑在接種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 tests at each inoculum concentration: 5 108 CFU/ml (3 108 CFU/gram soil), and 1 107CFU/ml (6 106 CFU/gr

12、am soil). 每個處理10個重復(fù)，用R1B1菌株，進(jìn)行了兩個接種測試：5 108 CFU/ml (3 108 CFU/gram soil), 1 107CFU/ml (6 106 CFU/gram soil). 讀書報告12磷酸對天竺葵青枯病的控制11 Two additional controls (five plants each) were added at the lower inoculum rate to monitor concentrations of R.solanacearum within the potting medium. These controls con

13、sisted 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 other treatments. 在接種較低的盆栽介質(zhì)中增加了兩個處理，每個處理5株，只有盆栽 介質(zhì)(盆栽介質(zhì)+天竺葵，盆栽

14、介質(zhì)+接種)。與其它處理一樣澆水。 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 replicaplated onto modified semiselective medium. 讀書報告12磷酸對天竺葵青枯

15、病的控制12 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 (8.5 g/liter NaCl) and inject

16、ed into parenchymatous tissue of tobacco for hypersensitive response (HR) following protocols of Lozano and Sequeira (12). 在28C下培養(yǎng)48小時后計算青枯菌群，接種6周后從天竺葵中分離細(xì)菌， 如前面所述得到的懸浮液注入馬鈴薯的薄壁組織中來測其敏感性。 分離莖和土壤，把馬鈴薯種到每個盆子中來驗證青枯菌是否足夠病害 發(fā)展。馬鈴薯對于R1B1 和R3B2菌屬是十分敏感的，最少4周后就會出 現(xiàn)癥狀，與前面相同，出現(xiàn)癥狀就進(jìn)行分離。 讀書報告12磷酸對天竺葵青枯病的控制13 The

17、 R3B2 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. R3B

18、2在環(huán)境檢測箱內(nèi)放12h后測定，兩個實驗的接種濃度為5 108CFU/ml 其它的實驗參數(shù)和控制條件都與前面相同。 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-pota

19、ssium. We tested the efficacy for controlling bacterial wilt by P2O5, H3PO3, H3PO4, and KCl. 很多含磷產(chǎn)品在工廠都以肥料或菌劑來買，一般有P2O5, H3PO3, kH2PO4， k2HPO4。我們用P2O5, H3PO3, H3PO4,和 KCl來測它們對青枯菌的控制效率。 讀書報告12磷酸對天竺葵青枯病的控制14 Reagent grade KCl and the three P chemicals buffered with CaCO3 were used in the first test, a

20、nd the P chemicals only buffered with KOH were used in the sec-ond test. We used four concentrations of P, 0.032, 0.063, 0.095, and 0.127%, for each of the three chemicals in the two tests; and the first four concentrations of K, 0.054, 0.108, 0.162, and 0.216%, were made from KCl. 第一個實驗中，用CaCO3來緩沖K

21、Cl和三種含磷化合物。第二個實驗中 用KOH來緩沖含磷化合物，四種P的濃度為 0.032, 0.063, 0.095, 0.127%, 兩個處理的三種含磷物質(zhì)濃度一樣，4種K的濃度為0.054, 0.108, 0.162, 0.216%。 As in the initial product, screening solutions were applied on a 7-day interval with four applications. Again, plants (10 per replication) were inoculated with strain R1B1 3 days a

22、fter the first compound application. Two weeks after last application, all plants were sampled for R. solanacearum as previously described. 在起始篩選中，篩選溶液將會以7天的間隔施用在四個處理中。作物接 種R1B1菌，三天后施用第一次，最后一次施用之后兩周收集所有植株 來詳細(xì)描述青枯菌。 讀書報告12磷酸對天竺葵青枯病的控制15 We measured the in vitro effect of phosphorous acid on the growt

23、h of both R1B1 and R3B2 strains. Flasks containing 40 ml of the following solutions were used: (i) SDW, (ii) SDW + P at 0.127% as H3PO3, (iii) SDW + K-Phite at 1%, (iv) Nutrient broth (Difco) containing 5 g sucrose (NB), (v) NB + P at 0.127% as H3PO3, and (vi) NB + K-Phite at 1%. 我們在試管中進(jìn)行了磷酸對 R1B1 和

