Allen Tamara R, Millar Tony, Berch Shannon M, Berbee Mary L
Department of Botany, The University of British Columbia, Vancouver BC, V6T 1Z4, Canada.
Ministry of Forestry, Research Branch Laboratory, 4300 North Road, Victoria, BC V8Z 5J3, Canada.
New Phytol. 2003 Oct;160(1):255-272. doi: 10.1046/j.1469-8137.2003.00885.x.
• This study compares DNA and culture-based detection of fungi from 15 ericoid mycorrhizal roots of salal (Gaultheria shallon), from Vancouver Island, BC Canada. • From the 15 roots, we PCR amplified fungal DNAs and analyzed 156 clones that included the internal transcribed spacer two (ITS2). From 150 different subsections of the same roots, we cultured fungi and analyzed their ITS2 DNAs by RFLP patterns or sequencing. We mapped the original position of each root section and recorded fungi detected in each. • Phylogenetically, most cloned DNAs clustered among Sebacina spp. (Sebacinaceae, Basidiomycota). Capronia sp. and Hymenoscyphus erica (Ascomycota) predominated among the cultured fungi and formed intracellular hyphal coils in resynthesis experiments with salal. • We illustrate patterns of fungal diversity at the scale of individual roots and compare cloned and cultured fungi from each root. Indicating a systematic culturing detection bias, Sebacina DNAs predominated in 10 of the 15 roots yet Sebacina spp. never grew from cultures from the same roots or from among the > 200 ericoid mycorrhizal fungi previously cultured from different roots from the same site.
• 本研究比较了从加拿大不列颠哥伦比亚省温哥华岛的15株越橘(Gaultheria shallon)石南型菌根根段中基于DNA和培养法检测真菌的情况。
• 从这15个根段中,我们通过PCR扩增真菌DNA,并分析了156个包含内转录间隔区2(ITS2)的克隆。从同一根段的150个不同亚段中,我们培养真菌,并通过限制性片段长度多态性(RFLP)模式或测序分析其ITS2 DNA。我们绘制了每个根段的原始位置,并记录了在每个根段中检测到的真菌。
• 在系统发育上,大多数克隆的DNA聚集在角担菌属(Sebacina spp.,角担菌科,担子菌门)中。卡普龙菌属(Capronia sp.)和石南状膜盘菌(Hymenoscyphus erica,子囊菌门)在培养的真菌中占主导地位,并在与越橘的再合成实验中形成细胞内菌丝线圈。
• 我们展示了单个根段尺度上的真菌多样性模式,并比较了每个根段中克隆的真菌和培养的真菌。表明存在系统性的培养检测偏差,角担菌属的DNA在15个根段中的10个中占主导地位,但角担菌属从未从相同根段的培养物中生长出来,也未从之前从同一地点不同根段培养的200多种石南型菌根真菌中生长出来。