Transgenic sugarcane for the sustainable management of the sugarcane borer Diatraea saccharalis

Sugar Industry Pub Date : 2024-01-29 DOI:10.36961/si30898
F. Budeguer, J. Racedo, R. Enrique, M.F. Perera, S. Ostengo, A.S. Noguera
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Abstract

The sugarcane borer Diatraea saccharalis is a major pest of sugarcane (Saccharum spp. hybrids) in the Americas. The insect is partially controlled by cultural, biological, and chemical methods but still causes significant economic losses to sugarcane growers and processors. Currently, one of the most efficient strategies for pest control in other crops is the use of transgenic plants harbouring genes from Bacillus thuringiensis (Bt), which encode crystalline proteins (Cry and/or Vip) with insecticidal activity, known as Bt genes/proteins. In sugarcane, the expression of individual Bt proteins has been previously reported, but not the stacking of two or more Bt proteins. In this study, Bt genes were incorporated by microprojectile bombardment into embryogenic sugarcane calli of the clones TUC 95-10 and TUC 03-12. The presence of the transgenes in 33 transgenic lines was verified by using PCR assays. Subsequently, transgenic lines were acclimated and multiplied in the greenhouse to generate vegetative material for their phenotypic and molecular assessments. The expression levels of transcripts in candidate lines were quantified by real time quantitative PCR (qRT-PCR) assays. In conclusion, transgenic sugarcane lines with a higher level of expression of Bt transcripts compared with the control were developed. These promising lines will be used for future phenotypic tests to determine their resistance against D. saccharalis.
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转基因甘蔗用于甘蔗螟虫的可持续管理
甘蔗螟 Diatraea saccharalis 是美洲甘蔗(蔗属杂交种)的主要害虫。这种昆虫可通过文化、生物和化学方法得到部分控制,但仍给甘蔗种植者和加工商造成重大经济损失。目前,其他作物中最有效的害虫控制策略之一是使用携带苏云金芽孢杆菌(Bt)基因的转基因植物,这些基因编码具有杀虫活性的结晶蛋白(Cry 和/或 Vip),即 Bt 基因/蛋白。以前曾报道过甘蔗中单个 Bt 蛋白的表达,但未报道过两个或多个 Bt 蛋白的叠加。在这项研究中,Bt 基因是通过微弹射轰击法植入 TUC 95-10 和 TUC 03-12 克隆甘蔗胚胎茧中的。通过 PCR 检测验证了 33 个转基因品系中转基因的存在。随后,在温室中对转基因品系进行适应性培养和繁殖,以产生无性系材料,对其进行表型和分子评估。通过实时定量 PCR(qRT-PCR)检测,对候选品系中转录本的表达水平进行了量化。总之,与对照相比,开发出的转基因甘蔗品系具有更高的 Bt 转录本表达水平。这些有希望的品系将用于未来的表型测试,以确定它们对糖核菌的抗性。
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