24、R3B2的控制實驗，分6組。分別 為滅菌水滅菌水+0.127% H3PO3滅菌水+1%磷酸鉀營養(yǎng)液 營養(yǎng)液+0.127% H3PO3營養(yǎng)液+1%磷酸鉀 Concentrations of viable cells in each flask were enumerated on 0, 1, 7, 14, 21, and 28 days.Counts were discontinued when concentrations of cells reached 109CFU/ml. 在適當(dāng)條件下進(jìn)行培養(yǎng)，在0, 1, 7, 14, 21, 和28 天計算其存活的細(xì)菌數(shù) 量。當(dāng)其濃度達(dá)到109CFU

25、/ml，不在計算。 讀書報告12磷酸對天竺葵青枯病的控制16 Safety of K-Phite to crop. To determine if phytotoxicity occurred with the K-Phite product, an experiment was set up as previously described using four concentrations (vol/vol, 0.50, 0.75, 1, and 2%) with 10 plants per treatment. Dry weights of both roots and foliage o

26、f all plants were determined at the end of the test.In addition, we measured medium pH and soluble salts for five plants of each treatment and performed a complete foliar analysis. 為了確定K-Phite 對作物是否安全，我們進(jìn)行了如前面所述4個濃度 （ 0.50, 0.75, 1, 和2%），每個處理10株。實驗最后測所有植株的根系 和葉片，每個處理測5株的介質(zhì)濃度和鹽溶液。對葉片進(jìn)行完整的分 析。 讀書報告12磷

27、酸對天竺葵青枯病的控制17 RESULTS Screening of products. Further testing of promising products. Efficacy of phosphorous compounds. Safety of K-Phite to crop. 讀書報告12磷酸對天竺葵青枯病的控制18 Screening of products. In the initial testing, nearly all the products slowed disease progress. However, they did not protect the pla

28、nts from infection, except for benzothiadiazole (Actigard), oxolinic acid (Starner), and potassium salts of phosphorous acid (K-Phite) (data not shown). Further testing of benzothiadia-zole was discontinued due to leaf abscission. This abscission occurred even at rates as low as 5 l/liter. 在開始的測試中，所

29、有的物質(zhì)都能減慢病害發(fā)生的過程，但不能保護(hù) 作物免受侵染，除了苯并噻二唑，奧利索酸和磷酸鉀鹽。而苯并噻二 唑即便在很低的濃度下也會損害葉片，所以不再繼續(xù)實驗。 Further testing of promising products. Low rates and intervals of Starner were ineffective in protecting plants from infection. High rates above 0.5% on a 7-day interval provided protection for the majority of the plants

30、. However, at inoculation rates: of 3 108 CFU/gram soil and 6 106 CFU/gram soil, systemic infections were occasionally observed in plants treated with 0.75 and 1% Starner. Due to the high rates and intervals required for disease protection, testing of Starner was discontinued. 低比率和間隔的Starner沒有保護(hù)作用，高

31、于0.5%雖有效，但作物還是 會出現(xiàn)偶然的系統(tǒng)性侵染，所以對Starner不再進(jìn)行實驗。 讀書報告12磷酸對天竺葵青枯病的控制19 K-Phite was fairly effective in protecting geranium plants from infection at 3 108CFU/gram soil (Fig. 1) and very effective in protecting plants at 6 106 CFU/gram soil. 在接種濃度為 3 108CFU/gram時，磷酸鉀的保護(hù)作用是有 效的，在 6 106 CFU/gram 時保護(hù)效果非常明顯。具體

32、情 況如圖一所示。 讀書報告12磷酸對天竺葵青枯病的控制20 讀書報告12磷酸對天竺葵青枯病的控制21 Populations of R. solanacearum were not detectable in the soil after potting medium was drenched with K-Phite for either two or four applications (Fig. 2).the R1B1 strain was more persistent than the R3B2 strain while the R3B2 strain was undetectab

