Combining nested PCR and restriction digest of the internal transcribed spacer region to characterize arbuscular mycorrhizal fungi on roots from the field

Mycorrhiza ◽  
2003 ◽  
Vol 13 (4) ◽  
pp. 191-198 ◽  
Author(s):  
Jochen Heinrichs ◽  
Michael Kaldorf ◽  
François Buscot ◽  
Carsten Renker
2012 ◽  
Vol 77 (1) ◽  
pp. 17-27 ◽  
Author(s):  
Anna Lisek ◽  
Lidia Sas Paszt ◽  
Beata Sumorok

Summary In organic farming, mineral fertilizers are replaced by various preparations to stimulate plant growth and development. Introduction of new biopreparations into horticultural production requires an assessment of their effects on the growth and yielding of plants. Among the important indicators of the impact on plants of beneficial microorganisms contained in bioproducts is determination of their effectiveness in stimulating the growth and yielding of plants. Moreover, confirmation of the presence of arbuscular mycorrhizal (AM) fungi in the roots and plant growth promoting rhizobacteria (PGPR) in the rhizosphere is also necessary. In addition to conventional methods, molecular biology techniques are increasingly used to allow detection and identification of AM fungi in plant roots. The aim of this study was identification and initial taxonomic classification of AM fungi in the roots of ‘Elkat’ strawberry plants fertilized with various biopreparations using the technique of nested PCR. Tests were performed on DNA obtained from the roots of ‘Elkat’ strawberry plants: not fertilized, treated with 10 different biopreparations, or fertilized with NPK. Amplification of the large subunit of ribosomal gene (LSU rDNA) was carried out using universal primers, and then, in the nested PCR reaction, primers specific for the fungi of the genera Glomus, Acaulospora, and Scutellospora were used. Colonization of strawberry roots by arbuscular mycorrhizal fungi was determined on the basis of the presence of DNA fragments of a size corresponding to the types of the fungi tested for. As a result of the analyses, the most reaction products characterizing AM fungi were found in the roots of plants treated with the preparation Florovit Eko. The least fragments characteristic of AM fungi were detected in the roots of plants fertilized with NPK, which confirms the negative impact of mineral fertilizers on the occurrence of mycorrhizal fungi in the roots of strawberry plants. The roots of plants fertilized with Tytanit differed from the control plants by the presence of one of the clusters of fungi of the genus Glomus and by the absence of a cluster of fungi of the genus Scutellospora. In the roots of plants treated with other biopreparations there were reaction products indicating the presence of fungi of the genera Glomus, Scutellospora and Acaulospora, like in the roots of the control plants. The results will be used to assess the suitability of microbiologically enriched biopreparations in horticultural production.


Phytotaxa ◽  
2021 ◽  
Vol 524 (3) ◽  
pp. 191-198
Author(s):  
RONG-JIAN HE ◽  
CHUN-LI LONG ◽  
LI-MEI YAO ◽  
LONG JIANG

Acaulospora fanjing is here described as a new species of arbuscular mycorrhizal fungi (AMF; Glomeromycota) based on morphological and phylogenetic analyses. It was associated with roots of Acer flabellatum colonizing of the massy dwarf forest to fanjing Mountain, China, and this is the only site of its occurrence known to date. A.fanjing is characterized by forming reddish orange to reddish brown globose spores of (109–)123(–138) µm diam and three layers of outer spore walls (SWL1–SWL3) and two layers of germinal walls (MW and IW). SWL1 hyaline, short–lived, evanescent, SWL2 laminate, orange to reddish orange, 2.0–3.5 μm thick and SWL3 thin, difficult to observe; MW is hyaline, bilayered (MWL1–MWL2), semiflexible; IW is hyaline, with three layers and not “beaded” structure, only the IWL2 were stained light pink in Melzer’s reagent. A.fanjing can be distinguished from all other Acaulospora species by spore size, color and spore wall structure. Phylogenetic analysis based on the AMF extended DNA barcode covering a 1.5-kb fragment of the small subunit (SSU), internal transcribed spacer region (ITS), and the large subunit (LSU) of rRNA genes places A.fanjing in the genus Acaulospora, separated from other described Acaulospora species. All available sequences in public databases suggest that this new fungal species has not yet been previously detected.


2008 ◽  
Vol 58 (2) ◽  
pp. 341-344 ◽  
Author(s):  
Adália Cavalcanti do Espírito Santo Mergulhão ◽  
Márcia Vanusa da Silva ◽  
Márcia do Vale Barreto Figueiredo ◽  
Hélio Almeida Burity ◽  
Leonor Costa Maia

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