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Bio-control bacteria strain 1JN2 for preventing and treating bacterial fruit blotches of watermelons and application thereof

A bio-control strain, 1JN2 technology, applied in the field of agricultural biology, can solve the problems of low control effect, pesticide residues, pathogen resistance, etc., and achieve the effect of being beneficial to export trade and saving expenses

Active Publication Date: 2013-05-29
NANJING AGRICULTURAL UNIVERSITY
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  • Summary
  • Abstract
  • Description
  • Claims
  • Application Information

AI Technical Summary

Problems solved by technology

Bacterial fruit spot of watermelon is a devastating bacterial disease. It is not easy to be found during the field growth period, and the onset occurs suddenly and develops rapidly. Since no better method for seed treatment has been found, the chemical agents currently used in production There are agricultural streptomycin, which can kill the disease, phytomycin, bactericide, Ye Shuangqing, Garenon, copper succinate, etc. Although the chemical agents have a certain control effect on the disease, the control effect is not high, and Extensive use of chemical fungicides often causes environmental pollution, pesticide residues, and pathogenic bacteria resistance

Method used

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  • Bio-control bacteria strain 1JN2 for preventing and treating bacterial fruit blotches of watermelons and application thereof
  • Bio-control bacteria strain 1JN2 for preventing and treating bacterial fruit blotches of watermelons and application thereof
  • Bio-control bacteria strain 1JN2 for preventing and treating bacterial fruit blotches of watermelons and application thereof

Examples

Experimental program
Comparison scheme
Effect test

Embodiment 1

[0017] 一种防治西瓜细菌性果斑病病害的生防菌株1JN2,为Bacillus axarquiensis,由山东省莱芜市生姜茎内分离得到。其具体分离、筛选、鉴定方法如下:

[0018] 1生防菌株1JN2的分离

[0019] 剪掉生姜茎部3g,用1%的次氯酸钠1mL浸5min,70%酒精浸2min,无菌水清洗3次。(取最后一次清洗的溶液0.1mL涂R2A,或取0.1mL最后一次清洗液加入R2A培养液,30℃振荡培养,若48h后无菌生长则认为表面消毒干净)。用灭菌的研砵研碎表面消毒好的茎,加3mL无菌水,无菌棉布过滤,用无菌水稀释10倍,取0.1mL涂LB平板,每个点重复3次,28℃培养48h,计数,挑取平板上所有的菌落分离纯化,-70℃、40%甘油保存备用。

[0020] 2生防菌株1JN2的筛选

[0021] 将西瓜细菌性果斑病病原菌燕麦嗜酸菌西瓜亚种(Acidovorax avenae)FC440(JianjunFeng,Erin L.Schuenzel,Jianqiang Li,and Norman W.Schaad.2009.Multilocus SequenceTyping Reveals Two Evolutionary Lineages of Acidovorax avenae subsp.Citrulli.Bacteriology.99(8):913-920)在LB培养液(氯化钠10g / L,胰蛋白胨10g / L,酵母浸粉5g / L,调节pH值为7.2)中培养16-24h至CFU为10 8 / ml),加入熔化冷却至40℃左右的固体培养基中,体积比1:200,将培养基每20mL倒入9cm直径的平板中。待冷却凝固后,牙签接种生防菌1JN2。置于28℃恒温培养箱培养,24h后记录抑菌圈环径,挑取抑菌圈最大的命名为1JN2,于2012年1月16日保藏于中国微生物菌种保藏管理委员会普通微生物中心,地址:北京市朝阳区北辰西路1号院中科院微生物研究所,菌种保藏号为CGMCC NO.5731。

[0022] 3生防菌株1JN2的鉴定

[0023] 3.1生防菌株1JN2的形态特征

[0024] 菌落扁平、灰白色,边缘波纹状,较粘稠,菌体直杆状,大小为2.0×0.5μm,单生、对生或成短链状、丝状,周生多根鞭毛,内生孢...

Embodiment 21

[0031] 实施例21JN2生防菌剂的制备

[0032] 将上述防治西瓜细菌性果斑病的生防菌株1JN2(CGMCC NO.5731)在LB培养液28℃180rpm振荡培养12-16h,然后6000rpm离心10min,用灭菌水进行稀释制成菌剂,菌剂成品中活菌总浓度为1×10 9 -1×10 10 CFU / mL,以下称该生防菌剂为1JN2生防菌剂。

Embodiment 31

[0033] 实施例31JN2温室促生试验

[0034] 将催芽后的西瓜种子播种于装有无菌土壤中盆砵中,每盆一粒,16 / 8h光周期。待西瓜长到6叶期时,将悬浮好的1JN2生防菌剂(实施例2制备,下同)以10 8The concentration of CFU / ml is sprayed on the surface of the leaves with a manual sprayer to make the leaves moist and the fungicide does not flow down. Place them in a greenhouse for cultivation at 28°C and a relative humidity above 70%. After 30 days of inoculation with biocontrol bacteria, take the plants from the treatment group and the control group, carefully remove the root soil, clean them, and let them dry naturally. , plant fresh weight. 60 DEG C, measure plant dry weight after drying 72h.

