Identification of Fusarium cf. Verticillioides as The Causal Agent of Pokka Boheng Disease in Sugarcane in the Department of Antioquia, Colombia

Juliana Giraldo-Arias, Santiago Celis-Zapata, Nicolás D. Franco-Sierra, Juan José Arroyave-Toro, Claudia Jaramillo-Mazo, Javier Correa-Álvarez
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引用次数: 1

Abstract

The cultivation of sugarcane represents an important part in the Colombian economy due to the diverse applications in industries like liquor, food, paper and biofuels. Sugarcane worldwide production is affected by the presence of phytopathogenic agents, mainly filamentous fungi such as Physalospora tucumanensis (red rot disease) and Fusarium spp. To date in Colombia, Pokka boheng disease whose causal agent is the fungus Fusarium verticillioides, has not been reported, which is why it is necessary to identify appropriately this microorganism, responsible for losses in productivity and food contamination. In order to isolate and identify the infectious agent from symptomatic tissues, disinfection and culture in liquid and solid culture mediums were performed in malt extract (2%) and yeast extract (0.2%) both liquid and solid, for 7 days. After several replicates in agar plate, a purification was made along with a morphological characterization based on the shape and color of the mycelium, as well as the type of spores generated. Additionally, the genetic material was extracted and gene markers (ITS, Elongation Factor 1-α (EF) and β-Tubulin (Btub)) were amplified by PCR. Then, DNA sequencing was used to obtain the data to make a phylogenetic reconstruction by probabilistic methods (Maximum Likelihood and Bayesian Inference). The isolated strain, named as EA-FP0013 was located in the Fujikuroi complex group, with high probable identity to Fusarium verticillioides. Thus, early and species-specific identification of these fungal isolates by molecular methods may allow the timely diagnosis of emerging pathophysiological diseases of interest in the region, and thus propose the respective control strategies.
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哥伦比亚安蒂奥基亚省鉴定了作为甘蔗Pokka-Boheng病病原体的黄腐镰刀菌
由于甘蔗在酒类、食品、造纸和生物燃料等行业的广泛应用,甘蔗的种植代表了哥伦比亚经济的重要组成部分。世界范围内的甘蔗生产受到植物病原体的影响,主要是丝状真菌,如红腐病和镰刀菌。迄今为止,在哥伦比亚,Pokka boheng病(其病原体是真菌Fusarium verticillioides)尚未报道,这就是为什么有必要适当识别这种导致生产力损失和食品污染的微生物。为了从有症状的组织中分离和鉴定感染原,在液体和固体培养基中分别用麦芽提取物(2%)和酵母提取物(0.2%)进行消毒和培养,培养时间为7 d。在琼脂板上进行多次复制后,进行纯化,并根据菌丝的形状和颜色以及产生的孢子类型进行形态学表征。提取遗传物质,通过PCR扩增ITS、延伸因子1-α (EF)和β-微管蛋白(Btub)等基因标记。然后,利用DNA测序获得数据,通过概率方法(极大似然和贝叶斯推理)进行系统发育重建。该分离株命名为EA-FP0013,属于Fujikuroi复合体群,极有可能为黄斑镰刀菌。因此,通过分子方法对这些真菌分离物进行早期和物种特异性鉴定,可以及时诊断该地区感兴趣的新出现的病理生理疾病,从而提出相应的控制策略。
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