WO2021180048A1 - 一种用于防治细菌性条斑病的组合物及其制备方法与应用 - Google Patents

一种用于防治细菌性条斑病的组合物及其制备方法与应用 Download PDF

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WO2021180048A1
WO2021180048A1 PCT/CN2021/079632 CN2021079632W WO2021180048A1 WO 2021180048 A1 WO2021180048 A1 WO 2021180048A1 CN 2021079632 W CN2021079632 W CN 2021079632W WO 2021180048 A1 WO2021180048 A1 WO 2021180048A1
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Prior art keywords
composition
rice
streak
parts
stirring
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PCT/CN2021/079632
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English (en)
French (fr)
Inventor
杨祁云
路征
林壁润
孙大元
张景欣
沈会芳
蒲小明
吕永华
邓海滨
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广东省农业科学院植物保护研究所
中国烟草总公司广东省公司
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Application filed by 广东省农业科学院植物保护研究所, 中国烟草总公司广东省公司 filed Critical 广东省农业科学院植物保护研究所
Priority to US18/548,989 priority Critical patent/US20240215587A1/en
Publication of WO2021180048A1 publication Critical patent/WO2021180048A1/zh
Priority to ZA2022/00294A priority patent/ZA202200294B/en

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    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N33/00—Biocides, pest repellants or attractants, or plant growth regulators containing organic nitrogen compounds
    • A01N33/02—Amines; Quaternary ammonium compounds
    • A01N33/12—Quaternary ammonium compounds
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N59/00—Biocides, pest repellants or attractants, or plant growth regulators containing elements or inorganic compounds
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01N—PRESERVATION OF BODIES OF HUMANS OR ANIMALS OR PLANTS OR PARTS THEREOF; BIOCIDES, e.g. AS DISINFECTANTS, AS PESTICIDES OR AS HERBICIDES; PEST REPELLANTS OR ATTRACTANTS; PLANT GROWTH REGULATORS
    • A01N59/00—Biocides, pest repellants or attractants, or plant growth regulators containing elements or inorganic compounds
    • A01N59/16—Heavy metals; Compounds thereof
    • A01N59/20—Copper
    • A—HUMAN NECESSITIES
    • A01—AGRICULTURE; FORESTRY; ANIMAL HUSBANDRY; HUNTING; TRAPPING; FISHING
    • A01P—BIOCIDAL, PEST REPELLANT, PEST ATTRACTANT OR PLANT GROWTH REGULATORY ACTIVITY OF CHEMICAL COMPOUNDS OR PREPARATIONS
    • A01P1/00—Disinfectants; Antimicrobial compounds or mixtures thereof

Definitions

  • the invention relates to the field of agricultural production, in particular to a composition for preventing and treating bacterial streak disease, and a preparation method and application thereof.
  • Rice bacterial streak caused by Xanthomonas oryzaes pv.oryzicola is one of the important phytosanitary diseases in China, and it is also an important disease in tropical and subtropical rice growing areas. Its occurrence is characterized by epidemic, fulminant and devastating characteristics. The disease was first discovered in Guangdong in 1957, and later confirmed by Fang Zhongda and others as bacterial streaks.
  • Bacterial Xanthomonas oryzae pv. oryzae mainly overwinters on diseased rice and straw. The pathogen infects the seeds and spreads long distances along with the seed transfer; the pathogens mainly contact the seedlings through irrigation water and rainwater, and invade from the stomata and wounds.
  • the primary purpose of the present invention is to overcome the shortcomings and deficiencies of the prior art and provide a composition for controlling rice bacterial streak.
  • Another object of the present invention is to provide a preparation method of the composition for controlling rice bacterial leaf streak.
  • Another object of the present invention is to provide an application of the composition for controlling rice bacterial leaf streak.
  • a composition for the prevention and treatment of rice bacterial streak including the following components in weight percentage: dodecyl dimethyl benzyl ammonium bromide 10-15 Parts, 15-20 parts of copper sulfate pentahydrate, 20-25 parts of 84 disinfectant, 40-55 parts of water.
