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智利由菜豆壳球孢菌引起的甜瓜严重炭腐病暴发。

A Severe Outbreak of Charcoal Rot in Cantaloupe Melon Caused by Macrophomina phaseolina in Chile.

作者信息

Jacob C J, Krarup C, Díaz G A, Latorre B A

机构信息

Pontificia Universidad Católica de Chile, Santiago, Chile.

出版信息

Plant Dis. 2013 Jan;97(1):141. doi: 10.1094/PDIS-06-12-0588-PDN.

Abstract

A severe outbreak of charcoal rot was observed in cantaloupe melon (Cucumis melo L.) in the summer of 2011 to 2012 in Curacaví Valley, Chile. Prior to harvest, of 72 plants per cultivar, charcoal rot prevalence varied from 32% to 82% in cvs. Colima, Charantias, Navigator, Origami, Otero, and Samoa. Symptoms were wilting and leaf browning associated with water-soaked lesions at the base of the crown with amber to dark brown exudates. Lesions dried out progressively, turned tan, and cracked. Affected plants declined and died before harvest. Reddish fruit decay was observed. Symptomatic stem and root samples (n = 97) were collected, surface disinfected (96% ethanol, 30 s), plated on PDA acidified with 0.5 ml/liter of 92% lactic acid (APDA), and incubated at 20 ± 1°C. A white, fast-growing mycelium was obtained that turned gray to black after 7 days due to the presence of spherical to oblong black microsclerotia 136 ± 52 μm (n = 80) in diameter. On the basis of colony morphology and microsclerotia, 57 isolates (59%), obtained from 97 melon samples, were tentatively identified as Macrophomina phaseolina (Tassi) Goid. (2,3). The morphological identification of four isolates M1HB-B, M2CO-B, M3CH-R, and M4OT-B (GenBank Accession Nos. JX203630, JX203631, JX203632, and JX203633) was confirmed by sequencing of the internal transcribed spacer region (ITS1-5.8S-ITS2) of rDNA, using primers ITS4 and ITS5, with >99% similarity with the sequences of M. phaseolina (GenBank Accession No. HQ660592) (4). Pathogenicity tests were conducted with isolates M1HB-B, M2CO-B, M3CH-R, and M4OT-B on melon fruits cvs. Colima, Origami, Charantias, and Diva. Four mature melon fruits per cultivar per isolate were surface disinfected with 0.5% sodium hypochlorite for 2 min before inserting a mycelium plug (19 mm) in a 6 mm diameter hole made with a sterile cork borer. An equal number of perforated fruits in which a sterile agar plug was inserted were left as non-inoculated controls. After 8 days of incubation at 20°C, inoculated fruits developed a spherical, reddish, soft necrotic lesion of 15 to 20 mm in diameter in the pulp. Non-inoculated fruits remained symptomless. The pathogenicity of the four isolates was also studied in 3-month-old melon plants (n = 4) cvs. Colima and Navigator. Plants were inoculated by inserting a mycelial plug (9 mm) underneath the epidermis of the crown, 5 cm above the soil level. The inoculation site was immediately wrapped with Parafilm to avoid dehydration. An equal number of non-inoculated, but injured plants, treated with a sterile agar plug, were left as controls. After 21 days of incubation under greenhouse conditions (17 ± 5.5°C), all inoculated plants developed water-soaked to dry necrotic lesions, 20 to 70 mm long, yellow to tan in color. No symptoms were obtained in non-inoculated controls. M. phaseolina was reisolated in 84% and 100% of the inoculated plants and fruits, respectively. To our knowledge, this study is the first report of charcoal rot in cantaloupe melon in Chile, previously found on watermelon and melon group inodorus (1). Charcoal rot appears as an emerging disease that aggressively affects current cantaloupe melon cultivars in central Chile. References: (1) G. Apablaza. Cien. Inv. Agr. 20:101, 1993. (2) B. D. Bruton and E. V. Wann. Charcoal rot. Page 9 in: Compendium of Cucurbit Diseases. T. A. Zitter, D. L. Hopkins, and C. E. Thomas, eds. APS, St. Paul, MN, 1996. (3) S. Kaur et al. Crit. Rev. Microbiol. 38:136, 2012. (4) J. Q. Zhang et al. Plant Dis. 95:872, 2011.

摘要

2011年夏到2012年期间,在智利库拉卡维山谷的甜瓜(Cucumis melo L.)上观察到严重的炭腐病爆发。收获前,每个品种取72株植株,炭腐病发病率在科利马、查兰蒂亚斯、航海家、折纸、奥泰罗和萨摩亚等品种中从32%到82%不等。症状包括萎蔫和叶片褐变,伴有冠基部水渍状病斑以及琥珀色至深褐色渗出物。病斑逐渐干枯,变为棕褐色并开裂。受影响的植株在收获前衰退并死亡。观察到果实有微红腐烂现象。采集了有症状的茎和根样本(n = 97),进行表面消毒(96%乙醇,30秒),接种到用0.5毫升/升92%乳酸酸化的马铃薯葡萄糖琼脂(APDA)上,并在20±1°C下培养。获得了一种白色、生长迅速的菌丝体,7天后由于存在直径为136±52微米(n = 80)的球形至椭圆形黑色微菌核而变为灰色至黑色。根据菌落形态和微菌核,从97个甜瓜样本中获得的57个分离物(59%)初步鉴定为菜豆壳球孢(Macrophomina phaseolina (Tassi) Goid.)。通过使用引物ITS4和ITS5对核糖体DNA的内部转录间隔区(ITS1 - 5.8S - ITS2)进行测序,确认了四个分离物M1HB - B、M2CO - B、M3CH - R和M4OT - B(GenBank登录号JX203630、JX203631、JX203632和JX203633)的形态鉴定,与菜豆壳球孢的序列相似度>99%(GenBank登录号HQ660592)。对分离物M1HB - B、M2CO - B、M3CH - R和M4OT - B在甜瓜品种科利马、折纸、查兰蒂亚斯和迪瓦上进行了致病性测试。每个品种每个分离物取四个成熟甜瓜果实,用0.5%次氯酸钠表面消毒2分钟,然后用无菌软木塞钻在果实上钻一个6毫米直径的孔,插入一个菌丝体菌块(19毫米)。同样数量的插入无菌琼脂块的穿孔果实作为未接种对照。在20°C下培养8天后,接种的果实果肉中出现直径为15至20毫米的球形、微红、软腐坏死病斑。未接种的果实无症状。还在3个月大的甜瓜植株(n = 4)品种科利马和航海家上研究了这四个分离物的致病性。通过在离土壤表面5厘米处的冠表皮下插入一个菌丝体菌块(9毫米)对接种植株进行接种。接种部位立即用保鲜膜包裹以避免脱水。同样数量的用无菌琼脂块处理的未接种但受伤的植株作为对照。在温室条件(17±5.5°C)下培养21天后,所有接种的植株都出现了水渍状至干腐坏死病斑,长20至70毫米,颜色从黄色到棕褐色。未接种对照未出现症状。在84%的接种植株和100%的接种果实中重新分离到了菜豆壳球孢。据我们所知,本研究是智利甜瓜上炭腐病的首次报道,此前在西瓜和甜瓜组的白兰瓜上发现过。炭腐病似乎是一种新出现的病害,对智利中部目前的甜瓜品种有严重影响。参考文献:(1) G. Apablaza. Cien. Inv. Agr. 20:101, 199

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