Wick R L, Dicklow M B
Department of Microbiology, University of Massachusetts, Amherst 01003.
Plant Dis. 2002 Sep;86(9):1050. doi: 10.1094/PDIS.2002.86.9.1050B.
From 1999 to 2001, a Massachusetts nursery received a number of shipments of Pothos, Epipremnum aureum (Lindl. & André) Bunting, with significant crown, petiole, and leaf rot. The plants were imported from Costa Rica. Sporangia were observed on diseased tissues, and five presumptive isolates of Phytophthora were recovered from infected petioles and stems for species identification. The five isolates were morphologically indistinguishable from each other. Sporangia were produced in water and on V8 juice agar under fluorescent light at 22°C. Mating type was determined by pairing isolates with A1 and A2 mating types of Phytophthora capsici Leonian. Sporangial measurements were taken from water cultures. Determination of caducity, and measurements of pedicels and oospores were taken from V8 agar cultures. Measurements represent an average of 50 observations a single isolate. In water culture, sporangia were borne in umbellate clusters. Sporangium length/breadth was 48.29 and 22.33 μm respectively; length/breadth ratio 2.16. On solid media, sporangia were upright and caducous. The bases of the sporangia were mostly tapered. Pedicel lengths were 22 to 49 μm (average 35 μm). Oogonia had amphigynous antheridia and developed only in the presence of an opposite mating type, and oospores measured 25.74 μm diameter. All five isolates were the A1 mating type. Chlamydospores were absent in V8 and corn meal agar (CMA) cultures. Metalaxyl sensitivity was determined at 0, 0.1, 0.5, and 5 ppm in CMA with five replications. The isolate was completely sensitive to 5 ppm metalaxyl, but grew as well as the controls at 0.1 ppm metalaxyl. Growth response to temperature was determined on V8 agar at 15, 20, 25, 30, and 35°C in five replications. After 4 days, colony diameters at 20, 25, and 30°C were not significantly different (P = 0.01) and colonies filled the 100-mm petri dishes. At 15 and 35°C, average colony diameter was 65.7 and 71.4 mm, respectively. Based on the above characteristics, the isolates were identified as P. capsici. Koch's postulates were carried out on pepper, Capsicum annuum 'Italia', squash, Cucurbita pepo 'Patty Pan' seedlings, and rooted cuttings of pothos. Pepper and squash seedlings and rooted pothos were transplanted in 4-in. (10 cm) pots containing a soilless growing medium (Metro Mix 360, W.R. Grace, Columbia, MD). Phytophthora cultures were grown on V8 juice agar for 4 days. An agar culture was added to 200 ml of sterile distilled water and briefly blended. Ten milliliters of the resulting mycelial slurry were pipetted in the soil one cm from the crown on two sides of the plant. Controls received no mycelial slurry. Petiole, leaf, and crown rot of pothos developed within 2 weeks following inoculation. Squash and pepper plants did not become diseased. In a second pathogenicity test, a 1-cm-diameter plug of mycelial growth from a V8 agar culture was placed between the stem and petiole of the lowest leaf of pothos cuttings directly after transplanting. Inoculated plants died within 3 days. The development of umbellate clusters of sporangia, sporangial shape, length/breadth ratio, and lack of pathogenicity to pepper suggest that the P. capsici isolated from pothos belong to the CAPB (tropical) subgroup of Mchau and Coffey (2). References: (1) S. S. A. Al-Hedaithy and P. H. Tsao. Mycologia 71:392, 1979. (2) G. R. Mchau. and M. D. Coffey. Mycol. Res. 99:89, 1995.
1999年至2001年期间,马萨诸塞州的一家苗圃收到了几批黄金葛(绿萝,学名Epipremnum aureum (Lindl. & André) Bunting),这些植株出现了严重的冠部、叶柄和叶片腐烂症状。这些植物是从哥斯达黎加进口的。在患病组织上观察到了孢子囊,并从受感染的叶柄和茎中分离出5株疑似疫霉菌株用于物种鉴定。这5个分离株在形态上彼此无法区分。孢子囊在水中以及在22°C荧光灯下的V8汁琼脂上产生。通过将分离株与辣椒疫霉(Phytophthora capsici Leonian)的A1和A2交配型配对来确定交配型。孢子囊的测量取自水培培养物。脱落性的测定以及梗和卵孢子的测量取自V8琼脂培养物。测量值代表单个分离株50次观察的平均值。在水培中,孢子囊呈伞状簇生。孢子囊长度/宽度分别为48.29和22.33μm;长宽比为2.16。在固体培养基上,孢子囊直立且易脱落。孢子囊基部大多呈锥形。梗的长度为22至49μm(平均35μm)。藏卵器有两性雄器,且仅在存在相反交配型时发育,卵孢子直径为25.74μm。所有5个分离株均为A1交配型。V8和玉米粉琼脂(CMA)培养物中没有厚垣孢子。在含有5个重复的CMA中,于0、0.1、0.5和5 ppm下测定甲霜灵敏感性。该分离株对5 ppm甲霜灵完全敏感,但在0.1 ppm甲霜灵下生长与对照一样好。在15、20、25、30和35°C下,在V8琼脂上以5个重复测定对温度的生长反应。4天后,20、25和30°C下的菌落直径无显著差异(P = 0.01),菌落布满100毫米的培养皿。在15和35°C下,平均菌落直径分别为65.7和71.4毫米。根据上述特征,这些分离株被鉴定为辣椒疫霉。对辣椒(Capsicum annuum 'Italia')、南瓜(Cucurbita pepo 'Patty Pan')幼苗以及绿萝的带根插条进行了柯赫氏法则验证。将辣椒和南瓜幼苗以及带根绿萝移植到装有无土生长介质(Metro Mix 360,W.R. Grace,哥伦比亚,马里兰州)的4英寸(10厘米)花盆中。疫霉菌培养物在V8汁琼脂上培养4天。将一个琼脂培养物加入200毫升无菌蒸馏水中并短暂搅拌。将10毫升所得的菌丝体悬浮液从植株冠部两侧距冠部1厘米处移液到土壤中。对照不接种菌丝体悬浮液。接种后2周内,绿萝出现叶柄、叶片和冠部腐烂。南瓜和辣椒植株未发病。在第二次致病性试验中,在移植后立即将一个直径1厘米的来自V8琼脂培养物的菌丝生长块放置在绿萝插条最低叶片的茎和叶柄之间。接种的植株在3天内死亡。孢子囊伞状簇生的发育、孢子囊形状、长宽比以及对辣椒无致病性表明,从绿萝中分离出的辣椒疫霉属于Mchau和Coffey(2)的CAPB(热带)亚组。参考文献:(1)S. S. A. Al-Hedaithy和P. H. Tsao。《真菌学》71:392,1979年。(2)G. R. Mchau和M. D. Coffey。《真菌学研究》99:89,1995年。