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炭疽菌对佛罗里达州入侵蕨类植物小叶海金沙和日本海金沙致病性的首次报道

First Report of the Pathogenicity of Colletotrichum gloeosporioides on Invasive Ferns, Lygodium microphyllum and L. japonicum, in Florida.

作者信息

Jones K A, Rayamajhi M B, Pratt P D, Van T K

机构信息

USDA/ARS, Invasive Plant Research Laboratory, 3205 College Ave., Ft. Lauderdale, FL 33314.

University of Florida, Ft. Lauderdale Research and Education Center, Ft. Lauderdale 33314.

出版信息

Plant Dis. 2003 Jan;87(1):101. doi: 10.1094/PDIS.2003.87.1.101B.

Abstract

Lygodium microphyllum (Cav.) R.Br. (Old World climbing fern) and L. japonicum (Thunb.) Sw. (Japanese climbing fern), in the family Schizaeaceae, are among the most invasive weeds in Florida (1). L. microphyllum invades fresh water and moist habitats in south Florida, while L. japonicum has spread in relatively well-drained habitats from Texas to North Carolina and central Florida. Some potted plants of both Lygodium spp. grown in shadehouse as well as in full sunlight developed discolored spots on pinnules (foliage), which coalesced and resulted in browning and dieback of severely infected vines. Symptomatic foliage obtained from these plants was surface-sterilized by immersing in a 15% solution of commercial bleach for 90 s, followed by a series of four rinses with sterile deionized distilled water. Disks (4 mm in diameter) of pinnules were cut from the junction of discolored and healthy tissues and placed on potato dextrose agar (PDA). A fungus, Colletotrichum gloeosporioides (Penz.) Penz. & Sacc. was consistently isolated from these disks. Fungal colonies produced abundant conidia on PDA. Conidia were hyaline, straight, cylindrical, averaging 14.7 μm (range 12.5 to 17.5 μm) × 5.0 μm (range 3.8 to 7.5 μm), and similar to those described for C. gloeosporioides (2). To confirm the pathogenicity of C. gloeosporioides on L. microphyllum and L. japonicum, Koch's postulates were performed. A fungal isolate was grown on PDA for 3 weeks, after which 10 ml of sterile deionized distilled water was added to the culture and agitated to dislodge conidia. The conidial suspension was strained through three layers of cheesecloth to remove hyphal fragments, and its concentration was adjusted to 1.7 × 10 conidia/ml. Foliage of healthy L. microphyllum and L. japonicum plants grown in 500-ml containers was sprayed with the conidial suspension until runoff. Plants were covered with plastic bags whose inner sides were misted with water to maintain high humidity and placed in a growth chamber under 12 h of fluorescent light per day. Temperature and relative humidity in the chamber ranged from 26 to 29°C and 44 to 73%, respectively. Plastic bags were removed after 3 days, and plants were further incubated for 3 weeks in the same growth chamber. Control plants were sprayed with sterile water, covered with plastic bags, and exposed to the same temperature, light, and humidity regime as those of the fungus-inoculated plants. Small, discolored foliar spots appeared 3 days after fungus inoculation. These spots were similar to those observed on pinnules of potted plants that originated from shadehouse and outdoor environments. Within 3 weeks after inoculation, the foliage of L. japonicum developed abundant discolored spots that led to edge browning and wilting of the pinnules. L. microphyllum had similar but more severe symptoms, with plants suffering as much as 50% dieback. C. gloeosporioides was consistently reisolated from the symptomatic tissues of both fern species. No symptoms appeared on the water-inoculated plants. To our knowledge, this is the first record of C. gloeosporioides pathogenicity on L. microphyllum and L. japonicum. References: (1) R. W. Pemberton and A. P. Ferriter. Am. Fern J. 88:165, 1998. (2) B. C. Sutton. Colletotrichum: Biology, Pathology and Control. CAB International, Wallingford, Oxon, UK, 1992.

摘要

小叶海金沙(Lygodium microphyllum (Cav.) R.Br.)(旧世界攀援蕨)和日本海金沙(L. japonicum (Thunb.) Sw.)属于莎草蕨科,是佛罗里达州最具入侵性的杂草之一(1)。小叶海金沙侵入佛罗里达州南部的淡水和潮湿栖息地,而日本海金沙已从得克萨斯州蔓延至北卡罗来纳州以及佛罗里达州中部相对排水良好的栖息地。在温室以及全光照条件下种植的这两种海金沙属植物的一些盆栽植株,其羽片(叶子)上出现了变色斑点,这些斑点融合在一起,导致严重感染的藤蔓褐变和枯死。从这些植株上获取的有症状叶片通过浸入15%的市售漂白剂溶液中90秒进行表面消毒,随后用无菌去离子蒸馏水进行一系列四次冲洗。从变色组织与健康组织的交界处切下直径4毫米的羽片圆盘,置于马铃薯葡萄糖琼脂(PDA)上。一种真菌,胶孢炭疽菌(Colletotrichum gloeosporioides (Penz.) Penz. & Sacc.)始终能从这些圆盘上分离出来。真菌菌落在PDA上产生大量分生孢子。分生孢子透明、直、圆柱形,平均大小为14.7微米(范围为12.5至17.5微米)×5.0微米(范围为3.8至7.5微米),与已描述的胶孢炭疽菌的分生孢子相似(2)。为证实胶孢炭疽菌对小叶海金沙和日本海金沙的致病性,进行了柯赫氏法则验证。将一种真菌分离株在PDA上培养3周,之后向培养物中加入10毫升无菌去离子蒸馏水并搅拌以释放分生孢子。分生孢子悬浮液通过三层粗棉布过滤以去除菌丝片段,其浓度调整至1.7×10个分生孢子/毫升。将生长在500毫升容器中的健康小叶海金沙和日本海金沙植株的叶片喷洒分生孢子悬浮液直至径流。植株用塑料袋覆盖,塑料袋内侧喷水以保持高湿度,并置于每天有12小时荧光光照的生长室中。生长室内的温度和相对湿度分别为26至29°C和44至73%。3天后移除塑料袋,植株在同一生长室中进一步培养3周。对照植株喷洒无菌水,用塑料袋覆盖,并暴露于与接种真菌的植株相同的温度、光照和湿度条件下。接种真菌3天后出现小的变色叶斑。这些斑点与在源自温室和室外环境的盆栽植株羽片上观察到的斑点相似。接种后3周内,日本海金沙的叶片出现大量变色斑点,导致羽片边缘褐变和枯萎。小叶海金沙有类似但更严重的症状,植株枯死率高达50%。始终能从这两种蕨类植物有症状的组织中重新分离出胶孢炭疽菌。在接种水的植株上未出现症状。据我们所知,这是胶孢炭疽菌对小叶海金沙和日本海金沙致病性的首次记录。参考文献:(1)R. W. 彭伯顿和A. P. 费里特。《美国蕨类植物杂志》88:165,1998年。(2)B. C. 萨顿。《炭疽菌:生物学、病理学与防治》。CAB国际出版社,英国牛津郡沃灵福德,1992年。

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