[0035] Biomass increase (%) = (dry weight of plants in treatment group - dry weight of control plants) / dry weight of control plants × 100%

[0036] Table 2. Growth-promoting effect of Bacillus axarquiensis 1JN2 on watermelon seedling stage

[0037]

[0038] The results of the growth-promoting test in the greenhouse showed that Bacillus 1JN2 can significantly promote...

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Abstract

The invention belongs to the technical field of crop disease prevention and treatment and discloses a bio-control bacteria strain 1JN2 for preventing and treating bacterial fruit blotches of watermelons and application thereof. The bio-control bacteria strain 1JN2 is named as Bacillus axarquiensis in classification, and is filed at General Microbiological Center of China General Microbiological Culture Collection Committee with CGMCC No.5731 on January 16, 2012. The bio-control bacteria strain 1JN2 is cultured in LB (luria-bertani) broth at the temperature of 28 DEG C by vibration of 180rpm for 12-16 hours, centrifuged by 6000rpm for 10 minutes, diluted by sterilization water to prepare bactericide, and total concentration of viable bacteria in the finished bactericide is 1X10<9>-1X10<10>CFU / ml. The experiment results in a greenhouse indicates that the bio-control bactericide with the 1JN2 has evident growth acceleration effect to watermelons, biomass live weight is increased to 55.92% and evident prevention effect to disease of fruit blotches of the watermelons, disease incidence can be reduced by 50.13%.

Description

technical field [0001] 本发明属于农业生物技术领域,涉及防治西瓜细菌性果斑病生防菌株1JN2及其应用。 Background technique [0002] 西瓜细菌性果斑病(Bacterial fruit blotch of watermelon,BFB)是西瓜上的一种危险性检疫病害,又称细菌性斑点病,该病最早于1969年由Crall和Schenck在美国佛罗里达州发现,当时只是对果斑病症状进行了描述,未做病原菌的鉴定;目前为止,在我国许多省市,像新疆,陕西,河北,山西,吉林,福建,甘肃等地都已有果斑病的报道。该病害主要危害果实,引起西瓜果实腐烂等,可造成毁灭性损失,一旦发病用药防治基本没有效果。而且带菌种子将成为下一年重要的初侵染源。该病的发生严重影响了西瓜的产量和品质,成为西瓜生产上亟待解决的难题之一。 [0003] 该病害是由燕麦嗜酸菌西瓜亚种(Acidovorax avenae subsp.citrulli,Aac)引起的,西瓜果斑目前在瓜类作物中无免疫品种。西瓜细菌性果斑病是一种毁灭性的细菌性病害,大田生长期不易被发现,且发病突然、发展迅速,由于目前还没有发现种子处理的更好方法,目前生产中所采用的化学药剂有农用链霉素,可杀得,新植霉素,细菌杀,叶双青,加瑞农,琥胶肥酸铜等,虽然化学药剂对该病有一定的防治效果,但是防治效果不高,而且大量使用化学杀菌剂往往会造成环境污染、农药残留,以及引起病原菌抗药性等问题。生物防治是一种安全有效的防治手段,符合农业持续发展的方向。 Contents of the invention [0004] 本发明的目的针对西瓜产业潜在危害较大的西瓜细菌性果斑病,提供开发一种防治西瓜细菌性果斑病的单一生防菌菌株及其应用。 [0005] 本发明的目的可通过如下技术方案实现: [0006] 一种防治西瓜细菌性果斑病病害的生防菌株1JN2,分类命名为Bacillus axarquiensis,2012年1月16日保藏于中国微生物菌种保藏管理委员会普通微生物中心,地址北京市朝阳区北辰西路1号院3号,中国科学院微生物研究所,菌种保藏号为CGMCC NO.5731。 [0007] 用所述的保藏号为CGMCC NO.5731的生防菌株1JN2制备的生防菌剂。 [0...

Claims

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Application Information

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Patent Type & Authority Patents(China)
IPC IPC(8): C12N1/20A01N63/02A01P1/00C12R1/07
Inventor 郭坚华蒋春号武芳
Owner NANJING AGRICULTURAL UNIVERSITY