Thakur Kshema, Kaur Manpreet, Kaur Satwinder, Kaur Amritpal, Kamboj Sukhdev Singh, Singh Jatinder
Department of Molecular Biology and Biochemistry, Guru Nanak Dev University, Amritsar-143 005, India.
J Environ Biol. 2013 Jan;34(1):31-6.
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.
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。