Chitosan/DsiRNA nanoparticle targeting identifies AgCad1 cadherin in Anopheles gambiae larvae as an in vivo receptor of Cry11Ba toxin of Bacillus thuringiensis subsp. jegathesan.
Chitosan/DsiRNA nanoparticle targeting identifies AgCad1 cadherin in Anopheles gambiae larvae as an in vivo receptor of Cry11Ba toxin of Bacillus thuringiensis subsp. jegathesan.
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DOI：
10.1016/j.ibmb.2015.03.001
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发表时间：
2015-05
影响因子：
3.8
通讯作者：
Qi Zhang;G. Hua;M. Adang
中科院分区：
文献类型：
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作者：
Qi Zhang;G. Hua;M. Adang
苏云金芽孢杆菌 jegathesancrystals 亚种的 Cry11Ba 蛋白对多种蚊子具有独特的高毒性。 Cry11Ba 对冈比亚按蚊的高效力是通过识别多种中肠蛋白引起的，包括糖基磷脂酰肌醇锚定的碱性磷酸酶 AgALP1、氨肽酶 AgAPN2、α-淀粉酶 AgAmy1 和 α-葡萄糖苷酶 Agm3，这些蛋白以高亲和力与 Cry11Ba 结合并充当推定受体。 An 中的钙粘蛋白 AgCad2。冈比亚幼虫还以高亲和力 (Kd= 12 nM) 结合 Cry11Ba，并被认为是一种推定受体，而钙粘蛋白 AgCad1 以低亲和力 (Kd= 766 nM) 结合 Cry11Ba，这种特性不支持 Cry11Ba 受体的作用。在这里，我们展示了 AgCad1 在 An 体内参与 Cry11Ba 毒性。冈比亚幼虫使用壳聚糖/DsiRNA 纳米粒子抑制幼虫中的 AgCad 表达。 Cry11Ba 对 AgCad1 沉默幼虫的毒性明显低于对照幼虫。由于 AgCad1 被 AgCad2 DsRNAi 共同抑制，因此无法确定 AgCad2 是否参与 Cry11Ba 毒性。 4龄幼虫肠道组织中AgCad1:AgCad2转录水平的比例为36:1。沉默 AgCad 表达对 Cry11Ba 其他结合受体的转录水平没有影响。我们得出结论，An 中存在 AgCad1 和可能的 AgCad2。冈比亚幼虫是体内 Cry11Ba 毒素的功能受体。
The Cry11Ba protein of Bacillus thuringiensis subsp.jegathesancrystals has uniquely high toxicity against a spectrum of mosquito species. The high potency of Cry11Ba against Anopheles gambiae is caused by recognition of multiple midgut proteins including glycosyl phosphatidylinositol-anchored alkaline phosphatase AgALP1, aminopeptidase AgAPN2, α-amylase AgAmy1 and α-glucosidase Agm3 that bind Cry11Ba with high affinity and function as putative receptors. The cadherin AgCad2 inAn. gambiaelarvae also binds Cry11Ba with high affinity (Kd= 12 nM) and is considered a putative receptor, while cadherin AgCad1 bound Cry11Ba with low affinity (Kd= 766 nM), a property not supportive for a Cry11Ba receptor role. Here, we show thein vivoinvolvement of AgCad1 in Cry11Ba toxicity inAn. gambiaelarvae using chitosan/DsiRNA nanoparticles to inhibit AgCad expression in larvae. Cry11Ba was significantly less toxic to AgCad1-silenced larvae than to control larvae. Because AgCad1 was co-suppressed by AgCad2 DsRNAi, the involvement of AgCad2 in Cry11Ba toxicity could not be ascertained. The ratio of AgCad1:AgCad2 transcript level is 36:1 for gut tissue in 4th instar larvae. Silencing AgCad expression had no effect on transcript levels of other binding receptors of Cry11Ba. We conclude that AgCad1 and possibly AgCad2 inAn. gambiaelarvae are functional receptors of Cry11Ba toxinin vivo.