宁夏枸杞内生真菌的分离及多样性分析

Separation and Diversity Analysis of Endophytic Fungi From Lycium barbarum L.in Ningxia

以枸杞为试材,采用组织分离法从不同品种、不同部位的枸杞组织中进行内生真菌的分离,根据培养性状、菌落、孢子等的形态特征和rDNA-ITS序列分析对分离菌株进行鉴定;根据枸杞内生真菌的相对频率、物种多样性指数、丰富度指数和相似性系数分析其组成特点。结果表明:21份枸杞样品中共分离得到363株内生真菌,这些菌株分属于链格孢属(Alternaria)、曲霉属(Aspergillus)、双极霉属(Bipolaris)、毛壳菌属(Chaetomium)、枝孢属(Cladosporium)、镰刀菌属(Fusarium)、青霉菌属(Penicillium)、光黑壳属(Preussia)、裂壳菌属(Schizothecium)、粪壳菌属(Sordaria)、戴氏霉属(Taifanglania)、梭孢壳属(Thielavia)、火丝菌属(Tricharina)和炭角菌属(Xylaria)14个属,其中链格孢属(Alternaria)、梭孢壳属(Thielavia)和曲霉属(Aspeirgllus)为优势属,分别占分离菌数的46%、14%和13%。枸杞植株各器官都有内生真菌分布,其中,茎部内生真菌多样性指数最大,为0.86,且果与茎部的相似性系数最大。不同枸杞品种间内生真菌分布存在差异,"宁杞1号""宁杞2号"内生真菌多样性指数和丰富度指数远远高于黄果枸杞和黑果枸杞。内生真菌群落组成的相似性结果表明,部分内生真菌具有一定的宿主和组织偏好性。枸杞体内含有丰富的内生真菌资源,其内生真菌具有很高的宿主特异性,而且其分布受生境影响。

Lycium barbarum L.was used as test material，endophytic fungi were separated from different varieties and different parts of Lycium barbarum L.by tissue separation.They were classified according to their morphological characteristics of culture，colonies and spores and rDNA-ITS sequences analysis.The composition characteristics of endophytic fungi were analyzed according to their colonization rate（CR），relative frequency（RF），shannon-weiner index（H），margalef index（D） and Sorenson′s similarity coefficient（CS）.The results showed that 363 endophytic fungi were isolated from L.barbarum L.，and they belonged to 14 genus，Alternaria，Aspergillus，Bipolaris,Chaetomium,Cladosporium,Fusarium,Penicillium,Preussia,Schizotheciu,Sordaria,Taifanglania,Thielavia,Tricharina,Xylaria，respectively. Alternaria，Thielavia and Aspeirgllus were dominant genus and their relative frequency（RF） were 46%，14% and 13% respectively.These endophytic fungi were distributed in various organs of L.barbarum L.The types of endophytic fungi in the stems were the most 0.86 in these organs of L.barbarum L.Sorenson′s similarity coefficient（CS）of leaf and fruit was the largest.The distribution of endophytic fungi was different between different varieties of Lycium barbarum L.Shannon-wiener index（H） and margalef index（D） of endophytic fungi from cultivated variety of L.barbarum L.，‘Ningqi 1’ ‘Ningqi 2’ were significantly higher than wild varieties，Lycianthes barbatum L.var.auranticarpum K.F.Ching and Lycium ruthenicum Murr.The community composition similarity of endophytic fungi showed that some endophytic fungi had the preference of a certain host and tissue.The endophytic fungi were rich in the resources of endophytic fungi，the endophytic fungi had high host specificity，and their distribution was influenced by the environment.