  • composition for preventing and treating rice bacterial streak disease preferably includes the following components in weight percentage: 10-13 parts of dodecyl dimethyl benzyl ammonium bromide, 15 ⁇ copper sulfate pentahydrate 18 parts, 20 to 23 parts of 84 disinfectant, 46 to 55 parts of water.
  • the composition for preventing and treating rice bacterial streak more preferably includes the following components in weight percentage: 10 to 12.5 parts of dodecyl dimethyl benzyl ammonium bromide, 15 parts of copper sulfate pentahydrate ⁇ 17.5 parts, 20 ⁇ 22.5 parts of 84 disinfectant, 47.5 ⁇ 55 parts of water.
  • composition for controlling rice bacterial streak disease most preferably comprises the following components in weight percentage: 12.5 parts of dodecyl dimethyl benzyl ammonium bromide, 17.5 parts of copper sulfate pentahydrate, 22.5 parts of 84 disinfectant and 47.5 parts of water.
  • the dodecyl dimethyl benzyl ammonium bromide can be commercial dodecyl dimethyl benzyl ammonium bromide with a content of more than 95%.
  • the copper sulfate pentahydrate is copper sulfate pentahydrate crystals.
  • the preparation method of the composition for controlling rice bacterial streak disease includes the following steps:
  • step (2) Add dodecyl dimethyl benzyl ammonium bromide to the copper sulfate solution obtained in step (1), stir and mix uniformly to obtain a transparent liquid A;
  • step (3) Add the 84 disinfectant solution to the transparent liquid A obtained in step (2), stir and mix uniformly, to obtain a composition for preventing and treating rice bacterial streak.
  • the stirring conditions described in step (1) are: stirring at 100-200 rpm for 40-50 minutes; preferably: stirring at 200 rpm for 40-50 minutes.
  • the stirring conditions described in step (2) are: 100-200 rpm stirring for 15-30 minutes; preferably, 100-200 rpm stirring at room temperature for 15-30 minutes.
  • the stirring conditions described in step (3) are: stirring at 100-200 rpm for 15-30 minutes; preferably: stirring at 100-200 rpm at 25°C-30°C for 15-30 minutes.
  • composition for preventing and treating rice bacterial leaf streak in the preparation of pesticide formulations for preventing and treating rice bacterial leaf streak.
  • Said pesticide preparations include liquids and the like.
  • composition for preventing and curing rice bacterial leaf streak in the prevention and treatment of rice bacterial leaf streak.
  • the application of the composition for preventing and curing rice bacterial leaf streak in the prevention and treatment of rice bacterial leaf streak is to dilute the composition for preventing and curing rice bacterial leaf streak by 500-2000 times, It is used for soaking rice seeds or spraying on rice leaves to prevent and control rice bacterial streak.
  • the dilution factor of the composition for controlling rice bacterial streak disease is preferably 500-1500 times; more preferably 1000-1500 times; most preferably 1500 times.
  • the time for soaking rice seeds is preferably 24 hours.
  • the dosage of the diluted composition for controlling rice bacterial streak disease is preferably 50 kg/mu.
  • the present invention has the following advantages and effects:
  • the present invention provides a composition that can be used for the prevention and treatment of rice bacterial streak.
  • the composition is an aqueous solution consisting of water, dodecyl dimethyl benzyl ammonium bromide, copper sulfate pentahydrate and 84 disinfectant. It is prepared according to a certain ratio and method; a drug with better control effect on rice bacterial stripe disease is developed, which solves the problem of shortage of pesticides for control of rice bacterial stripe disease, and the preparation and application method of the composition is simple and easy to operate. , Low input cost, quick effect, can be applied to many fields such as agricultural production, scientific research, etc. It has great application value in actual production, and it can reduce the food loss caused by disease more economically and environmentally.
  • the present invention will be further described in detail below in conjunction with examples, but the implementation of the present invention is not limited thereto.
  • the reagents and methods used in the present invention are conventional reagents and methods in the technical field.
  • all reagents used in the present invention are commercially available.
