中国农业科学Issue(1)：53-60,8.DOI:10.3864/j.issn.0578-1752.2014.01.006
草莓根颈腐烂病的病原鉴定
Identification of Pathogen Causing Crown and Root Rot on Strawberry
摘要
Abstract
[Objective]The objective of this study is to identify the pathogen causing crown rot on strawberry in Wuhan, and to provide a basis for disease prevention and resistance breeding.[Method]Diseased samples from Wuhan, Hubei Province in central China were collected, pieces of infected tissue from the crowns were surface disinfested and plated on acidified potato dextrose agar (PDA). The growing edges of any fungal hyphae developing from the tissues were then transferred aseptically to another PDA plate. Isolates were characterized by colony morphology, conidial characteristics and mycelia growth rate under 18℃ in dark. One-way analyses of variance (ANOVA) were conducted to determine the significance of differences in growth rates of different isolates. According to the morphological characteristics, isolates Zhd-3, Zhd-4-1 and Zhd-5 were selected as representatives for next analysis. Pathogenicity test was carried out by inoculating conidia of the three isolates to crown of strawberry with toothpick, respectively. Conidia produced on petioles were observed by optical microscope and scanning electron microscopy. The length and width of 20 conidia of each isolate were measured. Partial actin (ACT), beta-tubulin (TUB2) and calmodulin (CAL) genes of Zhd-3, Zhd-4-1 and Zhd-5 were amplified and sequenced. The ACT, TUB2 and CAL sequences of Zhd-3, Zhd-4-1, Zhd-5, along with the 23 different representative isolates in C. gloeosporioides species complex were used for multigene loci analyses with Mega 4.1 and Neighbour-Joining method.[Result]Fifteen isolates were morphologically identified as Colletotrichum spp.. Colony morphology of these isolates on the above of PDA plate appeared similar but significantly different on the back. The 15 isolates were divided into three groups based on the colony morphology, and isolates Zhd-3, Zhd-4-1 and Zhd-5 belonged to each group, respectively, were selected for follow-up tests and analysis. The pathogenicity tests showed that the symptoms identical to those observed in field were produced on all inoculated plants. Control plants remained symptomless. The same pathogen was reisolated from the diseased tissues. Biological tests showed that mycelial growth rates of Zhd-3, Zhd-4-1 and Zhd-5 were 0.82, 0.68 and 0.88 cm∙d-1, respectively. The average size of 20 conidia was 11 μm × 3.8 μm. The biological characteristics indicated these isolates were C. gloeosporioides species complex. The result of multigene loci analysis based on ACT, TUB2 and CAL indicated that isolates Zhd-3, Zhd-4-1 and Zhd-5 were divided into one group with C. siamense strains ICMP12567, ICMP17795, ICMP18121, CBS113199 and CBS112983.[Conclusion]The results of biological characteristics and multigene loci analysis based on the sequences of TUB2, ACT and CAL are concluded that the crown rot disease of strawberry occurred in Wuhan is caused by C. siamense.关键词
草莓/根颈腐烂病/炭疽病/ColletotrichumsiamenseKey words
strawberry/crown rot/anthracnose/Colletotrichum siamense引用本文复制引用
韩永超,向发云,曾祥国,张鹏,顾玉成..草莓根颈腐烂病的病原鉴定[J].中国农业科学,2014,(1):53-60,8.基金项目
国家科技支撑计划项目（2013BAD02B04-02-04）、湖北省农业科技创新中心资助项目（2007-620-006-003）、湖北省农业科学院青年科学基金项目（2013NKYJJ04）、湖北省自然科学基金项目（2011CDB124）、国家质检总局科技项目 （）
