Li Y, Chi L D, Mao L G, Yan D D, Wu Z F, Ma T T, Guo M X, Wang Q X, Ouyang C B, Cao A C
Department of Pesticides, Key Laboratory of Pesticide Chemistry and Application, State Key Laboratory for Biology of Plant Diseases and Insect Pests, Institute of Plant Protection, Chinese Academy of Agricultural Sciences, Beijing, 100193, China.
Plant Dis. 2014 Feb;98(2):282. doi: 10.1094/PDIS-07-13-0729-PDN.
Ginger (Zingiber officinale Roscoe) is an important commercial crop that is planted in 60,000 to 70,000 ha every year in Shandong Province, China. In 2010, rotted rhizomes of cultivar Laiwu Big Ginger were reported on 20 ha in Anqiu, Shandong Province, and yield losses of up to 70% were reported. The aboveground symptoms were the water-conducting portion of symptomatic rhizomes was discolored brown and had a black dry rot of the cortex tissues (3). Thirty symptomatic rhizomes were sampled from six fields in six farms. Komada's method (1) was used to isolate the pathogen. Ten pieces from each rhizome were washed with sterile distilled water and plated on Komada selective medium at 25°C. White fungal colonies turned orchid after 7 days of incubation. Two types of asexual spores were associated with the colonies: microconidia and macroconidia. The microconidia were the most abundantly produced spores and were oval, elliptical or kidney shaped, and produced on aerial mycelia. Macroconidia had three to five cells and gradually pointed or curved edges, varied in size from 3 to 5 × 19 to 36 μm. The rDNA of the internal transcribed spacer regions 1 and 2 and the 5.8S gene in five isolates were amplified using primers ITS1 and ITS4, and the nucleotide sequence was the same as isolate no. 3, which was deposited in GenBank (Accession No. KC594035). A BLAST search showed 99% identity with the strain Z9 of Fusarium oxysporum (EF611088). Pathogenicity tests of five isolates were carried out in a greenhouse and the pathogenicity test of isolate no. 3 was selected for the method description. Ten 1-month-old ginger plants (cv. Laiwu Big Ginger) were grown in plastic pots (diameter 20 cm) with sandy soil and inoculated. Ten plants were used as untreated controls. Isolate no. 3 was grown on casein hydrolysate medium (4) for 72 h and the spores were harvested in sterile distilled water. Aqueous spore suspensions of isolate no. 3 were adjusted with deionized water to 1 × 10 CFU/ml as the inoculum. The prepared inoculum was injected with a syringe into the soil around the rhizome of ginger plants. Inoculated plants were placed in the greenhouse at 24 to 26°C and assessed for rhizome rot on the 14th day after inoculation. Disease severity was recorded based on a scale in which - = no symptoms; 1 = small lesions on seedlings, no rot; 2 = seedling rot; and 3 = plant dead. Similar rhizome rot symptoms were observed after inoculation. The inoculated isolate was re-isolated from diseased rhizomes, confirming its pathogenicity. To our knowledge, this is the first report of rhizome rot of ginger caused by F. oxysporum in China. Rhizome rot of ginger caused by Fusarium spp. is well known in Asian countries such as India (2). References: (1) H. Komada. Rev. Plant Prot. Res. 8:114, 1975. (2) V. Shanmugam et al. Biol Control. 66:1, 2013. (3) E. E. Trujillo. Diseases of Ginger (Zingiber officinale) in Hawaii, Circular 62, Hawaii Agricultural Experiment Station, University of Hawaii, December, 1964. (4) G. E. Wessman. Appl. Microbiol. 13:426, 1965.
生姜(姜科姜属植物)是一种重要的经济作物,在中国山东省,每年种植面积达6万至7万公顷。2010年,山东省安丘市20公顷的莱芜大姜种植区出现了根茎腐烂现象,据报道产量损失高达70%。地上部分症状为,发病根茎的输水组织变为褐色,皮层组织出现黑色干腐(3)。从六个农场的六个地块采集了30个发病根茎样本。采用小田法(1)分离病原菌。每个根茎取10片,用无菌蒸馏水冲洗后,接种于小田选择性培养基上,在25°C下培养。白色真菌菌落培养7天后变为淡紫色。菌落产生两种无性孢子:小分生孢子和大分生孢子。小分生孢子是产生量最多的孢子,呈椭圆形、椭圆状或肾形,着生于气生菌丝上。大分生孢子有3至5个细胞,边缘逐渐变尖或弯曲,大小为3至5×19至36μm。使用引物ITS1和ITS4对五个分离株的内部转录间隔区1和2以及5.8S基因的rDNA进行扩增,核苷酸序列与保藏于GenBank(登录号KC594035)的3号分离株相同。BLAST搜索显示,其与尖孢镰刀菌菌株Z9(EF611088)的同源性为99%。在温室中对五个分离株进行了致病性测试,并选择3号分离株的致病性测试进行方法描述。将10株1月龄生姜植株(品种为莱芜大姜)种植于直径20 cm的塑料盆中,盆内装有沙土,进行接种处理。10株植株作为未处理对照。3号分离株在酪蛋白水解物培养基(4)上培养72小时,然后将孢子收获于无菌蒸馏水中。用去离子水将3号分离株的孢子悬浮液调整至1×10 CFU/ml作为接种物。用注射器将制备好的接种物注射到生姜植株根茎周围的土壤中。接种后的植株置于24至26°C的温室中,接种后第14天评估根茎腐烂情况。根据以下分级标准记录病害严重程度:- = 无症状;1 = 幼苗上有小病变,无腐烂;2 = 幼苗腐烂;3 = 植株死亡。接种后观察到类似的根茎腐烂症状。从发病根茎中再次分离出接种的分离株,证实了其致病性。据我们所知,这是中国首次关于尖孢镰刀菌引起生姜根茎腐烂的报道。镰刀菌属引起的生姜根茎腐烂在印度等亚洲国家较为常见(2)。参考文献:(1)H. Komada. Rev. Plant Prot. Res. 8:114, 1975.(2)V. Shanmugam等. Biol Control. 66:1, 2013.(3)E. E. Trujillo. Diseases of Ginger (Zingiber officinale) in Hawaii, Circular 62, Hawaii Agricultural Experiment Station, University of Hawaii, December, 1964.(4)G. E. Wessman. Appl. Microbiol. 13:426, 1965.