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大豆胰蛋白酶抑制剂N对南方玉米根虫幼虫的转录调控

Transcriptional regulation in southern corn rootworm larvae challenged by soyacystatin N.

作者信息

Liu Yilin, Salzman Ron A, Pankiw Tanya, Zhu-Salzman Keyan

机构信息

Department of Entomology, Texas A&M University, College Station, TX 77843, USA.

出版信息

Insect Biochem Mol Biol. 2004 Oct;34(10):1069-77. doi: 10.1016/j.ibmb.2004.06.010.

Abstract

When fed on a diet containing a soybean cysteine protease inhibitor soyacystatin N (scN), southern corn rootworm larvae exhibited increased mortality and reduced growth rate. scN impacted mortality in a dose-dependent manner, and its effect on insect growth was more severe at early developmental stages. Insects that survived from continuous exposure to the inhibitor at doses ranging from 0.1% to 0.5% had less reduction in body weight during later developmental stages. This insensitivity as insects developed was not observed in the insect group fed on diet containing 0.05% scN, the lowest dose tested. Thus, individuals that survived the higher dose treatments may have had higher fitness under dietary inhibitory challenge. Subtractive hybridization and cDNA microarray analyses identified 29 transcript species responsive to scN. Southern corn rootworm larvae over-expressed cysteine and aspartic proteases to compensate for inhibition of digestion. Induction of a peritrophin gene suggested that strengthening the peritrophic membrane plays a role in coping with protease inhibitors. scN down-regulated genes encoding proteins involved in insect metabolism and development, reflecting the insect's ability to reallocate resources to prioritize its defense response. Further, protease and the peritrophin genes were also developmentally regulated, which may explain the lower toxicity in older larvae than in neonates when first encountering dietary scN. Multiple regulatory mechanisms of counter defense-related genes may allow insects to evade the effect of plant defense proteins, and impose an obstacle to biotechnology-based insect control.

摘要

以含有大豆半胱氨酸蛋白酶抑制剂大豆胱抑素N(scN)的饲料喂养时,南方玉米根虫幼虫的死亡率增加,生长速率降低。scN以剂量依赖的方式影响死亡率,并且其对昆虫生长的影响在发育早期更为严重。在0.1%至0.5%剂量范围内持续接触该抑制剂后存活下来的昆虫,在后期发育阶段体重减轻较少。在以含有0.05% scN(测试的最低剂量)的饲料喂养的昆虫组中,未观察到随着昆虫发育而出现的这种不敏感性。因此,在较高剂量处理下存活下来的个体在饮食抑制挑战下可能具有更高的适应性。消减杂交和cDNA微阵列分析鉴定出29种对scN有反应的转录本。南方玉米根虫幼虫过表达半胱氨酸和天冬氨酸蛋白酶以补偿消化抑制。一种围食膜蛋白基因的诱导表明,强化围食膜在应对蛋白酶抑制剂方面发挥作用。scN下调了编码参与昆虫代谢和发育的蛋白质的基因,这反映了昆虫重新分配资源以优先进行防御反应的能力。此外,蛋白酶和围食膜蛋白基因也受到发育调控,这可能解释了老龄幼虫在首次接触饲料中的scN时比新生幼虫毒性更低的原因。与防御相关基因的多种调控机制可能使昆虫逃避植物防御蛋白的影响,并给基于生物技术的昆虫控制带来障碍。

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