华盛顿州生菜(Lactuca sativa)上由三色轮枝菌引起的黄萎病首次报道

First Report of Verticillium Wilt on Lettuce (Lactuca sativa) in Washington Caused by Verticillium tricorpus.

作者信息

Powell M, Gundersen B, Miles C, Coats K, Inglis D A

机构信息

Departments of Plant Pathology and Horticulture, Washington State University Mount Vernon NWREC, 16650 State Route 536 Mount Vernon, 98273.

Washington State University Puyallup REC 2606 West Pioneer, Puyallup, 98371.

出版信息

Plant Dis. 2013 Jul;97(7):996. doi: 10.1094/PDIS-12-12-1166-PDN.

Abstract

Symptoms of Verticillium wilt were observed on lettuce (Lactuca sativa L.) harvested from high tunnel and open field experimental plots in annual, consecutive spring plantings in western Washington from 2010 to 2012. Leaves had v-shaped, chlorotic lesions, and yellow or brown vascular tissue was noted in the crowns. Total disease incidence increased from 0.2% in 2010 to 1.9% in 2011 and to 14.4% in 2012. Verticillium spp. obtained from infected crown tissues and cultured on half-strength potato dextrose agar medium produced yellow pigment, black microsclerotia, white mycelia, tan chlamydospores, and uniseptate conidia averaging 10.6 × 3.7 μm. Isolates were identified tentatively as Verticillium tricorpus I. (3). Three isolates, Vt.Ls.2010, Vt.Ls.2011-1, and Vt.Ls.2011-2, were evaluated for pathogenicity on 4-week-old 'Coastal Star' seedlings in two greenhouse trials. In Trial I, four replicates of two duplicate plants per each isolate, and in Trial II, five replicates of one plant per each isolate were inoculated with conidial suspensions adjusted to 2.0 × 10 and 5.0 × 10 conidia/ml, respectively. Additionally, in each trial, two sets of control treatments of five plants each were inoculated with either an isolate of V. dahliae at the same conidial concentration or with sterile water. Root tips were cut and exposed to the suspensions for 5 s, then seedlings were transplanted into Sunshine Mix #1 (SunGro Horticulture Distribution Inc., Bellevue, WA), and kept in a greenhouse at 17.7 ± 3.4°C. Plants were harvested 8 to 9 weeks post-inoculation, and symptoms were rated visually. Vt.Ls.2010, Vt.Ls.2011-1, and Vt.Ls.2011-2 caused chlorosis and vascular discoloration on 25, 13, and 13% of the plants in Trial I; and 40, 60, and 20% of plants in Trial II, respectively. V. dahliae caused similar symptoms on 25 and 40% of the plants in the two trials, respectively, but these plants had greater intensity and length of vascular discoloration compared with the three test isolates. None of the water control plants were symptomatic. All V. tricorpus isolates were recovered from inoculated plants, and colony morphologies were similar to the original isolates. The internal transcribed spacer (ITS) rDNA of isolate Vt.Ls.2010 was amplified with ITS4 and ITS6 primer sets. ITS rDNA sequences between Vt.Ls.2010 and two isolates of V. tricorpus in GenBank (Accession Nos. FJ900211 and AB353343) were 100% identical. V. tricorpus is considered a weak pathogen of lettuce crops in California (2), but authors in Japan recently reported pathogenic isolates of V. tricorpus on lettuce (4). To our knowledge, this is the first report of Verticillium wilt caused by V. tricorpus in Washington. Lettuce is the number two crop grown in high tunnels in the United States (1), and cropping lettuce continuously in them can increase the risk of this and other soilborne pathogens. References: (1) E. E. Carey et al. HortTechnology 19:37, 2009. (2) Q.-M. Qin et al. Plant Dis. 92:69, 2008. (3) H. C. Smith. N. Z. J. Agric. Res. 8:450, 1965. (4) T. Usami et al. J. Gen. Plant Pathol. 77:17, 2010.

摘要

2010年至2012年期间,在华盛顿州西部连续多年春季种植的生菜(Lactuca sativa L.)上观察到了黄萎病症状,这些生菜分别采自温室和露地试验田。叶片上出现V形褪绿病斑,在叶冠处可见黄色或褐色维管组织。总发病率从2010年的0.2%增加到2011年的1.9%,并在2012年达到14.4%。从受感染的叶冠组织中分离得到的黄萎病菌株,在半强度马铃薯葡萄糖琼脂培养基上培养,产生黄色色素、黑色微菌核、白色菌丝体、褐色厚垣孢子和平均大小为10.6×3.7μm的单隔膜分生孢子。分离物初步鉴定为三色黄萎菌I.(3)。在两个温室试验中,对三个分离物Vt.Ls.2010、Vt.Ls.2011 - 1和Vt.Ls.2011 - 2在4周龄的“海岸之星”幼苗上进行致病性评估。在试验I中,每个分离物接种两份重复植株,共四个重复;在试验II中,每个分离物接种一份植株,共五个重复,接种的分生孢子悬浮液浓度分别调整为2.0×10和5.0×10个分生孢子/毫升。此外,在每个试验中,两组对照处理各有五株植株,分别接种相同分生孢子浓度的大丽轮枝菌分离物或无菌水。将根尖切断并在悬浮液中浸泡5秒,然后将幼苗移植到Sunshine Mix #1(SunGro Horticulture Distribution Inc.,华盛顿州贝尔维尤)中,并置于17.7±3.4°C的温室中。接种后8至9周收获植株,并对症状进行目测评分。在试验I中,Vt.Ls.2010、Vt.Ls.2011 - 1和Vt.Ls.2011 - 2分别导致25%、13%和13%的植株出现褪绿和维管组织变色;在试验II中,分别导致40%、60%和20%的植株出现上述症状。大丽轮枝菌在两个试验中分别导致25%和40%的植株出现类似症状,但与三个测试分离物相比,这些植株的维管组织变色强度更大、长度更长。所有水对照植株均无症状。所有三色黄萎菌分离物均从接种植株中重新分离得到,菌落形态与原始分离物相似。用ITS4和ITS6引物对分离物Vt.Ls.2010的内部转录间隔区(ITS)rDNA进行扩增。Vt.Ls.2010与GenBank中两个三色黄萎菌分离物(登录号FJ900211和AB353343)的ITS rDNA序列100%相同。三色黄萎菌在加利福尼亚州被认为是生菜作物的弱病原菌(2),但日本的作者最近报道了生菜上的三色黄萎菌致病分离物(4)。据我们所知,这是华盛顿州首次报道由三色黄萎菌引起的黄萎病。生菜是美国温室种植的第二大作物(1),在温室中连续种植生菜会增加这种病害和其他土传病原菌的发生风险。参考文献:(1)E. E. Carey等人,《园艺技术》19:37,2009年。(2)Q.-M. Qin等人,《植物病害》92:69,2008年。(3)H. C. Smith,《新西兰农业研究杂志》8:450,1965年。(4)T. Usami等人,《植物病理学报》77:17,2010年。

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