  • the dodecyl dimethyl benzyl ammonium bromide (95%, V/V) involved in the present invention was purchased from Shandong Lanying Chemical Technology Co., Ltd.; 84 disinfectant (effective chlorine content 5% to 7%) is Medical disinfectant was purchased from Shantou Yaqi Trading Co., Ltd.; copper sulfate was a crystal of copper sulfate pentahydrate and was purchased from Guangzhou Donghui Trading Co., Ltd.
  • the preparation method is:
  • the first step place 5500ml of water and 1500g of copper sulfate pentahydrate in a stirring device and disperse evenly (at 50°C, at a speed of 200 rpm, stirring for 40 minutes) to obtain a copper sulfate pentahydrate solution;
  • composition A In the third step, add the solution obtained in the second step into the reaction kettle, add 2000 ml of 84 disinfectant solution, and stir at 200 rpm at 25° C. for 30 minutes to obtain composition A.
  • the preparation method is:
  • composition B In the third step, add the solution obtained in the second step into the reaction kettle, add 2250 ml of 84 disinfectant solution, and stir for 15 minutes at 100 rpm at 30°C to obtain composition B.
  • the preparation method is:
  • the first step place 4000ml of water and 2000g of copper sulfate pentahydrate in a stirring device to disperse evenly, and stir for 50 minutes at a speed of 200 rpm at 60°C to obtain a copper sulfate pentahydrate solution;
  • the second step add 1250 g of dodecyl dimethyl benzyl ammonium bromide to the copper sulfate pentahydrate solution prepared in the first step, and stir for 15 minutes at 150 rpm at 30°C, and continue to stir until The solution becomes a transparent liquid;
  • composition C In the third step, add the solution obtained in the second step into the reaction kettle, add 2250 ml of 84 disinfectant solution, and stir at 100 rpm for 20 minutes at 28°C to obtain composition C.
  • composition of the present invention can be used for soaking seeds (rice seeds) or spraying rice leaves after being diluted with water, for preventing and treating rice bacterial leaf streak.
  • composition A, B, C and 84 disinfectant, dodecyl dimethyl benzyl ammonium bromide, copper sulfate pentahydrate were respectively used at 1:500, 1:1000, 1:1500, 1:2000 ( v/v) Dilute with water, and use sterile water as a control.
  • the bacterial streaks of rice cultured on PSA slant medium 200g potato, 20g sucrose, 20g agar and 1000mL distilled water, pH 5.6 ⁇ 6.6) for 2 days
  • Germs Xanthomonas oryzaes pv.oryzicola; Institute of Plant Protection, Guangdong Academy of Agricultural Sciences
  • the formula for bacterial growth inhibition rate is as follows:
  • Bacterial growth inhibition rate % [(control inhibition zone diameter-treatment inhibition zone diameter)/control inhibition zone diameter] ⁇ 100.
  • the treated bacterium loses its vitality and is difficult to grow on the PSA medium. Therefore, the stronger the bactericidal power of the medicament, the greater the inhibitory dilution concentration.
  • compositions A, B, and C have strong bactericidal activity against rice bacterial leaf streak, and when the composition B and composition C are diluted 1500 times, the growth inhibition rate of rice bacterial leaf streak reaches 100. %, is the best of the three compositions (see Table 1).
  • 84 disinfectant, dodecyl dimethyl benzyl ammonium bromide, copper sulfate (the raw material is copper sulfate pentahydrate crystals) have a certain inhibitory effect on pathogens, but the effect is poor, not as good as composition A, B, C, description After they are combined, there is a certain synergistic effect, which may be related to the production of copper complexes by the three compounds.
  • compositions A, B, and C Dilute 1500 and 2000 times with the compositions A, B, and C respectively, and use them for soaking rice seeds for 24 hours.
  • compositions A and B have no inhibitory effect on rice seed germination and seedling growth, indicating that the combination of the three compounds is relatively safe for rice, and can be used for seed soaking, seedling stage and field use;
  • 84 disinfectant, dodecyl Dimethyl benzyl ammonium bromide and copper sulfate have inhibitory effects on rice seed germination and seedling growth, and
  • 84 disinfectant, dodecyl dimethyl benzyl ammonium bromide and copper sulfate are prone to phytotoxicity caused by pesticides. Suitable for use on rice alone.
