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A new observation on feeding behaviour of pink bollworm and its application in screening Bt-resistant population

机译:粉红棉铃虫摄食行为的新观察及其在抗Bt人群中的应用

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摘要

The pink bollworm (PBW), Pectinophora gossypiella (Saund.) does not feed on leaves in natural circumstances. We made an attempt with the first instar larva (5 days old) and found it eats the leaf in the absence of other foods. Utilizing this new feeding behaviour, a simple methodology was developed for screening of resistance in PBW against Bt cotton plants. The PBW collected from BGII Bt cotton fields (BGII-resistant population) and NBAIR culture (susceptible population) were reared under laboratory conditions for two generations. Laboratory reared 5-day old larvae of PBW were released on cotton leaf discs individually for screening. The BGII-resistant and susceptible larvae fed an average leaf area of 75.52 ± 16.68 and 5.95 ± 0.93 mm2 with survival rate of 90 and 4% respectively, in BGII Bt cotton leaves. Whereas in case of non-Bt cotton (MCU13), the BGII-resistant and susceptible larvae consumed average leaf area of 114.84 ± 23.70 and 116.80 ± 24.14 mm2 with survival rate of 93.34 and 95.33%, respectively. In addition to the screening process, the larval survivors were transferred to an artificial diet after 7 days of experiment and observed up to their emergence as adults. To confirm the development of resistance in PBW, sequencing of larval DNA amplicons was carried out and it revealed mutation in the cadherin gene of the BGII-resistant PBW population. Hence, the detached leaf bit feeding assay described here could be used in a simple manner for screening of resistance developed by PBW against Bt cotton and also for evaluating Bt cotton plants for their inherent resistance to PBW. This method could also be used for studying toxicity of Bt isolates by coating spore-crystal mixture on non-transgenic cotton leaf discs.Electronic supplementary materialThe online version of this article (10.1007/s13205-018-1262-7) contains supplementary material, which is available to authorized users.
机译:在自然情况下,粉红色的棉铃虫(PBW)棉铃虫(Pectinophora gossypiella(Saund。))不会以叶子为食。我们尝试了第一只幼龄幼虫(5天大),发现它在没有其他食物的情况下吞噬了叶子。利用这种新的喂养行为,开发了一种简单的方法来筛选PBW中对Bt棉株的抗性。从BGII Bt棉田(抗BGII的种群)和NBAIR培养物(易感种群)收集的PBW在实验室条件下饲养了两代。将实验室饲养的5天大的PBW幼虫分别释放到棉叶圆盘上进行筛选。在BGII Bt棉花叶片中,抗BGII的易感幼虫的平均叶面积分别为75.52±16.68和5.95±0.93 mm 2 ,存活率分别为90%和4%。而在非Bt棉花(MCU13)的情况下,抗BGII的和易感幼虫的平均叶面积为114.84±23.70 mm和116.80±24.14 mm 2 ,成活率分别为93.34和95.33%。 。除了筛选过程外,在7天的实验后,将幼虫存活者转入人工饮食,并观察其成年后的存活情况。为了证实PBW耐药性的发展,对幼虫DNA扩增子进行了测序,揭示了BGII耐药PBW人群钙粘蛋白基因的突变。因此,本文所述的分离的叶位进食测定法可以以简单的方式用于筛选PBW对Bt棉花产生的抗性,并且还可以评估Bt棉花植物对PBW的固有抗性。该方法还可以用于通过在非转基因棉叶圆片上包被孢子晶体混合物来研究Bt分离物的毒性。电子补充材料本文的在线版本(10.1007 / s13205-018-1262-7)包含补充材料,其中适用于授权用户。

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