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新墨西哥州发现菜豆荚孢菌引起巴伦西亚花生茎枯病的首次报道

First Report of Phomopsis longicolla Causing Stem Blight of Valencia Peanut in New Mexico.

作者信息

Sanogo S, Etarock B F

机构信息

Department of Entomology, Plant Pathology, and Weed Science, New Mexico State University, Las Cruces 88003.

出版信息

Plant Dis. 2009 Sep;93(9):965. doi: 10.1094/PDIS-93-9-0965A.

DOI:10.1094/PDIS-93-9-0965A
PMID:30754544
Abstract

Wilted plants of Valencia market-type peanut (Arachis hypogaea L.) were found in two fields in August 2006 and three fields in September 2007 in Curry County, New Mexico. Plants had extensive, light brown discoloration and interstices of greenish tissue on blighted stems and branches across plant canopy levels. Disease incidence was less than 1% with infected plants in groups of two to five within each field. Five 4-cm stem segments were taken from each of five diseased plants in each field, submerged for 5 min in 0.5% NaOCl, rinsed in sterile distilled water, cut into 0.5-cm pieces, and plated on acidified potato dextrose agar (APDA). Mycelial colonies, recovered from plant tissues and incubated on APDA at 25°C under a 12-h photoperiod, were white and floccose with light green-yellow areas becoming visible within 7 to 10 days of incubation. Black stromata formed, spreading in a concentric pattern or scattered as large masses on APDA. Ostiolate and rostrate pycnidia with long beaks more than 500 μm were observed. Alpha conidia exuded from pycnidia in creamy-to-yellowish drops and were ellipsoid and biguttulate with an average length of 6.6 μm and width of 2.10 μm. Colonies were identified as Phomopsis longicolla Hobbs (1). PCR amplification of the internal transcribed spacer (ITS) region of rDNA of three isolates using primer pair ITS4/ITS5 (3) was followed by sequencing and BLAST analysis and showed a 95% homology with the sequence of the ITS region of rDNA of P. longicolla (1). Digestion of PCR-amplified DNA with AluI yielded two restriction fragments of sizes consistent with those reported for P. longicolla (2). Koch's postulates were established with three isolates tested for pathogenicity on Valencia peanut cv. Val-C at the four- to six-leaf stage using stem and root inoculations. Stems were injected with conidial suspension (10 conidia/ml) with a hypodermic needle or stabbed at the cotyledon axils with a sterile toothpick dabbed into an exuded conidial drop. Control plants were stem injected with distilled water or stabbed with a sterile toothpick. For root inoculation, plants were uprooted, washed free from soil, and inserted up to the crown into a 50-ml plastic test tube containing 40 ml of conidial suspension (25,000 conidia/ml) or sterile distilled water. For each method, eight plants were inoculated with each isolate, and four plants served as control. All inoculation methods were performed on the same day and repeated three times. Inoculated plants were covered with a clear plastic bag that was removed after 4 days. Plants were placed at 30°C under a 14-h photoperiod for 2 weeks. On stem-inoculated plants, light-to-dark brown discoloration formed at the sites of inoculation and expanded up and down the stems, which became brown, resulting in plant death within 10 to 14 days. On root-inoculated plants, browning of crown areas progressed up the stems, followed by plant death. P. longicolla was recovered from all inoculated plants. To our knowledge, this is the first report of P. longicolla on peanut in New Mexico and the United States. This report demonstrates the association of P. longicolla with peanut and its ability to cause stem blight. The occurrence and extent of this disease may be of a concern, because on other crops, Phomopsis diseases can cause significant reduction in seed germination, plant vigor, and yield. References: (1) T. W. Hobbs et al. Mycologia 77:535, 1985. (2) A. W. Zhang et al. Plant Dis. 81:1143, 1997. (3) A. W. Zhang et al. Phytopathology 88:1306, 1998.

摘要

2006年8月,在新墨西哥州库里县的两块田地中发现了瓦伦西亚市场型花生(Arachis hypogaea L.)的萎蔫植株,2007年9月在三块田地中也发现了此类情况。植株茎和枝出现大面积浅褐色变色,整个植株冠层的枯萎茎和枝上有绿色组织间隙。发病率低于1%,每块田地中受感染植株以两到五株为一组。从每块田地的五株患病植株上各取5个4厘米长的茎段,在0.5%次氯酸钠中浸泡5分钟,用无菌蒸馏水冲洗,切成0.5厘米的小段,接种到酸化马铃薯葡萄糖琼脂(APDA)上。从植物组织中分离得到的菌丝菌落,在25°C、12小时光周期条件下于APDA上培养,起初为白色絮状,培养7至10天后可见浅绿色至黄色区域。形成黑色子座,在APDA上呈同心状扩展或聚集成大片状散布。观察到具孔口和喙状的分生孢子器,其长喙超过500微米。分生孢子从分生孢子器中以乳黄色至淡黄色液滴形式渗出呈椭圆形,具双油球,平均长度6.6微米,宽度2.10微米。菌落被鉴定为长喙茎点霉(Phomopsis longicolla Hobbs)(1)。使用引物对ITS4/ITS5(3)对三个分离株的核糖体DNA内部转录间隔区(ITS)进行PCR扩增,随后进行测序和BLAST分析,结果显示与长喙茎点霉核糖体DNA的ITS区域序列有95%的同源性(1)。用AluI对PCR扩增的DNA进行酶切,产生了两个大小与长喙茎点霉报道一致的限制性片段(2)。利用三个分离株对处于四至六叶期的瓦伦西亚花生品种Val-C进行致病性测试,通过茎部和根部接种确立了柯赫氏法则。用皮下注射针向茎部注射分生孢子悬浮液(10个分生孢子/毫升),或用无菌牙签蘸取渗出的分生孢子滴在子叶腋处穿刺。对照植株用蒸馏水进行茎部注射或用无菌牙签穿刺。对于根部接种,将植株连根拔起,洗净泥土,将根部以上部分插入装有40毫升分生孢子悬浮液(25000个分生孢子/毫升)或无菌蒸馏水的50毫升塑料试管中。对于每种方法,每个分离株接种八株植物,四株作为对照。所有接种方法均在同一天进行,并重复三次。接种后的植株用透明塑料袋覆盖,4天后移除。将植株置于30°C、14小时光周期条件下培养2周。在茎部接种的植株上,接种部位形成浅褐色至深褐色变色,并在茎部上下扩展,茎部变为褐色,导致植株在10至14天内死亡。在根部接种的植株上,冠部区域褐化向上蔓延至茎部,随后植株死亡。从所有接种的植株中都分离到了长喙茎点霉。据我们所知,这是长喙茎点霉在新墨西哥州和美国花生上的首次报道。本报告证明了长喙茎点霉与花生的关联及其引起茎枯病的能力。这种病害的发生和程度可能令人担忧,因为在其他作物上,茎点霉病害会导致种子发芽率、植株活力和产量显著降低。参考文献:(1)T. W. Hobbs等人,《真菌学》77:535,1985年。(2)A. W. Zhang等人,《植物病害》81:1143,1997年。(3)A. W. Zhang等人,《植物病理学》88:1306,1998年。