  • Test location conducted in Congress, Institute of Plant Protection, Guangdong Academy of Agricultural Sciences.
  • the rice variety Shuiyuan 290 was sown in the experimental Wimbledon room. At the three-leaf stage of the seedlings, the seedlings were inoculated by spray inoculation with Bacterial Xanthomonas oryzae (concentration of 10 9 CFU/ml). After moisturizing for 12 hours, use Composition A respectively. , B, C add water and spray with 1:1500, 1:2000 (v/v) diluted medicament and clear water (the amount of diluted medicament is 50kg/mu), after 10 days, use the same medicament and concentration for the second spray The diseased leaf rate was investigated 3 days after the last spray (that is, the second), and the average value was taken for three repetitions.
  • Diseased leaf rate% number of diseased leaves/total number of leaves under investigation ⁇ 100.
  • Control effect (%) [(control leaf diseased leaf rate-treatment leaf diseased leaf rate)/control leaf diseased leaf rate] ⁇ 100.
  • compositions A, B, and C In the early stage of rice bacterial streak disease, use the compositions A, B, and C to dilute 1500 times and 2000 times, respectively, and spray (the dosage of the diluted medicament is 50kg/mu), spray once every 7 days for a total of sprays Three times, 7 days after the last spray, the morbidity and disease index were investigated, and the control effect was calculated.
  • Control effect (%) [(control rice disease index-treated rice disease index)/control rice disease index] ⁇ 100.
  • composition B has the best control effect on rice bacterial streak, reaching 78.5% (see Table 4).
  • Table 4 The control effect of spray composition A, B, C on rice bacterial streak in the field of rice

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  • Plant Pathology (AREA)
