Reduced levels of membrane-bound alkaline phosphatase are common to lepidopteran strains resistant to Cry toxins from Bacillus thuringiensis.
Reduced levels of membrane-bound alkaline phosphatase are common to lepidopteran strains resistant to Cry toxins from Bacillus thuringiensis.
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DOI：
10.1371/journal.pone.0017606
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发表时间：
2011-03-01
期刊：
影响因子：
3.7
通讯作者：
Adang M
中科院分区：
文献类型：
--
作者：
Jurat-Fuentes JL;Karumbaiah L;Jakka SR;Ning C;Liu C;Wu K;Jackson J;Gould F;Blanco C;Portilla M;Perera O;Adang M
昆虫抗性的发展是使用表达来自细菌苏云金芽孢杆菌的Cry毒素的转基因作物的主要关注点之一。生物标志物的鉴定将有助于发展灵敏的基于DNA的方法来监测自然种群对Bt毒素的抗性进化。我们报告的蛋白质组学和基因组检测中肠膜结合碱性磷酸酶（mALP）的水平降低作为一个共同的特点，在菌株抗冷烟夜蛾，棉铃虫和草地贪夜蛾相比，敏感的幼虫。降低H。采用双向差异凝胶电泳（2D-DIGE）技术检测了抗虫幼虫和感虫幼虫中Virescens mALP蛋白（HvmALP）的表达，碱性磷酸酶（ALP）活性测定和Western blotting进一步验证了该方法的有效性。通过定量实时聚合酶链反应（qRT-PCR），我们证明了抗性幼虫中HvmALP蛋白水平的降低是转录本数量减少的结果。在耐Cry 1Ac的H. armigera和S.对Cry 1Fa有抗性。考虑到这项工作中使用的昆虫品系的独特抗性和交叉抗性表型，我们的数据表明，减少的mALP表达应该被靶向用于开发鳞翅目害虫中对Cry毒素的抗性的有效生物标志物。
Development of insect resistance is one of the main concerns with the use of transgenic crops expressing Cry toxins from the bacterium Bacillus thuringiensis. Identification of biomarkers would assist in the development of sensitive DNA-based methods to monitor evolution of resistance to Bt toxins in natural populations. We report on the proteomic and genomic detection of reduced levels of midgut membrane-bound alkaline phosphatase (mALP) as a common feature in strains of Cry-resistant Heliothis virescens, Helicoverpa armigera and Spodoptera frugiperda when compared to susceptible larvae. Reduced levels of H. virescens mALP protein (HvmALP) were detected by two dimensional differential in-gel electrophoresis (2D-DIGE) analysis in Cry-resistant compared to susceptible larvae, further supported by alkaline phosphatase activity assays and Western blotting. Through quantitative real-time polymerase chain reaction (qRT-PCR) we demonstrate that the reduction in HvmALP protein levels in resistant larvae are the result of reduced transcript amounts. Similar reductions in ALP activity and mALP transcript levels were also detected for a Cry1Ac-resistant strain of H. armigera and field-derived strains of S. frugiperda resistant to Cry1Fa. Considering the unique resistance and cross-resistance phenotypes of the insect strains used in this work, our data suggest that reduced mALP expression should be targeted for development of effective biomarkers for resistance to Cry toxins in lepidopteran pests.
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