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ZJUS-B
Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology)  2014, Vol. 15 Issue (11): 966-978    DOI: 10.1631/jzus.B1400063
Articles     
Designing a SCAR molecular marker for monitoring Trichoderma cf. harzianum in experimental communities
Gabriel Pérez, Valentina Verdejo, Clarissa Gondim-Porto, Julieta Orlando, Margarita Carú
Department of Ecological Sciences, Faculty of Sciences, University of Chile, Santiago, Chile
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Abstract  Several species of the fungal genus Trichoderma establish biological interactions with various micro- and macro-organisms. Some of these interactions are relevant in ecological terms and in biotechnological applications, such as biocontrol, where Trichoderma could be considered as an invasive species that colonizes a recipient community. The success of this invasion depends on multiple factors, which can be assayed using experimental communities as study models. Therefore, the aim of this work is to develop a species-specific sequence-characterized amplified region (SCAR) marker to monitor the colonization and growth of T. cf. harzianum when it invades experimental communities. For this study, 16 randomly amplified polymorphic DNA (RAPD) primers of 10-mer were used to generate polymorphic patterns, one of which generated a band present only in strains of T. cf. harzianum. This band was cloned, sequenced, and five primers of 20鈥?3 mer were designed. Primer pairs 2F2/2R2 and 2F2/2R3 successfully and specifically amplified fragments of 278 and 448 bp from the T. cf. harzianum BpT10a strain DNA, respectively. Both primer pairs were also tested against the DNA from 14 strains of T. cf. harzianum and several strains of different fungal genera as specificity controls. Only the DNA from the strains of T. cf. harzianum was successfully amplified. Moreover, primer pair 2F2/2R2 was assessed by quantitative real-time polymerase chain reaction (PCR) using fungal DNA mixtures and DNA extracted from fungal experimental communities as templates. T. cf. harzianum was detectable even when as few as 100 copies of the SCAR marker were available or even when its population represented only 0.1% of the whole community.

Key words： Trichoderma cf. harzianum      Sequence-characterized amplified region (SCAR)      Molecular marker      Figs. S1-S5')" href="#">Experimental fungal communities

The online version of this article contains supplementary materials Figs. S1-S5      FASTAs S1-S2      Table S1

     
Received: 03 March 2014      Published: 04 November 2014
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Articles by authors
Gabriel Pérez
Valentina Verdejo
Clarissa Gondim-Porto
Julieta Orlando
Margarita Carú
Cite this article:

Gabriel Pérez, Valentina Verdejo, Clarissa Gondim-Porto, Julieta Orlando, Margarita Carú. Designing a SCAR molecular marker for monitoring Trichoderma cf. harzianum in experimental communities. Journal of Zhejiang University-SCIENCE B (Biomedicine & Biotechnology), 2014, 15(11): 966-978.

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http://www.zjujournals.com/xueshu/zjus-b/10.1631/jzus.B1400063     OR     http://www.zjujournals.com/xueshu/zjus-b/Y2014/V15/I11/966

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