海南白沙和儋州油梨叶部及果实炭疽病菌的鉴定  

Identification of pathogen species of avocado(Persea americana Mill.)leaf and fruit anthracnose in Baisha and Danzhou,Hainan

作　　者：李晶[1] 仇芳 张超[1] 王玉梓 谢昌平[1] LI Jing;QIU Fang;ZHANG Chao;WANG Yuzi;XIE Changping(School of Plant Protection,Hainan University/Key Laboratory of Green Prevention and Control of Tropical Plant Diseases and Pests,Ministry of Education,Haikou 570228,Hainan,China)

机构地区：[1]海南大学植物保护学院·热带农林生物灾害绿色防控教育部重点实验室,海口570228

出　　处：《果树学报》2023年第7期1443-1454,共12页Journal of Fruit Science

基　　金：2021年海南省研究生创新科研课题(Qhys2021-57)。

摘　　要：【目的】明确海南省白沙县和儋州市油梨种植基地的油梨叶片和果实的病原菌种类,为该病害的准确鉴定和田间防治措施的制定提供理论依据。【方法】在海南省白沙县和儋州市的油梨种植基地调查炭疽病的发生情况并采集病叶和病果,通过组织分离法和单孢分离法获得纯化分离株,并对纯化菌株进行致病性测定,结合形态学和分子生物学对所得菌株进行病原菌种类鉴定。【结果】从采自白沙县的14份病叶种分离的得到3株菌株,儋州市的10份病叶中分离获得6株分离物以及自白沙市采集的5份病果中分离出的1株真菌,经致病性测定,菌株HNBSL01、HNDZL02和HNBSF03为致病菌,且均与田间症状一致。根据3种致病菌的菌落、分生孢子和附着胞的形态特征可初步判断引起油梨叶片和果实炭疽病的病原菌均为炭疽菌属(Colletotrichum sp.);多基因(ITS-ACT-TUB2-CHS-1-GAPHD-HIS3)联合分析构建系统发育树分析结果显示,病原菌HNBSL01与暹罗炭疽菌(C.siamense)的同源性为81%、HNDZL02与果生炭疽菌(C.fructicola)的同源性为100%、HNBSF03与长直孢炭疽菌(C.gigasporum)的相似性达100%。【结论】引起海南白沙县和儋州市油梨种植基地叶片和果实炭疽病的病原菌为暹罗炭疽菌(C.siamense)、果生炭疽菌(C.fructicola)和长直孢炭疽菌(C.gigasporum),其中,C.gigasporum为国内首次报道引起油梨果实炭疽病的病原菌。【Objective】Avocado(Persea americana Mill.)is a tropical and subtropical evergreen fruit crop.As the demand for avocado increases year by year,the planting area has expanded.The incidence of anthracnose is increasing in the planting area.In 2021,the avocado planting base in Baisha and Danzhou of Hainan province found that the symptoms of anthracnose appeared on the leaves of large trees,and more than 50%trees were affected,which seriously affected the yield and quality of avocado.Danzhou Avocado Planting Base,found the dark brown disease spot on the fruit during the fruit harvest and storage,and a sticky orange-red conidiomata appeared,so that the heavy loss was caused.This experiment was conducted to describe and identify the pathogen causing leaf and fruit anthracnose of avocado,so as to provide a theoretical basis for the accurate identification of the disease and the development of control measures in the field.【Methods】The disease incidence of Baisha and Danzhou Avocado Planting Bases were investigated,and samples of anthracnose disease were collected.Tissue isolation method and single spore isolation method were used to isolate and purify the strains.To confirm the pathogenicity,the wounded and unwounded leaves and fruits of the avocad were inoculated by stem cake and conidial suspension inoculation method.The pathogen was re-isolated from the inoculated sites,and the morphological characteristics of the reisolated strains were observed and recorded.The purified strains were transferred onto PDA medium and incubated at 25℃and 12 h light/12 h dark.After 7 d,the morphology,color and growth rate of the colonies were recorded.After sporulation,the morphology and size of conidia and appressoria were observed and recorded under the optical microscope to clarify their morphological characteristics.Genomic DNA was extracted using the Fungal Genomic DNA Rapid Extraction Kit(OMEGA BIO-TEK).The six target gene sequences,ITS(ITS1/ITS4),TUB2(T1/βt2b),ACT(ACT-512F/ACT-783R),HIS3(CYLH3F/CYLH3R),CHS-1(CHS-79F/CHS

关 键 词：油梨 炭疽菌 致病性 多基因联合分析 

分 类 号：S667.9[农业科学—果树学]