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陕西周至杏树流胶病病原菌鉴定及其对3种药剂的敏感性
许 静, 李 晶, 张雨溪
西安文理学院 生物与环境工程学院
摘要:
【目的】分离纯化陕西周至地区杏树流胶病病原菌,筛选对其菌丝生长有显著抑制作用的杀菌剂,为其田间防控提供参考。【方法】采集陕西周至地区杏树流胶病发病枝条,表面消毒后用组织块分离法分离、纯化、培养病原菌,并进行回接试验,对能引起相同病症的病原菌菌株进行菌落形态和孢子形态观察,并利用ITS通用引物进行PCR扩增,测序后采用MEGA软件通过邻接法(Neighbor joining)建立系统发育树,鉴定致病菌株的分类地位。采用生长速率法测定该病原菌对70%甲基硫菌灵可湿性粉剂、2.6%靓果安水剂和2%溃腐灵水剂3种药剂的敏感性。【结果】陕西周至地区杏树流胶病病原菌分离物(编号为SXZZ)培养初期为白色,随后逐渐变为黑色;菌丝无色有隔;分生孢子无色单孢纺锤形。对该病原菌的ITS序列进行Blast比对后发现,其与葡萄座腔菌的同源性高达99%;结合形态学鉴定结果,确定陕西周至杏树流胶病病原菌为葡萄座腔菌(Botryosphaeria dothidea)。药剂敏感性测定结果表明,该病原菌对甲基硫菌灵非常敏感,1 μg/mL甲基硫菌灵即可明显抑制其菌丝生长, EC50为2.831 5 μg/mL;但其对靓果安和溃腐灵不敏感,二者在最低质量浓度分别为86.7和20.83 μg/mL 时,才能抑制其菌丝生长,且二者的EC50非常高。【结论】陕西周至地区杏树流胶病病原菌为葡萄座腔菌,该菌株对甲基硫菌灵极其敏感,但对靓果安和溃腐灵不敏感。
关键词:  杏树  流胶病  葡萄座腔菌  病原鉴定  陕西周至
DOI:
分类号:
基金项目:陕西省教育厅专项科研计划项目(20JK0878);西安市科技局科技计划创新基金文理专项(2020KJWL20);陕西省自然科学基金青年项目(2017JQ3031)
Identification of apricot gummosis pathogen in Zhouzhi,Shaanxi and its sensitivity to three fungicides
XU Jing, LI Jing, ZHANG Yuxi
Abstract:
【Objective】This study aimed to identify causal agent of apricot gummosis in orchards in Zhouzhi,Shaanxi and thereby apply effective chemical control methods.【Method】Gummosis twigs were sampled and surface sterilized,followed by pathogen isolation using the tissue block isolation method.The pathogen was thereafter confirmed by its inoculation on healthy twigs and re-isolation from the inoculated twigs.To classify the isolated pathogen,its colony and conidia morphology was observed,combined with phylogenetic analysis by MEGA software using Neighbor-joining method after its ITS PCR amplification and sequencing.Moreover,three fungicides including 70% Thiophanate Methyl wetable powder,2.6% Liangguoan aqueous solution and 2% Kuifuling aqueous solution were respectively added into plates to culture the pathogen,followed by colony diameter evaluation and calculation of EC50.【Result】The colony of pathogen causing apricot gummosis in orchards in Zhouzhi,Shaanxi was white at the beginning of incubation and gradually turned into black afterwards.Its mycelium was colorless and septal,while its condium was fusiform and colorless.After amplification using PCR,its ITS sequence showed more than 99% homology towards Botryosphaeria dothidea by Blast.Thus,it was confirmed that the apricot pathogen was Botryosphaeria dothidea.Thiophanate Methyl significantly depressed Botryosphaeria dothidea growth at the minimum concentration of 1 μg/mL,the EC50 of Thiophanate-Methyl against our isolate was 2.831 5 μg/mL.While Liangguoan and Kuifuling showed a weak inhibition at least at the concentration of 86.7 and 20.83 μg/mL,respectively,the EC50 values of the two chemicals were very high.【Conclusion】Botryosphaeria dothidea induced apricot gummosis in Zhouzhi,Shaanxi and it was sensitive to Thiophanate-Methyl but not to Liangguoan and Kuifuling.
Key words:  apricot tree  gummosis  Botryosphaeria dothidea  pathogen identification  Zhouzhi,Shaanxi