  • Health & Medical Sciences (AREA)
  • Agronomy & Crop Science (AREA)
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Abstract

本发明公开了一种用于防治细菌性条斑病的组合物及其制备方法与应用。该组合物包括如下重量百分数计的组分：十二烷基二甲基苄基溴化铵10～15份，五水硫酸铜15～20份，84消毒液20～25份，水40～55份。将本发明中的组合物稀释后，可以用于水稻浸种以预防水稻细菌性条斑病，或将其喷施到水稻叶面上用于防治水稻细菌性条斑病，其应用方法操作简便，解决了水稻细菌性条斑病防治药剂紧缺难题，本发明为农业生产、科研等众多领域提供了技术支持。

Description

一种用于防治细菌性条斑病的组合物及其制备方法与应用 技术领域
本发明涉及农业生产领域，特别涉及一种用于防治细菌性条斑病的组合物及其制备方法与应用。
背景技术
随着种植业结构的调整和外来有害生物的入侵，近年来农作物重大病害频繁暴发。其中粮食作物中的水稻细菌性条斑病、水稻白叶枯病，果树中的柑橘溃疡病，瓜菜类中的茄科蔬菜青枯病等都给作物造成了严重损失。目前，生产上防治植物病害主要还是依靠化学农药，与细菌性病害防治难、危害普遍形成鲜明对比的是，目前国内用于防治细菌性病害的药剂登记数量还比较有限，截至2018年10月底，中国登记的杀菌剂产品约有1.09万个，其中防治细菌性病害的药剂仅280余个，占比2.6％。面对细菌性病害的威胁，药剂防治是不可替代的。未来作物细菌性病害的发生面积可能继续增加，专业防治药剂发展前景好，尤其是硫酸链霉素等的退出给市场留下了更大的空间。
由Xanthomonas oryzaes pv.oryzicola引起的水稻细菌性条斑病是国内重要的植物检疫病害之一，也是热带和亚热带稻作区的重要病害，其发生具有流行性、暴发性和毁灭性等特点，我国于1957年在广东首次发现该病，后由方中达等确认为细菌性条斑病。水稻细菌性条斑病菌主要在病稻谷和病稻草上越冬。病原菌侵染种子，随着种子调运而远距离传播；病原菌主要通过灌溉水、雨水接触秧苗，从气孔和伤口侵入，侵入后在气孔下繁殖扩展到薄壁组织细胞间隙并纵向扩展，形成条斑；田间病株病斑上溢出的菌脓，主要通过风雨和水传播，进行再侵染，引起病害扩展蔓延。近年来,随着杂交稻的大面积推广,种子的南繁北调,使得水稻细菌性条斑病的发病面积不断扩大，危害逐年加重，现在整个南方稻区均有发生，对水稻的高产稳产造成严重威胁。对水稻细菌性条斑病的防治，目前尚缺乏特效药剂，随着原来使用的农用链霉素和叶青双(叶枯净)等产品因国家禁用退出市场，因此研制新型高效低毒低残留的药剂是十分必要的。
发明内容
本发明的首要目的在于克服现有技术的缺点与不足，提供一种用于防治水稻细菌性条斑病的组合物。
本发明的另一目的在于提供所述用于防治水稻细菌性条斑病的组合物的制备方法。
本发明的再一目的在于提供所述用于防治水稻细菌性条斑病的组合物的应用。
本发明的目的通过下述技术方案实现：一种用于防治水稻细菌性条斑病的组合物，包括如下重量百分数计的组分：十二烷基二甲基苄基溴化铵10～15份，五水硫酸铜15～20份，84消毒液20～25份，水40～55份。
所述的用于防治水稻细菌性条斑病的组合物，优选为包括如下重量百分数计的组分：十二烷基二甲基苄基溴化铵10～13份，五水硫酸铜15～18份，84消毒液20～23份，水46～55份。
所述的用于防治水稻细菌性条斑病的组合物，更优选为包括如下重量百分数计的组分：十二烷基二甲基苄基溴化铵10～12.5份，五水硫酸铜15～17.5份，84消毒液20～22.5份，水47.5～55份。
所述的用于防治水稻细菌性条斑病的组合物，最优选为包括如下重量百分数计的组分：十二烷基二甲基苄基溴化铵12.5份，五水硫酸铜17.5份，84消毒液22.5份，水47.5份。
所述的十二烷基二甲基苄基溴化铵可采用市售含量95％以上的溴化十二烷基二甲基苄基铵。
所述的五水硫酸铜为五水硫酸铜结晶。
所述的用于防治水稻细菌性条斑病的组合物的制备方法，包括如下步骤：
