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台湾地区由立枯丝核菌AG-IV引起的海岸红杉组培育苗立枯病的首次报道

First Report of Rhizoctonia Blight of a Coastal Redwood Tissue-Culture-Derived Saplings Caused by Rhizoctonia solani AG-IV in Taiwan.

作者信息

Hsiao W W, Wu Y S, Wang Y N, Huang B L, Huang L C

机构信息

The Experimental Forest, College of Bio-Resource and Agriculture, National Taiwan University, Taiwan R.O.C.

Department of Forestry and Resource Conservation, College of Bio-Resource and Agriculture, National Taiwan University, Taiwan R.O.C.

出版信息

Plant Dis. 2008 Apr;92(4):655. doi: 10.1094/PDIS-92-4-0655A.

Abstract

Coastal redwood (Sequoia sempervirens (D. Don) Endl.) is native to North America. This tall tree species is used for forestation and lumber; its wood is also used for furniture, its burls for art ware, and its bark for fuel, insulation, and mulch. In August 2005, an instance of wilt was observed among 2-year-old tissue-culture-cloned plants (2) in the Sitou Forest of central Taiwan. Essentially, all plants were infected. The leaves or stems near the ground were affected first, but the wilt soon spread over the entire plant with the leaves becoming grayish brown and water soaked, and then wilting, drying, and finally defoliation occurred. Aerial hyphae were present over the affected areas, aerial mycelium was cob-web-like, hyaline, later becoming slightly brown. Hyphae were 6.5 to 10.4 μm wide with right-angle branching and septal constriction at their bases. Sclerotia were hemispherical, subglobose, to irregular in shape, 1 to 2 mm, and brown. The perfect stage of the fungus was not found. The fungus was identified as Rhizoctonia solani Kühn (3). Vegetative cells were stained with alkaline safranin solution and identified as multinucleate (1). Portions of the stem that displayed symptoms, together with adjacent healthy tissue, were disinfested for 1 min in 0.5% NaOCl and plated on to potato-dextrose agar (PDA) (Merck, Darmstadt, Germany) supplemented with 100 mg/L of ampicillin (Sigma, St. Louis, MO). Single hyphal tips were transferred to PDA and two isolates were established as pure cultures. On the basis of hyphal anastomosis with AG-IV tester isolates (exfop234, exfop241, and exfop250) (1), the fungus was identifed as R. solani AG-IV. Pathogenicity of the fungal isolates was confirmed by inoculating 2-month-old tissue-culture-derived S. sempervirens plants that were grown in pots and incubated in a growth chamber maintained at 28°C with a relative humidity above 95%. Inoculum consisted of a single mycelial 5-day-old 0.5-cm disc grown on PDA of the pathogen placed on the soil surface touching the base of each plant. Four plants were inoculated with mycelium and the four control plants were noninoculated. Inoculated plants wilted gradually over 4 days and all plants developed severe stem rot and were dead in 6 days, whereas control plants remained symptomless. The Rhizoctonia solani AG-IV was reisolated from all inoculated plants. This fungus has been observed to cause disease in many species of plants (4), but to our knowledge, this is the first report of Rhizoctonia blight of coastal redwood tissue-culture-derived saplings caused by Rhizoctonia solani AG-IV in Taiwan. References: (1) T. T. Chang. Taiwan J. For. Sci. 12:47, 1997. (2) L. C. Huang et al. Plant Physiol. 98:166, 1992. (3) B. Sneh et al. Identification of Rhizoctonia species. The American Phytopathological Society, St. Paul, MN, 1991. (4) S. T. Su et al. List of Plant Diseases in Taiwan. The Phytopathological Society of the Republic of China, 2002.

摘要

海岸红杉(Sequoia sempervirens (D. Don) Endl.)原产于北美洲。这种高大的树种用于造林和木材生产;其木材还用于制作家具,树瘤用于制作工艺品,树皮用于燃料、隔热材料和覆盖物。2005年8月,在台湾中部溪头森林中,2年生组织培养克隆植株(2)中出现了枯萎病实例。基本上,所有植株都受到了感染。靠近地面的叶片或茎首先受到影响,但枯萎病很快蔓延到整株植物,叶片变为灰棕色并出现水渍状,随后枯萎、干枯,最终落叶。在受影响区域出现气生菌丝,气生菌丝呈蛛网状,透明,后期略带褐色。菌丝宽6.5至10.4μm,呈直角分支,基部有隔膜缢缩。菌核呈半球形、近球形至不规则形,直径1至2mm,褐色。未发现该真菌的有性阶段。该真菌被鉴定为立枯丝核菌(Rhizoctonia solani Kühn)(3)。营养细胞用碱性番红溶液染色,鉴定为多核(1)。将表现出症状的茎段连同相邻的健康组织在0.5%次氯酸钠中消毒1分钟,然后接种到添加了100mg/L氨苄青霉素(Sigma,圣路易斯,密苏里州)的马铃薯葡萄糖琼脂(PDA)(默克,达姆施塔特,德国)上。将单个菌丝尖端转移到PDA上,建立了两个纯培养物分离株。根据与AG-IV测试分离株(exfop234、exfop241和exfop250)(1)的菌丝融合,该真菌被鉴定为立枯丝核菌AG-IV。通过接种在花盆中生长并在保持在28°C、相对湿度高于95%的生长室中培养的2个月大的组织培养来源的海岸红杉植株,证实了真菌分离株的致病性。接种物由在PDA上生长5天的单个菌丝体0.5cm圆盘组成,放置在接触每株植物基部的土壤表面。四株植物接种了菌丝体,四株对照植物未接种。接种的植物在4天内逐渐枯萎,所有植物在6天内都出现严重的茎腐病并死亡,而对照植物没有症状。从所有接种的植物中重新分离出立枯丝核菌AG-IV。据观察,这种真菌会在许多植物物种中引起病害(4),但据我们所知,这是台湾首次报道由立枯丝核菌AG-IV引起的海岸红杉组织培养来源树苗的立枯病。参考文献:(1)T. T. Chang。台湾林业科学杂志12:47,1997。(2)L. C. Huang等人。植物生理学98:166,1992。(3)B. Sneh等人。立枯丝核菌物种的鉴定。美国植物病理学会,圣保罗,明尼苏达州,1991。(4)S. T. Su等人。台湾植物病害名录。中华民国植物病理学会,2002。

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