干旱区土壤中西瓜专化型尖孢镰刀菌拮抗菌的筛选及分类
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摘要
针对宁夏中部干旱带压砂西瓜田长期连作种植所产生的土壤退化、土壤养分含量降低、土壤真菌性病害增加、西瓜减产、品质下降等问题,从患病的西瓜植株上分离出致病菌-尖孢镰刀菌西瓜专化型(Fusarium oxysporum f.sp.niveum),从宁夏环香山地区压砂瓜集中连片种植区采集健康土壤中分离筛选对西瓜枯萎病致病菌有抑菌效果的拮抗菌株,并对拮抗菌株的防病效果以及分类鉴定进行了研究。通过采用平板稀释法筛选出对目标靶具有拮抗作用的拮抗细菌。通过初筛选取8株对目标靶具有拮抗作用的细菌进一步进行复筛试验。经过数据分析,G-1的抑菌率最大,且与其他处理间存在显著性差异,筛选出对目标靶拮抗最强的拮抗菌为G-1。根据形态学观察、细菌生理生化鉴定和16 S r DNA序列的同源性分析,确定拮抗菌株G-1为萎缩芽孢杆菌(Bacillus atrophaeus)。
Bottleneck problems such as soil nutrients exhaustion,soil fungal diseases prevalent,soil structure degradation,low production and quality of water melon,already severely restricted Gravel-covered watermelon industry in middle arid area of Ningxia. This experiment is based on the production practice of Grovel-covered Watermellon in Ningxia. Separation of Fusarium oxysporum f. sp. niveum from the incidence of serious disease of watermelon,and the control effect of Fusarium wilt was screened from different types of healthy soil,the effect of antagonistic strain on disease control and classification and identification were studied. During dilution plate method screened with antagonism of antagonistic bacteria to target. Through the screening of 8 strains of bacteria which have antagonistic effect on the target,the screening test was carried out. After data analysis,G-1 inhibitory rate is maximum,and there was a significant difference compared with other treatments,it was screened on the target antagonistic strongest antagonistic to the G-1. According to its morphological,physiological and biochemical characteristics and 16 S r DNA sequence homology analysis can determine the antagonistic strain G-1 is Bacillus atrophaeus.
Bottleneck problems such as soil nutrients exhaustion,soil fungal diseases prevalent,soil structure degradation,low production and quality of water melon,already severely restricted Gravel-covered watermelon industry in middle arid area of Ningxia. This experiment is based on the production practice of Grovel-covered Watermellon in Ningxia. Separation of Fusarium oxysporum f. sp. niveum from the incidence of serious disease of watermelon,and the control effect of Fusarium wilt was screened from different types of healthy soil,the effect of antagonistic strain on disease control and classification and identification were studied. During dilution plate method screened with antagonism of antagonistic bacteria to target. Through the screening of 8 strains of bacteria which have antagonistic effect on the target,the screening test was carried out. After data analysis,G-1 inhibitory rate is maximum,and there was a significant difference compared with other treatments,it was screened on the target antagonistic strongest antagonistic to the G-1. According to its morphological,physiological and biochemical characteristics and 16 S r DNA sequence homology analysis can determine the antagonistic strain G-1 is Bacillus atrophaeus.
引文
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[3]柳春燕.郭敏,林学政,等.拟康氏木霉和枯草芽孢杆菌对黄瓜枯萎病的协同防治作用[J].中国生物防治,2005,21(3):206-208.NIU C Y.GUO M,LIN X Z,et al.The synergistic efficacy of trichoderma pseudokoningii and bacillus subtilis against cucumber wilt[J].Chinese Journal of Biological Control,2005,21(3):206-208.
[4]程亮,游春平,肖爱萍.拮抗细菌的研究进展[J].江西农业大学学报,2003,25(5):732-738.CHENG L,YOU C P,XIAO A P.Advance in the study on antagonistic bacteria[J].Acta Agriculturae Universitis Jiangxiensis,2003,25(5):732-738.
