摘要
目的:研发安全有效的抗人参根腐病微生物菌剂。方法:平板对峙试验筛选三七内生菌,形态学及分子生物鉴定拮抗菌株,单因素试验优选菌株发酵条件,响应面法优化微生物菌剂的载体及助剂配比制定生产工艺,通过对峙及盆栽试验验证菌剂验证防治效果。结果:平板对峙试验筛选出一株具有显著抗根腐病的菌株Fusarium pseudoanthophilum,其抑菌率为53.33%。单因素试验结果表明F.pseudoanthophilum的发酵条件为发酵时间60 h,培养温度26℃,最适摇瓶发酵转速120 r·min^(-1),pH 6.5。响应面优化试验结果表明,草炭60.00 g,羧甲基纤维素钠3.50 g,海藻酸钠4.76 g时,活菌数达到最大为5.23×10^(9) cfu·g^(-1),载体及助剂对活菌数的影响程度为草炭>羧甲基纤维素钠>海藻酸钠。菌剂对峙试验结果表明,菌剂质量浓度>1.00 g·L^(-1)时,对根腐病原菌尖孢镰刀菌具有显著的抑制作用且抑菌率>42.3%。盆栽试验结果表明,生防菌剂接种28 d后,显著降低了人参幼苗根腐病的发病率(66.99%)和病情指数(61.69%),且显著促进了人参幼苗的叶长(33.04%)和鲜重(34.48%)。结论:F.pseudoanthophilum菌剂具有较好的根腐病防治效果,值得进一步开发与利用。
Objective:To develop safe and effective microbial agents against Panax ginseng root rot.Method:P.notoginseng endophytes were screened in plate confrontation tests,followed by morphological and molecular biological identification of antagonistic strains,optimization of strain fermentation conditions in a single factor test,and determination of optimal carriers and auxiliary agents of the microbial agent and their ratio using response surface methodology for formulating the production process.The prevention and control effects of the microbial agent were verified in the confrontation and pot culture experiments.Result:The plate confrontation test yielded a strain named Fusarium pseudoanthophilum with significant resistance to root rot,and its antibacterial rate was 53.33%.According to the single factor test,the fermentation conditions of F.pseudoanthophilum were determined to be fermentation time 60 h,fermentation temperature 26℃,speed 120 r·min^(-1),and pH 6.5.The response surface optimization results showed that the number of viable bacteria reached the maximum(5.23×10^(9) cfu·g^(-1))when the peat was 60.00 g,sodium carboxymethylcellulose 3.50 g,and sodium alginate 4.76 g.The influences of carriers and auxiliary agents on the number of viable bacteria were sorted by degree in a descending order as follows:peat>sodium carboxymethylcellulose>sodium alginate.The confrontation test results showed that when the microbial agent concentration was greater than 1.00 g·L^(-1),it had a significant inhibitory effect on the root rot pathogen F.oxysporum and the inhibitory rate was more than 42.3%.As demonstrated by the pot culture experiment,the inoculation of biocontrol agent for 28 d significantly reduced the incidence(66.99%)of root rot in P.ginseng seedlings and disease index(61.69%)and increased their leaf length(33.04%)and fresh weight(34.48%).Conclusion:F.pseudoanthophilum inoculant is efficient in preventing and controlling the root rot,making it worthy of further development and utilization.
作者
黄贻杰
董林林
尉广飞
张小会
HUANG Yi-jie;DONG Lin-lin;WEI Guang-fei;ZHANG Xiao-hui(Dali University,Dali 671003,China;Institute of Chinese Materia Medica,China Academy of Chinese Medical Sciences,Beijing 100700,China;Institute of Medical Information,Chinese Academy of Medical Science&Peking Union Medical College,Beijing 100020,China)
出处
《中国实验方剂学杂志》
CAS
CSCD
北大核心
2022年第2期182-190,共9页
Chinese Journal of Experimental Traditional Medical Formulae
基金
北京市科技新星项目(Z181100006218020)
国家中医药管理局项目(GZYYGJ2020013)。
关键词
人参
三七
内生菌
根腐病
发酵
生防菌剂
Panax ginseng
P.notoginseng
endophytes
root rot
fermentation
biocontrol agent
