Cry10Aa过表达的稳定性和组织特异性提高了棉花对棉铃象鼻虫的抗性

Thuanne Pires Ribeiro , Marcos Fernando Basso , Mayara Holanda de Carvalho , Leonardo Lima Pepino de Macedo , Dagna Maria Laurindo da Silva , Isabela Tristan Lourenço-Tessutti , Osmundo Brilhante de Oliveira-Neto , Eduardo Romano de Campos-Pinto , Wagner Alexandre Lucena , Maria Cristina Mattar da Silva , Bruna Mendes Diniz Tripode , Tatianne Piza Ferrari Abreu-Jardim , José Ednilson Miranda , Marcio Alves-Ferreira , Carolina Vianna Morgante , Maria Fatima Grossi-de-Sa
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引用次数: 13

摘要

棉铃象鼻虫(Anthonomus grandis)是影响棉花作物的最具破坏性的棉花害虫。为了克服这一问题,成功地获得了过量表达苏云金芽孢杆菌昆虫毒素的转基因棉花植株。先前的研究结果表明,Cry10Aa原蛋白的过表达导致了温室条件下CBW幼虫的高死亡率。在本研究中,我们将Cry10Aa原蛋白再向前推进三代(T2至T4),并对几个棉花事件进行了组成性过表达,并对转基因稳定性和农艺性能进行了研究。此外,还生成了稳定的过表达棉花花芽特异性启动子驱动的Cry10Aa活性(缺乏α-螺旋n末端的Cry10Aa原蛋白)的转基因棉花,并进行了鉴定。在温室条件下,棉花植株组成性或组织特异性过表达Cry10Aa蛋白(原生蛋白或活性蛋白),导致CBW幼虫死亡率高达85%。在花芽特异性启动子控制下过表达Cry10Aa活性的事件,在雄蕊和心皮中的蛋白质积累量高于组成型表达事件。研究结果表明,Cry10Aa基因的高稳定性和花芽组织(主要是雄蕊和心皮)中表达水平和蛋白积累的增加有助于提高对CBW幼虫的抗性。最后,选择了一些值得注意的事件,这些事件有可能在巴西不同的棉花产区进行未来的田间试验。因此,棉花花芽组织中高表达Cry10Aa蛋白的事件可能在CBW综合管理中具有很强的商业应用潜力。
本文章由计算机程序翻译,如有差异,请以英文原文为准。
Stability and tissue-specific Cry10Aa overexpression improves cotton resistance to the cotton boll weevil

The cotton boll weevil (CBW, Anthonomus grandis) is the most destructive cotton insect pest affecting cotton crops. To overcome this problem, CBW-resistant genetically modified cotton plants overexpressing Bacillus thuringiensis entomotoxins were successfully obtained. Previous results showed that the overexpression of Cry10Aa protoxin led to high mortality of the CBW larvae in greenhouse conditions. In this study, we advanced three more generations (T2 to T4), with several cotton events constitutively overexpressing the Cry10Aa protoxin, and the transgene stability and agronomic performance were investigated. In addition, stable transgenic cotton overexpressing the Cry10Aa active (Cry10Aa protoxin lacking the α-helix N-terminal) driven by cotton flower bud-specific promoters were generated and characterized. Cotton events constitutively or tissue-specifically overexpressing the Cry10Aa protein (protoxin or active) represented mortality percentages of the CBW larva of up to 85 % in plants under greenhouse conditions. Events overexpressing the Cry10Aa active under control of the flower bud-specific promoter showed higher protein accumulation in stamens and carpels compared to the events with constitutive expression. Our findings suggested that the high stability of the Cry10Aa transgene and the elevated expression level and protein accumulation in flower bud tissues (primarily in stamen and carpels) contribute to improved resistance to CBW larvae. Finally, some notable events were selected with potential for future field trials in different cotton-producing regions of Brazil. Therefore, cotton events overexpressing high levels of the Cry10Aa protein in flower bud tissue may have a strong potential for commercial use in the integrated management of CBW.

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