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芋凝集素的纯化及其对瓜实蝇的抗虫潜力测定。

Purification of Colocasia esculenta lectin and determination of its anti-insect potential towards Bactrocera cucurbitae.

作者信息

Thakur Kshema, Kaur Manpreet, Kaur Satwinder, Kaur Amritpal, Kamboj Sukhdev Singh, Singh Jatinder

机构信息

Department of Molecular Biology and Biochemistry, Guru Nanak Dev University, Amritsar-143 005, India.

出版信息

J Environ Biol. 2013 Jan;34(1):31-6.

Abstract

The present study reports the purification of a lectin from Colocasia esculenta (L.) Schott corms and evaluation of its anti-insect potential towards Bactrocera cucurbitae (Coquilett). The lectin was found to be specific towards N-acetyl-D-lactosamine (LacNac), a disaccharide and asialofetuin, a desialylated serum glycoprotein in hemagglutination inhibition assay. Asialofetuin was used as a ligand to purify Colocasia esculenta agglutinin (CEA) by affinity chromatography. The purity of CEA was ascertained by the presence of a single band in reducing SDS-PAGE at pH 8.3. The affinity purified CEA was employed in artificial diet bioassay of second instar larvae (64-72 hr old) of the B. cucurbitae at concentrations ranging between 10-160 microg ml(-1). The lectin significantly (p < 0.01) decreased the percent pupation and emergence with respect to control. Effect on various enzymes was studied by employing LC50 (51.6 microg ml(-1)) CEA in the artificial diet bioassay of second instar larvae. All the enzymes tested namely esterases, phosphatases (acid and alkaline), superoxide dismutases, catalase and glutathione-S-transferase showed a significant (p < 0.01, p < 0.05) increase in their enzyme and specific activities. These results showed that CEA affected normal growth and development and presented stress to the larvae, activating their detoxification and anti-oxidant systems. Thus, the lectin seems to be a useful candidate for the control measures of B. cucurbitae under the integrated pest management (IPM) system.

摘要

本研究报道了从芋(Colocasia esculenta (L.) Schott)球茎中纯化一种凝集素,并评估其对南瓜实蝇(Bactrocera cucurbitae (Coquilett))的抗虫潜力。在血凝抑制试验中发现该凝集素对二糖N-乙酰-D-乳糖胺(LacNac)和去唾液酸胎球蛋白(一种去唾液酸化的血清糖蛋白)具有特异性。以去唾液酸胎球蛋白为配体,通过亲和色谱法纯化芋凝集素(CEA)。通过在pH 8.3的还原SDS-PAGE中出现单一条带确定了CEA的纯度。将亲和纯化的CEA用于南瓜实蝇二龄幼虫(64 - 72小时龄)的人工饲料生物测定,浓度范围为10 - 160微克/毫升。与对照相比,该凝集素显著(p < 0.01)降低了化蛹率和羽化率。在二龄幼虫的人工饲料生物测定中,采用LC50(51.6微克/毫升)的CEA研究其对各种酶的影响。所测试的所有酶,即酯酶、磷酸酶(酸性和碱性)、超氧化物歧化酶、过氧化氢酶和谷胱甘肽-S-转移酶,其酶活性和比活性均显著(p < 0.01,p < 0.05)增加。这些结果表明,CEA影响了正常的生长发育,并给幼虫带来压力从而激活其解毒和抗氧化系统。因此,在综合虫害管理(IPM)系统下,该凝集素似乎是控制南瓜实蝇的一种有用候选物。

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