(1)将五水硫酸铜加入到水中，于50℃～60℃下搅拌分散均匀，得到硫酸铜溶液；
(2)将十二烷基二甲基苄基溴化铵加入到步骤(1)中得到的硫酸铜溶液中，搅拌混合均匀，得到透明液体A；
(3)将84消毒液加入到步骤(2)中得到的透明液体A中，搅拌混合均匀，得到用于防治水稻细菌性条斑病的组合物。
步骤(1)中所述的搅拌的条件为：100～200rpm搅拌40～50分钟；优选为：200rpm搅拌40～50分钟。
步骤(2)中所述的搅拌的条件为：100～200rpm搅拌15～30分钟；优选为：常温下、100～200rpm搅拌15～30分钟。
步骤(3)中所述的搅拌的条件为：100～200rpm搅拌15～30分钟；优选为：25℃～30℃下、100～200rpm搅拌15～30分钟。
所述的用于防治水稻细菌性条斑病的组合物在制备防治水稻细菌性条斑病的农药制剂中的应用。
所述的农药制剂包括水剂等。
所述的用于防治水稻细菌性条斑病的组合物在防治水稻细菌性条斑病中的应用。
所述的用于防治水稻细菌性条斑病的组合物在防治水稻细菌性条斑病中的应用，为将所述的用于防治水稻细菌性条斑病的组合物稀释500～2000倍，用于水稻浸种或喷施到水稻叶面上，用于防治水稻细菌性条斑病。
所述的用于防治水稻细菌性条斑病的组合物的稀释倍数优选为500～1500倍；更优选为1000～1500倍；最优选为1500倍。
所述的用于水稻浸种的时间优选为24小时。
所述的稀释后的用于防治水稻细菌性条斑病的组合物的用量优选为50kg/亩。
本发明相对于现有技术具有如下的优点及效果：
本发明提供了一种可作为防治水稻细菌性条斑病的组合物，该组合物是水剂由水、十二烷基二甲基苄基溴化铵、五水硫酸铜和84消毒液，按一定配比和方法制备而成；研制出对水稻细菌性条斑病有较好防治效果的药剂，解决水稻细菌性条斑病防治药剂紧缺难题，且该组合物的制备和应用方法操作简便，投入成本低，见效快，可应用于农业生产、科研等众多领域，在实际生产中很有应用价值，更加经济环保地降低病害造成的粮食损失。
具体实施方式
下面结合实施例对本发明作进一步详细的描述，但本发明的实施方式不限于此。除非特别说明，本发明采用的试剂和方法为本技术领域常规试剂和方法除非特别说明，本发明所用试剂均可通过市售获得。
本发明中涉及的十二烷基二甲基苄基溴化铵(95％，V/V)，购于山东蓝鹰化工科技有限公司；84消毒液(有效氯含量5％～7％)为医用消毒液，购于汕头市亚奇商贸有限公司；硫酸铜为采用五水五水硫酸铜结晶，购于广州市东晖贸易有限公司。
实施例1
称取十二烷基二甲基苄基溴化铵1000g，五水硫酸铜1500g，84消毒液 2000ml，清水5500ml。
制备方法为：
第一步，将水5500ml和五水硫酸铜1500g置于搅拌设备中分散均匀(50℃下，按200转/分转速，搅拌40分钟)，得到五水硫酸铜溶液；
第二步，在第一步制得的五水硫酸铜溶液中加入十二烷基二甲基苄基溴化铵1000g，常温下，按200转/分转速，搅拌30分钟，继续搅拌至溶液变为透明液体；
第三步，将第二步所得溶液加入反应釜中，加入84消毒液2000ml，25℃下，按200转/分转速，搅拌30分钟，即得组合物A。
实施例2
称取十二烷基二甲基苄基溴化铵1250g，五水硫酸铜1750g，84消毒液2250ml，清水4750ml。
制备方法为：
第一步，将水4750ml和五水硫酸铜1750g置于搅拌设备中分散均匀，55℃下，按200转/分转速，搅拌45分钟，得到五水硫酸铜溶液；
第二步，在第一步制得的五水硫酸铜溶液中加入十二烷基二甲基苄基溴化铵1250g，常温下，按100转/分转速，搅拌20分钟，继续搅拌至溶液变为透明液体；
第三步，将第二步所得溶液加入反应釜中，加入84消毒液2250ml，30℃下，按100转/分转速，搅拌15分钟，即得组合物B。
实施例3
称取十二烷基二甲基苄基溴化铵1500g，五水硫酸铜2000g，84消毒液2500ml，水4000ml。
制备方法为：
第一步，将水4000ml和五水硫酸铜2000g置于搅拌设备中分散均匀，60℃下，按200转/分转速，搅拌50分钟，得到五水硫酸铜溶液；
