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路易斯安那州向日葵链格孢叶斑病和茎枯病的首次报道

First Report of Stem and Foliar Blight of Sunflower Caused by Alternariaster helianthi in Louisiana.

作者信息

Singh R, Ferrin D M

机构信息

Department of Plant Pathology and Crop Physiology, Louisiana State University Agricultural Center, Baton Rouge 70803.

出版信息

Plant Dis. 2012 May;96(5):761. doi: 10.1094/PDIS-01-12-0107-PDN.

DOI:10.1094/PDIS-01-12-0107-PDN
PMID:30727532
Abstract

During the fall of 2009, sunflower (Helianthus annuus L.) planted at the LSU AgCenter's Burden Center in Baton Rouge, LA exhibited severe stem and foliar blight symptoms. Symptoms on stems and petioles included elongated, slightly sunken lesions with dark brown margins. Leaf symptoms included irregular to circular, dark brown lesions with white centers and surrounded by a yellow halo. Several spots often coalesced to form large, blighted areas, and severely affected leaves turned yellow, followed by defoliation. The corolla and calyx exhibited similar lesions except for the yellow halo. Disease developed rapidly and the whole (100% disease incidence) field was blighted within a week following a rain (4 mm). Infected leaf and stem tissue was surface disinfested and plated on ¼-strength potato dextrose agar (PDA). Both leaf and stem tissue consistently produced dark olivaceous-to-black fungal colonies at room temperature under 12 h of fluorescent light per day. Conidia were 53 to 128 × 10 to 26 μm, borne singly on the conidiophores, hyaline to dark olivaceous, cylindrical, rounded at both ends, and with 6 to 10 transverse and 0 to 2 longitudinal septa. Conidiophores were single, unbranched, septate, hyaline to dark olivaceous, and measured 77 to 128 × 7 to 13 μm. Morphologically, the fungus was identified as Alternariaster helianthi (Hansf.) E.G. Simmons (= Alternaria helianthi [Hansf.] Tubaki & Nishih) (1). A single-spore isolate (PDC-4291) was obtained from the original culture and DNA from this isolate was extracted with a DNeasy Plant Mini Kit (Qiagen Inc., Valencia, CA). Primers ITS1 and ITS4 were used to amplify and sequence the internal transcribed spacer regions 1 and 2, and NCBI blast analysis of the 552-bp sequence (GenBank Accession No. JN208925) resulted in 100% homology with Alternaria helianthi isolated from sunflower infected with leaf spot and blight disease in India (GenBank Accession No. DQ156343). Pathogenicity was determined by inoculating 20 potted sunflower plants (Full Sun Improved TD, Fred C. Gloeckner and Company, Inc., Harrison, NY) with conidia from a 2-week-old culture of isolate PDC-4291. Each plant was sprayed with 25 ml of suspension containing 10 conidia/ml. Twenty control plants were sprayed with 25 ml of sterile distilled water. Inoculated and control plants were covered with plastic bags and maintained in a greenhouse at 28 ± 2°C. Plastic bags were removed 72 h after inoculation. Leaf spots similar to the original symptoms appeared on all 20 inoculated plants 5 days after inoculation. A few stem lesions were observed on 13 plants. Two weeks after inoculation, infected leaves turned yellow and blighted. Alternariaster helianthi (= Alternaria helianthi) was reisolated from the leaf spots and stem lesions. No symptoms developed on any of the 20 control plants. On the basis of morphology and sequence data, this pathogen was identified as A. helianthi, and to our knowledge, this is the first report of sunflower stem and foliar blight caused by A. helianthi in Louisiana. In Louisiana, sunflower is a popular ornamental that is grown in landscapes and gardens and by commercial flower growers who grow it for cut flower arrangements. Louisiana's hot, humid weather is ideal for disease development, which may discourage gardeners and commercial growers from planting sunflower. Reference: (1) E. G. Simmons. Alternaria: An Identification Manual. CBS Fungal Biodiversity Center, Utrecht, the Netherlands, 2007.

摘要

2009年秋季,种植于路易斯安那州巴吞鲁日市LSU农业中心负担中心的向日葵(Helianthus annuus L.)出现了严重的茎部和叶片枯萎症状。茎部和叶柄上的症状包括细长、略有凹陷的病斑,边缘呈深褐色。叶片症状包括不规则至圆形、深褐色病斑,中央为白色,周围有黄色晕圈。多个病斑常融合形成大的枯萎区域,严重受害的叶片变黄,随后脱落。花冠和花萼也出现类似病斑,但没有黄色晕圈。病害发展迅速,一场降雨(4毫米)后的一周内,整块田地(发病率100%)都出现了枯萎症状。对感染的叶片和茎组织进行表面消毒后,接种到1/4强度的马铃薯葡萄糖琼脂(PDA)培养基上。在室温下,每天12小时荧光光照条件下,叶片和茎组织均持续产生深橄榄色至黑色的真菌菌落。分生孢子大小为53至128×10至26微米,单生于分生孢子梗上,透明至深橄榄色,圆柱形,两端圆形,有6至10个横向隔膜和0至2个纵向隔膜。分生孢子梗单生,无分支,有隔膜,透明至深橄榄色,大小为77至128×7至13微米。从形态学上看,该真菌被鉴定为向日葵链格孢(Alternariaster helianthi (Hansf.) E.G. Simmons (= Alternaria helianthi [Hansf.] Tubaki & Nishih))(1)。从原始培养物中获得了一个单孢分离株(PDC - 4291),使用DNeasy植物小提试剂盒(Qiagen公司,加利福尼亚州瓦伦西亚)从该分离株中提取DNA。使用引物ITS1和ITS4扩增并测序内部转录间隔区1和2,对552碱基对序列(GenBank登录号JN208925)进行NCBI比对分析,结果显示与从印度感染叶斑病和枯萎病的向日葵中分离得到的向日葵链格孢(GenBank登录号DQ156343)具有100%的同源性。通过用分离株PDC - 4291两周龄培养物的分生孢子接种20株盆栽向日葵(Full Sun Improved TD,Fred C. Gloeckner and Company, Inc.,纽约州哈里森)来测定致病性。每株植物喷洒25毫升含有10个分生孢子/毫升的悬浮液。20株对照植物喷洒25毫升无菌蒸馏水。接种和对照植物用塑料袋覆盖,置于28±2°C的温室中。接种72小时后移除塑料袋。接种后5天,所有20株接种植物均出现了与原始症状相似的叶斑。13株植物上观察到少数茎部病斑。接种两周后,感染的叶片变黄并枯萎。从叶斑和茎部病斑中重新分离出向日葵链格孢(= Alternaria helianthi)。20株对照植物均未出现症状。基于形态学和序列数据,该病原菌被鉴定为向日葵链格孢,据我们所知,这是路易斯安那州首次关于向日葵链格孢引起向日葵茎部和叶片枯萎病的报道。在路易斯安那州,向日葵是一种受欢迎的观赏植物,种植于景观、花园以及为切花生产而种植的商业花农处。路易斯安那州炎热潮湿的气候非常适合病害发展,这可能会使园丁和商业种植者不愿种植向日葵。参考文献:(1)E.G. Simmons. Alternaria: An Identification Manual. CBS Fungal Biodiversity Center, Utrecht, the Netherlands, 2007.