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Community analysis of arbuscular mycorrhizal fungi in a warm-temperate deciduous broad-leaved forest and introduction of the fungal community into the seedlings of indigenous woody plants

机译:温带落叶阔叶林中丛枝菌根真菌的群落分析及将真菌群落引入本土木本植物的幼苗中

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A community of arbuscular mycorrhizal (AM) fungi was investigated in a warm-temperate deciduous broad-leaved forest using a molecular analysis method. Root samples were obtained from the forest, and DNA was extracted from the samples. Partial 18S rDNA of AM fungi were amplified from the extracted DNA by polymerase chain reaction using a universal eukaryotic primer NS31 and an AM fungal-specific primer AM1. After cloning the PCR products, 394 clones were obtained in total, which were divided into five types by restriction fragment length polymorphism (RFLP) with HinfI, RsaI, and Hsp92II. More than 20% of the clones were randomly selected from each RFLP type and sequenced. Phylogenetic analysis showed that all the obtained clones belonged to Glomus but could not be identified at species level. Topsoil of the forest containing plant roots was inoculated to nonmycorrhizal seedlings of indigenous woody plants, Rhus javanica var. roxburghii and Clethra barvinervis, to introduce the community of AM fungi into the seedlings. Among these five RFLP types, four types were detected from both seedlings, which indicates that the AM fungal community in the forest root samples was introduced at least partly into the seedlings. Meanwhile, an additional four types that were not found in the forest root samples were newly detected in the seedlings, these types were closely related to one another and close to G. fasciculatum or G. intraradices. It is expected that a community of indigenous diverse AM fungi could be introduced into target fields by planting these mycorrhizal seedlings.
机译:使用分子分析方法,在温带落叶阔叶林中调查了丛枝菌根(AM)真菌群落。从森林中获取根样品,并从样品中提取DNA。使用通用的真核引物NS31和AM真菌特异性引物AM1,通过聚合酶链反应从提取的DNA中扩增出AM真菌的部分18S rDNA。克隆PCR产物后,总共获得394个克隆,通过具有HinfI,RsaI和Hsp92II的限制性片段长度多态性(RFLP)将其分为五种类型。从每种RFLP类型中随机选择20%以上的克隆并进行测序。系统发育分析表明,所有获得的克隆均属于Glomus,但无法在物种水平上鉴定。将含有植物根的森林表土接种到本地木本植物Rhus javanica var的非菌根幼苗上。 roxburghii和Clethra barvinervis,将AM真菌群落引入幼苗。在这五种RFLP类型中,从两种幼苗中均检测到四种类型,这表明森林根部样品中的AM真菌群落至少部分引入了幼苗。同时,在幼苗中新发现了在森林根样品中未发现的另外四种类型,这些类型彼此密切相关并且与fasciculatum或G. intraradices接近。期望通过种植这些菌根苗,可以将土著的多种AM真菌群落引入目标田地。

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