Li B-J, Liu Y, Shi Y-X, Xie X-W, Guo Y-L
Institute of Vegetables and Flowers, Chinese Academy of Agricultural Sciences, Beijing, 100081, China.
Institute of Microbiology, Chinese Academy of Sciences, Beijing, 100101, China.
Plant Dis. 2010 Nov;94(11):1377. doi: 10.1094/PDIS-03-10-0217.
Grafting has been widely and effectively used in cucumber (Cucumis sativus) cultivation for approximately 30 years in China to avoid Fusarium wilt caused by Fusarium oxysporum Schl. f. sp. cucumerinum Owen. In greenhouses, 90% of cucumbers are grafted onto pumpkin (Cucurbita moschata) rootstock. However, in March 2009, a severe crown rot causing yellowing and wilting of the leaves was observed on grafted cucumber in a large number of greenhouses in Lingyuan, western Liaoning Province in China. Symptoms consisted of dark brown, water-soaked lesions and a dense, white mycelial mat at the base of the stem. Lingyuan is the largest district for cucumber cultivation using grafting techniques in solar greenhouses in China. In 30 surveyed greenhouses in Sanshijiazi Village in the city of Lingyuan, the incidence of affected plants ranged from 10 to 40%, which caused serious economic losses. Fusarium spp. were isolated from the surface-sterilized basal stems of symptomatic plants on potato dextrose agar and incubated on potato sucrose agar for 4 days at 25°C. Colonies of the isolates produced a brown pigmentation and sparse, aerial mycelia, with a cream color on the underside. Conidiophores were elongated and branched or unbranched. Microconidia were abundant, hyaline, ellipsoid to ovoid, and 6 to 14 × 2.5 to 3.5 μm. Macroconidia were cylindrical, abundant, mostly two to six septate, 22 to 63 × 3.2 to 5.0 μm, with the apical cell rounded and blunt, and the basal cell rounded. On the basis of morphological characteristics, the fungus was identified as F. solani (C. Booth). For confirmation, the internal transcribed spacer region of rDNA was amplified and sequenced. A 449-bp sequence shared 99% homology with that of a F. solani GenBank accession previously reported from Japan (No. AF150473.1). The new sequence was deposited in GenBank (Accession No. HM015882). Pathogenicity of three isolates was determined in two experiments using different methods of inoculation. In one, 30 seedlings of pumpkin (C. moschata) with one true leaf each were inoculated by dipping their roots in a suspension of 10 spores ml, while control plants were mock inoculated with sterile water. Plants were then potted in a sterile mix of peat moss and vermiculite (2:1 vol/vol). In the other, pregerminated pumpkin seeds were sown in the same medium with a conidial suspension added at a rate of 10 spores ml, while other seeds were sown in sterile soil as controls. Plants for both experiments were maintained in a greenhouse at 25°C. Twelve days after inoculation, inoculated plants in both experiments showed a cortical rot on the crown and stem base with a brown, water-soaked appearance. Twenty-one days later, inoculated plants developed wilting and yellowed leaves. Disease incidence was 100%. No symptoms occurred on the control plants. Both experiments were repeated once with the same results. The pathogen was recovered from symptomatic tissue, confirming Koch's postulates. F. solani has been previously reported to cause root rot on cucurbit in California (2) and crown rot on grafted cucumber in the Netherlands (1). To our knowledge, this is the first report of crown rot of grafted cucumber caused by F. solani in China. References: (1) L. C. P. Kerling and L. Bravenboer. Neth. J. Plant Pathol. 73:15, 1967. (2) T. A. Tousson and W. C. Snyder. Phytopathology 51:17, 1961.
在中国,嫁接技术已在黄瓜(Cucumis sativus)栽培中广泛且有效地应用了约30年,以避免由尖孢镰刀菌黄瓜专化型(Fusarium oxysporum Schl. f. sp. cucumerinum Owen)引起的枯萎病。在温室中,90%的黄瓜被嫁接到南瓜(Cucurbita moschata)砧木上。然而,2009年3月,在中国辽宁省西部凌源市的大量温室中,观察到嫁接黄瓜出现严重的茎基腐病,导致叶片发黄和枯萎。症状包括深褐色、水渍状病斑以及茎基部浓密的白色菌丝层。凌源是中国日光温室黄瓜嫁接栽培面积最大的地区。在凌源市三十家子镇的30个被调查温室中,发病植株的发生率在10%至40%之间,造成了严重的经济损失。从症状明显植株的表面消毒茎基部在马铃薯葡萄糖琼脂培养基上分离出镰刀菌属真菌,并在马铃薯蔗糖琼脂培养基上于25°C培养4天。分离菌株的菌落产生褐色色素,气生菌丝稀疏,背面呈奶油色。分生孢子梗细长,有分枝或无分枝。小型分生孢子丰富,透明,椭圆形至卵形,大小为6至14×2.5至3.5μm。大型分生孢子圆柱形,丰富,大多有2至6个隔膜,大小为22至63×3.2至5.0μm,顶端细胞圆形且钝,基部细胞圆形。根据形态特征,该真菌被鉴定为茄腐镰刀菌(F. solani (C. Booth))。为进行确认,扩增并测序了rDNA的内部转录间隔区。一个449 bp的序列与先前从日本报道的茄腐镰刀菌GenBank登录号(编号AF150473.1)的序列具有99%的同源性。新序列已存入GenBank(登录号HM015882)。通过两种不同接种方法在两个实验中测定了三个分离菌株的致病性。在一个实验中,将30株各有一片真叶的南瓜(C. moschata)幼苗的根部浸入每毫升含10个孢子的悬浮液中进行接种,而对照植株用无菌水进行模拟接种。然后将植株种植在泥炭藓和蛭石的无菌混合物(体积比2:1)中。在另一个实验中,将预先萌发的南瓜种子播种在相同培养基中,并以每毫升含10个孢子的速率添加分生孢子悬浮液,而其他种子播种在无菌土壤中作为对照。两个实验的植株均在25°C的温室中培养。接种12天后,两个实验中的接种植株在茎基部和冠部均出现皮层腐烂,呈褐色、水渍状外观。21天后,接种植株出现萎蔫和叶片发黄。发病率为100%。对照植株未出现症状。两个实验均重复一次,结果相同。从有症状的组织中重新分离到该病原菌,证实了柯赫氏法则。此前曾报道茄腐镰刀菌在加利福尼亚州引起葫芦科植物根腐病(2),在荷兰引起嫁接黄瓜茎基腐病(1)。据我们所知,这是中国首次报道茄腐镰刀菌引起嫁接黄瓜茎基腐病。参考文献:(1)L. C. P. Kerling和L. Bravenboer。《荷兰植物病理学杂志》73:15,1967。(2)T. A. Tousson和W. C. Snyder。《植物病理学》51:17,1961。