紫花苜蓿炭疽病病原新记录种(Colletotrichum liriopes)的鉴定及生物学特性分析

doi:10.6048/j.issn.1001-4330.2024.03.019

新疆农业科学 ›› 2024, Vol. 61 ›› Issue (3): 690-698.DOI: 10.6048/j.issn.1001-4330.2024.03.019

紫花苜蓿炭疽病病原新记录种(Colletotrichum liriopes)的鉴定及生物学特性分析

柳丰¹(), 祖丽皮耶·安外尔¹, 李克梅¹(), 托伦巴特·毕亚洪²

1.新疆农业大学农学院/农业农村部西北荒漠绿洲农林外来入侵生物防控重点实验室(部省共建)/农林有害生物监测与安全防控重点实验室,乌鲁木齐 830052
2.塔城地区塔城市二工镇农业(畜牧业)发展中心,新疆塔城 834300

收稿日期:2023-08-03 出版日期:2024-03-20 发布日期:2024-04-19
通信作者: 李克梅(1972-),女,江苏人,教授,博士,硕士生导师,研究方向为植物病理学,(E-mail)likemei@xjau.edu.cn
作者简介:柳丰(1998-),女,新疆人,硕士研究生,研究方向为植物病理学,(E-mail)1968484375@qq.com
基金资助:
国家重点研发计划项目(2022YFD1401101)

Identification of new pathogens of alfalfa anthracnose (Colletotrichum liriopes) and preliminary study on biological characteristics

LIU Feng¹(), Zulipiye Anwaier¹, LI Kemei¹(), Tuolunbate Biyahong²

1. College of Agronomy,Xinjiang Agricultural University/Key Laboratory of Prevention and Control of Invasive Organisms in Agriculture and Forestry in Northwest Desert Oases,Ministry of Agriculture and Rural Affairs(co-constructed by Ministry and Province)/Key Laboratory of Monitoring and Safety Prevention and Control of Agricultural and Forestry Pests,Urumqi 830052,China
2. Agriculture (animal husbandry) development center,Ergong town,Tacheng,Tacheng Xinjiang 834300,China

Received:2023-08-03 Published:2024-03-20 Online:2024-04-19
Supported by:
Nationaal Key R&D Program of China(2022YFD1401101)

摘要/Abstract

摘要：

【目的】 从炭疽病症状的紫花苜蓿病株分离得到一种炭疽菌,与此前报道的三叶草炭疽菌(Colletotrichum trifolii)、北美炭疽菌(C.americae-borealis)形态上略有差异,研究该菌致病性、分类地位及其生物学特性并鉴定。【方法】 采用组织分离法获得炭疽菌菌株,经柯赫氏法则证明其致病性,并运用形态学观察结合多基因片段PCR扩增法鉴定病原菌种类,测定不同培养基、碳源、氮源、温度、pH值、光照对该菌菌落生长的影响。【结果】 从紫花苜蓿炭疽病病部分离获得一种新的致病炭疽菌,为麦冬炭疽菌(Colletotrichum liriopes)。该菌在5~35℃,pH值4~11的多数供试培养基、氮源、碳源条件下均能生长。【结论】 麦冬炭疽菌(C.liriopes)可侵染紫花苜蓿引起炭疽病,其菌落最适生长温度为28℃,最佳氮源、碳源分别是酵母和可溶性淀粉,最适pH为7,菌丝致死温度61℃,12 h光暗交替对菌落生长有利。

关键词: 苜蓿炭疽病; 麦冬炭疽菌; 鉴定; 生物学特性

Abstract:

【Objective】 This project aims to isolate an anthrax strain from alfalfa with the symptoms of anthracnose,which would be slightly different from the previously reported Colletotrichum trifolii and C.americae borealis in morphology in order to clarify the pathogenicity,taxonomic status and biological characteristics of this strain through a series of studies.【Methods】 Anthrax strain was obtained by tissue isolation,and its pathogenicity was proved by Koch's rule and morphological observation combined with PCR amplification of polygene fragments was used to identify the pathogenic bacteria.Meanwhile,the effects of different culture media,carbon source,nitrogen source,temperature,pH value and light on the colony growth of anthrax strain were determined.【Results】 A new pathogenic anthrax strain was isolated from alfalfa,and identified as Colletotrichum liriopes.The strain could grow under the conditions of 5-35℃ and pH 4-11,and most of the test medium,nitrogen source and carbon source conditions.【Conclusion】 C.liriopes can infect alfalfa and cause anthracnose.The optimum growth temperature of C.liriopes is 28℃,the optimum nitrogen source and carbon source are yeast and soluble starch respectively,the optimum pH is 7,the lethal temperature of mycelium is 61℃,and the alternation of light and dark for 12 h is favorable to the growth of the colony.

