Institute of Plant Protection, Shandong Agricultural University, Taian 271018, China.
Biotechnol Lett. 2012 Oct;34(10):1921-8. doi: 10.1007/s10529-012-0989-1. Epub 2012 Jul 4.
Chitinases produced by mycoparasites play an important role in disease control in plants. To explore the functions of chitinases in Trichothecium roseum, we cloned a new chitinase gene named Trchi1 from T. roseum by RT (reverse transcription)-PCR techniques. The T. roseum gene, Trchi1, contains an 1278-bp ORF that shares 76 % similarity with chitinase from Bionectria ochroleuca (ABV57861 3G6L_A). A plant expression vector, containing the Trchi1 gene driven by the CaMV35S promoter, was constructed and transformed into tobacco via Agrobacterium tumefaciens. Southern blot analysis showed that Trchi1 was integrated into the tobacco genome. Total chitinase activity in Trchi1-transgenic tobacco leaves was enhanced 2.2- to 5.8- times with respect to non-transgenic leaves. Transgenic tobacco plants transformed with the Trchi1 gene had increased resistance to Alternaria alternata and Colletotrichum nicotianae.
真菌寄生物产生的几丁质酶在植物疾病控制中发挥着重要作用。为了探索玫瑰座腔菌（Trichothecium roseum）中几丁质酶的功能，我们通过 RT-PCR 技术从该菌中克隆了一个名为 Trchi1 的新几丁质酶基因。该基因 Trchi1 含有一个 1278bp 的开放阅读框，与层出镰刀菌（Bionectria ochroleuca，ABV57861 3G6L_A）的几丁质酶有 76%的相似性。我们构建了一个植物表达载体，其中包含由 CaMV35S 启动子驱动的 Trchi1 基因，并通过根癌农杆菌（Agrobacterium tumefaciens）将其转化为烟草。Southern blot 分析表明，Trchi1 已整合到烟草基因组中。与非转基因叶片相比，转 Trchi1 基因烟草叶片中的总几丁质酶活性提高了 2.2-5.8 倍。转 Trchi1 基因的烟草植株对Alternaria alternata 和 Colletotrichum nicotianae 的抗性增强。
