Differential effect of sample preservation methods on plant and arbuscular mycorrhizal fungal DNA

被引:45
|
作者
Bainard, L. D. [1 ]
Klironomos, J. N. [2 ]
Hart, M. M. [2 ]
机构
[1] Univ Guelph, Dept Integrat Biol, Guelph, ON N1G 2W1, Canada
[2] Univ British Columbia, Biol & Phys Geog Unit, Kelowna, BC V1V 1V7, Canada
基金
加拿大自然科学与工程研究理事会;
关键词
Arbuscular mycorrhizal fungi; DNA preservation; Real-time PCR; Glomus intraradices; FIELD PRESERVATION; COMMUNITY STRUCTURE; MOLECULAR-IDENTIFICATION; GLOMUS-INTRARADICES; DIVERSITY; ROOTS; GLOMEROMYCOTA; COLONIZATION; EXTRACTION; SOILS;
D O I
10.1016/j.mimet.2010.05.001
中图分类号
Q5 [生物化学];
学科分类号
071010 ; 081704 ;
摘要
A wide range of methods are commonly used for preserving environmental samples prior to molecular analyses. However, the effect of these preservation methods on fungal DNA is not understood. The objective of this study was to test the effect of eight different preservation methods on the quality and yield of DNA extracted from Bromus inermis and Daucus carota roots colonized by the arbuscular mycorrhizal (AM) fungus, Glomus intraradices. The total DNA concentration in sample extracts was quantified using spectrophotometry. Samples that were frozen (-80 degrees C and -20 degrees C), stored in 95% ethanol, or silica gel dried yielded total (plant and fungal) DNA concentrations that were not significantly different from fresh samples. In contrast, samples stored in CTAB solution or freeze-dried resulted in significantly reduced DNA concentrations compared with fresh samples. The preservation methods had no effect on the purity of the sample extracts for both plant species. However, the DNA of the dried samples (silica gel dried, freeze-dried, heat dried) appeared to be slightly more degraded compared with samples that remained hydrated (frozen, stored in ethanol or CTAB solutions) during storage when visualized on a gel. The concentration of AM fungal DNA in sample extracts was quantified using TaqMan real time PCR. Methods that preserved samples in hydrated form had similar AM fungal DNA concentrations as fresh samples, except D. carota samples stored in ethanol. In contrast, preservation methods that involved drying the samples had very low concentrations of AM fungal DNA for B. inermis, and nearly undetectable for D. carota samples. The drying process appears to be a major factor in the degradation of AM fungal DNA while having less of an impact on plant DNA. Based on these results, samples that need to be preserved prior to molecular analysis of AM fungi should be kept frozen to minimize the degradation of plant and AM fungal DNA. (C) 2010 Elsevier B.V. All rights reserved.
引用
收藏
页码:124 / 130
页数:7
相关论文
共 50 条
  • [1] Sample preservation methods impact arbuscular mycorrhizal DNA recovery from sugarcane root tissue
    Claassens, Anders
    Nock, Catherine J.
    Rose, Michael T.
    Van Zwieten, Lukas
    Rose, Terry J.
    RHIZOSPHERE, 2022, 22
  • [2] Quantification of arbuscular mycorrhizal fungal DNA in roots: how important is material preservation?
    Janouskova, Martina
    Pueschel, David
    Hujslova, Martina
    Slavikova, Renata
    Jansa, Jan
    MYCORRHIZA, 2015, 25 (03) : 205 - 214
  • [3] Quantification of arbuscular mycorrhizal fungal DNA in roots: how important is material preservation?
    Martina Janoušková
    David Püschel
    Martina Hujslová
    Renata Slavíková
    Jan Jansa
    Mycorrhiza, 2015, 25 : 205 - 214
  • [4] Differential decomposition of arbuscular mycorrhizal fungal hyphae and glomalin
    Steinberg, PD
    Rillig, MC
    SOIL BIOLOGY & BIOCHEMISTRY, 2003, 35 (01): : 191 - 194
  • [5] Independent and differential effects of arbuscular mycorrhizal fungal composition and plant pathogens on plant traits and nitrogen uptake
    Zhang, Li
    Rong, Yizhong
    Zhang, Peng
    Lin, Ziyuan
    Hu, Kui
    Wang, Xingxing
    Liu, Xiang
    Liu, Mu
    PROCEEDINGS OF THE ROYAL SOCIETY B-BIOLOGICAL SCIENCES, 2024, 291 (2037)
  • [6] Fungal and plant gene expression in arbuscular mycorrhizal symbiosis
    Balestrini, Raffaella
    Lanfranco, Luisa
    MYCORRHIZA, 2006, 16 (08) : 509 - 524
  • [7] Fungal and plant gene expression in arbuscular mycorrhizal symbiosis
    Raffaella Balestrini
    Luisa Lanfranco
    Mycorrhiza, 2006, 16 : 509 - 524
  • [8] Arbuscular mycorrhizal fungal communities in plant roots are not random assemblages
    Davison, John
    Oepik, Maarja
    Daniell, Tim J.
    Moora, Mari
    Zobel, Martin
    FEMS MICROBIOLOGY ECOLOGY, 2011, 78 (01) : 103 - 115
  • [9] The Costs and Benefits of Plant-Arbuscular Mycorrhizal Fungal Interactions
    Bennett, Alison E.
    Groten, Karin
    ANNUAL REVIEW OF PLANT BIOLOGY, 2022, 73 : 649 - 672
  • [10] Host plant height explains the effect of nitrogen enrichment on arbuscular mycorrhizal fungal communities
    Cheng, Yikang
    Rutten, Gemma
    Liu, Xiang
    Ma, Miaojun
    Song, Zhiping
    Maaroufi, Nadia I. I.
    Zhou, Shurong
    NEW PHYTOLOGIST, 2023, 240 (01) : 399 - 411