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末龄棉铃虫幼虫对摄入Bt毒素的反应:发育及CYP6AE14、CYP6B2和CYP9A12基因表达的变化

Response of last instar Helicoverpa armígera larvae to Bt toxin ingestion: changes in the development and in the CYP6AE14, CYP6B2 and CYP9A12 gene expression.

作者信息

Muñoz Pilar, López Carmen, Moralejo Marian, Pérez-Hedo Meritxell, Eizaguirre Matilde

机构信息

Agrotecnio Center, Universitat de Lleida, Lleida, Spain.

Department of Chemistry, Universitat de Lleida, Lleida, Spain.

出版信息

PLoS One. 2014 Jun 9;9(6):e99229. doi: 10.1371/journal.pone.0099229. eCollection 2014.

Abstract

Bt crops are able to produce Cry proteins, which were originally present in Bacillus thuringiensis bacteria. Although Bt maize is very efficient against corn borers, Spanish crops are also attacked by the earworm H. armigera, which is less susceptible to Bt maize. Many mechanisms could be involved in this low susceptibility to the toxin, including the insect's metabolic resistance to toxins due to cytochrome P450 monooxygenases. This paper examines the response of last instar H. armigera larvae to feeding on a diet with Bt and non-Bt maize leaves in larval development and in the gene expression of three P450 cytochromes: CYP6AE14, CYP6B2 and CYP9A12. Larvae fed on sublethal amounts of the Bt toxin showed reduced food ingestion and reduced growth and weight, preventing most of them from achieving the critical weight and pupating; additionally, after feeding for one day on the Bt diet the larvae showed a slight increase in juvenile hormone II in the hemolymp. Larvae fed on the non-Bt diet showed the highest CYP6AE14, CYP6B2 and CYP9A12 expression one day after feeding on the non-Bt diet, and just two days later the expression decreased abruptly, a finding probably related to the developmental programme of the last instar. Moreover, although the response of P450 genes to plant allelochemicals and xenobiotics has been related in general to overexpression in the resistant insect, or induction of the genes when feeding takes place, the expression of the three genes studied was suppressed in the larvae feeding on the Bt toxin. The unexpected inhibitory effect of the Cry1Ab toxin in the P450 genes of H. armigera larvae should be thoroughly studied to determine whether this response is somehow related to the low susceptibility of the species to the Bt toxin.

摘要

转基因抗虫作物能够产生Cry蛋白,该蛋白最初存在于苏云金芽孢杆菌中。尽管转基因抗虫玉米对玉米螟非常有效,但西班牙的作物也会受到棉铃虫的侵害,而棉铃虫对转基因抗虫玉米的敏感性较低。这种对毒素低敏感性可能涉及多种机制,包括昆虫因细胞色素P450单加氧酶对毒素产生代谢抗性。本文研究了棉铃虫末龄幼虫取食含转基因抗虫玉米叶片和非转基因抗虫玉米叶片的饲料后,在幼虫发育以及三种P450细胞色素(CYP6AE14、CYP6B2和CYP9A12)基因表达方面的反应。取食亚致死量转基因抗虫毒素的幼虫食物摄入量减少,生长和体重下降,大多数幼虫无法达到临界体重并化蛹;此外,在取食转基因抗虫饲料一天后,幼虫血淋巴中的保幼激素II略有增加。取食非转基因抗虫饲料的幼虫在取食非转基因抗虫饲料一天后CYP6AE14、CYP6B2和CYP9A12的表达最高,而仅两天后表达就突然下降,这一发现可能与末龄幼虫的发育程序有关。此外,虽然一般认为P450基因对植物化感物质和外源化合物的反应与抗性昆虫中的过表达或取食时基因的诱导有关,但在取食转基因抗虫毒素的幼虫中,所研究的三个基因的表达受到抑制。应深入研究Cry1Ab毒素对棉铃虫幼虫P450基因的意外抑制作用,以确定这种反应是否与该物种对转基因抗虫毒素的低敏感性有关。

https://cdn.ncbi.nlm.nih.gov/pmc/blobs/bc2f/4049824/c5086d85ae4b/pone.0099229.g001.jpg

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