[5]王小慧,张国漪,李蕊,等.拮抗菌强化的生物有机肥对西瓜枯萎病的防治作用[J].植物营养与肥料学报,2013,9(1):223-231.WANG X H,ZHANG G Q,LI X,et al.Control of watermelon fusarium wilt by using antagonist-enhanced biological organic fertilizers[J].Plant Nutrition and Fertilizer Science,2013,9(1):223-231.
[6]岳思君,郑蕊,张谋东,等.硒砂瓜枯萎病生防细菌的筛选与初步鉴定[J].江西农业大学学报,2013,35(6):1168-1172.YUE S J,ZHENG X,ZHANG M D,et al.Screening and Identification of antagonistic bacteria against Fusarium oxysporum in rhizosphere of Xisha melon[J].Acta Agriculturae Universitis Jiangxiensis,2013,35(6):1168-1172.
[7]王晓晓.西瓜枯萎病拮抗菌的筛选.生物学特性及发酵条件和抗生物质的研究[D].南京:南京农业大学,2012.WANG X X.Research on screening of antagonistic bacteria against fusarium wilt of watermelon,biological characteristics and fermentation conditions and antibiotic[D].Nanjing:Nanjing Agricultural University,2012.
[8]岳菊.西瓜枯萎病生防细菌筛选及其防治作用的研究[D].南京:南京农业大学,2010.YUE J.Research on screening of bacterial wilt of watermelon fusarium wilt and its prevention and treatment[D].Nanjing:Nanjing Agricultural University,2010.
[9]李敏.AM真菌对西瓜抗枯萎病的效应及其机制[D].中国农业大学,2005.LI M.Effects of AM fungi on fusarium wilt of watermelon and its mechanism[D].Beijing:China Agricultural University,2005.
[10]张璐.黄瓜枯萎病病原拮抗菌的筛选鉴定及其生防效果研究[D].泰安:山东农业大学,2007.ZHANG L.Research on screening and identification of Cucumber Wilt Pathogen antagonistic and anti effect[D].Taian:Shandong Agricultural University,2007.
[11]BENDING G D,TYURNER M K,JONES J E.Interactions between crop residue and soil organic matter quality and the functional diversity of soil microbial communities[J].Soil Biol Biochem.2002,34:1073-1082.
[12]于天祥,张明方.西瓜枯萎病研究进展[J].中国西瓜甜瓜,2004(1):17-19.YU T X,ZHANG M F.Research progress of fusarium wilt of watermelon[J].China Watermelon and Muskmelon,2004,1:17-19.
[13]曹明慧,冉炜,杨兴明,等.烟草黑胫病拮抗菌的筛选及其生物效应[J].土壤学报,2011,48(1):151-157.CAO M H,RUI W,YANG X M,et al.Screening of antagonist against tobacco black shank and effect of the strain[J].Acta Pedologica Sinica,2011,48(1):151-157.
[14]刘冬梅,李理,杨晓泉,等.用牛津杯法测定益生菌的抑菌活力[J].食品研究与开发,2006,27(3):110-111.LIU D M,LI L,YANG X Q,et al.Determination of the antimicrobial activity of probiotic by oxford plate assay system[J].Food Research and Development,2006,27(3):110-111.
[15]刘冰,黎园,丁莎,等.柑橘病害生防放线菌的分离筛选及抑菌作用研究[J].江西农业大学学报,2011,33(5):889-892.LIU B,LI Y,DING S,et al.Isolation and inhibitory effect of actinomyce strain against orange diseases[J].Acta Agriculturae Universitis Jiangxiensis,2011,33(5):889-892.
[16]JENG R S,SVIRCEV A M,MYERS A L,et al.The use of 16S and 16S-23 S r DNA to easily detect and differentiate common gram-negative orchard epiphytes[J].Microbiol Methods,2001,44:69-77.