| Title: | PCR primers comparisons for a successful Tuber spp. DNA region amplification in routine identifications |
|---|
| Authors: | ID Unuk Nahberger, Tina (Author)
ID Kraigher, Hojka (Author)
ID Grebenc, Tine (Author) |
|---|
| Files: |  PDF - Presentation file, download (6,81 MB)
MD5: 329A33D79FD56D910BC9412CC63BE443
 URL - Source URL, visit https://ojs.sazu.si/folia_bio_geo/article/view/7983
|
|---|
| Language: | English |
|---|
| Typology: | 1.01 - Original Scientific Article |
|---|
| Organization: |  SciVie - Slovenian Forestry Institute
|
|---|
| Abstract: | Since late 20th century DNA sequencing became the method of choice method in precision species identification. The ITS region is one of the official fungal barcoding DNA markers, although in some cases sequencing of the ITS re-gion may, due to misidentification, mislabeling or nomen-clature errors in public databases, lead to incorrect or insuf-ficient identification, as is currently a case in the genus Tu b e r. The aim of this study was to test, which ITS primer pairs are most appropriate and optimal for Tu b e r species DNA region amplification. Thereby we (1) compared ampli-fication success for different Tu b e r species using fungal spe-cific primer pair ITS1f and ITS4 and (2) compared amplifi-cation success using different ITS primer pair combinations in amplifying DNA region an example species Tuber aesti-vum. Based on results, Tuber aestivum was one of the most reluctant Tu b e r species in this study and in most cases failed to amplify with the above primer pair. After comparing dif-ferent ITS primer pairs, we conclude that the primer pair ITS5 and ITS7 is the most appropriate primer pair for ampli-fication DNA region of T. ae stiv um as it resulted in high am-plification success from ectomycorrhizal root tips. Based on sequences, gained from public databases, we found that ITS1f and ITS6 primers have a mismatch in one base pair compared to the target sequence of Tuber aestivum, thus re-sulting in poor or no amplification success. Although prim-er pair ITS5 and ITS7 in our study was proven to be the most appropriate primer pair in amplifying DNA region Tu b e r aestivum species, further analysis about appropriateness of it for a general barcoding and identification of ectomycorrhiza in complex community samples is needed. |
|---|
| Keywords: | Tuber spp., ITS region, PCR amplification, ITS primers |
|---|
| Publication status: | Published |
|---|
| Publication version: | Version of Record |
|---|
| Year of publishing: | 2020 |
|---|
| Number of pages: | str. 229-238 |
|---|
| Numbering: | Vol. 61, iss. 2 |
|---|
| PID: | 20.500.12556/DiRROS-12296  |
|---|
| UDC: | 630*17 |
|---|
| ISSN on article: | 1855-7996 |
|---|
| DOI: | 10.3986/fbg0076 |
|---|
| COBISS.SI-ID: | 21646339 |
|---|
| Publication date in DiRROS: | 30.07.2020 |
|---|
| Views: | 1610 |
|---|
| Downloads: | 1183 |
|---|
| Metadata: |     |
|---|
|
:
|
Copy citation |
|---|
| | | | Share: |  |
|---|
Hover the mouse pointer over a document title to show the abstract or click
on the title to get all document metadata. |
