Molecular Characterization of Fungal Communities in Non-Tilled, Cover-Cropped Upland Rice Field Soils
Molecular Characterization of Fungal Communities in Non-Tilled, Cover-Cropped Upland Rice Field Soils
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DOI：
10.1264/jsme2.me10108
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发表时间：
2010-09-03
影响因子：
2.2
通讯作者：
Ohta, Hiroyuki
中科院分区：
文献类型：
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作者：
Nishizawa, Tomoyasu;Zhaorigetu;Ohta, Hiroyuki
本研究旨在利用基于PCR的分子生物学方法，对翻耕/免耕和冬季覆盖栽培（毛箭豌豆和黑麦）条件下高地田土壤真菌群落特征进行研究。研究地块保持5年的高地和土壤采样在第二和第五年进行了分析，使用T-RFLP（末端限制性片段长度多态性）分析和克隆库的内部转录间隔区（ITS）区域和结构域1（D1）的真菌大亚基（fLSU）rRNA（D1（fLSU））作为目标DNA序列。从第二年的样本中，获得了372个真菌ITS-D1（fLSU）克隆序列，并在4个真菌门中聚类成80个非冗余真菌OTU（操作分类单位）。对第2年和第5年的样本进行T-RFLP分析，并使用ITS-D1（fLSU）克隆的理论片段分析鉴定主要T-RF（末端限制性片段）。这些分子生物学分析表明，覆盖种植比耕作措施对真菌群落的影响更大。此外，非耕作，覆盖种植土壤的特点是隐球菌属的优势。在门担子菌。我们提供了一个遗传数据库的真菌ITS-D1（fLSU）在不同管理的高地田土壤。
This study aimed to characterize soil fungal communities in upland rice fields managed with tillage/non-tillage and winter cover-cropping (hairy vetch and cereal rye) practices, using PCR-based molecular methods. The study plots were maintained as upland fields for 5 years and the soils sampled in the second and fifth years were analyzed using T-RFLP (terminal restriction fragment length polymorphism) profiling and clone libraries with the internal transcribed spacer (ITS) region and domain 1 (D1) of the fungal large-subunit (fLSU) rRNA (D1(fLSU)) as the target DNA sequence. From the 2nd-year-sample, 372 cloned sequences of fungal ITS-D1(fLSU) were obtained and clustered into 80 non-redundant fungal OTUs (operational taxonomic units) in 4 fungal phyla. The T-RFLP profiling was performed with the 2nd- and 5th-year-samples and the major T-RFs (terminal restriction fragments) were identified using a theoretical fragment analysis of the ITS-D1(fLSU) clones. These molecular analyses showed that the fungal community was influenced more strongly by the cover-cropping than tillage practices. Moreover, the non-tilled, cover-cropped soil was characterized by a predominance of Cryptococcus sp. in the phylum Basidiomycota. We provided a genetic database of the fungal ITS-D1(fLSU)s in the differently managed soils of upland rice fields.