P. P. Li and
Z. Y. Cao, Mycotoxin and Molecular Plant Pathology Laboratory, Agricultural University of Hebei, Lekai South Road 2596, Baoding 071001, P. R. China;
J. G. Dong, Mycotoxin and Molecular Plant Pathology Laboratory, Agricultural University of Hebei, Lekai South Road 2596, Baoding 071001, P. R. China and The National Maize Industry Technology RD Center, MOA; and
L. H. Zhang,
S. H. Li,
Z. M. Hao,
S. Q. Gu, and
X. Y. Wang, Mycotoxin and Molecular Plant Pathology Laboratory, Agricultural University of Hebei, Lekai South Road 2596, Baoding 071001, P. R. China
Corn is the most important cereal crop in China, with over 34.94 million ha being cultivated in the country annually. However, fungal diseases are a major limiting factor in corn production. In August 2012, 20 ha of corn fields in Anhui Province were found to be heavily infected by fungi. The margin of the lesion was achlorotic, and the middle was yellowish white or off-white, which was similar to the corn Curvalaria leaf spot. The oval lesions were approximately 5 to 7 mm. Lesion tissue was removed from the border between symptomatic and healthy tissue. The surface was sterilized in 75% ethanol for 30 s and 0.1% HgCl2 for 1 min, after which the sample was washed three times in sterile distilled water. The isolate was purified and subcultured on potato dextrose agar (PDA) at 25 ± 2°C. The initial color of the colony was light brown, turning dark brown after being cultured for 7 days. The conidia were boat-shaped or inverted pear-shaped and were clearly bent to one side. The cells of both ends were slightly lighter and respectively ranged from 34.5 to 44.0 μm and 12.0 to 21.0 μm away from the base, with the second cell as the widest. The majority conidia had three or four false septates; isolates produced light brown to medium brown conidiophore, scattered or clustered, often branching, and exhibited bending. These morphological characteristics matched with the description of Bipolaris papendorfii reported by Zhang (3). A pathogenicity test was conducted with the two isolates on each of the 36 corns by spraying 2 ml spore suspension (106 conidia/ml). For the control treatment, 36 corns were inoculated with an equal volume of sterilized water. Inoculated plants were placed in a greenhouse from 29 to 33°C and 95% relative humidity. The typical 5 to 7 mm oval lesions were observed 7 days after inoculation, except on the control samples. Three replications of 36 corns were used for each treatment. The isolate was consistently 100% reisolated from the diseased tissue according to Koch's postulate. The isolate was found to be morphologically similar to B. papendorfii. Preliminary morphological identification of the fungus was confirmed by PCR assay using genomic DNA extracted from the mycelium of a 7-day-old culture on PDA at 25 ± 2°C. A 550-bp amplified region of the internal transcribed spacer (ITS) of rDNA was generated using ITS1 (5′-TCCGTAGGTGAACCTGCGG-3′) and ITS4 (5′-TCCTCCGCTTATTGATATGC-3′) universal primers (1). The ITS region (GenBank Accession No. KC592365) was then sequenced by Sangon Biotech (Shanghai, China), and displayed 99% nucleotide similarity with the rDNA-ITS of B. papendorfii (JQ753972.1) separately after BLASTn research in GenBank. Based on the symptoms, fungal morphology, ITS sequence, and pathogenicity testing, this fungus was identified as B. papendorfii. The pathogen could reportedly infect tobacco and cotton (2). To our knowledge, this is the first study to report that B. papendorfii can infect corn in China. This report will establish a foundation for the further study of B. papendorfii to address the disease effectively. Further studies will be conducted to determine the incidence of the disease and the severity of damage caused by B. papendorfii as well as determine a possible mode for controlling the spread of the disease.
References: (1) Y. J. Cao et al. Chin. J. Trop. Crops 31:1098, 2010. (2) H. Deng et al. Mycosystema 21:327, 2002. (3) T. Y. Zhang. Chin. Fungi Chi. 30:21, 2010.