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棕榈疫霉引起的哥伦比亚非洲油棕芽腐病首次报道

First Report of Bud Rot Caused by Phytophthora palmivora on African Oil Palm in Colombia.

作者信息

Torres G A, Sarria G A, Varon F, Coffey M D, Elliott M L, Martinez G

机构信息

Colombian Oil Palm Research Center-Cenipalma, Bogota, Colombia.

University of California, Riverside.

出版信息

Plant Dis. 2010 Sep;94(9):1163. doi: 10.1094/PDIS-94-9-1163A.

Abstract

A bud rot disease, referred to as "pudricion del cogollo" or PC (for its name in Spanish), is a major disease of oil palm in four production areas of Colombia. Even though the disease has been studied for more than 40 years in Central and South America, the causal agent remained unknown (1). Infection first develops in the palm heart (cogollo) above the meristematic zone. Initial external symptoms appear as necrotic lesions on the sides of the spear leaf (youngest, unexpanded leaf) and the sizes of the lesions remain static as tissues emerge, expand, and mature. Under favorable environmental conditions (temperature around 26°C and relative humidity above 80%), further rotting of the heart eventually destroys the bud. A baiting technique with papaya and cocoa fruits was successfully used to trap the causal agent from symptomatic spear leaf tissue collected from the provinces of Nariño and Santander (2), where the disease is severe. Small, triangular-shaped pieces were removed from healthy fruits that had been surface sterilized, and a corresponding piece of diseased oil palm tissue was inserted into the hole and sealed with the removed fruit tissue. The whole fruit was wrapped with plastic and incubated at 25°C. Once the fruit traps showed initial necrotic symptoms, fruit tissue from the advancing point of the lesions was plated on Phytophthora-selective media. Four isolates of Phytophtora palmivora, all A2 mating type, were obtained during this study and each presented similar white and depressed mycelium when grown on oatmeal agar (OMA) medium. Papillated, ellipsoidal sporangia with short pedicels had an average size of 37.1 (breadth) × 52.8 μm (length). Chlamydospores averaged 35.1 μm in diameter and were similar to those observed in diseased palm tissue. DNA from the Phytophthora isolates was used as a template for PCR using ITS1/ITS4 primers. BLASTn analysis of the ITS sequences (GenBank Accession No. GQ398157) showed 99.9% homology with P. palmivora. The sequences of the four isolates were identical. For pathogenicity testing, cultures were grown on OMA. Two milliliters of distilled water containing 40,000 sporangia were injected into the base of the spear leaf of 105 nursery palms; the same number of palms was injected with 2 ml of distillated water as controls. Initial PC symptoms were observed 3 to 4 days after inoculation on 85% of inoculated palms and 15% of these palms developed typical bud rot symptoms under favorable conditions. No symptoms were observed on the controls. The presence of Phytophthora was confirmed by light microscopy and P. palmivora was reisolated from symptomatic palms with the fruit-baiting technique. The presence of Phytophthora in the four Colombian growing areas was also confirmed using Agdia (Elkhart, IN) immunostrips followed by DNA extraction from the Immunostrip wick and ITS sequencing to confirm the species. To our knowledge, this is the first report of P. palmivora causing PC on oil palm in Colombia. References: (1) H. de Franqueville. Exp. Agric. 39:225, 2003. (2) G. Sarria et al. Rev. Palmas, Colombia. 29(3):31, 2008.

摘要

一种芽腐病,在哥伦比亚的四个产区被称为“pudricion del cogollo”或PC(西班牙语名称),是油棕的一种主要病害。尽管在中美洲和南美洲对该病已研究了40多年,但致病因子仍不清楚(1)。感染首先在分生组织区上方的棕榈心(cogollo)中发展。最初的外部症状表现为枪形叶(最幼嫩、未展开的叶)两侧的坏死病变,随着组织的出现、扩展和成熟,病变大小保持不变。在适宜的环境条件下(温度约26°C,相对湿度高于80%),心部的进一步腐烂最终会破坏芽。用木瓜和可可果实的诱饵技术成功地从纳里尼奥省和桑坦德省(该病严重)采集的有症状枪形叶组织中捕获了致病因子(2)。从经过表面消毒的健康果实上切下小的三角形块,将相应的一块患病油棕组织插入孔中,并用切下的果实组织密封。整个果实用塑料包裹,在25°C下培养。一旦果实诱捕器出现最初的坏死症状,将病变进展部位的果实组织接种到疫霉选择性培养基上。在本研究中获得了4株棕榈疫霉分离株,均为A2交配型,在燕麦粉琼脂(OMA)培养基上生长时,每株都呈现出类似的白色凹陷菌丝体。具乳头状、椭圆形的孢子囊,具短梗,平均大小为37.1(宽)×52.8μm(长)。厚垣孢子平均直径为35.1μm,与在患病棕榈组织中观察到的相似。以疫霉分离株的DNA为模板,使用ITS1/ITS4引物进行PCR。对ITS序列(GenBank登录号GQ398157)的BLASTn分析显示与棕榈疫霉的同源性为99.9%。这4个分离株的序列相同。为了进行致病性测试,在OMA上培养菌株。将含有40000个孢子囊的2毫升蒸馏水注射到105株苗圃棕榈的枪形叶基部;同样数量的棕榈注射2毫升蒸馏水作为对照。接种后3至4天,在85%的接种棕榈上观察到最初的PC症状,在适宜条件下,其中15%的棕榈出现典型的芽腐症状。对照未观察到症状。通过光学显微镜确认了疫霉的存在,并使用果实诱饵技术从有症状的棕榈中重新分离出棕榈疫霉。还使用Agdia(印第安纳州埃尔克哈特)免疫试纸条,随后从免疫试纸条芯提取DNA并进行ITS测序以确认物种,从而证实了疫霉在哥伦比亚四个种植区的存在。据我们所知,这是棕榈疫霉在哥伦比亚导致油棕芽腐病的首次报道。参考文献:(1)H. de Franqueville。《实验农业》39:225,2003。(2)G. Sarria等人。《哥伦比亚棕榈》29(3):31,2008。

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