First Report of Soybean Neocosmospora Stem Rot Caused by Neocosmospora vasinfecta var. vasinfecta in China.

Abstract

During October 2009, the occurrence of a disease on soybean (Glycine max) was observed in several fields in Boluo County and Zengcheng City, Guangdong Province. Top leaves of infected plants initially turned yellow and plants eventually were defoliated, while stems and roots became black and rotted. The stem lesions sometimes extended 10 to 15 cm upward from the soil surface. Orange-to-brown spherical fruiting bodies, which were very similar with those of the soybean red crown rot pathogen, scattered or congregated on the stem lesions. Plants with symptoms were sampled from fields. Fruiting bodies were excised from diseased tissues. Microscopic examination revealed that they were perithecia, globose to pyriform, ostiolate with a short neck, and measured 160 to 298 × 151 to 235 μm. Under gentle pressure, asci and ascospores were exuded from these perithecia. Asci were eight spored, narrowly cylindrical to clavate, thin walled, with a short stalk, and measured 58 to 124 μm long and 8 to 15 μm in diameter. Ascospores were uniseriately arranged, globose to ellipsoid, thick walled, one celled, hyaline to pale, and measured 14 to 17 × 8 to 12 μm. Isolation was made from stem tissues at the edge of disease lesions on potato dextrose agar (PDA) amended with streptomycin sulfate and incubated at 25°C. Mycelia were white and floccose. Conidia were cylindrical to oblong-ellipsoidal, hyaline, one celled, and measured 6 to 22 × 2 to 5 μm, aggregating in a slimy mass on the apex of the conidiogenous cell. Abundant orange-to-brown spherical perithecia were produced on the colony. Ascospores had walls with a rugose ornamentation that could be clearly seen under a scanning electron microscope. The fungus was identified as Neocosmospora vasinfecta var. vasinfecta (anamorph Acremonium sp.) (1). The internal transcribed spacer (ITS) region of rDNA of two isolates were amplified with universal primers ITS1/ITS4 and sequenced (GenBank Accession No. JF705861 and JF705862), and comparisons with GenBank accessions showed 99% similarity with N. vasinfecta strain Pec070 (Accession No. FJ940902) and strain NRRL22497 (Accession No. AY381142). Pathogenicity tests were conducted. Five, 3-week-old seedlings of soybean cv. Huaxia No. 3 planted in plastic pots (20 cm in diameter) were wounded with a needle at the base of the stem below the soil line and near the root system, and then inoculated by drenching the soil with a conidial suspension (105 per ml). Control plants were inoculated with sterile water. There were six replicates for each treatment. The treated plants were incubated at 25 ± 2°C in a greenhouse. All inoculated plants exhibited symptoms of leaf yellowing and black rot of stems and roots 3 weeks after inoculation. N. vasinfecta var. vasinfecta was reisolated from the diseased plants. All control plants remained healthy. To our knowledge, this is the first observation of Neocosmospora stem rot of soybean in China. The pathogen could pose a threat to soybean, which is a major crop in China. This disease has been previously reported in the United States though the anamorph of the pathogen has either not been identified or has been identified as a Cylindrocarpon sp. (2,4). This fungus is also associated with human infections (3).