中文摘要：

采集巨桉林下马勃子实体,培养其菌丝体,提取子实体及菌丝体基因组DNA,进行rDNA-ITS区序列的扩增、克隆和序列测定,并对rDNA-ITS不同区域作序列分析,首次构建马勃的系统发育树。结果表明：野外采集获得马勃子实体10种,其中成功培养6种,测序结果表明马勃rDNA-ITS区长度在607～766bp之间,系统发育分析表明硬皮马勃属（Scleroderma）与豆包菌（Pisolithus）亲缘关系较近,秃马勃属（Calvatia）、马勃属（Lycoperdon）及横膜马勃属（Vascellum）之间亲缘关系较近,ITS1-5.8S rDNA-ITS2区可建立马勃类真菌属间系统发育树,ITS2区可用于建立马勃类真菌属内系统发育树,3个待定种硬皮马勃Scleroderma sp.11-1,Scleroderma sp.2-2和Scleroderma sp.5-2为金黄硬皮马勃（S.aurantium）的可能性较大。此研究可为探讨巨桉人工林下外生菌根种类与作用机制、马勃分类系统学及菌丝体的开发研究奠定基础。

英文摘要：

Mycelia were cultivated from a fruiting body of puffballs which were collected from the Eucalyptus grandis forest,and the genomic DNA was extracted as template.Subsequently,the internal transcribed spacer（ITS） region in rDNA gene was amplified,cloned and sequenced.The phylogenetic tree of puffballs was first built based on different part of ITS region.The results showed that six species of puffballs were cultivated successfully using a PDA medium after comparing the sequences of puffballs fruiting body and mycelium ITS regions.The rDNA-ITS region of different puffballs was 607 bp to 766 bp in length.Analysis of phylogenesis revealed that the Scleroderma had close genetic relationships with Pisolithus,Calvatia,Lycoperdon and Vascellum.The three undetermined species,Scleroderma sp.11-1,Scleroderma sp.2-2 and Scleroderma sp.5-2,could probably be S.aurantium.The sequence of ITS1-5.8S rDNA-ITS2 can be used as an evolutionary marker between different genus,and ITS2 as an evolutionary marker in genus.Thus,this work laid a foundation for studying the ectotrophic mycorrhiza,the mechanisms of action,the classification of puffballs and its exploitation and utilization.