Yang Yan-kun, Cai Quan-xin, Cai Ya-jun, Yan Jian-ping, Yuan Zhi-ming
Wuhan Institute of Virology, Chinese Academy of Sciences, Wuhan 430071, China.
Wei Sheng Wu Xue Bao. 2007 Jun;47(3):456-60.
Mosquitocidal toxin 1 (Mtx1) was synthesized during vegetative phase of Bacillus sphaericus and it had been proved to have higher activity to Aedes spp. larvae and Binary toxin (Bin) resistance Culex larvae. The truncated 97 kDa Mtx1 with a deletion of the signal peptide and the Cyt1 Aa crystal protein, a 27.3 kDa delta-endotoxin from Bacillus thuringiensis subsp. israelensis (Bti), were purified from Escherichia coli and B. thuringiensis recombinant strains respectively. Both purified toxins had high toxicity against Culex quinquefasciatus larvae. Bioassay result revealed the purified Mtx1 toxin had high toxicity against the target mosquito larvae, with LCso of 45.2 ng/mL. However, the mixture of Mtx1 and Cytl Aa exhibited higher toxicity against the mosquito larvae, with a lowest LC50 value of 20.19 ng/mL at the ratio of 1:3. (Mtx1:Cyt1 Aa). The calculated synergistic factor of different mixtures suggested a strong synergistic effect between Cyt1 Aa toxin and Mtx1. Furthermore, the presence of Cyt1Aa in the mixture could induce early larval mortality, enhancing the activity of Mtx1 to the target mosquito larvae. The synergistic effect of Cyt1 Aa on mortality of Mtx1 to mosquito larvae might be caused by the damage of the larval midgut-hemocoel barrier induced by the activated CytlAa toxin, which enhanced the specific pathogenesis of Mtx1 on mosquito larvae. It is suggested that the co-application of Mtx1 and Cyt1 Aa in future will be integrated for mosquito management.
球形芽孢杆菌在营养生长阶段合成杀蚊毒素1(Mtx1),已证明其对伊蚊幼虫和对二元毒素(Bin)具有抗性的库蚊幼虫具有更高的活性。从大肠杆菌和苏云金芽孢杆菌重组菌株中分别纯化出缺失信号肽的截短型97 kDa Mtx1和来自苏云金芽孢杆菌以色列亚种(Bti)的27.3 kDa δ-内毒素Cyt1Aa晶体蛋白。两种纯化毒素对致倦库蚊幼虫均具有高毒性。生物测定结果显示,纯化的Mtx1毒素对目标蚊虫幼虫具有高毒性,LC50为45.2 ng/mL。然而,Mtx1和Cyt1Aa的混合物对蚊虫幼虫表现出更高的毒性,在1:3(Mtx1:Cyt1Aa)比例下最低LC50值为20.19 ng/mL。不同混合物的计算协同因子表明Cyt1Aa毒素与Mtx1之间存在强协同效应。此外,混合物中Cyt1Aa的存在可诱导幼虫早期死亡,增强Mtx1对目标蚊虫幼虫的活性。Cyt1Aa对Mtx1致蚊虫幼虫死亡的协同效应可能是由活化的Cyt1Aa毒素诱导幼虫中肠-血腔屏障损伤所致,这增强了Mtx1对蚊虫幼虫的特异性致病作用。建议未来将Mtx1和Cyt1Aa联合应用于蚊虫防治。