Ding X, Gopalakrishnan B, Johnson L B, White F F, Wang X, Morgan T D, Kramer K J, Muthukrishnan S
Department of Plant Pathology, Kansas State University, Manhattan 66506-5502, USA.
Transgenic Res. 1998 Mar;7(2):77-84. doi: 10.1023/a:1008820507262.
Chitinase expression in the insect gut normally occurs only during moulting, where the chitin of the peritrophic membrane is presumably degraded. Thus, insects feeding on plants that constitutively express an insect chitinase gene might be adversely affected, owing to an inappropriately timed exposure to chitinase. This hypothesis was tested by introducing a cDNA encoding a tobacco hornworm (Manduca sexta) chitinase (EC 3.2.1.14) into tobacco via Agrobacterium tumefaciens-mediated transformation. A truncated but enzymatically active chitinase was present in plants expressing the gene. Segregating progeny of high-expressing plants were compared for their ability to support growth of tobacco budworm (Heliothis virescens) larvae and for feeding damage. Both parameters were significantly reduced when budworms fed on transgenic tobacco plants expressing high levels of the chitinase gene. In contrast, hornworm larvae showed no significant growth reduction when fed on the chitinase-expressing transgenics. However, both budworm and hornworm larvae, when fed on chitinase-expressing transgenic plants coated with sublethal concentrations of a Bacillus thuringiensis toxin, were significantly stunted relative to larvae fed on toxin-treated non-transgenic controls. Foliar damage was also reduced. Plants expressing an insect chitinase gene may have agronomic potential for insect control.
几丁质酶在昆虫肠道中的表达通常仅在蜕皮期间发生,在此期间围食膜的几丁质可能会被降解。因此,取食组成型表达昆虫几丁质酶基因的植物的昆虫,可能会因不恰当地接触几丁质酶而受到不利影响。通过根癌农杆菌介导的转化将编码烟草天蛾(烟草天蛾)几丁质酶(EC 3.2.1.14)的cDNA导入烟草,对这一假设进行了验证。在表达该基因的植物中存在一种截短但具有酶活性的几丁质酶。比较了高表达植物的分离后代支持烟草夜蛾幼虫生长的能力和取食损伤情况。当烟草夜蛾取食表达高水平几丁质酶基因的转基因烟草植株时,这两个参数均显著降低。相比之下,烟草天蛾幼虫取食表达几丁质酶的转基因植株时,生长没有显著减少。然而,当烟草夜蛾和烟草天蛾幼虫取食涂有亚致死浓度苏云金芽孢杆菌毒素的表达几丁质酶的转基因植物时,相对于取食毒素处理的非转基因对照的幼虫,它们明显发育不良。叶片损伤也减少了。表达昆虫几丁质酶基因的植物可能具有用于昆虫防治的农艺潜力。