The Transformation in <i>Trichoderma viride</i> Protoplasts by Restriction Enzyme Mediated Integration (REMI)

Abstract

<jats:p>The transformation conditions of the protoplasts from Trichoderma viride mediated by restriction enzyme were studied in this paper. The optimum generation conditions of protoplasts were as followed: 8 mg/ml glucanex was added into the phosphate buffer (pH 6.98), the mycelial that cultured for 24 hr was hydrolyzed for 4 hr at 30°C under 40 r/min shaking speed. The protoplast yield was 4.7×107 cfu/mg. The regeneneration rate of protoplast was 14.5% on CM medium contained 0.3 mol/L KCl and 0.3 mol/L Inositol. Transformants were obtained by transfering hygromycin B resistance gene into T. viride by restriction enzyme mediated integration (REMI), The preliminary identification of the transformants indicated that the exogenous gene had been integrated into T. viride genome.</jats:p>

Construction of new GFP-tagged fusants for Trichoderma harzianum with enhanced biocontrol activity

“…The same finding has also been reported by Naseema et al (2008). The use of Novozym L1412 was the most advantageous and yields were higher than that reported elsewhere for Trichoderma species (Savitha et al 2010;Balasubramanian et al 2012;You et al 2012). However, by using this lytic system, the period required for protoplast isolation (2 h) showed a result lower or similar to that reported for the same and other related species (ranging from 2 to 4 h) (Balasubramanian et al 2003;Balasubramanian et al 2012).…”