第二步，在第一步制得的五水硫酸铜溶液中加入十二烷基二甲基苄基溴化铵1250g，30℃下，按150转/分转速，搅拌15分钟，继续搅拌至溶液变为透明液体；
第三步，将第二步所得溶液加入反应釜中，加入84消毒液2250ml，28℃下， 按100转/分转速，搅拌20分钟，即得组合物C。
本发明所述的组合物水剂稀释后可用于浸种(水稻种子)或喷施水稻叶面，用于防治水稻细菌性条斑病。
效果实施例
1、室内抑菌试验：
分别将组合物A、B、C和84消毒液、十二烷基二甲基苄基溴化铵、五水硫酸铜，分别以1：500、1：1000、1：1500、1：2000(v/v)加水进行稀释，以无菌水做对照，对在PSA斜面培养基(200g马铃薯、20g蔗糖、20g琼脂和1000mL蒸馏水，pH5.6～6.6)上培养2天的水稻细菌性条斑病菌(Xanthomonas oryzaes pv.oryzicola；广东省农业科学院植物保护研究所)浸渍处理24小时后,制成细菌悬浮液,离心(4500转/分)20分钟,取沉淀,用无菌水洗涤,再离心(4500转/分)20分钟,取沉淀在PSA培养基平面上划线培养,3次重复，3天后观察记录水稻细菌性条斑病菌在PSA培养基上的生长情况。细菌生长抑制率的公式如下：
细菌生长抑制率％＝[(对照抑菌圈直径-处理抑菌圈直径)/对照抑菌圈直径]×100。
如药剂对水稻细菌性条斑病菌生长有抑制作用,则处理后的病菌失去生命力,在PSA培养基上难以生长，因此药剂的杀菌力越强，则其有抑制作用的稀释浓度也越大。
试验结果表明，组合物A、B、C对水稻细菌性条斑病菌的杀菌力均较强，其中组合物B和组合物C稀释1500倍时对水稻细菌性条斑病的生长抑制率达到100％，是三个组合物中最好的(见表1)。84消毒液、十二烷基二甲基苄基溴化铵、硫酸铜(原料为五水硫酸铜结晶)对病原菌有一定抑制作用，但是效果较差，不如组合物A、B、C，说明他们组合在一起后有一定的增效作用，这可能是三个化合物一起产生铜络合物有关。
表1 组合物A、B、C对水稻细菌性条斑病菌的生长抑制效果
Figure PCTCN2021079632-appb-000001
Figure PCTCN2021079632-appb-000002
2、药剂对水稻种子和秧苗生长的安全性试验：
用组合物A、B、C分别稀释1500和2000倍液，用于水稻种子浸种24小时，用无菌水作为对照，将浸泡过的种子分别播在培养皿中，每皿2g，每个处理3次重复，培养6天后(温度28℃、光照12小时/天)，测定发芽率、芽长、根长(邵志慧等,生态学杂志2005,24(12):1440～1443)。试验结果表明，组合物A、B对水稻种子发芽和秧苗生长没有抑制作用，说明三个化合物组合后对水稻比较安全，可以用于浸种、秧苗期和大田使用；84消毒液、十二烷基二甲基苄基溴化铵、硫酸铜对水稻种子发芽和秧苗生长有抑制作用，且84消毒液、十二烷基二甲基苄基溴化铵、硫酸铜容易产生农药的药害，不适合单独在水稻上 使用。
表2 组合物A、B、C用于水稻种子浸种的安全性试验结果
Figure PCTCN2021079632-appb-000003
3、秧苗期施药试验：
试验地点：在广东省农业科学院植物保护研究所温网室内进行。
试验品种：水源290(水稻感病品种，由广东省农业科学院植物保护研究所提供)。
在试验温网室里播种水稻品种水源290，在秧苗3叶期用人工繁殖的水稻细菌性条斑病菌(浓度为10 9CFU/ml)进行喷雾接种,保湿12小时后,分别用组合物A、B、C加水以1：1500、1：2000(v/v)的稀释药剂和清水进行喷雾(稀释后的药剂的用量为50kg/亩)，10天后用同样药剂和浓度进行第二次喷雾，最后一次(即第二次)喷雾3天后调查病叶率，三次重复取平均值。
病叶率％＝发病叶片数/调查总叶片数×100。
防治效果(％)＝[(对照叶片病叶率-处理叶片病叶率)/对照叶片病叶率]×100。
水稻苗期喷雾组合物A、B、C对水稻细菌性条斑病的防治效果试验结果表明，组合物B的1500倍稀释液对水稻细菌性条斑病的防治效果最好，达到98.5％ (见表2)。
表3 水稻苗期喷雾组合物A、B、C对水稻细菌性条斑病的防治效果
Figure PCTCN2021079632-appb-000004
4、大田试验：
