Bandani A R, Alizadeh M, Talebi K
Plant Protection Department, Faculty of Agriculture, University of Tehran 31587-11167, Karaj, Iran.
Commun Agric Appl Biol Sci. 2005;70(4):875-8.
A key constraint on increasing wheat production in Iran and some neighbouring countries is Sunn pest which cause severe damage to vegetative growth stage of plant in the early season. It also feeds on wheat grain in late growth stage of plants thus damaged wheat grains loose their bakery properties. Because of injecting protease enzymes into the grain during feeding, enzymes degrade gluten proteins and cause rapid relaxation of dough which results in the production of bread with poor volume and texture. Organophosphorus insecticides are the main pesticides used to control the insect pest. However, suitable reduction in pest population has not been achieved partly due to resistance to pesticides. Esterase plays crucial roles in insect physiology and detoxifies a broad range of xenobiotics including insecticides. Enhanced esterase activity is a major mechanism if insecticide resistance and has been detected in a number of insects. To evaluate esterase activity adult bugs were collected from wheat field in Karaj area of Iran and transferred to the laboratory. For biochemical assay, two adult bugs (either males or females) were homogenized in 500 microl Na-phosphate buffer pH 7.2. The homogenates were centrifuged at 14000 g for 10 minutes at 4 degrees C. The supernatants as the enzyme source were pooled and stored at -20 degrees C for later use. For enzyme assay, 300 microl of supernatant was mixed with equal volume of substrates (30 mM alpha-naphthyl acetate or 30 mM beta-naphthyl acetate) and incubated at 30 degrees C for 30 minutes. Then, 50 microl of fast blue solution (150 mg fast blue B in 15 ml distilled water plus 35 ml 5% SDS) was added and esterase activity was determined in a spectrophotometer at 595 nm. Data showed that there are no differences in esterase activity between male and female. However, There was significant differences between hydrolysis of substrates, alpha-naphthyl acetate and beta-naphthyl acetate. Insect esterase hydrolyzes alpha-naphthyl acetate much more than beta-naphthyl acetate.
伊朗及一些邻国小麦增产的一个关键制约因素是麦长管蚜,它会在生长季早期对植物的营养生长阶段造成严重损害。它还会在植物生长后期取食小麦籽粒,从而使受损的小麦籽粒失去烘焙特性。由于在取食过程中向籽粒中注入蛋白酶,这些酶会降解面筋蛋白,导致面团迅速松弛,从而生产出体积和质地不佳的面包。有机磷杀虫剂是用于控制这种害虫的主要农药。然而,由于对杀虫剂产生抗性,害虫数量并未得到适当减少。酯酶在昆虫生理中起着关键作用,能使包括杀虫剂在内的多种异源生物解毒。酯酶活性增强是昆虫抗药性的主要机制,并且已在多种昆虫中被检测到。为了评估酯酶活性,从伊朗卡拉季地区的麦田收集成年蚜虫并转移到实验室。用于生化分析时,将两只成年蚜虫(雄性或雌性)在500微升pH 7.2的磷酸钠缓冲液中匀浆。匀浆物在4℃下以14000g离心10分钟。将上清液作为酶源合并,并在-20℃下储存以供后续使用。用于酶分析时,将300微升上清液与等体积的底物(30 mM α-萘乙酸或30 mM β-萘乙酸)混合,并在30℃下孵育30分钟。然后,加入50微升固蓝溶液(150毫克固蓝B溶于15毫升蒸馏水中,再加35毫升5% SDS),并在分光光度计上于595纳米处测定酯酶活性。数据表明,雄性和雌性之间的酯酶活性没有差异。然而,底物α-萘乙酸和β-萘乙酸的水解存在显著差异。昆虫酯酶对α-萘乙酸的水解比对β-萘乙酸的水解多得多。