Alkaline phosphatases and aminopeptidases are altered in a Cry11Aa resistant strain of Aedes aegypti.
Alkaline phosphatases and aminopeptidases are altered in a Cry11Aa resistant strain of Aedes aegypti.
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DOI：
10.1016/j.ibmb.2014.09.004
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发表时间：
2014-11
影响因子：
3.8
通讯作者：
Gill SS
中科院分区：
文献类型：
--
作者：
Lee SB;Aimanova KG;Gill SS
苏云金芽孢杆菌亚种israelensis (Bti) 已广泛用于蚊子种群的生物控制。然而，Bti毒素的机制仍不完全清楚。为了进一步阐明 Bti 毒素的机制，我们开发了一种埃及伊蚊抗性菌株，该菌株对 Cry11Aa 毒素表现出高水平的抗性。经过 27 次 Cry11Aa 毒素筛选后，幼虫对 Cry11Aa（G30 株）的耐药性提高了 124 倍。 G30幼虫对Cry4Aa表现出交叉抗性（66倍抗性），对Cry4Ba（13倍）较少，但对Cry11Ba（2倍）没有交叉抗性。与易感幼虫 (WT) 相比，这些抗性幼虫的中肠在处理 Cry11Aa 毒素方面没有显示出可检测到的差异。与 WT BBMV 相比，来自抗性幼虫的刷状缘膜囊泡 (BBMV) 结合的 Cry11Aa 略少。为了鉴定与 Cry11A 抗性相关的潜在蛋白质，不仅使用 Illumina 测序和 qPCR 分析了幼虫中肠中的转录物变化，还使用免疫印迹研究了先前鉴定的受体蛋白质的变化。与WT相比，抗性幼虫中肠中有375个基因的转录显着增加，208个基因的转录下调。在这些分析中，先前鉴定的 Cry11Aa 受体（伊蚊钙粘蛋白、ALP1、APN1 和 APN2）的转录本均未发生改变。与 WT 相比，抗性幼虫中肠中已鉴定的功能受体的基因在其序列中不包含任何突变，其转录物表达水平也没有任何变化。然而，在抗性菌株 BBMV 中，ALP 蛋白的表达水平降低 (~40%)。 APN 蛋白及其活性在抗性菌株中也略有降低。 ALP（AAEL013330 和 AAEL015070）和 APN（AAEL008158、AAEL008162）的转录水平显着降低。这些结果强烈表明 ALP 和 APN 可能与 Ae 的 Cry11Aa 抗性有关。埃及伊蚊。
Bacillus thuringiensis subsp. israelensis (Bti) has been widely for the biological control of mosquito populations. However, the mechanism of Bti toxins is still not fully understood. To further elucidate the mechanism of Bti toxins, we developed an Aedes aegypti resistant strain that shows high-level resistance to Cry11Aa toxin. After 27 selections with Cry11Aa toxin, the larvae showed a 124-fold resistance ratio for Cry11Aa (strain G30). G30 larvae showed cross-resistance to Cry4Aa (66-fold resistance), less to Cry4Ba (13-fold), but not to Cry11Ba (2-fold). Midguts from these resistant larvae did not show detectable difference in the processing of the Cry11Aa toxin compared to that in susceptible larvae (WT). Brush border membrane vesicles (BBMV) from resistant larvae bound slightly less Cry11Aa compared to WT BBMV. To identify potential proteins associated with Cry11A resistance, not only transcript changes in the larval midgut were analyzed using Illumina sequencing and qPCR, but alterations of previously identified receptor proteins were investigated using immunoblots. The transcripts of 375 genes were significantly increased and those of 208 genes were down regulated in the resistant larvae midgut compared to the WT. None of the transcripts for previously identified receptors of Cry11Aa (Aedes cadherin, ALP1, APN1, and APN2) were altered in these analyses. The genes for the identified functional receptors in resistant larvae midgut did not contain any mutation in their sequences nor was there any change in their transcript expression levels compared to WT. However, ALP proteins were expressed at reduced levels (~40%) in the resistant strain BBMV. APN proteins and their activity were also slightly reduced in resistance strain. The transcript levels of ALPs (AAEL013330 and AAEL015070) and APNs (AAEL008158, AAEL008162) were significantly reduced. These results strongly suggest that ALPs and APNs could be associated with Cry11Aa resistance in Ae. aegypti.