Zottich Umberto, Da Cunha Maura, Dias Germana B, Rabelo Guilherme R, Oliveira Antonia Elenir A, Carvalho André O, Fernandes Kátia Valevski S, do Nascimento Viviane V, Gomes Valdirene M
Laboratório de Microbiologia Ambiental e Biotecnologia, Complexo Biopráticas, Universidade Vila Velha, Vila Velha, ES, Brazil.
Protein J. 2014 Oct;33(5):422-31. doi: 10.1007/s10930-014-9575-9.
In this work, we analyzed the effects of coffee seed proteins, especially Cc-LTP1 on the larval development of Callosobruchus maculatus (F.) (Coleoptera: Bruchidae), a bruchid pest of beans and the most important insect pest of Vigna unguiculata (L.) Walp. Artificial seed assay, which incorporated the F/0-90 fraction from Coffea canephora seeds, resulted in the reduction of oviposition and caused an inhibition of C. maculatus larval development in a dose-dependent manner. The F/0-90 fraction used at a 4 % concentration resulted in the survival of no larvae. The purified Cc-LTP1, at a concentration of 0.5 %, also demonstrated effective inhibition of larval development, reducing both females oviposition and the weight and number of larvae. Cc-LTP1 was also able to inhibit the C. maculatus gut α-amylase activity, and immunolabeling by an anti-LTP serum was observed in the midgut tissues of the C. maculatus larvae. Cc-LTP1 has shown binding affinity towards microvillar cells, endoplasmic reticulum and mitochondria, as demonstrated by micrographic images taken by a transmission electron microscope. The results from this study indicate that Cc-LTP1 has insecticidal actions toward C. maculatus and exerts anti-nutritional effects with direct actions on intestinal tissues.
在本研究中,我们分析了咖啡种子蛋白,特别是Cc-LTP1对黄斑豆象(Callosobruchus maculatus (F.),鞘翅目:豆象科)幼虫发育的影响。黄斑豆象是豆类的一种豆象害虫,也是豇豆(Vigna unguiculata (L.) Walp.)最重要的害虫。人工种子试验中加入了卡内弗拉咖啡种子的F/0-90组分,结果导致产卵量减少,并以剂量依赖的方式抑制了黄斑豆象幼虫的发育。以4%浓度使用的F/0-90组分导致无幼虫存活。浓度为0.5%的纯化Cc-LTP1也显示出对幼虫发育的有效抑制作用,减少了雌虫的产卵量以及幼虫的重量和数量。Cc-LTP1还能够抑制黄斑豆象肠道α-淀粉酶活性,并且在黄斑豆象幼虫的中肠组织中观察到抗LTP血清的免疫标记。如透射电子显微镜拍摄的显微图像所示,Cc-LTP1对微绒毛细胞、内质网和线粒体具有结合亲和力。本研究结果表明,Cc-LTP1对黄斑豆象具有杀虫作用,并通过对肠道组织的直接作用发挥抗营养效应。