Aragon-Caballero L M, Hurtado-Gonzales O P, Flores-Torres J G, Apaza-Tapia W, Lamour K H
National Agricultural University, La Molina, Lima, Peru.
University of Tennessee, Knoxville 37996.
Plant Dis. 2008 Jun;92(6):982. doi: 10.1094/PDIS-92-6-0982B.
During 2006, spears, roots, and crowns of asparagus (Asparagus officinalis) exhibiting brown necrotic lesions with water soaking were collected from several sites across Peru (Ica, Lima, and Trujillo). Small infected tissue sections were washed thoroughly with tap and sterile distilled water and transferred to corn meal agar plates (CMA) amended with PARP (100 ppm of pimaricin, 100 ppm of ampicillin, 30 ppm of rifampicin, and 100 ppm of pentachloronitrobenzene) and incubated for five days at 25°C. Hyphal tips were subcultured from actively expanding mycelium. Sporangia produced on CMA were papillate and averaged 38 μm long × 29 μm wide. Chlamydospores were terminal or intercalary and averaged 35 μm in diameter. Isolates incubated in the dark for more than 3 weeks did not produce oospores in single culture. Mating with Phytophthora capsici tester isolates CBS 121656 = A1 and CBS 121657 = A2 indicate that all five isolates were A2. For pathogenicity tests, inoculum was generated by incubating 300 g of autoclaved wheat seeds with four agar plugs (7 mm) of expanding mycelium in polyethylene bags for 1 month at 25°C. Nine-week-old asparagus plants (UC151 F1) were transferred into pots containing autoclaved substrate (1 part sand, 1 part potting soil, and 1 part peat). Inoculum was added as 1 g of inoculum per kilogram of substrate. Plants were maintained in the greenhouse at 23°C and watered daily. Decline symptoms as well as root and spear rot were observed after 7 days and a Phytophthora sp. was reisolated from infected tissue. No symptoms were observed on asparagus plants inoculated with sterile inoculum. DNA was isolated from two representative isolates, and the nuclear ribosomal internal transcribed spacer (ITS) region was amplified with ITS4 and ITS6 primers and sequenced. ITS sequence was submitted for a BLAST search in the NCBI database, showing Phytophthora nicotianae strain UQ848 Accession No AF266776 as the closest match with 99% sequence similarity (1). The consensus ITS sequence was deposited in NCBI (Accession No. EU433396). These results, together with the morphological characteristics, indicate that the Phytophthora sp. isolated from asparagus in Peru is P. nicotianae (Breda de Haan) (2). To our knowledge, this is the first report of P. nicotianae infecting asparagus and represents a new threat for asparagus growers in Peru. Control methods such as moderate watering and metalaxyl application are being applied to reduce Phytophthora outbreaks. References: (1) D. E. Cooke et al. Fungal Genet. Biol. 30:17, 2000. (2) D. C. Erwin and O. K. Ribeiro. Phytophthora Diseases Worldwide. The American Phytopathological Society. St Paul, MN, 1996.
2006年期间,从秘鲁各地(伊卡、利马和特鲁希略)的多个地点采集了芦笋(石刁柏)的茎、根和根颈,这些部位出现了带有水渍的褐色坏死病斑。将感染的小组织切片用自来水和无菌蒸馏水彻底冲洗,然后转移到添加了PARP(100 ppm匹马霉素、100 ppm氨苄青霉素、30 ppm利福平、100 ppm五氯硝基苯)的玉米粉琼脂平板(CMA)上,于25℃培养5天。从活跃生长的菌丝体上切取菌丝尖端进行继代培养。在CMA上产生的孢子囊有乳头状突起,平均长38μm×宽29μm。厚垣孢子为顶端着生或间生,平均直径35μm。在黑暗中培养超过3周的分离物在单培养中未产生卵孢子。与辣椒疫霉测试分离株CBS 121656 = A1和CBS 121657 = A2进行交配表明,所有五个分离物均为A2。进行致病性测试时,将300 g经高压灭菌的小麦种子与四个(7 mm)扩展菌丝体的琼脂块在聚乙烯袋中于25℃培养1个月以制备接种物。将9周龄的芦笋植株(UC151 F1)转移到装有经高压灭菌基质(1份沙子、1份盆栽土和1份泥炭)的花盆中。按每千克基质添加1 g接种物的量加入接种物。植株置于23℃的温室中,每天浇水。7天后观察到衰退症状以及根腐和茎腐,并且从感染组织中再次分离出一种疫霉属真菌。接种无菌接种物的芦笋植株未观察到症状。从两个代表性分离物中提取DNA,用ITS4和ITS6引物扩增核糖体DNA内部转录间隔区(ITS)并进行测序。将ITS序列提交到NCBI数据库进行BLAST搜索,结果显示烟草疫霉菌株UQ848登录号AF266776为最相似匹配,序列相似性为99%(1)。ITS一致序列保存在NCBI(登录号EU433396)。这些结果连同形态特征表明,从秘鲁芦笋中分离出的疫霉属真菌是烟草疫霉(布雷德·德·哈恩)(2)。据我们所知,这是烟草疫霉感染芦笋的首次报道,对秘鲁的芦笋种植者构成了新的威胁。目前正在采用适度浇水和施用甲霜灵等控制方法来减少疫霉病的爆发。参考文献:(1)D. E. Cooke等人,《真菌遗传学与生物学》30:17,2000年。(2)D. C. Erwin和O. K. Ribeiro,《世界疫霉病》,美国植物病理学会,明尼苏达州圣保罗,1996年。