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2025年8月8日 星期五
  2015, Vol. 23 Issue (1): 107-117    
  研究论文与报告 本期目录 | 过刊浏览 | 高级检索 |
五种抑菌物质对采后病原菌根霉菌生长发育的影响
汪莹1,张弦1,周佳一2,李秀超1,闵杰1,赖童飞1,周婷1
1. 杭州师范大学
2. 杭州师范大学 生命与环境科学学院
Inhibitory Effects of Five Antifungal Substances on Development of Postharvest Pathogen Rhizopus oryzae
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摘要 根霉菌(Rhizopus oryzae)是热带果蔬采后主要病原真菌之一,不仅引起果实腐烂,影响果实风味,还能分泌对人体有毒害作用的麦角类生物碱,对果蔬贮藏具有极大的危害。本研究从腐烂的草莓(Fragaria ananassa L. cv. Zhangji)中分离主要致病菌,经核糖体DNA内转录间隔区(rDNA-ITS)序列分析辅以传统的形态学观察法,鉴定其遗传背景,然后利用碳酸氢钠(SB)、硼酸(BA)、肉桂油(Co)、硝普钠(SNP)、亚磷酸盐(Phi)等5种已知抑菌物质处理病原菌。通过检测处理前后病原菌生理、生化指标的改变以及番茄(Solanum lycopersicum cv. Ailsa Cragi)果实接种实验,确定5种抑菌物质对病原菌发育及致病力的影响。结果表明,分离的病原菌通过回接试验证实为草莓的主要致病菌,致病力强,果实发病部位出现明显的褐化、软化以及水化,病征与匍枝根霉(Rhizopus stolonifer)引起的软腐病表型相似。同时,利用ITS通用引物扩增得到大小约600 bp的产物,经比对后与Rhizopus oryzae的rDNA-ITS序列完全一致,因此判断该病原菌为R. oryzae。此外,本研究选用的5种物质对R. oryzae都有明显的抑制作用,且抑菌效果均与浓度成正比。当SB、BA、SNP、Co以及Phi的浓度分别达到0.2%、0.2%、0.5%、0.02%及5 mmol/L时,培养6 h后对照病原菌的孢子萌发率超过90%,而抑菌物质处理的孢子萌发率均低于10%。因此,这些浓度被确定为5种物质的最低抑菌浓度。在最低抑菌浓度下,尽管抑制效力有所不同,但5种抑菌物质均能明显延缓芽管伸长及菌丝扩展速度,降低病原菌生物量积累,引起病原菌糖吸收障碍,并对R. oryzae诱发的番茄软腐病有明显的防治作用。其中BA、Co和Phi在体外抑菌实验中效果优于SB和SNP;SB、BA以及Phi对番茄R. oryzae软腐病的防治效果优于Co与SNP。本研究为果蔬采后病原菌R. oryzae的防治提供了新的思路,为开发SB、BA、Co、SNP以及Phi等5种抑菌物质的应用潜力提供了理论基础。
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汪莹
张弦
周佳一
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闵杰
赖童飞
周婷
关键词 根霉菌内转录间隔区(ITS)抑菌物质贮藏保鲜    
Abstract:Rhizopus oryzae is a major postharvest pathogen of tropical fruits and vegetables, and quite harmful to storage and preservation wihch result in huge economic losses. It can cause Rhizopus soft rot in fruit, destroy fruit quality and flavor, and also produce the ergot alkaloid agroclavine which is toxic to humans and animals. In the present study, we isolated the main pathogen from naturally infected strawberries (Fragaria ananassa L. cv. Zhangji), and identified it by morphological observation and internal transcribed spacer region of ribosomal DNA (rDNA-ITS) analysis. After that, five exogenous antifungal substances including sodium bicarbonate (SB), boric acid (BA), cinnamon oil (Co), sodium nitroprusside (SNP) and phosphite (Phi) were selected for known antifungal activities and commercially availability. And then, inhibitory effects of five substances on pathogen development and pathogenicity were evaluated via assessment of physiological properties, biochemical indexes and inoculation experiments on tomato (Solanum lycopersicum cv. Ailsa Cragi) fruits. Results indicated that the fungus isolated from strawberry was demonstrated as the main pathogen through an artificial infection test and it was highly pathogenic. The infected position of fruit appeared obvious browning, softening and hydration, and the symptoms were similar to those of soft rot caused by Rhizopus stolonifer. Meanwhile, after amplification using ITS universal primers, an approximately 600 bp fragment of ITS rRNA gene sequence was obtained and perfectly matched with that of the R. oryzae strains. In addition, antifungal activities of five substances were positive and dose-dependent. When the concentration of SB, BA, SNP, Co and Phi reached 0.2%, 0.2%, 0.5%, 0.02% and 5 mmol/L, respectively, spore germination rates of R. oryzae were all reduced to below 10% after 6 h of incubation, versus the germination rate was over 90% in the control at the same time. Therefore, these concentrations were considered to be the minimum inhibitory concentrations (MICs) of five substances. Under the MICs, despite the difference of antifungal efficacies, all of five exogenous antifungal could significantly delay germ tube elongation and mycelium extension of R. oryzae, lower fungal biomass accumulation, disturb carbohydrate absorption of pathogen, and effectively control soft rot of tomato caused by R. oryzae. Among five substances, BA, Co and Phi showed more inhibitory efficacies than those of SB and SNP in in vitro experiments; SB, BA and Phi worked better than Co and SNP in in vivo experiments. Our results explored the possibility of employing those antifungal substances as part of general program for management of postharvest diseases caused by fungal pathogens, and will provide some theoretical basis for integrated control of R.oryzae.
Key wordsRhizopus oryzae    Internal transcribed spacer(ITS)    Antifungal substance    Storage and preservation
收稿日期: 2014-06-16      出版日期: 2015-01-06
基金资助:国家级大学生创新创业训练计划 项目;浙江省自然科学基金项目;浙江省自然科学基金项目
通讯作者: 赖童飞     E-mail: laitongfei@163.com
引用本文:   
汪莹 张弦 周佳一 李秀超 闵杰 赖童飞 周婷. 五种抑菌物质对采后病原菌根霉菌生长发育的影响[J]. , 2015, 23(1): 107-117.
链接本文:  
http://journal05.magtech.org.cn/Jwk_ny/CN/     或     http://journal05.magtech.org.cn/Jwk_ny/CN/Y2015/V23/I1/107
 
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