生物光照对pcr法研究尖孢镰刀菌侵染基质微生物种群的影响。

M Pugliese, I Ferrocino, G Gilardi, M L Gullino, A Garibaldi
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引用次数: 0

摘要

生物光疗是将光疗与有机质施用相结合,以控制土传病原体。这种控制策略对微生物群落的影响几乎是未知的,需要用分子工具来研究。该研究的目的是研究生物光照如何影响微生物群体的结构,该方法采用非培养方法,使用直接从受感染底物中提取的DNA中pcr扩增的18S-ITS基因编码片段的DGGE进行评估。采用人工侵染法对基质样品进行粘接镰刀菌(Fusarium oxysporum f.sp. conglutinans, FOC)和尖孢镰刀菌(f.s oxysporum f.sp)侵染。采用非培养方法评价巴西耳霉(basilici, FOB)真菌种群的变化。夏季期间,在意大利北部的一个温室中,用透明聚乙烯薄膜进行了日光照射,并与无脂肪的油菜籽粕和/或堆肥结合或不结合。在生长室中进行生物光照处理,在最优温度(55-52℃6小时,50-48℃8小时,47-45℃10小时/天)和次优温度(50-48℃20小时,45-43℃8小时,40-38℃10小时/天)下加热底物7天和14天。通过平板计数和聚合酶链反应变性梯度凝胶电泳(PCR-DGGE)分析来评估生物光照对微生物种群的影响。由于生物光照,FOC和FOB的丰度降低,而实验期间细菌数量比对照样品高。从侵染底物样品中直接提取的DNA中获得子囊菌群落的PCR-DGGE指纹图谱,表明有机修饰的使用增加了真菌群体的相似性。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
EFFECT OF BIOSOLARISATION ON THE MICROBIAL POPULATIONS OF SUBSTRATES INFESTED WITH FUSARIUM OXYSPORUM BY PCR-DGGE.

Biosolarisation consists of combining solarisation and organic matter application for controlling soilborne pathogens. The effects of this control strategy on the microbial community is almost unknown and needs to be investigated with molecular tools. The aim of the research was to investigate how biosolarisation can affect the structure of the microbial populations evaluated by a culture independent method using DGGE of PCR-amplified 18S-ITS genes-coding fragments from DNA extracted directly from infested substrate. Substrate samples were artificially infested with Fusarium oxysporum f. sp. conglutinans (FOC) and F. oxysporum f.sp. basilici (FOB) in order to evaluate the shift in fungal population by using culture independent methods. Solarisation was carried out with transparent polyethylene film during the summer period in a greenhouse located in Northern Italy, in combination or not with Brassica carinata defatted seed meals and/or compost. Biosolarisation treatment was carried out in a growth chamber by heating the substrate for 7 and 14 days at optimal (55-52 degrees C for 6 h, 50-48 degrees C for 8 h and 47-45 degrees C for 10 h/day) and sub-optimal (50-48 degrees C for 20 h, 45-43 degrees C for 8 h and 40-38 degrees C for 10 h/day) temperatures. Plate counts and polymerase chain reaction denaturing gradient gel electrophoresis (PCR-DGGE) analyses were performed to evaluate the effect of biosolarisation on the microbial population. The abundance of FOC and FOB was reduced as a consequence of biosolarisation, while bacterial populations were higher compared to control samples during the experiment. PCR-DGGE fingerprints of the ascomycete community obtained from DNA directly extracted from infested substrate samples showed that the use of organic amendments increased the similarity of the fungal populations.

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