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Chilean Journal of Agricultural Research
Instituto de Investigaciones Agropecuarias, INIA
ISSN: 0718-5820
EISSN: 0718-5839
Vol. 71, No. 3, 2011, pp. 488-494
Bioline Code: cj11062
Full paper language: English
Document type: Note
Document available free of charge

Chilean Journal of Agricultural Research, Vol. 71, No. 3, 2011, pp. 488-494

 en MOLECULAR TOOLS FOR RAPID AND ACCURATE DETECTION OF BLACK TRUFFLE ( Tuber melanosporum check for this species in other resources Vitt.) IN INOCULATED NURSERY PLANTS AND COMMERCIAL PLANTATIONS IN CHILE
Cordero, Cecilia; Cáceres, Pablo; González, Gloria; Quiroz, Karla; Bravo, Carmen; Ramírez, Ricardo; Caligari, Peter D.S.; Carrasco, Basilio & García-Gonzales, Rolando

Abstract

Truffle ( Tuber melanosporum check for this species in other resources Vitt.) culture is an agroforestry sector in Chile of increasing interest due to the high prices that truffles fetch in the national market and the recent evidence that its commercial production is possible in Chilean climatic and soil conditions. In this study, the efficiency of three methods of DNA extraction from a mix of 5 g of soil and roots from both nursery and field plants of Quercus ilex check for this species in other resources L. mycorrhized with T. melanosporum were evaluated, and a simple and reproducible protocol was established. Detection of T. melanosporum was performed by the technique of cleaved amplified polymorphic sequence (CAPS) from amplicons generated with the primers ADL1 (5'-GTAACGATAAAGGCCATCTATAGG-3') and ADL3 (5'-CGTTTTTCCTGAACTCTTCATCAC-3`), where a restriction fragment of 160 bp specific for T. melanosporum was generated, which allows the discrimination of this species from the rest of the species belonging to the Tuber sp. genus. Direct detection of T. melanosporum in one step was also obtained by polymerase chain reaction (PCR) from total DNA isolated from mycorrhized roots and with the primers ITSML (5'-TGGCCATGTGTCAGATTTAGTA-3') and ITSLNG (5'-TGATATGCTTAAGTTCAGCGGG-3'), generating a single amplicon of 440 bp. The molecular detection of T. melanosporum by the methods presented here will allow the rapid and accurate detection of mycorrhization of trees, both under nursery and field conditions. This technology will also provide more security to farmers by controlling the quality of the mycorrhized trees they will plant and also by following the mycorrhization status of established orchards.

Keywords
Tuber melanosporum, ectomycorrhizas, molecular markers, PCR, CAPS

 
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