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小菜蛾对苏云金芽孢杆菌Cry1Acδ-内毒素抗性和敏感品系中刷状缘膜囊泡(BBMV)的脂质

Lipids of brush border membrane vesicles (BBMV) from Plutella xylostella resistant and susceptible to Cry1Ac delta-endotoxin of Bacillus thuringiensis.

作者信息

Kumaraswami N S, Maruyama T, Kurabe S, Kishimoto T, Mitsui T, Hori H

机构信息

Graduate School of Natural Science and Technology, Niigata University, 950-2181, Niigata, Japan.

出版信息

Comp Biochem Physiol B Biochem Mol Biol. 2001 May;129(1):173-83. doi: 10.1016/s1096-4959(01)00327-x.

Abstract

Plutella xylostella (PX) that were 130000-fold more resistant to Cry1Ac were selected from the susceptible strain and maintained in the laboratory. The LC50 of the susceptible strain (PXS) was 0.38 microg toxin/g diet, whereas that of the resistant strain (PXR) was 4800 microg toxin/g diet. Brush border membrane vesicles (BBMV) were prepared from both PXS and PXR. In ligand blot analysis, Cry1Ac bound to a 120-kDa protein of BBMV; however, the intensity of the band was almost equal in both strains of insect. Hence, we analyzed the lipid components of BBMV from PXS and PXR. BBMV lipids were fractionated into non-polar lipid, phospholipid, neutral glycolipid and acidic glycolipid. Neutral glycolipid content was substantially lower in the BBMV of PXR than of PXS. The same trend was observed when lipids were extracted from whole midgut instead of BBMV. Thin layer chromatography of midgut neutral glycolipids revealed the presence of more than seven components. Among the midgut neutral glycolipids, a possible hexasaccharylceramide and a possible trisaccharylceramide of PXR were less than half the level found in PXS. The other lipid fractions in PXR and PXS were similar to each other.

摘要

从敏感品系中筛选出对Cry1Ac抗性高130000倍的小菜蛾(PX),并在实验室中饲养。敏感品系(PXS)的LC50为0.38微克毒素/克饲料,而抗性品系(PXR)的LC50为4800微克毒素/克饲料。从PXS和PXR中均制备了刷状缘膜囊泡(BBMV)。在配体印迹分析中,Cry1Ac与BBMV的一种120 kDa蛋白结合;然而,该条带的强度在两种昆虫品系中几乎相等。因此,我们分析了PXS和PXR的BBMV的脂质成分。BBMV脂质被分离为非极性脂质、磷脂、中性糖脂和酸性糖脂。PXR的BBMV中中性糖脂含量显著低于PXS。当从整个中肠而非BBMV中提取脂质时,也观察到了相同的趋势。中肠中性糖脂的薄层色谱显示存在七种以上成分。在中肠中性糖脂中,PXR的一种可能的六糖基神经酰胺和一种可能的三糖基神经酰胺含量不到PXS中发现水平的一半。PXR和PXS中的其他脂质组分彼此相似。

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