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绿色木霉LTR-2是生物防治菌株。利用来自巨大芽胞杆菌Ap25的β-1,4-葡聚糖酶基因glu14构建木霉表达载体pSilent/glu14,利用限制性内切酶介导法(REMI)转化绿色木霉LTR-2。PCR扩增及Southern杂交证实目的基因已插入木霉转化子的染色体DNA上。转化子的β-1,4-葡聚糖酶水解活性,对小麦纹枯病菌的平板抑制作用及温室防治效果较原始菌株LTR-2明显提高(P<0.01),其中转化子L-10的效果最好,平板抑制率比LTR-2提高了27.0%,温室防治效果比LTR-2提高了26.7%。本试验表明,利用REMI技术,将β-1,4-葡聚糖酶基因重组到木霉染色体DNA上,是获得高效木霉工程菌株的有效手段。
Trichoderma viride LTR-2 is a biocontrol strain. The Trichoderma expression vector pSilent / glu14 was constructed using the β-1,4-glucanase gene glu14 from Bacillus megaterium Ap25 and the Trichoderma viride LTR-2 was transformed by restriction endonuclease-mediated method (REMI). PCR amplification and Southern blot confirmed that the target gene has been inserted into the chromosomal DNA of Trichoderma transformants. The β-1,4-glucanase hydrolyzate activity of transformant was significantly higher than that of LTR-2 (P <0.01), and the transformant L-10 The best effect was obtained. The inhibition rate of plate was 27.0% higher than that of LTR-2 and 26.7% higher than that of LTR-2. This experiment shows that the use of REMI technology, the β-1,4-glucanase gene recombined on the chromosome DNA of Trichoderma is to obtain efficient strains of Trichoderma effective means.