在水稻细菌性条斑病发生初期，用组合物A、B、C分别稀释1500倍和2000倍，进行喷雾(稀释后的药剂的用量为50kg/亩)，每隔7天喷雾一次，共喷雾三次，最后一次喷雾7天后调查发病率和病情指数，计算防治效果。
病情分级标准：0级，叶片无病斑；1级，病斑面积≤1％叶片面积；3级，1％叶片面积＜病斑面积≤5％叶片面积；5级，5％叶片面积＜病斑面积≤25％叶片面积；7级，25％叶片面积＜病斑面积≤50％叶片面积；9级，病斑面积＞50％叶片面积。
病情指数(表示病害危害的严重程度，根据计算公式计算出病害危害的严重程度)＝[Σ(病级数值×该病级病叶片数)×100]/(9×调查叶片数)。
防治效果(％)＝[(对照水稻病情指数-处理水稻病情指数)/对照水稻病情指数]×100。
试验结果表明组合物B的1500倍稀释药剂对水稻细菌性条斑病的防治效果最好，达到78.5％(见表4)。
表4 水稻大田期喷雾组合物A、B、C对水稻细菌性条斑病的防治效果
Figure PCTCN2021079632-appb-000005
Figure PCTCN2021079632-appb-000006
上述实施例为本发明较佳的实施方式，但本发明的实施方式并不受上述实施例的限制，其他的任何未背离本发明的精神实质与原理下所作的改变、修饰、替代、组合、简化，均应为等效的置换方式，都包含在本发明的保护范围之内。

Claims (10)

  1. 一种用于防治水稻细菌性条斑病的组合物，其特征在于，包括如下重量百分数计的组分：十二烷基二甲基苄基溴化铵10～15份，五水硫酸铜15～20份，84消毒液20～25份，水40～55份。
  2. 根据权利要求1所述的用于防治水稻细菌性条斑病的组合物，其特征在于，包括如下重量百分数计的组分：十二烷基二甲基苄基溴化铵10～13份，五水硫酸铜15～18份，84消毒液20～23份，水46～55份。
  3. 权利要求1～2任一项所述的用于防治水稻细菌性条斑病的组合物的制备方法，其特征在于，包括如下步骤：
    (1)将五水硫酸铜加入到水中，于50℃～60℃下搅拌分散均匀，得到硫酸铜溶液；
    (2)将十二烷基二甲基苄基溴化铵加入到步骤(1)中得到的硫酸铜溶液中，搅拌混合均匀，得到透明液体A；
    (3)将84消毒液加入到步骤(2)中得到的透明液体A中，搅拌混合均匀，得到用于防治水稻细菌性条斑病的组合物。
  4. 根据权利要求3所述的用于防治水稻细菌性条斑病的组合物的制备方法，其特征在于：
    步骤(1)中所述的搅拌的条件为：100～200rpm搅拌40～50分钟；
    步骤(2)中所述的搅拌的条件为：100～200rpm搅拌15～30分钟；
    步骤(3)中所述的搅拌的条件为：100～200rpm搅拌15～30分钟。
  5. 根据权利要求4所述的用于防治水稻细菌性条斑病的组合物的制备方法，其特征在于：
    步骤(1)中所述的搅拌的条件为：200rpm搅拌40～50分钟；
    步骤(2)中所述的搅拌的条件为：常温下、100～200rpm搅拌15～30分钟；
    步骤(3)中所述的搅拌的条件为：25℃～30℃下、100～200rpm搅拌15～30分钟。
  6. 权利要求1～2任一项所述的用于防治水稻细菌性条斑病的组合物在制备防治水稻细菌性条斑病的农药制剂中的应用。
  7. 权利要求1～2任一项所述的用于防治水稻细菌性条斑病的组合物在防治水稻细菌性条斑病中的应用。
  8. 根据权利要求7所述的应用，其特征在于：为将权利要求1～2任一项所述的用于防治水稻细菌性条斑病的组合物稀释500～2000倍，用于水稻浸种或喷施到水稻叶面上，用于防治水稻细菌性条斑病。
  9. 根据权利要求8所述的应用，其特征在于：
    所述的稀释后的用于防治水稻细菌性条斑病的组合物的用量为50kg/亩；
    所述的用于防治水稻细菌性条斑病的组合物的稀释倍数为500～1500倍。
  10. 根据权利要求8所述的应用，其特征在于：
    所述的用于防治水稻细菌性条斑病的组合物的稀释倍数为1000～1500倍。
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