Castillo J D, Lawrence K S
Department of Entomology and Plant Pathology, Auburn University, Auburn, AL 36849.
Plant Dis. 2011 Apr;95(4):490. doi: 10.1094/PDIS-07-10-0524.
A new fungal parasite of the reniform nematode has been observed parasitizing nematode populations that have increased on cotton in a sandy loam field soil in the greenhouse. When enumerated, 46% of the stock reniform nematode population was colonized by this fungus. Egg, vermiform, and adult stages of the reniform nematode were observed with light microscopy and scanning electron micrography (SEM). The nematophagous fungus, Catenaria auxiliaries, was identified morphologically. There are no sequences on the GenBank to achieve a molecular identification. This nematophagous fungus has been previously reported on the beet cyst nematode in Europe (1,2); however, to our knowledge there are no reports of this fungus parasitizing the reniform nematode. In vermiform life stages of the nematode, rhizomycelium is observed in the initial phase of infection and is characterized by ovoid cells, 9.5 to 13.5 × 17.0 to 24.5 μm in diameter, separated by septa. Usually 10 to 15 ovoid cells lacking intercellular hyphal filaments are produced within each vermiform body. Rhizoids 3.5 to 4.0 μm wide develop from the rhizomycelium. Mature swollen cells produce precursor sporangia that may mature into resting spores or zoosporangia. Resting spores are yellow-to-cream, 20 to 40 μm in diameter with a reticulate appearance, and are common in the vermiform nematode life stages. Zoosporangia are ovoid, 9.5 to 13.5 × 17.0 to 24.5 μm, and will erupt from the cuticle of the vermiform nematode releasing zoospores via papillae. Zoospores are 2.9 to 4.9 μm with visible globules in the anterior region and single flagella that are 9 to 11 μm long. The zoospores swim short distances, maneuvering in the direction of the flagellum. Adult reniform females observed through SEM exhibit zoospores encysted in the metacorpus region of the nematode. Parasitized eggs are internally colonized with zoosporangia that are 20 to 25 μm in diameter. In advanced stages of infection, the eggs darken in color and zoosporangia erupt through the cuticle of the egg. Reniform nematodes visibly colonized with zoosporangia and resting spores were placed on corn meal, water, and potato dextrose agars. None of these media supported growth of the fungus, supporting our theory that this organism appears to be an obligate parasite of the nematode. Koch's postulates was completed when eggs colonized with rhizomycelium and resting spores or zoosporangia were added to cotton plants in sterile soil previously inoculated with 2,000 healthy vermiform reniform life stages. Plants were allowed to grow for 30 days in the greenhouse after which the next generation of vermiform nematodes were extracted from the soil and examined under the microscope. Rhizomycelium, resting spores, and zoosporangia were present in 42% of the reniform vermiform life stages. Morphological comparisons of the rhizomycelium, resting spores or zoosporangia, and zoospores colonizing the reniform nematodes were similar to the initial observations. Thus to our knowledge, this is the first report of Catenaria auxiliaries parasitizing the reniform nematode. References: (1) B. Kerry. J. Nematol. 12:253, 1980. (2) H. T. Tribe. Trans. Br. Mycol. Soc. 69:367, 1977.
在温室的砂壤土田间土壤中,人们观察到一种新的肾形线虫真菌寄生虫寄生于在棉花上数量增加的线虫群体。经计数,46%的原肾形线虫群体被这种真菌定殖。通过光学显微镜和扫描电子显微镜(SEM)观察了肾形线虫的卵、蠕虫状和成虫阶段。从形态上鉴定出这种捕食线虫真菌为辅助链壶菌。GenBank上没有可用于进行分子鉴定的序列。此前在欧洲曾有关于这种捕食线虫真菌寄生于甜菜孢囊线虫的报道(1,2);然而,据我们所知,尚无该真菌寄生于肾形线虫的报道。在蠕虫状线虫的生活阶段,在感染初期可观察到根状菌丝体,其特征为卵形细胞,直径9.5至13.5×17.0至24.5μm,由隔膜分隔。通常每个蠕虫状体内会产生10至15个缺乏细胞间菌丝丝的卵形细胞。根状菌丝体上长出宽3.5至μm的假根。成熟的肿胀细胞产生前体孢子囊,其可能成熟为休眠孢子或游动孢子囊。休眠孢子呈黄色至奶油色,直径20至40μm,外观呈网状,在蠕虫状线虫生活阶段很常见。游动孢子囊呈卵形,9.5至13.5×17.0至24.5μm,会从蠕虫状线虫的角质层中破裂,通过乳头状突起释放出游动孢子。游动孢子直径为2.9至4.9μm,前部区域有可见的小球,有一条9至11μm长的单鞭毛。游动孢子短距离游动,朝着鞭毛的方向移动。通过扫描电子显微镜观察到的成年肾形雌虫在其线虫的中体区域有游动孢子包囊。被寄生的卵内部被直径为20至25μm的游动孢子囊定殖。在感染后期,卵颜色变深,游动孢子囊从卵的角质层中破裂。明显被游动孢子囊和休眠孢子定殖的肾形线虫被放置在玉米粉、水和马铃薯葡萄糖琼脂上。这些培养基均不支持该真菌生长,这支持了我们的理论,即这种生物似乎是线虫的专性寄生虫。当将带有根状菌丝体和休眠孢子或游动孢子囊的卵添加到先前接种了2000个健康蠕虫状肾形线虫生活阶段的无菌土壤中的棉花植株上时,科赫法则得以完成。植株在温室中生长30天后,从土壤中提取下一代蠕虫状线虫并在显微镜下检查。42%的肾形蠕虫状生活阶段中存在根状菌丝体、休眠孢子和游动孢子囊。定殖于肾形线虫的根状菌丝体、休眠孢子或游动孢子囊以及游动孢子的形态比较与最初的观察结果相似。因此,据我们所知,这是辅助链壶菌寄生于肾形线虫的首次报道。参考文献:(1)B. Kerry。《线虫学杂志》12:253,1980年。(2)H. T. Tribe。《英国真菌学会会刊》69:367,1977年。