Key words: alfalfa anthracnose; C.liriopes; identification; biological characteristics

中图分类号:

S435.5

柳丰, 祖丽皮耶·安外尔, 李克梅, 托伦巴特·毕亚洪. 紫花苜蓿炭疽病病原新记录种(Colletotrichum liriopes)的鉴定及生物学特性分析[J]. 新疆农业科学, 2024, 61(3): 690-698.

LIU Feng, Zulipiye Anwaier, LI Kemei, Tuolunbate Biyahong. Identification of new pathogens of alfalfa anthracnose (Colletotrichum liriopes) and preliminary study on biological characteristics[J]. Xinjiang Agricultural Sciences, 2024, 61(3): 690-698.

图/表 11

图1 病害田间症状及致病性测定注:a:苜蓿炭疽病田间症状;b、c:致病性测定

Fig.1 Determination of disease symptoms and pathogenicity Note:a:alfalfa anthracnose symptoms in field; b,c:pathogenicity determination results

图2 病原菌形态特征注:a:培养7 d菌落正反面;b:菌落正面的分生孢子堆;c:培养14 d菌落正面;d:分生孢子形态;e:分生孢子堆;f:刚毛;标尺:d=f=10 μm;e=0.02 mm

Fig.2 Pathogen morphology Note:a:Cultured for 7 days on both sides of the colony; b:The conidia pile on the front of the colony;c:Cultured for 14 days on the positive side of the colony; d:Conidia morphology; e:Conidia pile; f:The acervuli;Bar:d=f=10 μm; e=0.02 mm

图3 病原菌的PCR产物扩增电泳图注(Note):M:2000Marker;1,2:ITS基因:3,4:ACT基因;5,6:CHS-1基因; 7,8:HIS3基因:9,10:TUB2基因;11:双蒸水(CK)。M:2000 Marker; 1,2:ITS gene:3,4:ACT gene; 5,6:CHS-1 gene; 7,8: HIS3 gene:9,10:TUB2 gene; 11:Double distilled water(CK)

Fig.3 An amplified electrophoresis plot of the PCR products of the pathogens

图4 基于ITS、ACT、CHS-1、TUB2、HIS3基因构建的系统发育进化树注:OQ678010、OQ678011为C1-1、C1-2菌株的ITS基因序列;OQ659467、OQ659468为C1-1、C1-2菌株的ACT基因序列; OQ659469、OQ659470 C1-1、C1-2菌株的CHS-1基因序列;OQ659471、OQ659472为C1-1、C1-2菌株的TUB2基因序列;OQ659473、OQ659474为C1-1、C1-2菌株的HIS3基因序列

Fig.4 Phylogenetic tree constructed based on ITS, ACT, CHS-1, TUB2, and HIS3 genes Note:The OQ678010 and OQ678011 in the figure are the ITS gene sequences of C1-1 and C1-2 strains; OQ659467 and OQ659468 are the ACT gene sequences of C1-1 and C1-2 strains; OQ659469,OQ659470 is C1-1, C1-2 strain CHS-1 gene sequence; OQ659471,OQ659472 is C1-1, C1-2 strain TUB2 gene sequence; OQ659473,OQ659474 is C1-1, C1-2 strain HIS3 gene sequence

图5 10种培养基下麦冬炭疽菌菌丝生长变化

Fig.5 The effects of 10 kinds of medium on the growth of C.liriopes

图6 7种氮源下麦冬炭疽菌菌丝生长变化

Fig.6 The effects of 7 kinds of nitrogen sources on the growth of C.liriopes

图7 10种碳源下麦冬炭疽菌菌丝生长变化

Fig.7 The effects of 10 carbon sources on the growth of C.liriopes

图8 不同温度下麦冬炭疽菌菌丝生长变化

Fig.8 Effect of temperature on the growth of C.liriopes

图9 不同pH下麦冬炭疽菌菌丝生长变化

Fig.9 The effects pH on the growth of C.liriopes

图10 不同光照下麦冬炭疽菌菌丝生长变化

Fig.10 Effect of light on the growth of C.liriopes

图11 麦冬炭疽菌致死温度变化

Fig.11 Determination of lethal temperature of C.liriopes

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紫花苜蓿炭疽病病原新记录种(Colletotrichum liriopes)的鉴定及生物学特性分析

Identification of new pathogens of alfalfa anthracnose (Colletotrichum liriopes) and preliminary study on biological characteristics

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