摘要
近年来,由芸薹根肿菌(Plasmodiophora brassicae Woronin)引起的十字花科蔬菜根肿病发生严重,筛选生防菌并研究其对甘蓝根肿病的控制作用,为其生物防治应用提供科学依据。采用平板对峙生长法筛选拮抗细菌并进行拮抗细菌抑菌谱的测定,常规PCR方法扩增拮抗细菌16S rDNA序列,应用BLAST进行同源性搜索,送至中国科学院微生物研究所进行拮抗细菌的种属鉴定,用地衣红染色法测定拮抗细菌对根肿病菌休眠孢子萌发的抑制作用,盆栽试验测定生防菌对甘蓝根肿病的防治效果。结果表明:多黏芽胞杆菌N3-4和枯草芽胞杆菌PTS-394具有较好的抑菌谱,对多种病原真菌和病原细菌的生长有较强的抑制作用;对甘蓝根肿病菌休眠孢子萌发的抑制率分别为60.84%和53.78%。不同地区病土和不同甘蓝品种的盆钵条件下,多黏芽胞杆菌N3-4和枯草芽胞杆菌PTS-394防控根肿病效果不同,最高防效分别达71.09%和65.62%。表明,多黏芽胞杆菌N3-4和枯草芽胞杆菌PTS-394是2株对甘蓝根肿病有较好应用前景的生防菌。
Club root disease caused by Plasmodiophora brassicae Woronin is a common disease throughout the world.The aim of this study was to isolate the antagonistic bacteria and to evaluate the control efficiency of antagonistic bacteria against P.brassicae in vitro and in greenhouse.The antifungal and antibacterial spectrums of antagonistic bacteria were measured by co-culture with microbial phytopathogen on agar.Extraction of antagonistic bacterial genomic DNA was performed using the cetyl trimethylammonium bromide protocol（CTAB）.The 16 S rDNA of antagonistic bacteria was amplified from genomic DNA by a polymerase chain reaction（PCR）standard method using two universal primers： 27 f and 1541 r.A dendrogram was constructed based on the dissimilarity index by comparing with validly published 16 S rDNA sequences of related type strains from GenBank,using BLAST.The strains were identified by China General Microbiological Culture Collection Center（CGMCC）.The inhibition rate of antagonistic bacteria on the germination of resting spore of P.brassicae was measured by orcein staining method.The control effect on club root of cabbage with antagonistic bacteria in green house was studied in this paper.Antagonistic examination in vitro revealed that Paenibacillus polymyxa N3-4 and Bacillus subtilis PTS-394 showed strong and steady antagonism against the tested plant pathogen fungi and pathogen bacteria.P.polymyxa N3-4 and B.subtilis PTS-394 were identified and preserved by CGMCC,and they were given the sole number as CGMCC No.8084 and CGMCC No.8085,respectively.The inhibition rate of P.polymyxa N3-4 and B.subtilis PTS-394 on the germination of resting spore of P.brassicae was 60.84% and 53.78%,respectively.In the greenhouse experiment,the control effects of P.polymyxa N3-4 and B.subtilis PTS-394 against club root of cabbage were different according to the tested cabbage cultivars and the sick soil from different districts,with the maximum control effect 71.09% and 65.62%,respectively.The disease index of sick soil from Changyang district was higher than that from Lichuan district,increasing by 220.79% on tested resistant cultivar‘Sugan 26＇and 64. 45% on tested susceptible cultivar‘Qixia＇.The resistant cultivar‘Sugan 26＇showed the steady resistant ability both in sick soil from Lichuan district and from Changyang district. The disease index of susceptible cultivar‘Qixia＇was higher than resistant cultivar‘Sugan 26 ＇,increasing by 53.79% in sick soil from Changyang district and 200% from Lichuan district.The control effect of P.polymyxa N3-4 against club root of cabbage was higher than that of B. subtilis PTS-394,especially on susceptible cultivar‘Qixia＇in sick soil from Changyang district,the control effect of P.polymyxa N3-4 fermented liquid was increased by 59.93% with 10-fold dilution and 118.06% with 100-fold dilution compared with B.subtilis PTS-394.Therefore,P.polymyxa N3-4 and B.subtilis PTS-394 are prospective antagonistic bacteria against club root of cabbage.
出处
《南京农业大学学报》
CAS
CSCD
北大核心
2014年第4期83-90,共8页
Journal of Nanjing Agricultural University
基金
国家自然科学基金项目(31201555)
江苏省农业科技自主创新资金项目〔CX(12)3022〕