33、le at that time. 在兩次或四次施用磷酸鉀的處理中檢測部到青枯菌。R1B1 比 R3B2更持久，R1B1在接種后6周還可以檢測到青枯菌但 R3B2。 讀書報告12磷酸對天竺葵青枯病的控制22 Fig. 1. Effects of K-Phite application on control of Ralstonia solanacearum infection. 讀書報告12磷酸對天竺葵青枯病的控制23 Efficacy of phosphorous compounds.K-Phite effectively protected geranium plants from infe

34、ction with the R1B1 strain at all four applied rates (Fig. 3). Since it is a product of potassium salts of phosphorous acid, three P chemicals, P2O5, H3PO3, and H3PO4, were tested. 如圖3所示：磷酸鉀能的4個處理都能保護(hù)作物免受侵染， 而其它含磷物質(zhì)卻不可以。 讀書報告12磷酸對天竺葵青枯病的控制24 Fig. 3. Relative effects of K-Phite and phosphorus compoun

35、ds in control of Ralstonia solanacearum(strain P673, R1B1). 讀書報告12磷酸對天竺葵青枯病的控制25 The long-term in vitro growth curves were similar for both bacterial strains tested. In Figure 4, results are displayed for UW551. Phosphorous acid either in reagent grade or product form inhibited cell replication in N

36、B and SDW (Fig. 4). 兩種細(xì)菌的長期試管生長曲線都相似，R3B2菌屬結(jié)果如圖所示。 Safety of K-Phite to crop. There was a significant (P = 0.5) increase in the geranium shoot dry weight of 27% with the 1% K-Phite treatment relative to the water control. All other shoot dry weights were not significantly different from the water co

37、ntrol.Iron levels decreased from 122.5 ppm to 45.7 ppm (R2 0.58). Leaves were less green on plants treated with 1% K- Phite than on plants treated with waterIn order to maintain healthy geranium plants, the K-Phite product should not be drenched at rates above 1% on a prolonged basis。 與空白對照相比，用1%磷酸鉀

38、處理的天竺葵地上部干重明顯增加了 27%，別的處理與空白對照無顯著性差異。但磷酸鉀高于1%時，葉片 中鐵的含量從122.5 ppm減低到45.7 ppm，葉片發(fā)黃。所以磷酸鉀的比 率不能高于1%。 讀書報告12磷酸對天竺葵青枯病的控制26 Fig. 4. Growth of Ralstonia solanacearum UW551 in flasks containing 40 ml of the following solutions: ( ) sterile distilled water (SDW), () SDW + 0.127% P as H3PO3, () SDW + 1% K-

39、Phite (53% mono- and di-potassium salts of phosphorous acid), () Nutrient broth Difcocontaining 5 g sucrose (NB), () NB + 0.127% P as H3PO3, and () NB + 1% K-Phite. 讀書報告12磷酸對天竺葵青枯病的控制27 DISCUSSION The majority of the geraniums sold in the global north ,but where R. solanacearum is endemic.In order t

40、o exclude R3B2 from these geranium production acilities, rigorous sanitation measures are being implemented (24). However, complete eradication of bacterial wilt is difficult given an environment conducive to both host and pathogen. Thus, in addition o sanitation and resistant cultivars, growers nee

41、d other control options. 天竺葵的主要產(chǎn)地是北半球，但存在地方性青枯病，為了控制天竺葵 作物R3B2菌，對生產(chǎn)設(shè)施進(jìn)行了嚴(yán)格的控制，然而一般很難完全根除。 因此，除了控制環(huán)境和栽培抗性品種，種植者也需要其他控制。 讀書報告12磷酸對天竺葵青枯病的控制28 This study showed that most of the tested products slowed the progression of bacterial wilt on geranium, but did not protect the plants from infection and subse

42、quent death. Only phosphorous acid protected plants from infection.It appears that the protection occurs in the soil and root matrix, as pathogen cells could not be cultured from the potting medium of treatments with high frequency and application rates of K-Phite . 本實驗表明，大多數(shù)檢測產(chǎn)品都能減慢青枯菌在天竺葵中的傳播過程， 但只有磷酸才能保護(hù)天竺葵免受侵染。在高頻率和高比率施用磷酸鉀 的處理中，分離不出病菌，所以保護(hù)部位是在土壤和根系。 讀書報告12磷酸對天竺葵青枯病的控制29 The mechan